Food Safety and Quality-Based - Shelf Life

Food Microbiology and Food Safety
Practical Approaches
Peter J. Taormina
Margaret D. Hardin Editors
Food Safety and

Quality-Based
Shelf Life
of Perishable
Foods
LinkedIn: Emily Phuong

Series Editor:
Michael P. Doyle
Regents Professor of Food Microbiology (Retired)
Center for Food Safety
University of Georgia
Griffin, GA, USA
The Food Microbiology and Food Safety series is published in conjunction with
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Protection Trends and Journal of Food Protection), its educational Annual
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professionals.
More information about this series at http://www.springer.com/series/7131

Peter J. Taormina • Margaret D. Hardin
Editors
Food Safety
and Quality-Based Shelf
Life of Perishable Foods
Editors
Peter J. Taormina Margaret D. Hardin
Etna Consulting Group Vice President of Technical Services
Jacksonville, FL, USA IEH Laboratories and Consulting Group
Lake Forest Park
WA, USA

ISSN 2626-7578 ISSN 2626-7586 (electronic)
ISBN 978-3-030-54374-7 ISBN 978-3-030-54375-4 (eBook)
https://doi.org/10.1007/978-3-030-54375-4
© Springer Nature Switzerland AG 2021

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Preface
In the twenty-first century, a growing global population and development in formerly

underdeveloped nations has created a demand for food availability in unprecedented
amounts and in broad geographical and contextual locations. Such demand means
not only that food must be produced with optimal efficiency, but also that technologi-
cal advances must be utilized to ensure that the food that is produced remains edible
and wholesome for the longest amount of time possible in order to enable wide dis-
tribution and availability in places where people eat. For instance, the popularity of
food trucks, street vendors, and boat vendors has increased globally. The shelf life of
food is a key determining factor on how food is distributed, and consequently where
and when different food products are available for consumption. Therefore, shelf life
must be scientifically determined from both food safety and quality indicators.
Shelf life is determined by several factors, including microbiological, chemical,
physical, and organoleptic deterioration. Often these factors are interrelated and
interdependent, and the degree of influence of each upon shelf life varies with the
type of products. However, for most perishable foods, microbial deterioration is
usually the predominant factor. Depending on production process and packaging
formats, microbial safety or quality factors can determine the shelf life of a product.
Improvements in food processing, distribution, and storage techniques have led to
longer shelf life of food products. Examples include the use of antimicrobials and
modified atmosphere packaging and to delay or completely inhibit microbial growth
for the duration of shelf life. Microbial-based shelf life of perishable foods has long
been determined by quantitative plate count data, but now there is much promise in
the use of genomics to more fully understand how microbial communities change
over the course of shelf life.
This book focuses on microorganisms in perishable food products in keeping with
the series on food microbiology and food safety. It does not cover in-depth chemical
and physical degradation processes per se, but only as they are a result of microbial
growth. The techniques utilized for determination of shelf life, the frequency of shelf
life testing for different products, and the interpretation of data to make shelf life
determinations receive coverage. We also address newer techniques of analysis of
such metagenomics. The reader will gain information on how processing, packaging,
v
vi Preface
and formulation technologies can extend shelf life of perishable foods. Also, insights
into the science of shelf life determination of perishable foods is covered including
the microorganisms of concern for shelf life determination of major categories of
food products.
Jacksonville, FL, USA Peter J. Taormina

Acknowledgments
The editors thank the following people who have advised or assisted us through the
process of planning, writing, and editing this book:
Heather Hart
Jennifer Mason
Michael P. Doyle
Silembarasan Panneerselvam
Susan Safren
vii
Contents
1 Purposes and Principles of Shelf Life Determination�� 1

Peter J. Taormina
2 Food Safety Factors Determining Shelf Life �� 27
Margaret D. Hardin
3 Microbial Growth and Spoilage�� 41
Peter J. Taormina
4 Impact of Sanitation on Product Shelf Life �� 71
Steven Tsuyuki and Margaret D. Hardin
5 Advanced Processing Techniques for Extending
the Shelf Life of Foods �� 91
Sarah M. Hertrich and Brendan A. Niemira
6 Packaging of Perishable Food Products�� 105
Cynthia Ebner, Angela Morgan, and Clyde Manuel
7 Beyond the Standard Plate Count: Genomic Views into
Microbial Food Ecology�� 135
8 The Changing Shelf Life of Chilled, Vacuum-Packed Red Meat�� 145
John Summer, Paul Vanderlinde, Mandeep Kaur,
and Ian Jenson
Index�� 157
ix
Contributors
Cynthia Ebner Sealed Air Corporation, Charlotte, NC, USA

Margaret D. Hardin Vice President of Technical Services, IEH Laboratories and
Consulting Group, Lake Forest Park, WA, USA
Sarah M. Hertrich Food Safety and Intervention Technologies Research Unit,
Eastern Regional Research Center, USDA-ARS, Wyndmoor, PA, USA
Ian Jenson Meat & Livestock Australia, Sydney, NSW, Australia
Mandeep Kaur University of Tasmania, Hobar, TAS, Australia
Clyde Manuel GOJO Industries, Akron, OH, USA
Angela Morgan Aptar Group, Crystal Lake, IL, USA
Brendan A. Niemira Food Safety and Intervention Technologies Research Unit,
Eastern Regional Research Center, USDA-ARS, Wyndmoor, PA, USA
John Summer M&S Food Consultants, Deviot, TAS, Australia
Peter J. Taormina Etna Consulting Group, Jacksonville, FL, USA
Steven Tsuyuki Sanitary Design and Corporate Sanitation, Maple Leaf Foods,
Mississauga, ON, Canada
Paul Vanderlinde Vanderlinde Consulting, Carbrook, QLD, Australia
xi
Chapter 1
Purposes and Principles of Shelf Life
Determination
Peter J. Taormina
1.1 Purposes
Shelf life of food can be defined as the length of time that food can remain in a
wholesome, consumable state and retain acceptable quality under normal, expected
conditions of distribution and storage. Quality factors can include appearance, odor,
flavor, color, texture, nutritional properties, and microbiological populations. These
quality factors may diminish at differing or similar rates during shelf life, either
remaining within acceptable limit or not meeting quality limits resulting in an
unwholesome or nonconsumable state. The extent to which acceptable limits of
these factors can be defined empirically for a food product and accurately monitored
leads to precision in shelf life designation. Perishable foods require reduced tem-
perature in order to prolong the period for this state of acceptability. Three types of
food supply chains have been defined: frozen, chilled, and ambient (Akkerman et al.
2010). The frozen foods’ supply chain typically operates at −18 °C, with products
like ice cream requiring a frozen chain with an even lower temperature of −25 °C. For
the chilled food supply chain, temperatures range from 0 °C for fresh fish to 15 °C
for potatoes and bananas, for example. The ambient chain includes products that do
not require strict temperature control, such as canned goods. The focus of this book
is on the refrigerated shelf life of perishable food products, as that is where micro-
bial growth is of greatest importance.
Perishability is decay, damage, spoilage, evaporation, obsolescence, pilferage,
loss of utility, or loss of marginal value of a commodity that results in decreasing
usefulness from the original one (Wee, 1993). Certain perishable foods can also
support growth of bacterial pathogens during refrigerated storage and therefore
have a safety component to shelf life. Shelf life does not necessarily reflect the
P. J. Taormina (*)
Etna Consulting Group, Jacksonville, FL, USA
e-mail: peter@etnaconsulting.com
© Springer Nature Switzerland AG 2021 1

P. J. Taormina, M. D. Hardin (eds.), Food Safety and Quality-Based Shelf Life
of Perishable Foods, Food Microbiology and Food Safety,
https://doi.org/10.1007/978-3-030-54375-4_1
2 P. J. Taormina
physical state of a product, since many products deteriorate only a while after their
shelf life finishes; however, it may reflect its marketable life (Xu and Sarker 2003).
Shelf life determinations have an integral part in perishable food production,
processing, sale, distribution, and consumer use. The determination of shelf life of
perishable foods integrates food safety and quality factors, including microbiologi-
cal, chemical, and physical degradation. The sensory properties of foods such as
visible appearance, aroma or odor, flavor, and texture are the most obvious aspects
of shelf life. Yet the scientific determination of shelf life of perishable foods may
also involve tracking the behavior and growth of foodborne bacterial pathogens that
do not impart a noticeable sensory change. Ongoing measurement of organoleptic
properties of every production lot of food for the purposes of determining or mea-
suring shelf life is seldom feasible for food manufacturers, even if third-party labo-
ratories are employed to perform such work. Therefore, other factors are analyzed,
which are indicative of sensory breakdown of products over time such as the increase
in microbiological plate counts, increase in oxidative rancidity, or textural changes.
To understand the scientific principles of shelf life determination, it is first neces-
sary to outline the purposes for determining shelf life.
1.1.1 Stakeholders
The shelf life of perishable foods is a topic at which business, logistics, science,
regulatory compliance, and consumer interests intersect. For each of these stake-
holders, the amount of shelf life days assigned to a product is a very important factor
that impacts their roles and responsibilities. Commercialization of food products
requires collaboration between sales and marketing, business development, opera-
tions, and research and development (R&D) functions in order to design food prod-
ucts, scale up prototypes, implement processing and packaging on a larger scale,
and produce food products with reasonable consistency. The operations team would
be responsible for producing the product in the appropriate manner and under sani-
tary conditions that result in achievement of the targeted shelf life. Logistics and
transportation personnel react to the shelf life of the product and design supply
chain and workflows around the number of days allowed. Sales and business devel-
opment professionals negotiate with customers about the number of days of guaran-
teed shelf life remaining upon delivery, typically 30–45 days for perishable foods
with 3–4 months shelf life. Retail or foodservice end users must utilize the shelf life
of the product in the best possible way to maximize sales and minimize waste. The
regulator in the production facility or in the marketplace performing inspections
takes note of the shelf life of products and responds to consumer complaints.
Regulators are also responsible for preventing contaminated or unwholesome food
from reaching consumers. Consumers expect a reasonable number of days of use-
able shelf life and will ultimately validate or invalidate the effectiveness of each of
the above by making purchasing decisions en masse.
1 Purposes and Principles of Shelf Life Determination 3
1.1.1.1 Business Development
Food businesses are keenly interested in food product shelf life for several reasons.
Shelf life impacts the logistics and sale of foods because it sets limits on the amount
of time for brokering sales of bulk quantities. A short shelf life restricts the production
scheduling and impacts labor, raw material supply, transportation, and supply-chain
continuity. Fresh cut produce is one such example of a product with a relatively short
shelf life. Without antimicrobial interventions, typical fresh cut, bagged vegetables
have about a 12- to 15-day shelf life at refrigeration temperatures (Soliva-Fortuny and
Martı́n-Belloso 2003). Many fresh cut vegetable and fruit production plants process,
package, and ship on the same day to a distribution center or directly to foodservice or
retail customers. If product is shipped to a distribution center, those centers may
require 2 or 3 days to receive orders for vegetables and other products prior to loading
trucks and shipping to their customers. Allowing for a day or two of secondary distri-
bution, this leaves only about 10 days of shelf life remaining at the point of use or in
consumers’ possession. This means producers of products with short shelf life have
very little leeway and will typically work 7 days a week to fulfill orders. Any disrup-
tion in raw material supply, operations, or distribution can severely impact the ability
of processors to maintain the supply. Conversely, a long shelf life can allow processors
to capitalize on raw material downward price fluctuations and purchase large quanti-
ties of raw material when prices are low and store that raw material pending receipt of
orders for the product. Long shelf life enables more efficient temporary labor schedul-
ing because food processing plants can schedule long production runs leading to
inventory builds. In turn, these long production runs and inventory build-ups enable
sales professionals to broker better terms for customers in the marketplace. For exam-
ple, a producer of cooked ham products may have a validated food safety and quality-
based shelf life of 120 days. The processor may find that the raw material, frozen ham
muscles, can be purchased on the open market at favorable pricing. At that time, the
processor could hire temporary labor to work 14 days straight in order to build a large
inventory of fully cooked, vacuum-packaged, ready-to-eat ham with a long shelf life.
Further, storage of such products at just above its freezing point can extend the shelf
life even further and create more time to find buyers. Under this scenario, the producer
can permit necessary downtime in the production schedule, which enables time for
preventive maintenance and deep cleaning and sanitizing of the equipment and facil-
ity, while the workers can be provided training refreshers and education about health
and safety, food safety, and quality. This creates a virtuous cycle, whereby these activ-
ities lead to better quality performance, which results in better more consistent prod-
uct that meets or exceeds the designated shelf life.
1.1.1.2 Logistics
Well-managed food logistics can reduce the incidents of product reaching the end of
shelf life without a buyer and without reaching a consumer. The perishability of
fresh food products limits the opportunities to use inventories as a buffer against
4 P. J. Taormina
variability in demand and transportation (Ahumada and Rene Villalobos 2009).

Food processors must avoid over-ordering raw material supplies and packaging
materials and overproducing finished product beyond the actual demand. This pres-
ents a challenge to forecast accurately the demand. Conversely, retail or foodservice
customers must avoid overestimating consumer demand and thereby purchasing too
much product. Thus, efforts to successfully extend shelf life of perishable foods can
create more flexibility in the supply and demand planning, operations, and transpor-
tation dynamic, as well as potentially provide customers more time to display and
potentially sell food products.
Designing food distribution systems that are able to provide high-quality food in
a cost-efficient way is a challenge that requires collaboration between cross-
functional teams such as logistics, food engineering, and operations management
(Apaiah et al. 2005). Food produced by manufacturers, packers, and processors may
be shipped directly to retail or foodservice points or is often sent to distribution
centers that serve as intermediaries in logistical transport and delivery of food
(Fig. 1.1). While this general distribution structure remains the norm for a majority
of food, increasingly, food is being sent directly to consumers as online meal kit
delivery services rise in popularity. Direct-to-consumer food distribution models
drastically change the shelf life potential due to lack of consistently verifiable tem-
perature control.
The limited shelf life of perishable food products, requirements for strict tem-
perature and humidity control, possible interaction effects between products, time
windows for delivering the products, high customer expectations, and low profit
margins make food distribution management very challenging (Akkerman et al.
2010). On a strategic level, perishability may also play an important role by forcing
better relations and more integration between the supply chain networks of within
and between organizations (Amorim et al. 2013). For example, most of the food
industries rely on third-party logistic providers (3PL) to perform the distribution of
their products. A good relation between those companies can result in additional
gains in efficiency leading to lower costs of distribution and less waste. At the other
end of the supply chain is a vendor-managed inventory system, relating a supplier
of a perishable raw product and a company that processes this product.
Retail
Distribution
Food
centre / Consumer
manufacturing
wholesaler
Foodservice
Fig. 1.1 General structure of distribution within the food supply chain (from Akkerman et al. 2010)
Vendor-managed supply chains mean perishable foods produced on demand factor-

ing in expected shelf life. In either system, enhanced communication between these
entities that enables better supply and demand planning should potentially lead to
less spoilage and, hence, less costs that all parties may reap, including the consumer.
A forecast-based ordering model for Nordic supply chains for milk, fresh fish,
and poultry were studied by Kaipia et al. (2013) as a means to reduce waste and
increase sustainability. They concluded that supply chain structure should be
streamlined to avoid additional handling and delays and that the order penetration
point (OPP) location needs to support the specific features of fresh food supply
chains. Their initial findings indicated that the OPP should be moved as close to the
retail customer as possible. They also determined that an efficient forecasting pro-
cess becomes more important in this model because a larger share of the chain oper-
ates on the basis of forecasts. The demand data should be utilized to balance
operations between made to order (MTO) and made to stock (MTS) production.
They concluded that demand data form a good basis for forecasting and, on the
other hand, can trigger larger MTO production instead of producing to stock. The
way to utilize demand data needs to be selected in a way to support optimally the
specific product characteristics and demand patterns (Kaipia et al. 2013). Perishable
products characterized by a short shelf life are well suited to the MTO model, espe-
cially if raw materials can be on hand and ready for processing into finished prod-
uct. For foods like fresh fish or fresh cut produce, timing a harvest date to on-demand
production could be a more significant challenge.
Utilization of information technology can potentially enhance the efficiency of
food logistics. Radio frequency identification (RFID) has been researched as a
means to manage highly perishable food inventory distribution (Grunow and
Piramuthu 2013). When a food item is RFID-tagged, the item’s remaining shelf life
is known with certainty to the retailer, distributor, and customer, and the retailer
cannot sell an item beyond its “expiry date.” While RFID temperature and time
monitoring in trucks are not uncommon, monitoring at the pallet level is costlier,
and monitoring of individual units is cost-prohibitive in most cases. However, even
pallet-level RFID tags with sensors could enable distributors to send those pallets
with higher remaining shelf life to the more distant retailers. The value of real-time
access to product shelf life information will be realized by quality and food safety
professionals responsible for assuring safe, wholesome food is delivered and con-
sumed within the code date.
Technologies like Blockchain are now being used to facilitate the transactional
transfer of food through the supply chain (Tian 2016, 2017). The efficiencies gained
by use of Blockchain can benefit perishables with a relatively short shelf life.
Blockchain is now being used to establish within the supply chain a permission
access-based, viewable, and immutable history of network, which can be shared
among all nodes in the system (Tian 2017). With such incredible insight into the
history of food, there will be more insight into the actual shelf life of food products
for brokers, customers, and consumers.
6 P. J. Taormina
1.1.1.3 Science
Shelf life determination must be science-based and supportable using data. The
limitations of number of days of shelf life are a matter of what can be established
with scientifically accepted methods (Kilcast 2001; Man 2004). The development of
new technologies to prolong the shelf life of foods include improvements to raw
material harvesting and speed to processing, advanced food processing techniques,
packaging technology, formulation, and cold-chain integrity. Indeed, shelf life
extension has become a goal of many food producers in order to alleviate some of
the previously discussed business challenges with producing and distributing per-
ishables. Consumer and customer demands for “clean label” food and beverage
products have resulted in the expectation that traditional preservatives like benzo-
ate, sorbate, nitrite, and propionate (Brul and Coote 1999) be removed and replaced
with ingredients that are perceived as natural such as cultured antimicrobials, vine-
gar, bacteriocins, and plant extracts (David et al. 2013). The resulting span of shelf
life of so-called clean label products is seldom the extent possible by incorporation
of traditional preservatives. Scientists and research and development professional
must study these food systems and challenge them with worst-case conditions of
processing, packaging, storage, and use and must prove food products will hold-up
for the designated shelf life.
The research, development, and technology team of a food company or external
partners in academia or food laboratories could employ culinary and food science
techniques to create a product that meets the market demand but must also under-
stand the sales and marketing professional has a real need for competing in the
marketplace. This means that the product developers, processing engineers, packag-
ing engineers, and quality and food safety professionals must work together to
develop least cost options for formulation, process, package, and shelf life. The
shelf life has to be sufficient to enable production planning professionals to meet the
supply demand and for logistics professionals to deliver product to the customer in
a timely manner. The sales, marketing, and business development professionals
work with research and development scientists and technicians to craft a product
that meets the market needs. The formulation and processing techniques required to
meet shelf life targets must support the business and marketing objectives in terms
of costs of production and in nutritional labeling and ingredient usage that meet
consumer expectations. Operations personnel manufacture product according to the
specifications set by R&D professionals. R&D professionals design formulations
and usually outline target processing parameters for achievement of a desired shelf
life. Quality personnel are responsible for monitoring product quality and perfor-
mance and may utilize ongoing shelf life testing of representative samples of pro-
duction as one of a variety of metrics, albeit a lagging one. Growth of spoilage
microorganisms during shelf life can result in the development of an unwholesome
product over time, and therefore food safety and quality professionals and regula-
tors are concerned with the scientific aspects of shelf life determination. Pathogen
growth during refrigerated shelf life can cause foods to become unsafe for con-
sumption, and regulations exist concerning the suppression of growth of
psychrotrophic or psychrotolerant bacterial pathogens like Listeria monocytogenes,

non-proteolytic Clostridium botulinum, and Bacillus cereus (U.S. Department of
Agriculture 2015; U.S. Food and Drug Administration 2017). R&D professionals
oversee the performance of challenge studies to validate safety of perishable food
products throughout shelf life.
R&D professionals are often tasked with the job of collecting shelf life valida-
tion data and presenting it to foodservice or retail customers for approval.
Foodservice and retail customers review and accept shelf life validation data pro-
vided by their various manufacturing suppliers, and if such data are deemed accept-
able will allow these perishable food products into their inventory. Retailers must
have sufficient time to distribute the product through their network and to display
the product to the consumer. The consumer also must have a reasonable and accept-
able amount of time to see the product package in the retail environment, and store,
use, or reuse the product once it has been purchased.
Martins et al. (2008) reviewed computational methods for shelf life dating.
Computational methods use modeling techniques that bridge experimental data
such as quality and safety characterizations, stress level testing, accelerated shelf
life testing, and distribution chain data with information systems. This enables
multi-scale scenario analysis and interpretation in order to better understand the
complex food quality dynamics of shelf life of food. Certainly, a scientific process
of experimental data collection about the product coupled with real distribution
information offers promise as a means to accurately predict shelf life and set accu-
rate code dates.
1.1.1.4 Regulatory
Control of pathogen growth is often the most necessary part of shelf life determina-
tions and can be the limiting factor in setting shelf life of certain perishable foods.
Several foodborne pathogens have little to no bearing upon shelf life of refrigerated
foods due to the fact that they are mesophilic and growth is inhibited at refrigeration
temperatures. For example, pathogens within the family Enterobacteriaceae such as
Salmonella and pathogenic Escherichia coli are typically not able to grow at refrig-
eration temperatures (Roberts and Tompkin 1996). Even shelf stable products that
might support growth of xerotolerant yeasts and molds would not support growth of
most bacterial pathogens. Growth of psychrotrophic pathogens such as Listera
monocytogenes, Yersinia enterocolitica, and group II Clostridium botulinum must
be assessed on refrigerated perishable foods. These psychrotrophic bacterial patho-
gens of concern are reviewed in detail in Chap. 3.
Microbial spoilage of food and beverage products can be caused by a number of
factors, such as a loss of process control, post-processing contamination, inadequate
packaging performance, damage during distribution, or temperature abuse during
distribution, storage, or display. The causative microorganisms of food and bever-
age spoilage are usually not the same as those that are attributable to foodborne ill-
ness. However, in some instances in North America, spoiled products have been
8 P. J. Taormina
subjected to class II recalls (United States Department of Agriculture, Food Safety

and Inspection Service 2015), rather than a more discreet market withdrawal. With
the recent mandatory recall authorization provided to the U.S. Food and Drug
Administration in the Food Safety Modernization Act (Anonymous 2011), the dif-
ference between “market withdrawal” and “recall” is an important regulatory and
legal matter that impacts food brands and the bottom line for food businesses.
Psychrotrophic lactic acid bacterial growth and spoilage effects were responsible
for numerous recalls in Belgium of meat, dairy, vegetable, egg products, and com-
posite foods (Pothakos et al. 2014). Thus, spoilage and shelf life do become a matter
of regulatory concern.
The term “undesirable microorganisms“ as defined in the Hazard Analysis and
Risk-based Preventive Control Final Rule includes those microorganisms that are of
public health significance, that subject food to decomposition, that indicate that
food is contaminated with filth, or that otherwise may cause food to be adulterated
(U.S. Food and Drug Administration 2015). Spoilage of food prematurely before
the expiration date results in undesirable if not unwholesome food. However, ques-
tions remain as to what circumstances and which microorganism-product interac-
tions turn a food spoilage event into a food safety concern. Consumers are also
confused about the differences between true food safety hazards and undesirable,
but innocuous, food spoilage. Expert interpretation of the microbiological data, risk
assessments on the product, product status in the marketplace, and normal con-
sumer use and handling of the product must be evaluated on a case-by-case basis to
make decisions about potential public health impact of spoiled foods implications
on market withdrawal or recall.
1.1.1.5 Consumers
Consumers form quality expectations of food products based on perceived cues

prior to purchase, and post-purchase quality experiences will impact future pur-
chases (Grunert 2002). At retail markets, consumers primarily observe the visual
appearance of products and assess potential quality and solely this way for many
packaged (sealed) products. Consumers expect a reasonable number of days to store
and use perishable food products once purchased (Man and Jones 1994). Consumers
certainly use the sense of smell to assess food quality whenever that is possible.
Package labeling also weighs heavily into consumer perception of the food.
Consumers interpret the printed code date, or expiration date, on the package with
some flexibility, with variation depending upon the type of food product (Van
Boxstael et al. 2014). Consumers are less likely to eat expired raw products than
expired ready-to-eat products.
In terms of different date labelling, likelihood of consumers checking expiration
dates appears to depend on the food category (Aschemann-Witzel et al. 2015).
Consumers check expiration dates frequently for products in which a decrease in
quality is risked and for products with which they have experience of usage (as
measured by the household consumption rate for the category). Relatedly, the
willingness to pay for a perishable product decreases throughout its shelf life.
Freshness dating influences the acceptability of products in a discontinuous or non-
linear manner because it influences perceptions of freshness and of healthfulness,
not of safety (Wansink and Wright 2006). As a product approaches its “best if used
by” date, there may be more for a manufacturer to lose than to gain by having
decided to use “freshness dating” in the first place.
1.1.2 ssuring Quality and Wholesomeness of Food through

A
Code Dates
A code date is the date stamped on the product box and/or package that indicates to
the end user or intermediate user, the time for which the product can be safely and
acceptably used. Code dates vary with the product type and the intended customer.
For example, food products sold to foodservice distributors will typically end up in
institutional or restaurant kitchens. In those settings, food preparers and chefs will
likely see boxes and packages stamped with date codes that are somewhat cryptic.
This is referred to as open dating. Some date codes will indicate only the date of
production. Other date codes will be Julian dates. Julian dates (abbreviated JD) are
simply a continuous count of days and fractions since noon Universal Time on
January 1, 4713 BC (on the Julian calendar). Sometimes Julian dates are nested
within other codes that could relate to production line number, facility, or customer.
Foodservice workers therefore have considerable discretion about how long to use
such products, whether they are held frozen or refrigerated. If frozen product is
received, foodservice workers must decide how long the product is usable once
thawed, unless guidance has been provided by the manufacturer. In some cases,
government entities provide safe handling days for products. At the retail point of
purchase, consumers are presented with a variety of formats through which the shelf
life is communicated (Newsome et al. 2014; Wansink and Wright 2006). This has
become a point of confusion and misunderstanding with regard to safety and whole-
someness of foods in general, but it is of more importance with perishable foods
compared to shelf-stable foods due to the potential for microbial growth during
shelf life.
1.1.2.1 Use by, Sell by, Best by, Best Before, Best If Used by, and Enjoy by
Code dates formats at the retail point of purchase have well been researched. In a
Belgian study, it was found that 30.4% of the respondents declared they did not
know the difference of the meaning between the best before and use by date and
only about half of the respondents indicated to use the type of label (difference
between the labels) for assessing the edibility of a product (Van Boxstael et al.
2014). The authors surmised that part of the confusion lies in the fact that products
10 P. J. Taormina
that have only quality degradation as the limiting factor (e.g., mayonnaise) and per-
ishable products that have safety and quality factors determining the shelf life (e.g.,
meats), both have use by dates. They proposed to simplify this by having govern-
ment assign all refrigerated products a use by label (and the communication that use
by products are unsafe to consume after the shelf life date and that for best before
products the edibility can be judged by the consumer). The authors admitted the
difficulty in predicting the outcome of this simplified approach and noted there to be
advantages with respect to food safety and a mixed outcome with respect to
food waste.
The issue of date labelling was found to be a primary issue in consumer-related
food waste (Aschemann-Witzel et al. 2015). The terminology and use of date labels
vary widely, and this contributes to consumers’ confusion about the meaning
(Newsome et al. 2014). A more uniform and consistent date label display format
that gets away from sell by or display by has been proposed to reduce misunder-
standing for consumers (Aschemann-Witzel et al. 2015; Newsome et al. 2014).
For perishable foods, consumers search for visual and other cues of freshness,
such as expiration dates. The greater the risks associated with a product, the more
frequently consumers check expiration dates (Tsiros and Heilman 2005). If con-
sumers perceive risk associated with the quality of a perishable food product as it
approaches its expiration date and foresee possible health risks from consuming the
product, they are more likely to check and adhere to expiration dates. Consumer
willingness to pay (WTP) for perishable foods decreases markedly as products
approach expiration dates (Fig. 1.2). WTP for chicken and lettuce declines rapidly
even 6 days preceding expiration (Tsiros and Heilman 2005).
Expiration of perishable food accounts for about 20% of the total unsalable items
in grocery stores, drug stores, and wholesalers in the United States (Joint Industry
Unsaleables Leadership Team 2008). From 2005 to 2007, 56% of retailers and dis-
tributors surveyed indicated that unsaleable costs due to expired products have
increased. The report indicated that since the use of open date coding is diminishing
and more intuitive code date formats such as use by are more prevalent, consumers
are more aware of expiration dates, and therefore, manufacturers are more conser-
vative about shelf life days assigned to products. This has led to more product expir-
ing in the marketplace.
1.1.2.2 Food Waste
Expired products (past the sell by date) account for much of the losses at retail
supermarkets. It has been estimated that 12% of fruits and vegetables and 9.5% of
seafood is discarded at North American retail markets, and consumers discard
nearly 30% of seafood and produce (Gunders 2012). The most common reason for
waste at the retail store is that products’ expiry dates have passed (Kaipia et al.
2013). More than a quarter of food discarded from retail stores in Austria were had
no defect other than the sell by date had passed (Lebersorger and Schneider 2014).
The causes for this include ordering more than real demand or products reaching the
1
.9
WTP as a Percentage .8
.7
.6
of List Price
.5
.4
.3
.2
.1
0
7 6 5 4 3 2 1
Number of Days Before Product
Reaches Its Expiration Date
Lettuce Carrots Chicken

Yogurt Milk Beef
Fig. 1.2 Consumer willingness to pay (WTP) as a percentage of list price throughout a food prod-
uct’s perishable shelf life (from Tsiros and Heilman 2005 with permission)
store shelf too late and with a short remaining shelf life (Mena et al. 2011). Therefore,
inefficiencies or disorganization in perishable food supply chains can directly
impact the amount of food that is expired before consumption and ends up being
destroyed.
1.1.3 Expiration = Decision Time
1.1.3.1 Distressed Product
When food in distribution centers or cold-storage warehouse facilities near the end
of the stated shelf life, it is too late to change the code date. Often, product managers
or sales and marketing professionals might ask the scientists for extension of shelf
life after the product has been produced, packaged, shipped, and received. Apart
from the appearance of impropriety, there are inherent risks with addition of days to
product shelf life ex post facto. The proper food safety and quality culture should
influence the decision by the manufacturer to hold fast to the shelf life of the product
and not allow additional days to be added to the code date. This pertains to refriger-
ated, perishable foods, but the concept is important for shelf stable foods and frozen
foods as well. Even if those products do not deteriorate at a rate that would greatly
impact the addition of a few weeks of shelf life, the very practice of granting
12 P. J. Taormina
extensions can lead to dangerously false perceptions that quality and food safety is
negotiable, and allowance of other unacceptable quality practices. It could eventu-
ally overwhelm the technical quality assurance personnel, as they will be constantly
asked for approvals of extensions of shelf life. Rather, the safest, most appropriate
approach is to perform proper shelf life testing (i.e., validation) to establish the
maximum reasonable shelf life to assign to products. Inventory management sys-
tems can alert product managers, sales, and marketing professionals when food
product lots are close to reaching the expiration date. At such point, product can be
allocated as “distressed,” signaling to the business that it is time to unload this
inventory customers at a discount or risk not selling it at all. Strong inventory man-
agement systems should include an ongoing plan for discounted selling of distressed
product.
1.1.3.2 Donation
Donation is the second option when food is near the expiration date. If a manufac-
turer or distributor or retailer possesses food that is close to the expiration date, and
the product cannot be sold, donation is a sustainable option. In the U.S., the Bill
Emerson Act of 1996 (Anonymous 1996) encourages the donation of food and gro-
cery products to nonprofit organizations for distribution to needy individuals by
giving the Model Good Samaritan Food Donation Act the full force and effect of
law. The law states that “a person or gleaner shall not be subject to civil or criminal
liability arising from the nature, age, packaging, or condition of apparently whole-
some food or an apparently fit grocery product that the person or gleaner donates in
good faith to a nonprofit organization for ultimate distribution to needy individuals.”
There are a variety of organizations that receive and redistribute food donations on
a large scale, and so the option for donation of nearly expired food should be a fore-
thought for food producers, distributors, and retailers.
1.1.3.3 Diversion
Diversion is another possible choice for food close to the expiration date. Food
products can be diverted as an incoming raw material stream to other forms of food
processing, if it is compatible. This is an option for raw commodities, such as raw
meat being diverted to a cooking process, more so than for packaged foods. Besides
compatibility, feasibility becomes a factor as product is often in boxes with packag-
ing material. Businesses must consider the labor and other costs associated with
reprocessing and/or repackaging nearly expired food to divert it to another product
stream. Another option is to divert to animal food production. However, animal food
is somewhat precise in terms of desired proteins, carbohydrate, fat, and nutrient
content. Further, the food ingredients used for human food are not always approved
for animal food.
1.1.3.4 Destruction
Destruction should be the last option in dealing with expired food. It is an unfortu-
nate and sometimes unavoidable consequence of the food business. Sustainable
companies should predefine the proper channels for food donation and diversion,
and once those channels are exhausted, follow a destruction plan that does not
ignore sustainability. Companies should create operating procedures to assure
removal of packaging materials such as pallet wrapping, corrugated boxes, plastic
food packaging so that it may be recycled and so that the perishable food matter is
separate and the only part that is either incinerated or sent to a landfill. Ideally,
unless the food producer is at fault, it should not be always responsible for the entire
cost of destruction. In instances when product must be destroyed due to the end of
shelf life, a shared cost would be a reasonable way to assure that it is a last resort,
and that neither producer, broker, nor customer are compelled to force destruction
before a reasonable consideration of the other potential options.
1.1.3.5 Scandal
Unfortunately, incidents of food fraud concerning shelf life have occurred. The
owner and a top executive of US egg company were found guilty of distributing
adulterated eggs and were sentenced by the federal government to prison time and
fines (U.S. Department of Justice 2015). Among the various violations, they were
found guilty of instructing employees affix labels to egg shipments that indicated
false expiration dates with the intent to mislead state regulators and retail egg cus-
tomers regarding the true age of the eggs.
In Huanan province in China, several gangs smuggled frozen meat that was so
old that it was dubbed, “zombie meat” since it was up to 40 years old (Alice 2017).
The rotten meat included pork’s feet and chicken’s claws, chicken’s wings and other
meat products were moved to the mainland via Vietnam, with smugglers hiring resi-
dents of border areas to move the products to Chinese border cities and then on to
Changsha before the products were transported to several sites within China.
Importers soaked them in hydrogen peroxide, a banned food addictive, to make
them look healthy and fresh and to extend their shelf life (Alice 2017). Global, pub-
licly traded quick-serve restaurant chains were involved in this scandal, which led
to substantial loss in share price. This and other non-shelf life-related food safety
scandals in China prompted a comprehensive revision of the 2009 Food Safety Law
of China on April 24, 2015 (Geng et al. 2015).
Scandals related to shelf life fraud such as these are likely to diminish because
modern food and beverage production and brokering of food commodities have
reached unprecedented levels of transparency. Consumers have increasing concerns
about the safety, quality, and authenticity of foods as a result of various food scan-
dals, recalls, and outbreaks (Bánáti 2011; de Jonge et al. 2010). These concerns
have translated to the need for more visibility into food and agriculture production
systems, between businesses, and between business and consumers. Among those
14 P. J. Taormina
concerns, the shelf life of foods has emerged as one of the key areas. With the
intense scrutiny of food safety and quality systems through auditing and quality data
management and sharing systems, gone are the days when shelf life of perishable
foods can be arbitrarily set. Further, the unscrupulous practice of deeming shelf life
longer than technologically supported will no longer go unnoticed and without con-
sequence. Therefore, determination of shelf life of perishable foods must be a sci-
entific process following conventional principles.
1.2 Principles of Shelf Life Determination
1.2.1 Quality Deterioration Rates
The shelf life of foods is simply the duration of time from the point of production
that the product is safe, wholesome, and suitable for consumption. Determining
those factors, however, is not simple due to the variety of intrinsic properties that
interactively affect shelf life performance and the high likelihood of variability in
the external environment of temperature, humidity, light, and time (Office of
Technology Assessment 1979; Labuza 1984; Fu and Labuza 1993). Shelf life of
perishable foods is generally based upon the rate of microbial, chemical, and physi-
cal degradation, which work individually or, more often, in concert to speed the rate
of organoleptic deterioration of the product. Of each of the external factors, tem-
perature has the greatest impact on the rate of deterioration of most perishable
foods. In order to understand the quality deterioration of foods, a few mathematical
equations and concepts are presented, but this chapter does not provide a full picture
of the quality deterioration reaction kinetics and equations. A full review of such
information can be found from various other sources (Office of Technology
Assessment 1979; Singh 1994; Singh and Cadwallader 2004; Van Boekel 2008;
Labuza 1984; Fu and Labuza 1993). A brief overview of quality deterioration reac-
tion kinetics gleaned from these sources will be presented here in order to introduce
the concept of quality and shelf life.
The decrease in a quality attribute of food during storage can be defined in a first-
order reaction. The temperature-dependent general loss of food quality can be rep-
resented by a mathematical Eq. (1.1):
dA
Rate = = kAn (1.1)
dθ
where:
A = the quality factor to be measured
𝛳 = time
k = a constant that depends on temperature and other factors (e.g., water
activity, pH)
n = a power factor called the order of the reaction, which defines whether the
reaction rate is independent of the amount of quality remaining
dA
= the rate of change of A with time. A negative sign is used if the deteriora-
dθ
tion is a loss of A, and a positive sign is used if it is for production of an undesirable
end product of deterioration.
This general loss of quality equation results in a rate calculation, but in practice,
the results of shelf life studies are rather defined as the amount of A left or produced
as a function of time.
Based on Eq. (1.1), the order of the reaction (n) is often treated as if it is equal to
zero. This zero-order reaction assumes that the rate of loss is constant for some
constant temperature:
dA
Rate of loss = = k = Constant at some temperature (1.2)
dθ
If that equation is integrated, it becomes Eq. (1.3):
Amount left ( A ) = Initial amount ( A0 ) − kθ (1.3)
where:
A0 is the initial (time zero) value of the quality factor.
To put this in terms of shelf life, the equation becomes (Eq. 1.4)
A0 − As
θs = (1.4)
k
where:
ϴs is end of shelf life in time (in days, weeks, and months)
As is the maximum allowable loss value, or the threshold point at which quality
has deteriorated such that the product no longer has acceptable quality.
The rate of deterioration becomes the rate constant in Eq. (1.5):
100%
Rate of quality loss = k = = Constant % loss per time (1.5)
θs
This zero-order reaction means that n from Eq. (1.1) must equal 0. The constant, k,
could be different factors such as constant rate of development of oxidative rancid-
ity or vitamin loss. However, not every quality factor will deteriorate at a constant
rate, such as described by a zero-order reaction. Particularly, microbial growth tends
to follow a first-order, or exponential, reaction for which n = 1. Mathematically, the
rate of loss for first-order reactions is shown in Eq. (1.6):
16 P. J. Taormina
−dA
Rate of loss = = k ( A) (1.6)
dθ
Integrating Eq. (1.6) gives a logarithmic function of the first-order as shown in

Eq. (1.7):
A
1n = − kθ (1.7)
A0
Graphically, the zero- and first-order reactions with respect to shelf life of foods are
shown in Fig. 1.3.
Shelf life limit (ϴs) would be set at the point when the zero-order reaction (linear
decrease in quality over time) crosses the As. However, as mentioned many deterio-
ration scenarios follow the first-order reaction rate, and sometimes the As will not be
reached graphically as the quality tails off asymptotically. Rather than claiming a
length of shelf life days far beyond where that curve may extrapolate, a more rea-
sonable choice is to plot the quality factor as semi-log and select a higher As (i.e., a
higher quality standard) such that the first order quality deterioration curve plotted
logarithmically crosses As at some ϴs as shown in Fig. 1.4.
Fig. 1.3 Loss of food quality as a function of time showing difference between zero- and first-
order reactions. End of shelf life time (ϴs) occurs when quality reaches maximum allowable loss
value (As). (Adapted from Labuza 1984, with permission)
Fig. 1.4 Loss of food quality for a first-order reaction plotted as semilog. In this plot, the end of
shelf life (ϴs) occurs when quality reaches maximum allowable loss value (As). (Adapted from
Labuza 1984, with permission)
The term As could also be considered a percent of usable quality remaining. The
difficulty is in designating the criteria for As. This can be a complicated quality
metric based upon sensory research, or it can be an arbitrary value based upon popu-
lations of aerobic mesophilic microorganisms measured as the aerobic plate count
(APC) and/or lactic acid bacterial counts (“Lactics”) as measured by anaerobic,
mesophilic incubation on specific media. If only one factor can be used, it is best to
select the most pertinent quality metric that degrades the fastest. It may very well be
that for some products, APC or Lactics are indicative of organoleptic degradation
and would be ideal for As.
1.2.2 Defining Quality Factors
Food are inherently unstable, as each type of quality deterioration reaction will
proceed at some rate affected by intrinsic properties of the food, as well as the
extrinsic factors like light, temperature, humidity, and atmosphere. Food quality is
what is usually measured to determine the shelf life of foods, but the safety factors
like bacterial pathogen growth over time at the given storage temperature, take pre-
cedence if they are relevant. In food product development, the quality metric must
18 P. J. Taormina
first be defined for the food product based on specifications of microbial, chemical,
physical, and organoleptic attributes. The measurement of the quality of the food
pertains to the degree to which it meets these predetermined attributes. The ultimate
shelf of food is influenced by what the food composition is, how the food is pro-
cessed, how it is packaged, and how it is stored. Indicators of food quality change
over time and can be classified in four ways (Van Boekel 2008):
• Chemical reactions, principally from oxidation or Maillard reactions
• Microbial growth, leading to spoilage and, if pathogens grow, unsafe food
• Biochemical reactions: endogenous enzymes that catalyze reactions leading to
quality loss (enzymatic browning, lipolysis, proteolysis, etc.)
• Physical reactions: such as moisture migration leading to staling, softening, or
“freezer burn”; loss of heterogeneity of dispersed or suspended particles leading
to coalescence, aggregation, and sedimentation
1.2.2.1 Microbiological Profiles
Decades prior to the time of this publication, little was known about the relationship
between microbial activity and biochemical spoilage parameters of food under dif-
ferent packaging and storage conditions (in’t Veld 1996). Back then, the ability to
characterize the total microflora and metabolites developing during food spoilage,
but the ability to identify specific microorganisms in relation to food composition
was lacking (in’t Veld 1996). Now metagenomics techniques coupled with bioinfor-
matics have been applied to food shelf life research (Benson et al. 2014). Quantitative
PCR (qPCR) of 16S rRNA can be used to confirm dominant and subdominant spe-
cies of bacteria that are contributing the most to the overall microbial population, as
well as understand the role of bioprotective cultures in food systems and investigate
how environmental processing conditions impact these microbial communities
(Cauchie et al. 2017; Rouger et al. 2018). Advanced molecular techniques are
beginning to be applied within the food industry, but the standard plate count tech-
niques are by far more utilized and will continue to be into the future; plate count
limits are still ingrained in specifications within the food business and in regulatory
inspections.
Setting the minimum acceptable quality of foods requires attention to the scien-
tific requirements of maintaining wholesomeness, as well as regulatory limits and
customer required limits. For example, there may be a customer limit for Aerobic
Plate Count of 20,000 CFU/mL, and so that it would be at least one criterium for
end of shelf life. Microbial degradation at refrigeration temperatures is commonly
the limiting factor for shelf life as byproducts of microbial metabolism alter the
physiology and integrity of food systems (Gram et al. 2002; Remenant et al. 2015).
As mentioned previously, microbial growth curves follow first-order reaction kinet-
ics. Predictive microbiology models can account for microbial ecology and physiol-
ogy to accurately estimate the remaining shelf life of such food systems (McMeekin
and Ross 1996). One such example shown in Fig. 1.5 is the growth of Photobacterium
Fig. 1.5 Growth of

Photobacterium
phosphoreum in naturally
contaminated cod fillets
stored at 0 °C in an
atmosphere with 100% N2.
Error bars indicated
standard deviations (n = 2).
From Dalgaard et al.
(1997) with permission
phosphoreum in cod fillets stored at 0 °C (Dalgaard et al. 1997). Growth of the

spoilage bacterium was predicted and associated with organoleptic failure, which
goes beyond the arbitrary plate count limit of quality and shows how populations of
a specific spoilage bacterium correspond to real degradation. Shelf life prediction
based upon microbial growth at non-isothermal conditions simulating distribution
and storage variation has been researched for a wide variety of perishable foods
including fish (Koutsoumanis 2001) and ham (Kreyenschmidt et al. 2010) for exam-
ple. There have been many more applied research studies concerning predictive
microbial modeling for shelf life prediction.
1.2.2.2 Consumer Acceptance
There are different levels of quality, and interpretation of product quality involves
laboratory techniques, as well as sensory science. Otherwise, quality becomes
amorphous and/or subjective. While consumers are considered the most appropriate
gauge of determining the shelf life of food, assemblage of consumer panels for
multiple measurements necessary for shelf life studies is not feasible (Gámbaro
et al. 2004). Nonetheless, the performance of consumer sensory analysis can be
achieved for larger product groups, and data resulting from such work can form the
basis for shelf life prediction for product categories. For example, during the prod-
uct development phase for a line of Greek-style yogurts, scientists should conduct
consumer acceptance testing on a model product formulation to establish the likely
shelf life of the product category. With such data in hand, future product variations
(e.g., flavors, packaging types, portions) could reasonably be expected to perform
similarly over time. Consumer preference of one product over another is an impor-
tant determination that impacts the shelf life of products. Depending on the product
attributes, as the food ages, the degree of consumer acceptance can diminish. So, a
20 P. J. Taormina
decision must be made as to the acceptable quality attributes of the product during
shelf life. Consumer acceptance of food products varies with individuals, and so the
shelf life of foods depends not only on the food quality but also on the interaction of
the food with the consumer (Hough et al. 2003).
Survival analysis is a branch of statistics that has been applied to the study of the
shelf life of foods (Hough et al. 2003; Gámbaro et al. 2004). The focus of survival
analysis is on the risk of consumer rejection of the product and that impacts on shelf
life, rather than on the product deterioration. Another quality designation that helps
determine the shelf life of products is the degree to which a product meets customer
and consumer expectations at a base level. Determination that a product is suitable
or fit for purpose means that the product meets the minimum standards. The “Just
Noticeable Difference” (JND) concept applies to sensory evaluations performed by
trained panel and/or consumers. The shelf life may also be limited by consumer
complaints of some factor pertaining to not only the product itself, but also the way
the product performs in the package on display.
1.2.3 Shelf Life Testing
Assigning a shelf life to a product involves the research and development process of
formulation, processing parameters, packaging, and consumer acceptance testing.
The deterioration of food can be due to microbial, chemical, or physical factors, and
all of these can singly or in combinations affect the organoleptic properties of the
product. Organoleptic attributes of appearance, odor, taste, and texture are each
important and measurable. The factors that impact shelf life can be safety related,
such as growth of foodborne pathogens over time, or can be quality related, such as
growth of spoilage microorganism. While food safety implications are the limiting
factor in many perishable foods, the ways this is determined are much different than
the quality factors.
Shelf life testing occurs at various stages and involves differing approaches at
each stage of a product life cycle (Table 1.1). The process of establishing a product
shelf life occurs in the new product development phase or the research and develop-
ment phase. New product development entails something altogether new and differ-
ent from existing product lines. Such novelty necessitates relatively elaborate shelf
life testing in order to determine the usable days. Challenge studies are an important
aspect of measuring the potential for pathogen growth during shelf life (National
Advisory Committee on Microbiological Criteria for Foods 2010) and should be
conducted early enough in the process in order to obtain results prior to decisions to
launch new products. Routine shelf life testing involves collecting freshly manufac-
tured food products in their finished, packaged state, and storing them at a defined
refrigerated temperature and periodically analyzing samples for some of the afore-
mentioned factors. Changes such as microbial growth, oxidative rancidity develop-
ment, organoleptic degradation (e.g., color, flavor, texture), or increase in purge are
monitored over time until thresholds of acceptance are met or exceeded. At that
Table 1.1 Types of shelf life testing different stages of food product development and impact of
data generated
Stage of
product
development Type of shelf life testing Use of data
1. Ideation/ Predictive microbial modeling Go/no go decisions
proof of
concept
2. Prototype Chemistry (proximates, pH, aw); predictive Attain food safety and
development microbial modeling quality approval to develop
further
3. Pilot plant Shelf life testing at refrigeration and moderately Attain food safety and
testing abusive temperature; measuring general quality approval for plant
microflora, fat oxidation, texture, and production; set shelf life
organoleptic acceptance; microbial challenge target days (i.e., code date)
studya
4. Scaled-up Shelf life testing at refrigeration temperature; Food safety and quality
plant trials measuring general microflora, fat oxidation, validation
texture, and organoleptic acceptance; microbial
challenge studya
5. First Shelf life testing of representative samples; Quality validation
production run samples stored at refrigeration and analyzed for
organoleptic, general microflora, fat oxidation
6. Routine Shelf life testing of representative samples Quality monitoring
production retrieved at a necessary frequency in order to
assess quality; samples stored at refrigeration
and analyzed for organoleptic, general
microflora, fat oxidation
a
Microbial challenge testing will take 1.25–1.5 times the shelf life target. Therefore, challenge
study results for a product with 30-day target shelf life would not be completed until up to 45 days.
For products with a long, refrigerated shelf life (e.g., 120 days), there may be a need to perform a
challenge study earlier in the process, such as at the pilot plant stage
time, a shelf life test is deemed complete, and the number of days the product was
acceptable is considered the result. Alternatively, a pass or fail rating is given, and
those results are aggregated over time. Routine shelf life testing involves no intro-
duction of other microorganisms to the food product. It is simply measuring the
behavior of the autochthonous microflora over time, along with the other factors.
Shelf life testing is beneficial in the sense that it provides an actual profile of the
product as it changes over refrigerated storage. It is limited in the sense that the level
and type of autochthonous microflora present from production lot to production lot
can vary. Also, there are sometimes variations between samples collected from one
particular lot. Due to cost, limited refrigerated storage space for samples, and labo-
ratory labor required, shelf life testing is usually performed only periodically and
often involves analysis of only one packaged unit per time point. Indeed, if dupli-
cate or triplicate samples are not analyzed at each time point, the sample variability
can lead to inaccurate results and conclusions. It is often the case that one packaged
food product sample out of several will succumb to spoilage days or weeks before
22 P. J. Taormina
the other samples from the same lot exhibit signs of degradation. Data in Table 1.2
would indicate that everything is performing adequately. The starting populations
are low as expected, and even though the T14 sample shows markedly higher counts
for both APC and Lactics, the subsequent sample at T28 is comfortably back into
the acceptability limits. The final analytical unit sampled at T60 indicates that popu-
lations were approaching the limit but were still sufficiently below thresholds.
Therefore, by policy, the technician would deem the result a “pass.” Such data
would be aggregated into pass/fail shelf life trend reports, and the quality manager
would assume that the data are indicative of acceptable conditions in the production
environment and within the product. However, what if the analytical unit sampled at
T14 were to be instead sampled at T28 or T60? Would it have still had acceptable
plate counts or would that population have grown much more rapidly than within
other samples? Is this shelf life test based upon destructive sampling of only four
analytical units enough from which to draw conclusions, or does it only provide
misleading, anecdotal information? Table 1.3 demonstrates how inclusion of dupli-
cate samples might avoid instances of misinterpretation of microbial counts in shelf
life testing. Table 1.2 data represent a scenario where a sample set of four analytical
units were collected the beginning of the production run, while a second sample set
of four analytical units were collected from near the end of the production run. T0
samples from both sets were analyzed, and then the remaining units were stored at
4.4 °C. While data generated from the first sample set (Sample 1) show acceptable
results, the data from Sample Set 2, which were collected from the production line
later, surpassed the acceptable limits for APC at T28. Incidentally, the Lactics data
at T28 were still barely acceptable, while APC surpassed the limits. This is a com-
mon observation in plant-derived samples, as often the populations on APC are
inclusive of many of the same lactic acid bacteria that were enumerated on the
Lactics count. At any rate, the result is much different, deemed a “Fail” at T28. The
span of time between analyses of samples should factor into interpretation of results.
For example, a failure from at least one analytical unit occurred at T28, but the prior
sample was analyzed at T14. If data were being generated about the number of days
of shelf life achieved, then listing either T14 or T28 would falsely skew the result.
An arbitrary number half way between the two (i.e., T21) is a suitable way to esti-
mate days achieved.
Table 1.2 Routine shelf life data from plant-produced samples for which criteria for end of shelf
life is 1,000,000 CFU/g for either APC or Lactics
Day of Aerobic plate count Lactic acid bacteria count Interpretation of
sample (CFU/g) (CFU/g) result
T0 <1000 1000 Acceptable
T14 322,000 103,000 Acceptable
T28 15,000 24,000 Acceptable
T60 700,000 475,000 Acceptable
Table 1.3 Routine shelf life data from replicate plant produced samples for which criteria for end
of shelf life is 1,000,000 CFU/g for either APC or Lactics
Day of Aerobic plate count Lactic acid bacteria count Interpretation of
sample (CFU/g) (CFU/g) result
Sample Sample
Sample set 1 set 2 Sample set 1 set 2
T0 <1000 2000 1000 2000 Acceptable
T14 322,000 250,000 103,000 140,000 Acceptable
T28 15,000 1,075,000 24,000 920,000 Fail
T60 700,000 2,500,000 475,000 1,700,000 Fail
1.2.4 Summary
This book focuses on perishable foods, as there is little to no microbial degradation

on low-water activity foods. Indeed, the shelf life of low moisture foods involves
principally lipid oxidation (Hu 2016), physical decomposition such as staling, and
the resulting organoleptic failure over time. Also, this work is not the same as food
spoilage, as that was covered in another title in this series (Sperber and Doyle 2009).
Rather, the book focuses on the procedures utilized to establish and monitor shelf
life of various food types, with subsequent chapters covering in more detail the
metrics for analysis of shelf life of foods, the interpretation of data to set code dates,
the types of microorganisms that must be monitored during shelf life testing, and the
techniques that can be employed to extend shelf life of perishable foods.
References
Ahumada, Omar, and J. Rene Villalobos. 2009. Application of planning models in the Agri-Food
supply chain: A review. European Journal of Operational Research 196 (1): 1–20. https://doi.
org/10.1016/j.ejor.2008.02.014.
Akkerman, Renzo, Poorya Farahani, and Martin Grunow. 2010. Quality, safety and sustainability
in Food distribution: A review of quantitative operations management approaches and chal-
lenges. OR Spectrum 32 (4): 863–904. https://doi.org/10.1007/s00291-010-0223-2.
Alice, Giusto. 2017. “Language and Food Safety: The ‘Zombie Meat’ Scandal.” Вестник Санкт-
Петербургского Университета. Серия 13. Востоковедение. Африканистика 9 (1).
Amorim, P., H. Meyr, C. Almeder, and B. Almada-Lobo. 2013. Managing perishability in
production-distribution planning: A discussion and review. Flexible Services and Manufacturing
Journal 25 (3): 389–413. https://doi.org/10.1007/s10696-011-9122-3.
Anonymous. 1996. “Bill Emerson Good Samaritan Food Donation Act.” 42 U.S. Code § 1791.
United States of America.
———. 2011. Food safety modernization act domestic and foreign facility re-inspections, recall,
and importer re-inspection user fee rates for fiscal year 2012. Federal Register 76 (147):
45820–45825.
Apaiah, Radhika, K. Eligius, M.T. Hendrix, Gerrit Meerdink, and Anita R. Linnemann. 2005.
Qualitative methodology for efficient Food chain design. Trends in Food Science & Technology
16 (5): 204–214. https://doi.org/10.1016/j.tifs.2004.09.004.
24 P. J. Taormina
Aschemann-Witzel, Jessica, Ilona de Hooge, Pegah Amani, Tino Bech-Larsen, and Marije
Oostindjer. 2015. Consumer-related Food waste: Causes and potential for action. Sustainability
7 (6): 6457–6477. https://doi.org/10.3390/su7066457.
Bánáti, Diána. 2011. Consumer response to Food scandals and scares. Trends in Food Science &
Technology 22 (2): 56–60.
Benson, Andrew, K. Jairus, R.D. David, Stefanie Evans Gilbreth, Gordon Smith, Joseph Nietfeldt,
Ryan Legge, Jaehyoung Kim, et al. 2014. Microbial successions are associated with changes in
chemical profiles of a model refrigerated fresh pork sausage during an 80-day shelf life study.
Edited by D W Schaffner. Applied and Environmental Microbiology 80 (17): 5178–5194.
https://doi.org/10.1128/AEM.00774-14.
Brul, S., and P. Coote. 1999. Preservative agents in foods: Mode of action and microbial resis-
tance mechanisms. International Journal of Food Microbiology 50 (1–2): 1–17. https://doi.
org/10.1016/S0168-1605(99)00072-0.
Cauchie, Emilie, Mathieu Gand, Gilles Kergourlay, Bernard Taminiau, Laurent Delhalle, Nicolas
Korsak, and Georges Daube. 2017. The use of 16S RRNA gene Metagenetic monitoring of
refrigerated Food products for understanding the kinetics of microbial subpopulations at dif-
ferent storage temperatures: The example of white pudding. International Journal of Food
Microbiology 247: 70–78. https://doi.org/10.1016/j.ijfoodmicro.2016.10.012.
Dalgaard, Paw, Ole Mejlholm, and Hans Henrik Huss. 1997. Application of an iterative approach for
development of a microbial model predicting the shelf-life of packed fish. International Journal
of Food Microbiology 38 (2–3): 169–179. https://doi.org/10.1016/S0168-1605(97)00101-3.
David, Jairus R.D., Larry R. Steenson, and P. Michael Davidson. 2013. Expectations and appli-
cations of natural antimicrobials to foods. Food Protection Trends 33 (4): 238–247. http://
www.researchgate.net/profile/Jairus_David/publication/270273839_PEER-REVIEWED_
ARTICLE/links/54a4239e0cf257a636071d3b.pdf.
de Jonge, Janneke, Hans Van Trijp, Reint Jan Renes, and Lynn J. Frewer. 2010. Consumer con-
fidence in the safety of food and newspaper coverage of food safety issues: A longitudinal
perspective. Risk Analysis 30 (1): 125–142. https://doi.org/10.1111/j.1539-6924.2009.01320.x.
Fu, Bin, and Theodore P. Labuza. 1993. Shelf-life prediction: Theory and application. Food
Control 4: 125–133. https://doi.org/10.1016/0956-7135(93)90298-3.
Gámbaro, Adriana, S. Fiszman, A. Giménez, P. Varela, and A. Salvador. 2004. Consumer accept-
ability compared with sensory and instrumental measures of white Pan bread: Sensory shelf-
life estimation by survival analysis. Journal of Food Science 69 (9): S401–S405.
Geng, Shu, Xu Liu, and Roger Beachy. 2015. New Food safety law of China and the special issue
on Food safety in China. Journal of Integrative Agriculture 14 (11): 2136–2141. https://doi.
org/10.1016/S2095-3119(15)61164-9.
Gram, Lone, Lars Ravn, Maria Rasch, Jesper Bartholin Bruhn, Allan B. Christensen, and
Michael Givskov. 2002. Food spoilage-interactions between Food spoilage Bacteria.
International Journal of Food Microbiology 78 (1–2): 79–97. https://doi.org/10.1016/
S0168-1605(02)00233-7.
Grunert, Klaus G. 2002. Current issues in the understanding of consumer Food choice. Trends in
Food Science & Technology 13 (8): 275–285. https://doi.org/10.1016/S0924-2244(02)00137-1.
Grunow, Martin, and Selwyn Piramuthu. 2013. RFID in highly perishable Food supply chains -
remaining shelf life to supplant expiry date? International Journal of Production Economics
146 (2): 717–727. https://doi.org/10.1016/j.ijpe.2013.08.028.
Gunders, Dana. 2012. Wasted: How America is losing up to 40 percent of its Food from farm to
fork to landfill. Natural Resources Defense Council: 1–26.
Hough, G., K. Langohr, G. Gómez, and A. Uria. 2003. Survival analysis applied to sensory shelf life
of foods. Journal of Food Science 68 (1): 359–362. https://doi.org/10.1111/j.1365-2621.2003.
tb14165.x.
Hu, Min. 2016. Oxidative stability and shelf life of low-moisture foods. In Oxidative stability
and shelf life of foods containing oils and fats, ed. Min Hu and Charlotte Jacobson, 313–371.
Amsterdam: AOCS Press.
in’t Veld, Jos H. Huis. 1996. Microbial and biochemical spoilage of foods: An over-
view. International Journal of Food Microbiology 33 (1): 1–18. https://doi.
org/10.1016/0168-1605(96)01139-7.
Joint Industry Unsaleables Leadership Team. 2008. “Joint Industry Unsaleables Report: The Real
Causes and Actionable Solutions.”
Kaipia, Riikka, Iskra Dukovska-Popovska, and Lauri Loikkanen. 2013. Creating sustainable fresh
Food supply chains through waste reduction. International Journal of Physical Distribution
and Logistics Management 43 (3): 262–276. https://doi.org/10.1108/IJPDLM-11-2011-0200.
Kilcast, David. 2001. Shelf-life evaluation of foods (second edition). International Journal of Food
Science & Technology 36 (8): 856. https://doi.org/10.1046/j.1365-2621.2001.0530b.x.
Koutsoumanis, Konstantinos. 2001. Predictive modeling of the shelf life of fish under noniso-
thermal conditions. Applied and Environmental Microbiology 67 (4): 1821–1829. https://doi.
org/10.1128/AEM.67.4.1821-1829.2001.
Kreyenschmidt, J., A. Hübner, E. Beierle, L. Chonsch, A. Scherer, and B. Petersen. 2010.
Determination of the shelf life of sliced cooked ham based on the growth of lactic acid Bacteria
in different steps of the chain. Journal of Applied Microbiology 108 (2): 510–520. https://doi.
org/10.1111/j.1365-2672.2009.04451.x.
Labuza, T.P. 1984. Application of chemical kinetics to deterioration of foods. Journal of Chemical
Education 61 (4): 348. https://doi.org/10.1021/ed061p348.
Lebersorger, S., and F. Schneider. 2014. Food loss rates at the Food retail, influencing factors and
reasons as a basis for waste prevention measures. Waste Management (New York, NY) 34 (11):
1911–1919.
Man, C.M.D. 2004. Shelf-life testing. In Understanding and Measuring the Shelf-
Life of Food, 340–356. Sawston: Woodhead Publishing. https://doi.org/10.1016/
B978-1-85573-732-7.50019-6.
Man, C.M.D., and Adrian A. Jones. 1994. Shelf life evaluation of foods. New York: Springer.
Martins, R.C., V.V. Lopes, A.A. Vicente, and J.A. Teixeira. 2008. Computational shelf-life dating:
Complex systems approaches to Food quality and safety. Food and Bioprocess Technology 1
(3): 207–222. https://doi.org/10.1007/s11947-008-0071-0.
McMeekin, Thomas A., and Thomas Ross. 1996. Shelf life prediction: Status and future
possibilities. International Journal of Food Microbiology 33: 65–83. https://doi.
org/10.1016/0168-1605(96)01138-5.
Mena, Carlos, B. Adenso-Diaz, and Oznur Yurt. 2011. The causes of Food waste in the supplier–
retailer Interface: Evidences from the UK and Spain. Resources, Conservation and Recycling
55 (6): 648–658.
National Advisory Committee on Microbiological Criteria for Foods. 2010. Parameters for deter-
mining inoculated pack/challenge study protocols. Journal of Food Protection 73: 140–202.
https://doi.org/10.4315/0362-028X-73.1.140.
Newsome, Rosetta, Chris G. Balestrini, Mitzi D. Baum, Joseph Corby, William Fisher, Kaarin
Goodburn, Theodore P. Labuza, Gale Prince, Hilary S. Thesmar, and Frank Yiannas. 2014.
Applications and perceptions of date labeling of Food. Comprehensive Reviews in Food
Science and Food Safety 13 (4): 745–769.
Office of Technology Assessment, U.S. Government Printing Office. 1979. “Open Shelf-Life
Dating of Food.”
Pothakos, Vasileios, Bernard Taminiau, Geert Huys, Carine Nezer, Georges Daube, and Frank
Devlieghere. 2014. Psychrotrophic lactic acid Bacteria associated with production batch recalls
and sporadic cases of early spoilage in Belgium between 2010 and 2014. International Journal
of Food Microbiology 191: 157–163. https://doi.org/10.1016/j.ijfoodmicro.2014.09.013.
Remenant, Benoît, Emmanuel Jaffrès, Xavier Dousset, Marie France Pilet, and Monique Zagorec.
2015. Bacterial spoilers of food: Behavior, fitness and functional properties. Food Microbiology
45: 45–53. https://doi.org/10.1016/j.fm.2014.03.009.
Roberts, T.A., and R.B. Tompkin. 1996. Microorganisms in foods 5: Characteristics of microbial
pathogens. Vol. 5. New York: Springer Science & Business Media.
26 P. J. Taormina
Rouger, Amélie, Nicolas Moriceau, Hervé Prévost, Benoît Remenant, and Monique Zagorec. 2018.
Diversity of bacterial communities in French chicken cuts stored under modified atmosphere
packaging. Food Microbiology 70: 7–16. https://doi.org/10.1016/j.fm.2017.08.013.
Singh, R.P. 1994. Scientific principles of shelf life evaluation. In Shelf Life Evaluation of Foods,
3–26. Cham: Springer Nature. https://doi.org/10.1007/978-1-4615-2095-5_1.
Singh, T.K., and K.R. Cadwallader. 2004. Ways of measuring shelf-life and spoilage. In
Understanding and Measuring the Shelf-Life of Food, 165–183. Sawston: Woodhead
Publishing. https://doi.org/10.1016/B978-1-85573-732-7.50013-5.
Soliva-Fortuny, Robert C., and Olga Martı́n-Belloso. 2003. New advances in extending the shelf-
life of fresh-cut fruits: A review. Trends in Food Science & Technology 14 (9): 341–353.
Sperber, William H., and Michael P. Doyle. 2009. Compendium of the microbiological spoilage of
foods and beverages. New York: Springer.
Tian, Feng. 2016. An agri-food supply chain traceability system for China based on RFID &
Blockchain technology. In 2016 13th international conference on service systems and service
management (ICSSSM), 1–6. https://doi.org/10.1109/ICSSSM.2016.7538424.
———. 2017. “A supply chain traceability system for food safety based on HACCP, block-
chain & internet of things.” 14th International Conference on Services Systems and Services
Management, ICSSSM 2017-Proceedings. https://doi.org/10.1109/ICSSSM.2017.7996119.
Tsiros, Michael, and Carrie M. Heilman. 2005. The effect of expiration dates and perceived risk
on purchasing behavior in grocery store perishable categories. Journal of Marketing 69 (2):
114–129. https://doi.org/10.1509/jmkg.69.2.114.60762.
U.S. Department of Agriculture, and Food Safety and Inspection Service. 2015. “Kraft Heinz
Foods Company Recalls Turkey Bacon Products Due To Possible Adulteration.” https://
www.fsis.usda.gov/wps/portal/fsis/topics/recalls-and-public-health-alerts/recall-case-archive/
archive/2015/recall-113-2015-release.
———. 2015. “Control of Listeria Monocytogenes in ready-to-eat meat and poultry products.”
Vol. 80.
U.S. Department of Justice, and Office of Public Affairs. 2015. “Quality Egg, Company Owner
and Top Executive Sentenced in Connection with Distribution of Adulterated Eggs.” https://
www.justice.gov/opa/pr/quality-egg-company-owner-and-top-executive-sentenced-connec-
tion-distribution-adulterated.
U.S. Food and Drug Administration. 2015. “Current Good Manufacturing Practice, Hazard
Analysis, and Risk-Based Preventive Controls for Human Food.” 21 C.F.R. § 117.
———. 2017. “FDA Food Code.” https://doi.org/10.1016/j.parint.2011.08.011.
Van Boekel, M.A.J.S. 2008. Kinetic modeling of Food quality: A critical review.
Comprehensive Reviews in Food Science and Food Safety 7: 144–158. https://doi.
org/10.1111/j.1541-4337.2007.00036.x.
Van Boxstael, S., F. Devlieghere, D. Berkvens, A. Vermeulen, and M. Uyttendaele. 2014.
Understanding and attitude regarding the shelf life labels and dates on pre-packed Food
products by Belgian consumers. Food Control 37 (1): 85–92. https://doi.org/10.1016/j.
foodcont.2013.08.043.
Wee, H.M., 1993. Economic production lot size model for deteriorating items with partial back-
ordering. Computers & Industrial Engineering, 24(3), pp. 449–458.
Wansink, Brian, and Alan O. Wright. 2006. ‘Best if used by …’ how freshness dating
influences Food acceptance. Journal of Food Science 71 (4): S354–S357. https://doi.
org/10.1111/j.1750-3841.2006.00011.x.
Xu, Y. and Sarker, B.R., 2003. Models for a family of products with shelf life, and production and
shortage costs in emerging markets. Computers & Operations Research, 30(6), pp. 925–938.
Chapter 2
Food Safety Factors Determining
Shelf Life
Margaret D. Hardin
2.1 Introduction
The shelf life of food products is often defined as the recommended maximum
amount of time that food products can be stored under specified conditions of tem-
perature, humidity, and other external factors while maintaining acceptable quality
without exhibiting spoilage. Spoilage of perishable foods may be the result of
changes in the sensory characteristics of the product, such as the development of
off-flavors, off-colors, gas, or slime, which make the product undesirable or unac-
ceptable to the consumer, or by exceeding a certain level of indicator organisms
specified for the product. Many of these undesirable changes are the result of micro-
bial growth and the associated metabolic activity and by-products of the organism(s).
Food products naturally contain microorganisms that can include spoilage and/or
pathogenic microorganisms. However, the growth of pathogenic microorganisms in
food products seldom coincides with noticeable spoilage characteristics such as off-
odor, discoloration, gas, or slime. Microorganisms that spoil foods and those that
are of public health significance can do so at various times in the process including
before and during preparation and/or processing, under normal conditions of stor-
age and intended use, particularly if not destroyed or controlled by normal process-
ing techniques. While a range of methods such as salting, curing, smoking, freezing,
and canning have been successfully used over the years for extending the shelf life
of foods, consumers seem to prefer fresh foods over frozen or shelf-stable foods. In
addition, consumers are becoming more aware of product labels and are demanding
products that are preservative-free and have minimal processing. Consumers are
M. D. Hardin (*)
Vice President of Technical Services, IEH Laboratories and Consulting Group,
Lake Forest Park, WA, USA
e-mail: mh@iehinc.com

https://doi.org/10.1007/978-3-030-54375-4_2
28 M. D. Hardin
avoiding products containing ingredients that are not recognizable or perceived as

natural. Consumer demand for so-called clean label products is increasing. These
ready-to-eat (RTE) and perishable fresh foods that have an enhanced but limited
shelf life rely upon time and temperature as critical factors for maintaining micro-
biological quality and safety.
With few exceptions, refrigerated foods have had a very good history of safety.
However, over the past 20–30 years, innovations in refrigerated foods with an
extended shelf life have gained in popularity. Various methods of processing (e.g.,
flash pasteurization, high pressure processing) and packaging (e.g., vacuum packag-
ing and modified atmosphere packaging) have added extra days, and even weeks or
months, to the refrigerated shelf life of these products. The benefits of an extended
shelf life include products that can remain on the shelf at retail or in a consumer’s
refrigerator for longer periods of time; reduced waste; reduced product returned to
the manufacturer; increased distribution of perishable products over a wider geo-
graphical area; and the production and extended storage of seasonal products. These
refrigerated foods include conventional RTE products such as luncheon meats and
sausages (both cured and uncured), as well as refrigerated salads containing meat,
egg and/or seafood, fresh pasta and pasta sauces, soups, sauces, entrees, complete
meals, as well as minimally processed and novel products such as sous-vide type
foods (Marth 1998; Austin 2001).
2.2 Food Safety Concerns for Extended Shelf Life Foods
With the growing demand for minimally processed refrigerated foods with extended
shelf life, concern has increased related to the risk associated with RTE foods that
require refrigeration and support the growth of psychrotrophic pathogens (NACMCF
2005; Marth 1998; FDA 2003). Four pathogens of concern capable of growth at
refrigeration temperatures have been identified for this group of products (i.e.,
refrigerated RTE foods): Yersinia enterocolitica, Bacillus cereus, nonproteolytic
Clostridium botulinum, and Listeria monocytogenes. These four organisms were
identified by an Institute of Food Technologists (IFT) expert panel on food safety
and nutrition in 1998 (Marth 1998) and again in 2005 by US National Advisory
Committee on Microbiological Criteria in Foods (NACMCF), when they performed
a hazard analysis to identify microorganism of concern when establishing safety-
based date labeling (NACMCF 2005). While all of these pathogens are capable of
growth at refrigeration temperatures, many are underreported due to the self-limiting
nature of the disease, lack of routine laboratory testing, limited availability of rapid
methods, and/or lack of culture procedures available for isolating the organisms. In
addition, some of the illnesses, such as those associated with B. cereus, are not
reportable diseases and are therefore likely underestimated in official reporting
systems.
2 Food Safety Factors Determining Shelf Life 29
2.2.1 Yersinia enterocolitica
The genus Yersinia includes both pathogenic and nonpathogenic strains of the
organism. Of the pathogenic strains, Yersinia enterocolitica is the strain most com-
monly associated with outbreaks of foodborne illness. Y. enterocolitica is generally
ubiquitous in the environment and has been recovered from a wide variety of ani-
mals (dogs, cats, birds, monkeys, and shellfish), foods (milk and raw milk, raw
pork, prepared foods, vegetables, etc.) and water (Barton and Robins-Browne 2003;
Austin 2001; ICMSF 1996; Kapperud 1991; Robins-Browne 2001; Ackers et al.
2000). Low recovery rates are often attributable to a lack of routine testing in clini-
cal situations and during outbreak investigations and to the limited of sensitivity of
available culture methods (ICMSF 1996; Robins-Browne 2001; Barton and Robins-
Browne 2003). Although pets may be occasional carriers of Y. enterocolitica, pigs
are considered a major primary source of yersiniosis infections in humans particu-
larly those caused by bioserotype 4,O:3. Symptom of the illness, yersiniosis, include
abdominal pain (sometimes confused with appendicitis), headache, fever, diarrhea,
nausea, vomiting. Data has shown that Y. enterocolitica is able to grow over a wide
range of temperatures from temperatures near 0 °C to 42 °C, particularly when con-
ditions for growth are most favorable (ICMSF 1996; Kapperud 1991). The ability of
the organism to grow in foods when stored at very low temperatures will vary with
other factors including substrate, pH, gaseous atmosphere, salt, preservatives, or
competing flora (ICMSF 1996; Kapperud 1991; Barton and Robins-Browne 2003).
The true incidence of foodborne yersiniosis is uncertain for various reasons: few
outbreaks of foodborne illness are investigated; yersiniosis has only recently been
known to be food or water borne; long periods of time may be required using cold
enrichment to recover certain strains from food; and not all clinical laboratories
routinely test for the organism. In addition, there are limited methods available that
differentiate between pathogenic and nonpathogenic strains and species of the
organism, and serological and biochemical testing is outside of the scope for most
laboratories. Advances in molecular methods will likely improve detection of
Y. enterocolitica in foods. Although outbreaks of illness are uncommon, the foods
generally associated with illness outbreaks of yersiniosis include milk, water, pro-
duce, and undercooked pork products. Some person-to-person transmission of the
disease has also been reported (Todd et al. 2007). Pigs appear to be the major source
of Y. enterocolitica in foods. Some milk borne outbreaks have been reported in pas-
teurized milk; however, cross-contamination from milk crates used on pig farms
was identified as the source (Barton and Robins-Browne 2003). During an outbreak
investigation of illnesses due to contaminated tofu in Washington State, the same
strain of Y. enterocolitica serotype O:8 was isolated from both the tofu and the
plant’s untreated spring water (Tacket et al. 1985). Untreated water has been identi-
fied as a source of infection in other cases. Spring water was identified as the source
in a case of Yersinia enterocolitica septicemia in New York State (Keet 1974) and
untreated well water with a small outbreak of gastroenteritis in Canada (Thompson
and Gravel 1986). A more recent outbreak of yersiniosis in Norway in 2014
30 M. D. Hardin
i dentified salad mix containing imported radicchio rosso as a source of the illnesses
(MacDonald et al. 2016). Although the organism was never isolated from the salad
mix, numerous failures in hygiene were observed at the production facility, includ-
ing infrequent changing of the water used to rinse the produce. The longer shelf life
of the product was also cited as a contributing factor. This was not the first outbreak
associated with produce in Norway. In 2011, an outbreak of yersiniosis was associ-
ated with ingestion of ready-to-eat salad mix (MacDonald et al. 2012). In this case,
radicchio rosso was identified as the likely source of infection due to the fact that it
can be stored for several months and was the only ingredient included in the sus-
pected salad mix that had delivery, production, and storage dates consistent with the
outbreak. Although investigators were unable to conclusively link any of the iso-
lates from the salad ingredients to the human Y. enterocolitica isolates, they con-
cluded that finding nonpathogenic Yersinia in packaged salads reinforced that the
environment of the food product was processed in was conducive to the persistence
of the bacterium.
As previously mentioned, pigs are a major source of the organism. Outbreaks of
yersiniosis, in the US, primarily with infants and children, have been associated
with cross-contamination during the preparation of raw pork, specifically pork chit-
terlings (Abdel-Hag et al. 2000; Lee et al. 1990; MMWR 2003). Traditionally pre-
pared chitterlings are thoroughly cooked and infections have not been associated
with the consumption of the final product; however, preparation of chitterlings
involves a lengthy and wet process of cleaning and cooking large amounts of raw
pork intestines that may contain the organism. Therefore, during preparation of the
product, there is considerable opportunity for cross-contamination to occur, and
outbreaks have been reported to other prepared foods, people, baby bottles, and
toys, with infants and young children being particularly susceptible to the disease
(Abdel-Hag et al. 2000; Lee et al. 1990). Educational materials targeting consumers
have been developed focusing on preparation of chitterlings in the home (USDA
FSIS 2011). While education of the public on preparation of this product has had
some success, cases do still occur. Since Y. enterocolitica is capable of multiplying
a very low temperatures, refrigerated storage is not generally recommended a means
of preventing outbreaks; however, refrigeration temperatures do prolong lag periods
(ICMSF 1996). Prevention of cross-contamination from untreated water and raw
foods to cooked foods such as unpasteurized milk and raw pork is an essential
aspect of a control program (ICMSF 1996; Barton and Robins-Browne 2003; USDA
FSIS 2011).
2.2.2 Bacillus cereus
Bacillus cereus is a spore-forming bacterium that has been isolated from a wide
variety of products. Due to its widespread presence in nature, it is virtually impos-
sible to obtain raw product that are free of B. cereus spores. Milk products and
products of plant origin are the main sources of B. cereus (Granum 2001; Jenson
and Moir 2003). The bacterium may be transferred to other food products and may
survive, as spores, in heated products where competition for other bacteria is not
usually present. The growth and survival of B. cereus have been studied extensively
over a range of temperatures, pH values, salt concentrations, preservatives, and
other factors. Some strains are psychrotrophic being able to grow at
4–5 °C. Psychrotrophic strains have been a problem for the dairy industry particu-
larly in certain countries and for situations where maintaining low temperatures
(<7 °C) cannot always be assured (Notermans et al. 1997; Granum 2001).
B. cereus is the cause of two different types of food poisoning: an emetic type
and a diarrheal type. The emetic syndrome is caused by the ingestion of the heat-
stable emetic toxin produced in foods, and the diarrheal syndrome is mainly due to
the ingestion of B. cereus cells in foods followed by toxin production in the small
intestine (Granum and Lund 1997; Granum 2001). Carlin et al. (2006) evaluated
100 representative strains of B. cereus selected from a total collection of 372
B. cereus strains in order to investigate differences in the growth limits and heat-
resistance profiles of emetic toxin-producing and non-emetic toxin-producing
strains of B. cereus. Emetic toxin-producing strains were able to grow at 48 °C;
however, none of the emetic toxin-producing strains were able to grow below 10 °C
and spores from the emetic toxin-producing strains showed a higher heat resistance
at 90 °C and a lower germination, particularly at 7 °C, than spores from the other
strains. The authors concluded that while emetic toxin-producing strains of B. cereus
pose a particular risk in heat-processed or preheated foods that are kept warm (such
as hot-holding in restaurants), they will not pose a risk in refrigerated foods (Carlin
et al. 2006). Of the non-emetic toxin-producing strains, 50 strains (28 diarrheal and
28 food-environmental strains) out of 83 were able to grown at 4 °C or 7 °C. All of
the diarrheal strains showed lower D-values that emetic toxin-producing strains,
and none of the diarrheal strains produced the emetic toxin (Carlin et al. 2006).
Granum (1994) evaluated different strains of enterotoxigenic B. cereus and the inac-
tivation of toxin following exposure to the low pH and proteolytic enzymes of the
stomach. The researcher concluded that it is likely that in the case of the diarrheal
illness, food poisoning is caused by ingestion of cells or spores rather than by pre-
formed enterotoxin. In addition, the level of enterotoxin produced by different
strains of B. cereus varies, making it possible that only a few of the enterotoxigenic
strains are of public health significance. The author also concluded that ingestion of
104 to 107 cells or spores are the main cause of food poisoning associated with this
illness syndrome.
Outbreaks of foodborne illness have generally involved food that has been heat
treated and growth of surviving spores, in absence of any competitive flora, are the
source of the illness. B. cereus has frequently been isolated from a wide variety of
foods and in addition to products of plant origin such as rice, pasta and spices, dairy
products, including both raw and pasteurized milk, are the most common food vehi-
cles for B. cereus (Granum 2001; Granum et al. 1993). Although the actual number
of foodborne illness outbreaks associated with B. cereus is likely underestimated, a
few outbreaks of foodborne illness have been reported from psychrotrophic,
enterotoxin-producing strains of B. cereus. Psychrotrophic strains of B. cereus were
32 M. D. Hardin
recovered during outbreak investigations that occurred in Spain and The Netherlands
between 1986 and 1989. The food poisoning strains were recovered from various
dairy products including pasteurized milk and grew within a temperature range of
between 4 and 37 °C (van Netten et al. 1990). Spores of B. cereus in raw milk have
been reported to survive pasteurization and subsequently colonize production
equipment. Lin et al. (1998) isolated both spores and vegetative cells of B. cereus
from raw milk, pasteurized milk, and environmental samples. Most of the isolates
obtained from the pasteurized milk and final products belonged to the same sub-
groups as the strains germinated from spores in raw milk suggesting that spores in
raw milk were a major source in pasteurized milk. Strains of B. cereus isolated from
environmental samples came from HTST pipes, pasteurized milk tanks, and fillers,
suggesting that they may be a potential reservoir for B. cereus within the facility and
a potential contributing factor post-pasteurization contamination with the organism.
Since B. cereus is a spore-former and is ubiquitous in the environment, a low
level of contamination can be expected in most foods. However, low numbers of
B. cereus vegetative cells and/or its spores are not expected to cause problems unless
growth is permitted to occur. Foods implicated in cases of B. cereus associated ill-
ness usually contain at a range of 105 to 107–8 viable cells or spores due to differ-
ences in the amounts of enterotoxin produced by different strains (Granum 2001;
Granum 1994). Control in foods relies on complete destruction by heating or other
lethality treatment designed to control germination of spores or prevent multiplica-
tion to hazardous levels in foods. While psychrotrophic strains are capable of growth
during shelf life and have the potential to cause illness, few outbreaks have been
reported in refrigerated foods held under proper refrigeration. In addition, the rela-
tively short duration and mild nature of the illnesses caused by B. cereus have kept
this organism in the epidemiological background, particularly when compared to
other more prominent foodborne infections and intoxications (ICMSF 1996;
Granum 2001; NACMCF 2005).
2.2.3 Non-Proteolytic Clostridium botulinum
Clostridium botulinum is an anaerobic, spore-forming microorganism capable of

producing one or more biological neurotoxins (types A–H). Spores of C. botulinum
are widely distributed in nature and are commonly found in soil, sediments, and
water. The species is divided up into four groups (I–IV) based on DNA homology
and physiological differences. Most outbreaks of human botulism are caused by
Group I (proteolytic) strains which produce toxins of type A, B, or F and Group II
(non-proteolytic) strains which produce type B, E, or F. The optimal temperature for
growth of Group I proteolytic strains is between 35 °C and 40 °C with a minimum
growth temperature of 10 °C (ICMSF 1996; Austin 2001). Therefore, control of
growth of Group I strains can be achieved by storage of food products at tempera-
tures below 10 °C. Outbreaks of foodborne illness associated with Group I strains
have often involved incorrectly canned or retorted foods.
Non-proteolytic Group II strains of C. botulinum have an optimum temperature

for growth of 28–30 °C and will grow at temperatures as low as 3 °C (Graham et al.
1997). Growth is inhibited by 5% salt and a water activity of 0.97 in a NaCl solution
(Graham et al. 1997; Austin 2001). Outbreaks associated with non-proteolytic
C. botulinum often involve smoked, dried, or salted fish, and type E neurotoxin
(Peck 2006; EFSA 2016). Most recently, an outbreak of C. botulinum type E associ-
ated with dried and salted fish was reported in Germany and Spain, however, and as
is the case with many reported outbreaks, no details were provided regarding pack-
aging and storage conditions at the time of the reported illnesses (EFSA 2016).
While outbreaks of illness associated with Group II strains occur infrequently, the
severity of the illness associated with the neurotoxin produced by this organism
makes any outbreaks of botulism too many.
Non-proteolytic Group II strains are of particular concern for refrigerated prod-
ucts with an extended shelf life particularly when stored in MAP or vacuum packag-
ing. As the shelf life of refrigerated foods packaged in reduced oxygen packaging
(ROP) is increased, more time is available for C. botulinum growth and toxin forma-
tion. As mentioned previously, pathogenic psychrotrophic microorganisms do not
tend to impart a sensory defect that is perceived as spoilage. Group II C. botulinum
strains are non-proteolytic, and therefore, no off-odor or evidence of spoilage may
coincide with toxin development. Although storage at temperatures below 3 °C
would prevent the growth of the organism, these temperatures are difficult to achieve
on a consistent basis, if at all, particularly during distribution, storage, and display
at retail or in a home refrigerator (Lund and Peck 1994; ICMSF 1996; Audits
International 1999; USDA FSIS 2010; USDA CFSAN 2003). As storage tempera-
tures increase, the time required for toxin formation is significantly shortened, and
temperature abuse (>10 °C) could allow Group I strains to grow as well (Szabo and
Gibson 2003). Group II C. botulinum have also been identified as a particular risk
for minimally processed heated and chilled sous-vide products. These products are
cooked under vacuum in sealed pouches (oxygen barrier bags), at precise (and
sometimes low) temperatures, and often for long times. Sous-vide can be used to
prepare foods with an extended shelf life for retail sale or use in food service. For
some products, this results in minimally processed foods that may be undercooked
depending on recipe. The lack of heat treatment sufficient to destroy spores of
C. botulinum and sometimes lack of thorough heating prior to consumption, com-
bined with packaging in an atmosphere with reduced or no oxygen, increases the
risk of botulism from these foods. This places the reliance for the safety of these
products solely on adequate refrigeration and time. However, as was previously
mentioned above, the temperatures required to preclude the growth of, and toxin
production by, non-proteolytic C. botulinum are not necessarily achieved in the dis-
tribution, retailing, and storage of chilled foods.
Modified atmosphere packaging, once considered a new technology, is continu-
ally being applied to new and novel foods and food processes. The main purpose of
reduced oxygen/modified atmosphere packaging for processed foods is to inhibit
the growth of aerobic spoilage microbes and spoilage characteristics associated
with these types of proteolytic and lipolytic spoilage organisms, thereby increasing
34 M. D. Hardin
the shelf life of the product. However, reducing the levels of oxygen in a product
and inhibiting the growth of competing flora allow for the outgrowth of C. botulism
spores if temperature is not adequately controlled, particularly during extended
shelf life. Control measures for specific categories of products including meat and
meat products, fishery products, fruits and vegetables, dairy products, and refriger-
ated processed foods of extended shelf life with reduced oxygen packaging, also
known as refrigerated processed foods of extended durability (REPFEDs), have
been outlined in numerous publications (FSA 2017; NZFA 2005; Szabo and Gibson
2003). Although the minimum growth temperature for non-proteolytic strains of
C. botulinum recommended in most of these publications is 3.3 °C, Graham et al.
(1997) reported growth and toxin formation of non-proteolytic strains of C. botuli-
num in 5–6 weeks. Due to the risk for growth of Group II strains of C. botulinum at
refrigeration temperatures, several regulatory agencies and advisory committees
have that for prepared chilled foods with extended shelf life (>10 days), additional
controls such as pH, water activity, % salt, 6D heat treatment, antimicrobial addi-
tives, should also be used singly or in combination with temperature to preclude
growth and toxin production of psychrotrophic C. botulinum (FSA 2017; NZFA
2005; Szabo and Gibson 2003).
2.2.4 Listeria monocytogenes
Listeria is a ubiquitous organism, widely distributed in the environment including

soil, vegetation, silage, feces, sewage, and water. The genus Listeria contains sev-
eral species with Listeria monocytogenes being the one associated with human ill-
ness outbreaks. Listeria monocytogenes was first identified in 1923 and associated,
at the time, with zoonotic diseases including abortions in cattle and sheep. L. mono-
cytogenes was first recognized as a significant cause of human illness in 1981. Since
that time, both humans and animals have been recognized as asymptomatic carriers
of the organism. Clinical symptoms of the illness range, in healthy adults, from mild
flu-like symptoms of noninvasive gastroenteritis to more severe invasive cases of the
disease. In the more severe cases, symptoms may include septicemia and meningi-
tis. Infections may become life threatening particularly for individuals with sup-
pressed immune systems such as pregnant women, newborns, elderly, and
immunocompromised individuals such as HIV-positive patients, individuals receiv-
ing dialysis or chemotherapy. The high mortality rate of between 25 and 35% is a
major concern with listeriosis (Bean and Griffin 1990).
Lower growth limits for L. monocytogenes have been reported in sterile foods
(chicken broth and UHT milk) as low as 0 °C; however, growth at this temperature
is slow (Walker et al. 1990). L. monocytogenes also grows well under varying gas-
eous atmosphere aerobic, anaerobic, and microaerophilic conditions (Ingham et al.
1990). L. monocytogenes is not resistant to heat, and consequently, thorough cook-
ing will destroy the organism. Food safety concerns and public health implications
related to Listeria monocytogenes stem primarily from contamination of ready-to-eat
food products that receive no additional cooking by the food preparer or consumer.
Although outbreaks of foodborne illness have been reported in a wide variety of
RTE foods, some foods have been identified as posing a higher risk for L. monocy-
togenes. This includes RTE foods that have the potential to be contaminated with
L. monocytogenes and which support the growth of the organism to high numbers,
particularly due to storage for extended periods of time at refrigeration temperatures
(ILSI 2005; FDA 2003; USDA FSIS 2010). Some of the categories of foods that
have been identified to be high include unpasteurized milk products, smoked sea-
food, deli meats, and frankfurters (FDA 2003; USDA FSIS 2010). Differences in
risk for L. monocytogenes have been reported for products prepared at a commercial
production facility versus a retail delicatessen (Gombas et al. 2003; Pouillot et al.
2015; Pradhan et al. 2011). Higher prevalence rates and levels of L. monocytogenes
have been reported for RTE deli meats and salads packaged at retail as compared to
product packaged at a processing facility (Gombas et al. 2003). However, a more
recent survey found no significant differences between deli packaged and prepack-
aged seafood salads and deli-type salads without meat (Luchansky et al. 2017). This
multiyear Market Basket Survey also reported that the occurrence of L. monocyto-
genes in the RTE foods tested had decreased and that the prevalence was lower than
reported in the Gombas et al. study conducted in 2003.
While it is nearly impossible to eliminate L. monocytogenes from raw foods,
most measures to control L. monocytogenes in RTE products involve continual
management and a multiple-hurdle approach: Control measures focusing on the
hygienic design of the facility and equipment; good manufacturing practices
(GMPs); effective sanitation to avoid cross-contamination of RTE processed prod-
ucts; a stringent environmental sampling program; and when possible, prevention of
growth of the organism in the product through the use of antimicrobial ingredients
and/or processes, and temperature control. In the environment, limiting product
contamination and proper sanitation including employee hygiene, controlling traf-
fic, preventing establishment of the organism in coolers, cold rooms, air handling
units, on equipment, and conveying systems of post-lethality areas (ready-to-eat
areas) of a processing plant or in a food preparation kitchen in conjunction with a
focused and targeted environmental testing program will reduce the risk of product
contamination with this organism (Tompkin 2002; USDA 2010; FDA 2003;
Carpentier and Cerf 2011).
2.3 Challenge Studies and Shelf Life
While some publications refer to shelf life studies as challenge studies (NACMCF
2010), there are distinct differences between shelf testing and challenge studies.
Shelf life studies are an objective means to determine the time a product can be
expected to keep under specified storage conditions without appreciable changes in
product quality or safety. The end of shelf life is often based on one or more changes
in sensory, chemical, functional, physical, and microbiological characteristics of the
36 M. D. Hardin
product. They may be conducted for various reasons such as a new product launch;
for a change in product formulation, ingredients, supplier, or packaging; to validate
a process; or as part of an on-going QA/QC verification program. Microbiological
challenge studies are conducted to determine the ability of a food to support the
growth of spoilage organisms or pathogens. Microbiological challenge studies are
performed by inoculating a specific level of selected microorganisms into a food
product or ingredient to determine to the food safety risk or risk of spoilage.
Microbiological challenge studies can be designed and used for a variety of pur-
poses. They can determine if a particular microorganism will grow in a particular
food under a specified set of circumstances. Challenge studies may be conducted to
evaluate the effectiveness of a lethality step, or of an antimicrobial ingredient or
processes, or to simulate what happens to a product during processing, distribution,
and/or subsequent handling. The shelf life of a product is often determined during
the product development stage prior to any large-scale production or market testing.
At this time, the risk for contamination, survival, and growth of pathogenic organ-
isms during shelf life may also be evaluated, particularly for high risk food such as
food stored under reduced oxygen for extended periods of time at refrigeration tem-
peratures. When deciding whether a shelf life or challenge study needs to be done,
it is important to have the appropriate expertise available, such as an experienced
food microbiologist, to help determine the need for the study and assist in the design
and in the interpretation of results. An expert food microbiologist will also be able
to identify the appropriate target organism, inoculum type and level, and method of
inoculation. All known sources of expected and unexpected variability in the prod-
uct, batch, as well as with the equipment and manufacturing process, need to be
taken into consideration. Worst-case scenario conditions should be considered dur-
ing the study design process. While shelf life trials for refrigerated perishable foods
are often conducted at temperatures of 40 °F (4.4 °C) or lower, shelf life trials,
particularly those conducted for perishable refrigerated foods, should also include
temperatures of mild abuse (45–50 °F/7–10 °C) to reflect actual cold chain tempera-
tures that may occur during commercial distribution and storage or display. These
trials should be conducted for several days or weeks beyond the targeted shelf life
unless the product fails earlier. Products are then evaluated at set times throughout
the trial for physical and chemical changes in the product that may indicate spoil-
age, as well as conducting microbiological testing for spoilage and/or indicator
organisms, and sometimes for pathogens. Several publications are available to pro-
vide assistance in designing these types of studies (NACMCF 2010; Scott et al.
2005; Hardin 2012; FSA 2017).
2.4 Summary
As stated in the beginning of this chapter, the development and production of new
and novel refrigerated foods with an extended shelf life have gained in popularity
over the past 20–30 years.
These precooked, prepackaged convenience foods have increased in popularity

for both the foodservice and retail segments of the industry. The types of products
that are available are seemingly endless and range from soups and salads to sauces
and complete meals and may contain a variety of food ingredients from anywhere
in the world. These products are often packaged under vacuum or modified air, have
an extended refrigerated shelf life, are not protected by conventional preservation
systems such as reduced water activity or pH, and are intended to receive little or no
additional heating prior to consumption. This presents a public health concern with
respect to the growth of pathogens of concern outlined in this chapter. If present,
these organisms can multiply in low-oxygen packaging systems under extend stor-
age at refrigeration temperatures. Shelf life determination of these refrigerated
foods must involve an assessment of risk of bacterial pathogens that are capable of
growth at refrigeration temperatures. Challenge studies can be used to further eluci-
date the growth potential of these bacterial pathogens and risk of the product to
public health.
References
Abdel-Hag, N.M., B.I. Asmar, W.M. Abuhammour, and W.J. Brown. 2000. Yersinia enterocolitica
infection in children. Pediatric Infectious Disease Journal 19: 954–958.
Ackers, Marta-Louise, Susan Schoenfeld, Markman John, M. Geoffrey Smith, Mabel A. Nicholson,
Wallis DeWitt, Daniel N. Cameron, Patricia M. Griffin, and Laurence Slutsker. 2000. An out-
break of Yersinia enterocolitica O:8 infections associated with pasteurized milk. Journal of
Infectious Diseases 181: 1834–1837.
Audits International/FDA. 1999. “U.S. food temperature evaluation design and summary pages.”
http://foodrisk.org/files/Audits-FDA_temp_study.pdf. Accessed 28 December 2017.
Austin, John W. 2001. Clostridium botulinum. In Food Microbiology Fundamentals and Frontiers,
ed. Michael P. Doyle, Larry R. Beuchat, and Thomas Montville, 2nd ed., 329–349. Washington,
DC: ASM Press.
Barton, Mary D., and Roy M. Robins-Browne. 2003. Yersinia enterocolitica. In Foodborne
Microorganisms of Public Health Significance, ed. Alisa D. Hocking, 6th ed., 577–596.
Marrickville, NSW: Southwood Press.
Bean, N.H., and P.M. Griffin. 1990. Foodborne disease outbreaks in the United States, 1973–1987.
Pathogens, vehicles, and trends. Journal of Food Protection 53: 804–817.
Carlin, Frédéric, Martina Fricker, Annemarie Pielaat, Simon Heisterkamp, Ranad Shaheen, Mirja
Salkinoja Salonen, Birgitta Svensson, Christophe Nguyen-the, and Monika Ehling-Schulz.
2006. Emetic toxin-producing strains of Bacillus cereus show distinct characteristics within
the Bacillus cereus group. International Journal of Food Microbiology 109: 132–138.
Carpentier, Brigette, and Olivier Cerf. 2011. Review-persistence of Listeria monocytogenes in
food industry equipment and premises. International Journal of Food Microbiology 145: 1–8.
European Food Safety Authority (EFSA). 2016. Type E botulism associated with fish product
consumption—Germany and Spain. EFSA Technical Report 2016:EN-1157. http://www.efsa.
europa.eu/sites/default/files/1157e.pdf. Accessed 28 December 2017.
Food Standards Agency (FSA). 2017. The safety and shelf-life of vacuum and modified atmosphere
packed chilled foods with respect to non-proteolytic Clostridium botulinum. United Kingdom:
Vacuum Packaging Technical Guidance. https://www.food.gov.uk/sites/default/files/multime-
dia/pdfs/publication/vacpacguide.pdf. Accessed 28 December 2017.
38 M. D. Hardin
Gombas, D., Y. Chen, R.S. Clavero, and V.N. Scott. 2003. Survey of Listeria monocytogenes in
ready-to-eat foods. Journal of Food Protection 66: 559–569.
Graham, A.F., D.R. Mason, F.J. Maxwell, and M.W. Peck. 1997. Effect of pH and NaCl on growth
from spores of non-proteolytic Clostridium botulinum at chill temperature. Letters in Applied
Microbiology 24: 95–100.
Granum, P.E. 1994. Bacillus cereus and its toxins. Journal of Applied Bacteriology Symposium
Supplement 76: 61S–66S.
Granum, Per Einar. 2001. Bacillus cereus. In Food Microbiology Fundamentals and Frontiers, ed.
Michael P. Doyle, Larry R. Beuchat, and Thomas Montville, 2nd ed., 373–381. Washington,
DC: ASM Press.
Granum, Per Einar, and Terje Lund. 1997. Bacillus cereus and its food poisoning toxins. FEMS
Microbiology Letters 157: 223–228.
Granum, Per Einar, Sigrid Brynestad, and John M. Kramer. 1993. Analysis of enterotoxin produc-
tion by Bacillus cereus from dairy products, food poisoning incidents and non-gastrointestinal
infections. International Journal of Food Microbiology 17: 269–279.
Hardin, M.D. 2012. Food process validations. In Microbiological Research and Development for
the Food Industry, ed. Peter J. Taormina, 45–107. Boca Raton. FL: CRC Press.
Ingham, S.C., J.M. Escude, and P. McCowen. 1990. Comparative growth rates of Listeria mono-
cytogenes and Pseudomonas fragi on cooked chicken loaf stored under air and two modified
atmospheres. Journal of Food Protection 53: 289–291.
Institute of Life Sciences (ILSI). 2005. Achieving continuous improvement in reductions in food-
borne Listeriosis—A risk-based approach. Journal of Food Protection 68: 11932–11994.
International Commission on Microbiological Specifications for Food. 1996. Microorganisms in
Foods 5 Microbiological specifications of Food pathogens. London: Blackie Academic and
Professional.
Jenson, Ian, and Catherine J. Moir. 2003. Bacillus cereus and other Bacillus species. In Foodborne
Kapperud, Georg. 1991. Yersinia enterocolitica in food hygiene. International Journal of Food
Keet, E.E. 1974. Yersinia enterocolitica septicemia. Source of infection and incubation period
identified. New York State Journal of Medicine 74: 2226–2230.
Lee, Lisa A., A. Russell Gerber, David R. Lonsway, J. David Smith, Geraldine P. Carter, Nancy
D. Phur, Christine M. Parrish, R. Keith Sikes, Robert J. Finton, and Robert V. Tauxe. 1990.
Yersinia enterocolitica O:3 infections in infants and children, associated with the household
preparation of chitterlings. The New England Journal of Medicine 322: 984–987.
Lin, S., H. Schraft, J.A. Odumeru, and M.W. Griffiths. 1998. Identification of contamination
sources of Bacillus cereus in pasteurized milk. International Journal of Food Microbiology
43: 159–171.
Luchansky, John B., Yuhuan Chen, Anna C.S. Porto-Fett, Régis Pouillot, Bradley A. Shoyer,
Rachel Johnson-DeRycke, Denise R. Elben, Karin Hoelzer, William K. Shaw Jr., Jane M. Van
Doren, Michelle Catlin, Jeehyun Lee, Rohan Tikekar, Daniel Gallagher, James A. Lindsay,
and The Listeria Market Basket Survey Multi-Institutional Team, and Sherri Dennis. 2017.
Survey for Listeria monocytogenes in and on ready-to-eat foods from retail establishments in
the United States (2010 through 2013): Assessing potential changes of pathogen prevalence
and levels in a decade. Journal of Food Protection 80: 903–921.
Lund, Barbara M., and M.W. Peck. 1994. Heat resistance and recovery of spores of non-proteolytic
Clostridium botulinum in relation to refrigerated, processed foods with an extended shelf-life.
Journal of Applied Bacteriology Symposium Supplement 76: 115S–128S.
MacDonald, Emily, Berit Tafjord Heier, Karin Nygård, Torunn Stakheim Kofitsyo S. Cudjoe,
Taran Skjerdal, Astrid Louise Wester, Bjørn-Arne Lindstedt, Trine-Lise Stavnes, and Line
Vold. 2012. Yersinia enterocolitica outbreak associated with ready-to-eat salad mix, Norway,
2011. Emerging Infectious Diseases 18: 1496–1499.
MacDonald, E., M. Einöder-Moreno, K. Borgen, L. Thorstensen Brandal, L. Diab, Ø. Fossli,

B. Guzman Herrador, A.A. Hassan, G.S. Johannessen, E.J. Johansen, R. Jørgensen Kimo, T. Lier,
B.L. Paulsen, R. Popescu, C. Tokle Schytte, K. Sæbø Pettersen, L. Vold, Ø. Ørmen, A.L.Wester,
M. Wiklund, and K. Nygård. 2016. National outbreak of Yersinia enterocolitica infections in
military and civilian populations associated with consumption of mixed salad, Norway, 2014.
Eurosurveillance 21:1–9. https://doi.org/10.2807/1560-7917.ES.2016.21.34.30321. Accessed
28 December 2017.
Marth, Elmer H. 1998. Extended shelf life refrigerated food: Microbiological quality and food
safety. Food Technology. 52(2):57–62.
Morbidity Mortality Weekly Report (MMWR). 2003. “Yersinia enterocolitica gastroenteritis
among infants exposed to chitterlings—Chicago, Illinois, 2002.” Centers for Disease Control
and Prevention. https://www.cdc.gov/mmwr/preview/mmwrhtml/mm5240a2.htm. Accessed
28 December 2017.
National Advisory Committee on Microbiological Criteria for Foods (NACMCF). 2005.
Considerations for establishing safety-based consume-by date labels for refrigerated ready-to-
eat foods. Journal of Food Protection 68: 1761–1775.
National Advisory Committee on Microbiological Criteria for Foods (NACMCF). 2010.
Parameters for determining inoculated pack/challenge study protocols. Journal of Food
Protection 73: 140–202.
New Zealand Food Safety Authority (NZFSA). 2005. A Guide to Calculating the Shelf Life of
Foods. Wellington, NZ: New Zealand Food Safety Authority. http://blpd.dss.go.th/micro/A%20
Guide%20to%20Calculating%20the%20Shelf%20Life%20of%20Foods%20-%20New%20
Zealand.pdf. Accessed 28 December 2017.
Notermans, S., J. Dufrenne, P. Teunis, R. Beumer, M. te Giffel, and P. Peeters Weem. 1997. A
risk assessment study of Bacillus cereus present in pasteurized milk. Food Microbiology 14:
143–152.
Peck, M.W. 2006. Clostridium botulinum and the safety of minimally heated, chilled foods: An
emerging issue? Journal of Applied Microbiology 101: 556–570.
Pouillot, Regis, Daniel Gallagher, Jia Tang, Karin Hoelzer, Janell Krause, and Sherri B. Dennis.
2015. Listeria monocytogenes in retail delicatessens: An interagency risk assessment-model
and baseline results. Journal of Food Protection 78: 134–145.
Pradhan, Abani K., R. Enata Ivanek, Yrjö T. Gröhn, Robert Bukowski, and Martin Wiedmann.
2011. Comparison of public health impact of Listeria monocytogenes product-to-product and
environment-to-product contamination of deli meats at retail. Journal of Food Protection 74:
1860–1868.
Robins-Browne, Roy M. 2001. Yersinia enterocolitica. In Food Microbiology Fundamentals and
Frontiers, ed. Michael P. Doyle, Larry R. Beuchat, and Thomas Montville, 2nd ed., 215–245.
Washington, DC: ASM Press.
Scott, V.N., K.M.J. Swanson, T.A. Freier, W.P. Pruett, W.H. Sveum, P.A. Hall, L.A. Smoot, and
D.G. Brown. 2005. Guidelines for conducting Listeria monocytogenes challenge testing of
foods. Food Protection Trends 25: 818–825.
Szabo, Elizabeth, and Angela M. Gibson. 2003. Clostridium botulinum. In Foodborne
Tacket, C.O., J. Ballard, N. Harris, J. Allard, C. Nolan, T. Quan, and M.L. Cohen. 1985. An
outbreak of Yersinia enterocolitica infections caused by contamination tofu (soybean curd).
American Journal of Epidemiology 121: 705–711.
Thompson, J. Stephen, and Michael J. Gravel. 1986. Family outbreak of gastroenteritis due to
Yersinia enterocolitica serotype O:3 from well water. Canadian Journal of Microbiology 32:
700–701.
Todd, Ewen C.D., Judy D. Greig, Charles A. Bartleson, and Barry S. Michaels. 2007. Outbreaks
where food workers have been implicated in the spread of foodborne disease. Part 3. Factors
40 M. D. Hardin
contributing to outbreaks and description of outbreak categories. Journal of Food Protection

9: 2199–2217.
Tompkin, R.B. 2002. Control of Listeria monocytogenes in the food processing environment.
Journal of Food Protection 65: 709–725.
United States Department of Agriculture (USDA FSIS). 2011. Yersiniosis and Chitterlings: Tips
to Protect You and Those You Care for from Foodborne Illness. Washington, DC: USDA
FSIS. https://www.fsis.usda.gov/shared/PDF/Yersiniosis_and_Chitterlings.pdf. Accesses 28
December 2017.
United States Department of Agriculture, Food Safety and Inspection Service (USDA FSIS). 2010.
“FSIS Comparative Risk Assessment for Listeria monocytogenes In Ready-to-eat Meat and
Poultry Deli Meats.” https://www.fsis.usda.gov/shared/PDF/Comparative_RA_Lm_Report_
May2010.pdf. Accessed 28 December 2017.
United States Food and Drug Administration, Center for Applied Nutrition (FDA CFSAN). 2003.
“Quantitative assessment of relative risk to public health from foodborne Listeria monocyto-
genes among selected categories of ready-to-eat foods.” https://www.fda.gov/downloads/Food/
FoodScienceResearch/UCM197330.pdf. Accessed 28 December 2017.
van Netten, P., A. van de Moosdijk, P. van Hoensel, D.A.A. Mossel, and I. Perales. 1990.
Psychrotrophic strains of Bacillus cereus producing enterotoxin. Journal of Applied
Bacteriology 69: 73–79.
Walker, S.J., P. Archer, and J.G. Banks. 1990. Growth of Listeria monocytogenes at refrigeration
temperatures. Journal of Applied Bacteriology 68: 157–162.
Chapter 3
Microbial Growth and Spoilage
Peter J. Taormina
3.1 Introduction
Spoilage is characterized by any change in a food product that results in unacceptable

sensory perception. In refrigerated foods, microbial growth often precedes or induces
chemical and physical degradation. Hence, concern with microbial growth leading to
spoilage receives higher priority than chemically or physically degradative processes.
Growth of microorganisms in food systems is dependent on extrinsic and intrinsic
factors. Extrinsic factors include temperature, atmosphere, exposure to light, and
physical handling. Intrinsic factors include availability of nutrients, water activity
(aw), pH, presence of antimicrobial ingredients, competitive microflora, and endog-
enous enzymes. While chemical and physical degradation is certainly important and
influenced by these factors, the rate of microbial growth on refrigerated foods leading
to end of shelf life usually occurs more rapidly than autocatalytic chemical reactions.
Consequently, change in microbial population is often used to assess and monitor
shelf-life performance, with sensory evaluation always remaining the key aspect of
shelf-life evaluation and ongoing testing. Microbial growth and metabolism in foods
can impart biochemical changes (e.g., pH decline and production of acetoin) and pos-
sible formation of toxic metabolites, such as biogenic amines, or accumulation of
odoriferous compounds, such as H2S, byproducts of heterofermentative metabolism
like CO2, and exopolysaccharides (in’t Veld 1996). Temperature has the greatest
impact on the rate of deterioration of most perishable foods as increasing temperature
increases the rates of reactions (Chandler and McMeekin 1989; Jacxsens et al. 2002;
Ronsivalli and Charm 1975). A brief overview of quality deterioration reaction kinet-
ics as affected by temperature was presented in Chap. 1, and growth of psychrotro-
phic bacterial pathogens was covered in Chap. 2. This chapter continues exploring
P. J. Taormina (*)
Etna Consulting Group, Jacksonville, FL, USA
e-mail: peter@etnaconsulting.com

https://doi.org/10.1007/978-3-030-54375-4_3
42 P. J. Taormina
temperature-dependent rates of spoilage as well as growth temperature ranges of

various microorganisms, followed by a review of spoilage microflora in various food
categories.
3.2 Role of Temperature in Shelf Life
Temperature has a profound impact upon the rate of chemical, physical, and
microbial deterioration of foods. There are several mathematical models to esti-
mate the deterioration rate of foods during storage as affected by temperature as
Ea
well as other factors (Mizrahi 2004). The Arrhenius equation k A exp
RT
linearly describes many different types of reactions, including shelf life. The
approach recommended for shelf-life testing is to assume that certain principles
of chemical kinetics apply with respect to temperature acceleration, as described
by the Arrhenius relationship, and utilize kinetic design to make accurate shelf-
life predictions (Labuza 1984). Peleg et al. (2012) pointed out that the apparent
linearity of the Arrhenius plot in many food systems is due to a mathematical
property of the model’s equation rather than to the existence of a temperature-
independent “energy of activation” and proposed an exponential model that bet-
ter describes temperature dependencies traditionally described by the Arrhenius
equation without the assumption of a temperature-independent “energy of activa-
tion.” Stannard et al. (1985) studied the growth of psychrotrophic spoilage bacte-
ria in pure culture and determined that square root relationship (√r = b(T − To))
was better than the Arrhenius equation for the description of the microbial growth/
temperature relationship at chill temperatures and was also applicable to mix-
tures of organisms.
The complex interactions of microbial metabolism and impact on food biochem-
istry as it relates to temperature have been long studied (Mossel and Ingram 1955).
Foundational knowledge about microbial growth rates was advanced to the point of
predicting microbial growth as affected by temperature and other factors (Baranyi
and Roberts 1994; Wilson et al. 2002). Such predictions are useful especially when
temperature history of raw materials and ingredients, work-in-process food, and
finished food products varies. As such, the microbial profile of the raw materials and
finished product can change throughout the production process and impact the
degree of chemical and physical degradation and ultimately affect the shelf life.
Indeed, temperature changes infuse a high degree of complexity in predicting shelf
life. Such complexity lends itself to predictive modeling, but such models are based
on mathematical equations developed at many static temperatures that must be vali-
dated with experimental data.
Research on microbial metabolism and physiochemical breakdown of food sub-
strates was prompted by a goal to develop reliable time–temperature indicators for the
purpose of shelf-life prediction (Riva et al. 2001; Vaikousi et al. 2008). While applica-
tion of such predictive models in the day-to-day operations within the food industry
3 Microbial Growth and Spoilage 43
is not widespread, opportunities do occasionally arise for industry collaboration with

academia or government to utilize shelf-life prediction. For example, predictive
models for shelf life may be utilized for new product lines or for major initiatives to
extend shelf life of existing product lines.
3.3 Storage Temperatures and Microbial Growth
Often, the storage temperature for raw materials or finished products is based on the
minimum growth temperatures of microorganisms that are likely to grow and spoil
the food. The minimum growth temperatures of selected bacteria are displayed in
Table 3.1. As can be expected, the minimum growth temperatures widely range
based on bacterium and growing conditions, but notably Pseudomonas, Brochothrix,
Lactobacillus, Lactococcus, and Weissella have been demonstrated to grow
below 0 °C.
The minimum growth temperatures of selected fungi are displayed in Table 3.2.
Molds with ability to grow below 0 °C included Aureobasidium pullulans, Botrytis
cinerea, Cladosporium spp., Fusarium sporotrichioides, and Penicillium spp. Fewer
yeasts were found to grow at negative temperatures, but Rhodotorula glutinis was
found to grow on blanched peas at as low as −18 °C.
Bacterial pathogens tend to have higher minimum growth temperatures than
either spoilage bacteria or fungi (Table 3.3). This is of great practical importance as
growth of these pathogens will often lag that of the nonpathogenic, psychrotrophic
spoilage flora of foods. Of the pathogens, psychrotrophic Bacillus cereus, nonpro-
teolytic Clostridium botulinum, Listeria monocytogenes, and Yersinia enterocolit-
ica are capable of growth at the lowest temperatures. These pathogens were
addressed in detail in Chap. 2.
3.3.1 Freezing Temperatures
Microbes survive in frozen environments with little nutrient, like glacial and sea ice
and permafrost, at temperatures well below the freezing point of water (Mackelprang
et al. 2017). Microbial cells surviving in such harsh frozen environments are largely
dormant but can repair macromolecular damage by means of DNA-repair enzymes
and protein repair enzymes (Price and Sowers 2004). Microorganisms maintain via-
bility in frozen foods for long periods of time. When cells were suspended in 10%
glycerol and stored at −53 °C for 16 months, there were some species and genera
differences in survivability (Yamasato et al. 1973). Strains of coryneform bacteria,
genera of the family Enterobacteriaceae, and the genus Pseudomonas showed rela-
tively higher survivability, whereas organisms such as Pseudomonas putrefaciens
were sensitive. Lactic acid bacteria became sublethally injured and required special
resuscitation procedures following frozen storage, but all acetic acid bacteria sur-
Table 3.1 Minimum growth temperature (°C) of selected bacteria
44
Minimum Temperature
Organism temperature Substrate pH Atmosphere condition Refs.
Acetobacter 5 – – – – Kraft (1992)
Acinetobacter 4 – – – – Kraft (1992)
Aeromonas 5 – – – – Kraft (1992)
Aeromonas hydrophila Var., ≤0–10 Tryptic Soy Broth – Aerobic Isothermal Rouf and Rigney
(TSB) (1971)
Aeromonas salmonicida Var., ≤5–10 TSB – Aerobic Isothermal Rouf and Rigney
(1971)
Aeromonas shigelloides Var., ≤0–10 TSB – Aerobic Isothermal Rouf and Rigney
(1971)
Arthrobacter 5 – – – – Kraft (1992)
Brevibacterium 5 – – – – Kraft (1992)
Brochothrix thermosphacta −3.36 Modified Elliker broth Opt (ca – Isothermal Leroi et al. (2012)
7–7.4)
Chromobacterium 2 – – – – Kraft (1992)
Chromobacterium 4 – – – – Kraft (1992)
fluviatile
Chromobacterium lividum 2 – – – – Kraft (1992)
Clostridium putrefaciens 0 – – – – Kraft (1992)
Cytophaga xantha <0 – – – – Kraft (1992)
Escherichia coli 5–10 – – – – Kraft (1992)
Flavobacterium 5 – – – – Kraft (1992)
Gluconobacter oxydans 7 – – – – Kraft (1992)
Kurthia 5 – – – – Kraft (1992)
Lactobacillus 2 – – – – Kraft (1992)
Lactobacillusa <−1 DeMan-Rogosa-Sharpe – – Isothermal Korkeala et al. (1990)
(MRS) agar
P. J. Taormina
3
Minimum Temperature
Lactobacillus curvatus −3.27 ± 1.21 MRS broth 5.5–8.5b Aerobic – Wijtzes et al. (1995)
Lactobacillus pentosus >4 – – – – Lund et al. (2000)
Lactobacillus plantarum >4 – – – – Lund et al. (2000)
Lactobacillus plantarum 3.29 MRS broth – – – Zwietering et al. (1994)
Lactococcus piscium −4.8 Modified Elliker broth Opt (ca – Isothermal Leroi et al. (2012)
7–7.4)
Leuconostoc 5 – – – – Kraft (1992)
Microbial Growth and Spoilage
Leuconostoc mesenteroides 4, 3c MRS agar – – Isothermal Korkeala et al. (1990)

Microbacterium ≤10 – – – – Kraft (1992)
Moraxella 2 – – – – Kraft (1992)
Pediococcus >4 – – – – Lund et al. (2000)
Propionibacterium 2–3 – – – – Kraft (1992)
Pseudomonas −12.8, −11.8d Fish (gilthead – Aerobic Nonisothermal Koutsoumanis (2001)
seabream)
Pseudomonas 4 – – – – Kraft (1992)
Pseudomonas −0.08 Beef muscle – – – Zhang et al. (2011)
Pseudomonas fluorescens 0–4 – – – – Kraft (1992)
Pseudomonas putida ≤0 Histidine medium – – Isothermal Hug and Hunter (1974)
Serratia 4–5 – – – – Kraft (1992)
Staphylococcus 5–10 – – – – Kraft (1992)
Streptococcus faecalis 5–10 – – – – Kraft (1992)
Weissella cibaria 9.5 ± 1.5 Weissella medium 6.5 – – Ricciardi et al. (2009)
Weissella viridescens −1.33 ± 1.26e MRS broth – – Isothermal Longhi et al. (2016)
Weissella viridescens 0.12 ± 0.71f MRS broth – – Nonisothermal Longhi et al. (2016)
(continued)
45
Table 3.1 (continued)
46
Minimum Temperature
Weissella viridescens −1.57 ± 1.05g MRS broth – – Nonisothermal Longhi et al. (2016)
Xanthomonas >5 – – – – Kraft (1992)
a
Ropy slime-producing, homofermentative lactobacilli
b
Tmin was independent of pH
c
Continuous growth decreased at ca. 4 °C; microcolonies not observed microscopically below 3 °C
d
Tmin parameters for Belehradek models for tLag and μmax, respectively
e
Two-step modeling approach
f
Increasing temperature optimal experimental design approach
g
Decreasing temperature optimal experimental design approach
P. J. Taormina
Table 3.2 Minimum growth temperature (°C) of selected fungi

Minimum Water
Microorganism temperature Substrate pH activity Refs.
Molds
Alternaria alternata Var., −5–6.5 – – – Pitt and Hocking
(1997)
Aspergillus candidus 3–4 – – – Pitt and Hocking
(1997)
Aspergillus clavatus 5–6 – – – Pitt and Hocking
(1997)
Aspergillus flavipes 6–7 – – – Pitt and Hocking
(1997)
Aspergillus flavus Var., 10–12 – – – Pitt and Hocking
(1997)
Aspergillus fumigatus Ca. 12 – – – Pitt and Hocking
(1997)
Aspergillus niger 10.13 Malt extract 4.2 0.997 Gougouli and
agar Koutsoumanis
(2010)
Aspergillus niger 6–8 – – – Pitt and Hocking
(1997)
Aspergillus ochraceus 8 – – – Pitt and Hocking
(1997)
Aspergillus versicolor 9 – – 0.97 Pitt and Hocking
(1997)
Aureobasidium pullulans −5 – – – Pitt and Hocking
(1997)
Aureobasidium pullulans 4 – – – Samson et al.
var. pullulans (2010)
Aureobasidium pullulans 10 – – – Samson et al.
var. melanogenum (2010)
Absidia corymbifera 14 – – – Pitt and Hocking
(1997)
Botrytis cinerea Var., −2–12 – – – Pitt and Hocking
(1997)
Byssochlamys fulva 6.26 Solidified – – Longhi et al. (2014)
apple juice
Byssochlamys fulva 9.1 Malt extract – – Panagou et al.
agar (2010)
Byssochlamys nivea 10.5 Malt extract – – Panagou et al.
agar (2010)
Chaetomium globosum 4–10 – – – Pitt and Hocking
(1997)
Cladosporium −5 – – – Pitt and Hocking
cladosporioides (1997)
Cladosporium herbarum −10 – – – Pitt and Hocking
(1997)
(continued)
48 P. J. Taormina

Minimum Water
Cladosporium herbarum Ca. –5 Meat – – Pitt and Hocking
(1997)
Endomyces fibuliger Ca. 5 – – – Pitt and Hocking
(1997)
Emericella nidulans 6–8 – – – Pitt and Hocking
(1997)
Epicoccum nigrum Var., <5 – – – Pitt and Hocking
(1997)
Eupenicillium 4–6 – – – Pitt and Hocking
cinnamopurpureum (1997)
Eurotium repens 4–5 – – – Pitt and Hocking
(1997)
Eurotium rubrum Ca. 5 – – – Pitt and Hocking
(1997)
Fusarium culmorum 0 – – – Pitt and Hocking
(1997)
Fusarium moniliforme 2.5–5 – – – Pitt and Hocking
(1997)
Fusarium oxysporum >5 – – – Pitt and Hocking
(1997)
Fusarium poae Ca. 2.5 – – – Pitt and Hocking
(1997)
Fusarium sporotrichioides −2 – – – Pitt and Hocking
(1997)
Lasiodiplodia theobromae 15 – – – Pitt and Hocking
(1997)
Monascus ruber 14·61 ± 1·92 Malt extract – – Panagou et al.
agar (2003)
Monascus ruber 14.91 ± 1.85 Malt extract – – Panagou et al.
agar (2003)
Monascus ruber 15–18 – – – Pitt and Hocking
(1997)
Mucor hiemalis <0 – – – Pitt and Hocking
(1997)
Mucor piriformis Ca. 0 – – – Pitt and Hocking
(1997)
Mucor racemosus Ca. –4 – – – Pitt and Hocking
(1997)
Neosartorya fischeri 11–13 – – – Samson et al.
(2010)
Paecilomyces variotii Ca. 5 – – – Pitt and Hocking
(1997)
Penicillium Ca. −2 – – – Pitt and Hocking
aurantiogriseum (1997)
(continued)

Minimum Water
Penicillium –2 – – – Pitt and Hocking
brevicompactum (1997)
Penicillium chrysogenum 4 – – – Pitt and Hocking
(1997)
Penicillium citrinum Ca. 5 – – – Pitt and Hocking
(1997)
Penicillium crustosum Ca. –2 – – – Pitt and Hocking
(1997)
Penicillium expansum −5.74 Malt extract 4.2 0.997 Gougouli and
agar Koutsoumanis
(2010)
Penicillium expansum −6, −3, −2a – – – Pitt and Hocking
(1997)
Penicillium funiculosum Ca. 8 – – – Pitt and Hocking
(1997)
Penicillium glabrum ≤0 – – – Pitt and Hocking
(1997)
Penicillium hirsutum −5 – – – Pitt and Hocking
(1997)
Penicillium islandicum 10 – – – Pitt and Hocking
(1997)
Penicillium italicum −3, 0 – – – Pitt and Hocking
(1997)
Penicillium oxalicum 8 – – – Pitt and Hocking
(1997)
Penicillium purpurogenum 12 – – – Pitt and Hocking
(1997)
Penicillium variabile 12 – – – Pitt and Hocking
(1997)
Penicillium viridicatum <−2 – – – Pitt and Hocking
(1997)
Rhizomucor miehei 21 – – – Samson et al.
(2010)
Rhizomucor pusillus 20 – – – Pitt and Hocking
(1997)
Rhizopus oligosporus 12 – – – Samson et al.
(2010)
Rhizopus oryzae 7 – – – Pitt and Hocking
(1997)
Rhizopus oryzae 5–7 – – – Samson et al.
(2010)
Rhizopus stolonifer 4.5–5 – – – Pitt and Hocking
(1997)
Rhizopus stolonifer Ca. 5 – – – Samson et al.
(2010)
(continued)
50 P. J. Taormina

Minimum Water
Scopulariopsis candida 5 – – – Samson et al.
(2010)
Scopulariopsis fusca 5 – – – Samson et al.
(2010)
Thamnidium elegans ≤1 – – – Pitt and Hocking
(1997)
Wallemia sebi >5 – – – Samson et al.
(2010)
Yeasts
Candida krusei Ca. 8 – – – Pitt and Hocking
(1997)
Candida parapsilosis Ca. 8 – – – Pitt and Hocking
(1997)
Debaryomyces hansenii Ca. 2 YM broth – – Pitt and Hocking
(1997)
Kloeckera apiculata Ca. 8 – – – Pitt and Hocking
(1997)
Pichia membranaefaciens Ca. 5 – – – Pitt and Hocking
(1997)
Rhodotorula mucilaginosa 0.5–5 – – – Pitt and Hocking
(1997)
Rhodotorula glutinis <−18 Blanched Pitt and Hocking
peas (1997)
Saccharomyces cerevisiae Pitt and Hocking
(1997)
10% Glucose 4 – – –
50% Glucose 13 – – –
Trichoderma harzianum Ca. 5 – – – Pitt and Hocking
(1997)
Trichothecium roseum Ca. 5, 15b – – – Pitt and Hocking
(1997)
Zygosaccharomyces bailii Pitt and Hocking
(1997)
10% Glucose 6.5 – – –
30% Glucose 6.5 – – –
60% Glucose 13 – – –
Zygosaccharomyces rouxii Pitt and Hocking
(1997)
10% Glucose <4 – – –
60% Glucose 7 – – –
a
Minimum temperature reported as −6 °C (Brooks and Hansford 1923), −3 °C (Panasenko 1967),
and − 2 °C (Mislivec and Tuite 1970b)
b
Minimum temperature reported as ca. 5 (Pitt and Hocking 1997) and 15 (Domsch et al. 1980)
Table 3.3 Minimum growth temperature (°C) of selected pathogens

Minimum
Microorganism temperature Substrate Refs.
Bacillus cereus, 4 – ICMSF
psychrotrophic (1996)
Bacillus cereus, mesophilic 15 – ICMSF
(1996)
Campylobacter jejuni 32a Laboratory media ICMSF
(1996)
Clostridium botulinum, 3.3 – ICMSF
nonproteolytic (1996)
Clostridium botulinum, 10 – ICMSF
proteolytic (1996)
Clostridium perfringens 12 – ICMSF
(1996)
E. coli O157:H7 8 – ICMSF
(1996)
E. coli, pathogenic 7 – ICMSF
(1996)
Listeria monocytogenes Ca. 0 High nutrient content medium ICMSF
(1996)
Listeria monocytogenes −0.4 – ICMSF
(1996)
Salmonellab,c 5.2 – ICMSF
(1996)
Salmonella 5–10 – Kraft
(1992)
Shigella sonnei 6.1 – ICMSF
(1996)
Shigella flexneri 7.9 – ICMSF
(1996)
Staphylococcus aureus 7 – ICMSF
(1996)
Staphylococcus aureus, toxin 10 – ICMSF
production (1996)
Staphylococcus aureus 5–10 – Kraft
(1992)
Streptococcus pyogenes 10–15 – ICMSF
(1996)
Vibrio 4 – Kraft
(1992)
Vibrio cholerae 10 – ICMSF
(1996)
Vibrio parahaemolyticus 5 – ICMSF
(1996)
Vibrio vulnificus 8 – ICMSF
(1996)
(continued)
52 P. J. Taormina

Minimum
Microorganism temperature Substrate Refs.
Yersinia enterocolitica −1.3 Laboratory media,optimal ICMSF
nutrient conditions (1996)
Yersinia enterocolitica 0–4 – Kraft
(1992)
a
Microaerobic atmosphere
b
Most serotypes fail to grow at <7 °C. Growth rate substantially reduced at <15 °C
c
Ability of Salmonella to grow at temperatures <5 °C has been reported (D’Aoust 1991)
vived well in 10% glycerol and in 10% honey at −28 °C for 4.5 years. Interestingly,
the authors determined that 10% honey was a better cryoprotectant than 10%
glycerol. Frozen foods tend to be stored at −20 °C, which is below the minimum for
growth of the organisms reviewed in Tables 3.1, 3.2, and 3.3. However, temperature
settings on freezers utilized in the cold chain of food distribution, storage, and display
may cause temperatures to rise closer to 0 °C, thereby permitting slow metabolic
activity and some growth of certain microorganisms.
3.3.2 Refrigeration Temperatures
What is considered refrigerated storage temperatures for food also varies. Typically,
storage temperatures used for raw meats, poultry, and seafood are just below freez-
ing point of the muscle (ca. –7.2 °C). Food processing plants may, however, hold
raw proteins at higher temperatures such as 4–10 °C for short periods of time when
in-process. Refrigerated foods and ingredients are likely to be stored and distributed
at temperatures within a range of 1.67–4.4 °C. At retail and foodservice settings, the
maximum allowable temperature of storage is typically 5 °C. As can be seen from
Tables 3.1, 3.2, and 3.3, many types of bacteria and fungi will readily grow at these
temperatures. Refrigerated foods are sometimes subjected to moderately abusive
temperatures upward of 10–12 °C during distribution, storage, and display. Exposure
to such temperatures may be brief causing little to no change to the food tempera-
ture or can be long subjecting the food to more rapid microbial growth rates.
Temperature cycling occurs when food is moved from one area to another for the
purpose of distribution and/or display. The accuracy of shelf-life estimation is
diminished the more, these temperature cycling scenarios are in play.
Psychrotrophic microorganisms tend to dominate the microbiota of refrigerated
foods. Bacteria such as Brochothrix thermosphacta, Shewanella spp., Aeromonas,
and/or Enterobacteriaceae can grow under refrigeration temperatures (Gram et al.
2002; Holley et al. 2004). Pseudomonas spp. and a few other gram-negative psy-
chrotrophic organisms will dominate proteinaceous foods stored aerobically at chill
temperatures (Gram et al. 2002), including meat, poultry, milk, and fish. Gram-
negative bacteria like Pseudomonas, Hafnia, Shewanella, Photobacterium, and
Aeromonas grow in aerobically stored refrigerated products and degrade proteins,

leading to formation of off odors and flavors. Gram-positive bacteria like
homofermentative lactic acid bacteria, heterofermentative lactics, Carnobacterium,
and Brochothrix reach 106–108 CFU/g during refrigerated storage. While strict aer-
obes like Brochothrix thermosphacta can be controlled by packaging film with low-
oxygen permeability (Holley 1999), facultative anaerobes must be limited by
sanitation, temperature control, and other factors intrinsic to the food, such as
pH. Yeast and mold also readily grow on perishable foods, whether refrigerated or
shelf stable. Aspergillus, Penicillium, Mucor, and Rhizopus are molds commonly
causing food spoilage under refrigeration. Some of the more common yeasts genera
that spoil refrigerated foods are Candida, Dekkera, and Rhodotorula.
3.4 Shelf-Life Determination
Microbiological testing has historically been used in the food industry to assess
shelf life of raw materials and finished products. End of shelf life is often attributed
to the food attaining 106–107 CFU/g. To some degree, this is due to habit and con-
venience. It can be easier to train technicians to perform simple microbiological
assays than to train technicians to perform objective sensory evaluation of shelf-
life samples. The methods for microbiological data collection and reporting are
more streamlined compared to sensory analysis of products for shelf-life testing. In
food processing, the shelf-life measurement methods often rely on the total plate
count (TPC), which is essentially an aerobic mesophilic plate count. Other similar
assays are aerobic plate count (APC) and standard plate count (SPC). Typically,
agar media or film media are used for these purposes, and ISO methods for APC or
TPC call for incubation of plates at 35 ± 1 °C and counting within 48 ± 3 h (ISO
4833:2003 and ISO 15214:1998). However, these incubation conditions do not
permit maximum colony development for many psychrotrophic bacteria that are
the most abundant mid to late shelf life. Therefore, the term, “Total,” is a misno-
mer, and over reliance on this standard technique can result in incomplete data
concerning the true microbial profile of food samples. A considerable underestima-
tion (+0.5–3.2 log CFU/g) on the contamination levels of psychrotrophic lactic
acid bacteria (LAB) was observed for retail, packaged food products stored at
chilling temperature when the mesophilic enumeration technique was implemented
as reference shelf-life parameter (Pothakos et al. 2012; Pothakos et al. 2014). Most
of the isolates belonged to the genera Leuconostoc, Lactococcus, and Lactobacillus
and were unable to grow at 30 °C. However, several strains of Leuconostoc can
dominate the population of various packaged, refrigerated foods near the end of
shelf life (Pothakos et al. 2014). In an acetic acid herring preserve product, the
microbial diversity of the dominant psychrotrophic LAB recovered after incuba-
tion of plates at 22 °C for 5 days was determined using a polyphasic taxonomic
approach (Lyhs et al. 2004). A total of 212 LAB isolates were identified using a
combination of rep-PCR fingerprinting, amplified fragment length polymorphism
54 P. J. Taormina
(AFLP) analysis, and pheS gene sequencing. Leuconostoc gasicomitatum,

Leuconostoc gelidum, Leuconostoc spp., Lactococcus piscium, and Lactobacillus
algidus proved to be the most competent and predominant species that may go
undetected by the widely applied mesophilic enumeration protocols.
Microbiological data may be easy to gather and tabulate, but interpretation can be
difficult, and false assumptions are often made about the data. For example, a simple
threshold limit of population of APC such as 106 CFU/g may not correlate with sen-
sory unacceptance. As for ongoing quality-based shelf-life assessments, the APC
and/or lactic plate count data on samples will often exceed 106 or even 107, yet sam-
ples will look, smell, and taste acceptable. To avoid this confusion, some companies
have decided that shelf-life end-point determinations should be based solely on
visual/odor/flavor evaluations, knowing that from time to time, product could exceed
106 or even 107 of at around mid to late shelf life (e.g., 70–100 days). If reasonable
empirical data can be gathered to demonstrate that organoleptic evaluations will cap-
ture microbial spoilage and “undesirable” microorganisms, then it would lend cre-
dence to essentially ignoring (not collecting data on) APC and lactic even on
ready-to-eat (RTE) products that could support growth of psychrotrophic pathogens.
Beyond the standard plate counting techniques most used in industrial food micro-
biology laboratories, university and government laboratories often utilize techniques
like impedance, fluorometry, and RT-PCR (Guy et al. 2006; Martínez et al. 2011),
which are capable of rapid quantification in some food systems. Visible and short-
wavelength near-infrared (SW-NIR) diffuse reflectance spectroscopy (600–110 nm)
to quantify the microbial loads in chicken breast muscle and to develop a rapid meth-
odology for monitoring the onset of spoilage and in conjunction with principal com-
ponent analysis could detect APC increases slightly above 1-log (Lin et al. 2004).
These techniques can be utilized for more rapid quantification of microorganisms for
the purposes of measuring quality throughout shelf life but are typically cost prohibi-
tive, and so the plate count or version of the plate count often prevails.
3.5 Accelerated Shelf-Life Testing for Refrigerated Foods
Normal shelf-life testing must span the stated or potential usable time that a food
product is saleable and consumable, and ideally, the testing timeframe should
exceed that target shelf life so that the point at which the product loses acceptability
can be determined. However, the time from the genesis of an idea or stated goal to
launch a food product into the marketplace to the time that it is commercialized will
often exceed the desired shelf life of the product. When the ideation process has led
to a food product development project, there should be a general idea of the desired
number of days or weeks of shelf life for such product, but this number could be a
rough estimate. For truly innovative new product development, the shelf-life target
may be truly unknown. In either case, since products must have a stated shelf life
prior to commercialization, the answer as to what that shelf life might be is often
sought out as quickly as possible. Ergo, accelerated shelf-life testing has been
u tilized as means to predict the shelf life of perishable commodities such as refriger-
ated dairy or meat products (Corradini and Peleg 2007). Accelerated shelf-life
testing is based on measurements of the rate constant (k) at several different tem-
peratures, followed by extrapolation of the straight-line plot to lower temperatures
for predicted shelf life (Man 2002).
Although accelerated shelf-life testing is often used by food laboratories to pro-
vide customers with the much-desired early result for shelf-life testing, the assumed
relationship between temperature and reaction rate as it pertains to shelf life should
be validated per product type. There is a formula based on the Arrhenius equation,
which assumes a straight-line relationship between temperature and rate of degrada-
tion of product quality (i.e., end of shelf life). This is an equation based on “the rule
of 10s” which was proposed by Ted Labuza in the 1980s, and has caught on for use
by contemporary food microbiology testing labs:
Rate at temeprature T 10 C

Shelf liffe at T C
Q10
Shelf life at T 10 C

Rate at temperature T C
This equation works nicely for shelf life of foods that is determinative based on
some single chemical reaction like lipid oxidation or bread staling, but when applied
to complex microbial communities, it loses its applicability. This is because the
preferred growth temperatures of microorganisms are varied across genera. Indeed,
elevated storage temperature of food selects for mesophilic microorganisms, putting
psychrotrophs at a distinct disadvantage in competing for nutrients and physical
space within or on a food substrate. Therefore, incubating product samples at room
temperature (ca. 22 °C) and correlating the time to spoilage with an actual refriger-
ated product shelf life is dubious without rigorous data to establish the correlation
between shelf life at both elevated and refrigerated temperatures. A multivariate
accelerated shelf-life test for fresh-cut lettuce was developed using principal com-
ponent analysis and leading to shelf life of 12.4, 10.4, and 3.7 days at 0, 5, and 15 °C
(Derossi et al. 2016). However, only sensory, physical, and chemical attributes were
assessed, not microbial. A similar study design and outcome was reported for pine-
apple (Amodio et al. 2015). A study of accelerated shelf-life testing of ice cream
utilized total aerobic counts as well as sensory and pH in the Arrhenius equation to
estimate shelf life (Park et al. 2018), but the study did not explain growth of aerobic
counts at very low temperatures such as −6 °C, which is below the minimum growth
temperature for most microorganisms in foods (Tables 3.1, 3.2, and 3.3).
3.6 Chemical Indicators of Microbial Growth
Chemical changes to food during shelf life include oxidation of lipids and pigments,
protein denaturation, and fermentation of carbohydrates. A shift in titratable acidity or
pH can be indicative of growth of acid-producing microorganisms such as
56 P. J. Taormina
Lactobacillus, Acetobacter, or Gluconobacter. Chemical indicators of spoilage such

as D-lactate, in foods that do not contain that molecule, or pH reduction can be cor-
related with microbial growth and impact on shelf life. In vacuum-packaged pork,
D(−)lactic acid and acetoin/diacetyl concentrations increased progressively although
microbial counts stabilized from the 20th day of refrigerated storage (Pablo et al.
1989). Several vacuum-packed meat products from the market had D(−)lactic acid
concentrations that were indicative of the sensory and microbial end of shelf-life tar-
get. In raw pork stored under high-oxygen modified atmosphere at +4 °C levels of
acetoin, diacetyl and 3-methyl-1-butanol correlated with the sensory scores and bacte-
rial concentrations (Nieminen et al. 2016). Acetoin production can be associated with
growth of various types of bacteria including lactic acid bacteria, Listeria monocyto-
genes, and Bacillus spp. (El-Gendy et al. 1983; Xiao and Lu 2014; Zeppa et al. 2001).
Biogenic amines histamine, putrescine, cadaverine, spermine, spermidine, and
tyramine are toxic metabolites primarily produced by metabolically active
Enterobacteriaceae and enterococci. Lactic acid bacteria do not typically produce
significant levels of biogenic amines except during fermentation of meat and dairy
products. Biogenic amines are mainly formed by decarboxylation of amino acids.
Enterobacteriaceae, heterofermentative lactic acid bacteria, Enterococcus faeca-
lis, and Lactobacillus buchneri are among the bacteria capable of producing more
than 500 ppm of biogenic amines during meat or cheese fermentation (Janssen
et al. 1996). With respect to shelf-life, biogenic amine production would be a
concern for fresh meat, poultry, and seafood, and less so if temperature control is
maintained.
Trimethylamine oxide reductase enzymes are utilized by bacteria to reduce
trimethylamine oxide (TMAO) to the trimethylamine (TMA), an odorous sub-
stance. N-acyl homoserine lactones (AHL) are produced by a wide variety of bac-
teria growing in several different food substrates apparently as a means for
bacterial quorum sensing (Gram et al. 2002). Quantities of AHL may correlate
with the onset of sensory decomposition of foods and could therefore be a poten-
tial spoilage indicator. AHL can be extracted from food by homogenizing with
ethyl acetate and can be visualized by separation of extracted AHLs on thin-layer
chromatographic plates and subsequent development by AHL-monitor strains, for
example, Agrobacterium tumefaciens pZLR4 (Gram et al. 2002). Levels of vola-
tile organic compounds such as aldehydes, ketones, and alcohols can indicate
microbiological spoilage of beef (Saraiva et al. 2015), pork (Nieminen et al.
2016), poultry (Tománková et al. 2012), seafood (Mikš-Krajnik et al. 2016), dairy
products (Condurso et al. 2008), and tropical fruits (Taiti et al. 2015) and iceberg
lettuce (Ioannidis et al. 2018) to name a few.
Thiobarbituric reactive substances (TBARS) are commonly used in meat and
poultry to determine lipid oxidation, mostly not related to microbial growth.
However, a combination of microbiological and chemical spoilage markers can
lead to better shelf-life estimation. In cooked blood sausage, quantitative descrip-
tive analysis, Pseudomonas spp., lactic acid bacteria and Enterobacteriaceae
counts, TBARS, total basic volatile nitrogen, and lactic acid showed a better pre-
dictive ability of consumer acceptability than any single spoilage indicators

(Pereira et al. 2019).
3.7 Shelf Life of Specific Food Types
3.7.1 Fresh Produce
Fresh produce has a wide variety of intrinsic properties that have a bearing on
microbial growth rate. Many vegetables tend to have a near neutral pH, while many
fruits tend to be acidic. Most but not all produce has substantial moisture content
and carbohydrate. Most fresh produce contains only trace amounts of protein and
can vary in lipid content from significant (avocados) to trace (lettuce). Various raw
agricultural produce items possess a natural barrier in the form of a rind or “skin”
that protects the fleshy trace tissues from physical damage, oxidation of pigments
and nutrients, and microbial infiltration. Temperature and microflora are the key
factors affecting shelf life of produce (Brackett 1987). Microorganisms that typi-
cally grow on and eventually spoil whole fresh produce include Enterobacteriaceae,
pseudomonads, lactic acid bacteria, yeasts, and molds.
Once produce is cut and fleshy tissue is exposed, growth rates increase, and
shelf life shortens. A survey of 120 Australian retail samples of trimmed, cut let-
tuce usually with several days of remaining shelf life revealed most samples to
have APC in the range of 105 and 107 CFU/g (Szabo et al. 2000). Bacterial patho-
gens Listeria monocytogenes, Aeromonas spp., and Yersinia enterocolitica were
isolated from 3, 66, and 71 samples, respectively. These pathogens grow readily at
refrigeration temperatures (Tables 3.1 and 3.3). In a shelf-life study of cut fruit
pieces stored at 4 °C, it was found that the signs of spoilage differed among fruit
types (O’Connor-Shaw et al. 1994). Brown discoloration was observed only in
pineapple, while bitter flavor developed only in kiwifruit. Microbial growth did not
appear to contribute to spoilage in diced kiwifruit, papaya, and pineapple, but may
have affected honeydew melon and cantaloupe. No group of microorganisms (lac-
tobacilli, Enterobacteriaceae, yeasts, and molds) dominated the microflora of
fresh minimally processed fruit regardless of species of fruit (O’Connor-Shaw
et al. 1994). Kiwifruit, papaya, and pineapple pieces became softer during storage.
Undesirable textural changes in cut fruit are caused by enzymes such as
β-galactosidase and exo-polygalacturonase, which solubilize pectin in cell walls
(King and Bolin 1989).
Shelf life of fresh processed vegetables is increased with the use of modified
atmosphere packaging (MAP). Due to senescence, the modified atmospheres gener-
ated inside packages initially can change over the course of shelf life and can contain
moderate-to-high levels of CO2. While CO2 restricts the growth of aerobic microor-
ganisms, it can lead to safety concerns from anaerobic sporeforming bacteria such as
Clostridium botulinum. The lack of growth of aerobic microorganisms due to MAP
58 P. J. Taormina
coupled with potential C. botulinum growth could mean that the organoleptic
evidence of microbial growth could be lacking despite a pathogen prevalence (Farber
1991). MAP is widely used to extend the refrigerated shelf life of fresh cut produce.
Modified atmosphere stored mixed lettuce and cucumber slices had degraded sen-
sory properties that preceded microbial proliferation and achieved 7 days at 4 °C, but
microbial proliferation was the limiting factor for cut bell p eppers (Jacxsens et al.
2002). The product pH, starting microbial load, and storage temperature impacted
growth of L. monocytogenes and Aeromonas caviae on lettuce and cucumber.
3.7.2 Pasteurized Milk and Dairy Products
Milk is an ideal growth medium for microorganisms with ample protein, lipids, and
neutral pH. Consequently, shelf life of milk can be very short if microorganisms are
present and temperatures approach 10 °C. In Europe and North America for instance,
pasteurized milk must be heated at high temperature for a short time (at least 71.7 °C
for 15 s, or any equivalent combination) and must show a negative reaction to the
phosphatase test and a positive reaction to the peroxidase test. Immediately after
pasteurization, milk must be cooled to 6 °C or below. The shelf life of pasteurized
milk is dependent on raw milk quality, pasteurization conditions, contamination
from the food contact surface, contamination from environment, distribution
temperature, and effect of light (Rysstad and Kolstad 2006). Psychrotrophic micro-
organisms are the primary cause of pasteurized milk spoilage and the determining
factor in shelf life (Chandler and McMeekin 1989). Pseudomonads dominate the
spoilage flora of pasteurized, homogenized milk at 4–12 °C, and above 12 °C, the
spoilage flora is comprised of only 10–20% pseudomonads and the remainder are
mesophiles (Chandler and McMeekin 1989). It is long been generally recognized
that bacterial growth to a level of 7.5-log10 CFU/mL of refrigerated milk and dairy
products represents the end of shelf life (Griffiths et al. 1984). Shelf life of pasteur-
ized milk is greatly affected by the degree of sanitation and postpasteurization
recontamination by pseudomonads (Eneroth et al. 2000), but spoilage due to growth
of psychrotrophic Bacillus also occurs (Ternström et al. 1993) due to spore survival
at pasteurization temperatures. Therefore, microbial quality and temperature con-
trol of raw milk destined for pasteurization is of great importance and long-term
impact on pasteurized products.
Traditional extended shelf-life (ESL) technology in North America incorporates a
high heat treatment of the product, which provides normal pasteurized product
sensory characteristics, combined with ultraclean packaging, which includes a con-
trolled filling environment and container sterilization (Henyon 1999). This results in
product code dates for ESL dairy products in North America ranging from 45 to
60 days, and currently, the technology is expanding to include value-added products
along with regular drinking milks. ESL milk has become synonymous for a total sys-
tem process that encompasses everything from raw product quality to final distribution.
Generally, ESL milk has a shelf life longer than pasteurized milk. For ESL milks,
particularly if distributed at temperatures higher than 7 °C, the numbers of thermoduric

and aerobic sporeformers indicate spoilage. At 8–10 °C, aerobic sporeformers have
increased influence, and higher frequencies of psychrotrophic strains are reported. A
thermoduric count less than 1000 microorganisms/mL and an a erobic spore count less
than 10 microorganisms/mL are typical thresholds for microbial quality.
Butter is microbiologically more stable than other dairy-based products.
Pasteurization destroys microorganisms important in butter spoilage, but a variety
of these spoilage microorganisms may contaminate the product at some later stage
in production (Ayres et al. 1980). Proteolytic and lipolytic microorganisms can
bring about flavor defects in refrigerated and ambient stored butter. Pseudomonas
fluorescens can hydrolyze milk fat, liberating short-chain fatty acids that lead to
rancidity. Other bacteria like Flavobacterium maloloris and lactic acid bacteria can
cause surface taint, leading to putrid, decomposed, and cheesy flavor (Van Zijl and
Klapwijk 2000). Spoilage defects have also been caused by yeasts such as Candida
spp. and molds such as Aspergillus spp. Although butter-based products have low
aw, microorganisms can migrate to water droplets (usually <10 μm in size), and
there become metabolically active. In rare instances, bacterial pathogens like
Listeria monocytogenes and Salmonella have been isolated from butters and
vegetable oil products (Van Zijl and Klapwijk 2000).
Cheeses are also susceptible to microbial growth during shelf life. Microbial
growth on hard cheeses is restricted to molds and some yeasts due to an intermedi-
ate aw. Soft and semisoft cheeses are more susceptible to growth by yeast and molds,
in particular, and must be prepared and processed in high-care processing environ-
ments. Sliced, diced, and shredded cheeses were once limited to a shorter shelf life
due to eventual mold growth, until the widespread use of natamycin. Some yogurts,
cream cheeses, sour cream, and spreads are preserved with potassium sorbate to
inhibit mold and yeast growth and extend shelf life. Fermented products like sour
cream and yogurt contain organic acids and sustain a reduced pH that limits micro-
biological growth to lactic acid bacteria and yeast (Mataragas et al. 2011). Despite
high prevalence of organic acids in these products, they can succumb to fungal
spoilage before the end of shelf life.
3.7.3 Plant-Based Protein Products
Plant-based protein products meant to be analogous to meat products, like ham-

burgers, sausages, and roasts, contain proteins from sources such as wheat, soy,
pea, and lipids from canola, safflower, and avocado. These products are complex
matrices of proteins, lipids, starches, stabilizers, and seasonings designed to achieve
the texture and flavor like animal meat products. Due to a high proportion of sea-
sonings and other water-binding texture enhancers, the resulting matrix of such
products has a lower pH and aw than meat products from which they are modeled,
thereby inhibiting microbial growth during refrigerated shelf life. Many of these
products are refrigerated, vacuum packaged, and assigned a shelf life comparable
60 P. J. Taormina
to meat products, even though the pH and aw combination may impart additional
microbial hurdles compared to meat.
Nut- or legume-based alternatives to dairy, such as almond milk, cashew milk,
and soymilk, contain biochemical substrates that are supportive of microbial growth.
Just like in animal-derived milks, high-temperature short-time (HTST) pasteuriza-
tion of nut milk and soymilk reduces vegetative bacteria, yeast, and mold, but allows
sporeforming bacteria to survive. Typically, Bacillus and Paenibacillus dominate
pasteurized nut milks, and prolonged, slow cooling after pasteurization can enable
germination and proliferation and negatively impact shelf life. Like dairy-based
milks, plant-based milks are also susceptible to postprocessing recontamination and
will readily spoil if re-contaminated by Pseudomonas, lactic acid bacteria, yeast, or
mold. Fermented nut milk or soymilk products such as yogurts, sour cream, and
cream cheese contain with reduced pH can also succumb to microbial growth either
via postprocessing recontamination, failed fermentation, or due to survival and
growth of sporeforming bacteria.
3.7.4 Fresh Seafood
The rate of spoilage of fresh seafood is dependent on the number and the type of
bacteria, primarily Enterobacteriaceae and pseudomonads (Ronsivalli and Charm
1975). Temperature of storage affects the rate of spoilage reactions involving bacte-
rial and autolytic enzymes and affects the rate of bacterial multiplication. The
generation time for Pseudomonas fragi, a common fish-spoilage bacterium, is about
12 h at 0 °C and only about 2 h at 12.7 °C (Duncan and Nickerson 1961). Spencer
and Baines (1964) reported that the spoilage rate of white fish is related approxi-
mately linearly to the temperature at which the fish are stored in the range 1–25 °C. In
the range of 0–7.8 °C, the rates at which fish spoiled appeared to be linearly related
to the temperature at which they were stored (Ronsivalli and Charm 1975).
Pseudomonas spp. was found to be a major spoilage organism of raw Atlantic
Salmon fillets, and H2S-producing bacteria and Brochothrix thermosphacta were
also important part of the spoilage microflora (Mikš-Krajnik et al. 2016).
Temperature is a significant factor in the types of microorganisms that spoil fresh
shrimp, with H2S-producing bacteria dominating the spoilage microflora at 28 °C
and 7 °C and Pseudomonas spp. dominating at 0 °C (Dabadé et al. 2015).
As bacteria degrade proteins, the accumulation of ammonia and peptides leads to
off odors and flavors. Modified atmosphere packaging of fresh fish can prolong
shelf life compared to vacuum packaging. At 0 °C, cod fillets stored under 48% CO2
extended shelf life by 6–7 days compared to vacuum-packaged fillets, whereas an
atmosphere of pure CO2 only extended shelf life by 2–3 days (Dalgaard et al. 1993).
CO2-rich atmospheric conditions at 0 °C may enable microorganisms such as
Photobacterium phosphoreum to gain a selective advantage over the H2S-producing
bacteria like Shewanella putrefaciens.
Trimethylamine oxide (TMAO) is an important compound for maintenance of

the physiological functions in fish and shellfish, but it is also a key substance in the
spoilage of raw or processed seafood (Sotelo and Rehbein 2000). TMAO is present
at varying levels in fish and shellfish depending on factors such as diet, the age and
size of the fish, and environmental factors. Reduction of TMAO into trimethylamine
(TMA) is mainly responsible for the off-odor indicative of spoilage and has a very
low-odor threshold compared to ammonia. TMAO is reduced to TMA by the action
of trimethylamine oxide reductase enzyme, which is possessed by many bacteria
within Enterobacteriaceae such as Shewanella and by Alteromonas, Photobacterium,
and Vibrio. TMAO serves as a terminal electron acceptor in anaerobic growth. Many
types of bacteria found in spoiling fish are unable to reduce TMAO, and therefore,
the usefulness of TMA as a quality parameter is dependent on the storage conditions
of the product and the composition of the bacterial flora.
3.7.5 Raw Meat and Poultry
When refrigerated, raw ground poultry in reduced oxygen atmosphere packaging

becomes spoiled, microbial counts are at typically ≥107 CFU/g (O’Brien and Marshall
1996). However, ground turkey products typically have an order of magnitude higher
plate counts than chicken, often reaching 108 CFU/g without spoilage (Guthertz et al.
1976). Microorganisms responsible for spoilage of poultry can multiply at refrigera-
tion temperatures (e.g., 1.1–4.4 °C) (International Commission on Microbiological
Specifications for Foods [ICMSF] 1986). A criterion for APC in raw chicken of n = 5,
c = 3, m = 5 × 105, M = 107 has long been used, but the ICMSF cautioned that it is not
necessarily achievable for turkey even produced in modern processing systems due to
the differing microbial load and substrate differences. The ICMSF provides no strict
limits to the general microbial population (such as APC or lactic) on raw comminuted
meats, but rather suggest that samples of meat at various stages of processing can be
used to establish a baseline and understand changes in the microbial population during
processing (ICMSF 2011). Despite this, the Canadian Government set guidelines that
no sample of ground meat should exceed 7 log CFU/g for aerobic mesophilic counts
(Canadian Food Inspection Agency 1999). Culture-dependent and culture-indepen-
dent characterization of bacterial communities of raw pork stored under modified
atmosphere at 4 °C revealed that Brochothrix thermosphacta, lactic acid bacteria
(Carnobacterium, Lactobacillus, Lactococcus, Leuconostoc, and Weissella), and
Photobacterium spp. predominated in pork samples (Nieminen et al. 2016).
Yeasts occur in low numbers on freshly slaughtered cuts of poultry, but can pro-
liferate in minced or ground meats, and can reach 106 to 107 CFU/g. The yeasts most
frequently isolated from comminuted meats are Candida zeylanoides, Candida
famata (Debaryomyces hansenii), Candida sake, Yarrowia lipolytica, Rhodotorula,
and Cryptococcus laurentii (Pitt and Hocking 2009; Viljoen et al. 1998). Often,
growth by yeasts leads to the types of quality defects described as doughy or card-
board odor and discoloration.
62 P. J. Taormina
Modified atmosphere packaging of comminuted meat and poultry is commonly

used to extend shelf life and preserve color. Gas flush target compositions can range
from 100% N2 to 70% N2 with 30% CO2, to 100% CO2. Gas compositions with
carbon monoxide (CO) are sometimes utilized to fix heme pigments, thereby stabi-
lizing meat and poultry color. For example, a composition of 0.5% CO, 80% CO2,
and 19.5% N2 for ground poultry will fix the pigment to carboxymyoglobin, which
will remain for the duration of the nearly 3-week shelf life. The absence of O2 will
restrict bacterial growth to only facultative anaerobes or anaerobes, not the aerobes
like Pseudomonas, Brochothrix, or Hafnia. So, the microflora will shift toward lac-
tic acid bacteria, which will eventually sour the product (due to lactic and acetic
acid production). Human consumption of meat that has been packaged in a CO
mixture will result in only negligible levels of carboxyhemoglobin in the blood, and
it is highly improbable that the use of CO in the packaging of meat will present a
toxic threat to consumers (Sørheim et al. 1997). Technologically, there are few
drawbacks to the use of CO to form carboxymyoglobin. Some results indicated that
carboxymyoglobin is susceptible to lipid-oxidation-induced browning in a pH- and
temperature-dependent manner in vitro (not in meat) (Suman et al. 2006).
3.7.6 Further Processed Meat, Poultry, and Fish
Processing imparts changes to the substrate for microorganisms, which basically

prolongs the potential shelf life of food. Salting, smoking, and cooking reduce avail-
able moisture, alter proteins and fats, and cause lethality to the more sensitive por-
tion of the microbial population. For instance, the shelf life of lightly processed
seafood stored at 4 °C is dependent on initial pH and secondarily the NaCl content
and the microflora is dominated by lactic acid bacteria and yeasts (Boziaris et al.
2013). With prolonged shelf life comes the need to assure prevention of growth of
psychrotrophic bacterial pathogens, such as nonproteolytic Clostridium botulinum,
Listeria monocytogenes, Bacillus cereus, and Yersinia enterocolitica (covered in
Chap. 2). The pathogen of greatest concern on postlethality exposed ready-to-eat
(RTE) meats is Listeria monocytogenes. Cured RTE meat products, such as ham,
can either be formulated with antimicrobial agents to restrict growth of this human
pathogen during refrigerated shelf life, or products can be treated with surface
application of an antimicrobial agent. Surface application of antimicrobial agents
(i.e., postlethality treatments) can have immediate efficacy, resulting in a logarith-
mic reduction of L. monocytogenes, and may have a sustained, inhibitory effect
preventing growth throughout shelf life.
Microbial counts have long been used to measure general wholesomeness and
potential quality perception of meat products. Spoilage microorganisms are com-
mercially significant in meat products when their numbers reach around 107 CFU/g,
resulting in sensory changes limiting acceptability and shelf life (Holley et al. 2004;
Ingram and Dainty 1971; Mano et al. 1995).
The way processed ready-to-eat meats are formulated, packaged, and stored
inherently selects for the slow growth of lactic acid bacteria, which typically do not
impart negative sensory changes, and suppresses the growth of microorganisms that
can cause unpleasant sensory changes. For example, lactic acid bacteria cause the
normally acceptable souring of ham and other lightly cured meat products (Taormina
2014). Lactic acid bacteria are selected by levels of salt, nitrite, organic salts, and
other inhibitors, and by vacuum or modified-atmosphere packaging. During long-
term refrigerated storage, the lactic acid bacteria metabolize carbohydrates to acidic
products, thereby causing a drop in pH and accumulation of organic acids, chiefly
lactic acid. The production of acid and bacteriocins by certain species of lactic acid
bacteria prevents putrid spoilage, resulting from the ammoniacal products of micro-
bial protein and peptide degradation.
Lactic acid bacteria are ubiquitous and may be introduced to product via the
environment, personnel, and processing aids such as brine or lubricants like water,
and possibly through raw materials. For example, Weissella viridescens is a com-
mon spoilage bacterium on cured ham (Karpíšková et al. 2013). This bacterium,
formerly known as Lactobacillus viridescens, is in the order Lactobacillales, family
Leuconostocaceae, and causes known defects on cured ham such as slime formation
and greening. Control of this specific microorganism on RTE meat products would
be required to attain shelf-life targets. Once present in a package, bacteria are com-
peting for nutrients and opportunity to increase in population. Certain lactic acid
bacteria can hydrolyze sucrose and utilize an enzyme called dextran sucrose, to
build a large molecule called dextran. Dextran is the ropy, slimy material seen in
packages of cured and uncured luncheon meats and frankfurters. Figure 3.1 depicts
the biosynthesis of dextran. The enzyme dextran sucrase is involved in the polym-
erization reaction, and the energy needed for polymerization comes from the hydro-
lysis of sucrose (De Vuyst and Degeest 1999).
Although there is much to be explored with regard to the interaction of microbial
metabolism, chemical reactions, and bearing on shelf life at different temperatures,
some progress has been made to understand the microbiological and physiochemi-
cal changes occurring in a glucose-supplemented broth system with three
Lactobacillus sakei strains, two Lactobacillus curvatus strains, two Lactobacillus
plantarum strains, and two Lactobacillus paracasei strains (Vaikousi et al. 2008).
There was a temperature-dependent impact on the ultimate levels of lactic acid and
viable counts of lactic acid bacteria. After 450 h at 4 °C, lactic acid bacteria reached
8 log10 CFU/mL, pH declined from 6 down to 4, and lactic acid levels rose to
Fig. 3.1 Biosynthesis of the homopolysaccharide dextran

64 P. J. Taormina
8 mM. After 200 h at 8 °C, lactic acid bacteria reached 8 log10 CFU/mL, pH declined
from 6 down to 4, and lactic acid levels rose to nearly 10 mM. At 12 °C, similar
results were seen with plate counts and pH within 180 h, but the lactic acid levels
rose to 16 mM. These data indicate that temperature obviously has a bearing on rate
of microbial growth, but also upon the levels of lactic acid produced in the medium.
This could provide clues as to how food spoilage occurs and manifests differently at
different temperatures. The same microorganisms may be present, the same popula-
tions are ultimately reached, but abusive temperature would impart a much different
biochemical profile than slower growth at refrigeration temperature.
3.8 Conclusion
Microbial growth is often but not always the primary determinant of shelf life.
Temperature is often the primary determinant of microbial growth. Several micro-
organisms have minimum growth temperatures at or below the storage temperatures
of perishable foods and tend to outcompete other microorganisms and predominate
at the end of shelf life. In some food products processed in certain ways, innocuous
growth of homofermentative lactic acid bacteria does not correlate with sensory
loss. Standard plate count techniques are widely used for day-to-day shelf-life test-
ing, although more rapid and precise techniques are available and are used in
research and special projects within industry. Techniques that identify chemical
byproducts of microbial metabolism can serve as early warning of end of shelf life
of foods. Spoilage microflora varies with different food commodities, but certain
psychrotrophic bacteria and molds are comprised of the spoilage flora of multiple
categories of foods. Processing techniques, food ingredients, and packaging shift
eliminate or inhibit those microorganisms that have the greatest capability of spoil-
age, leading to extending shelf life of foods.
References
Amodio, M.L., A. Derossi, L. Mastrandrea, G.B. Martínez Hernández, and G. Colelli. 2015. The
use of multivariate analysis as a method for obtaining a more reliable shelf-life estimation of
fresh-cut produce: A study on pineapple. In III international conference on fresh-cut produce:
Maintaining quality and safety. Acta Horticulturae 1141: 131–136.
Ayres, J.C., J.O. Mundt, and W.E. Sandine. 1980. Microbiology of Foods, Chp 16—Dairy Products.
San Francisco: W.H. Freeman and Company.
Baranyi, J., and T.A. Roberts. 1994. A dynamic approach to predicting bacterial growth in food.
International Journal of Food Microbiology 23 (3–4): 277–294.
Boziaris, I.S., A.P. Stamatiou, and G.J.E. Nychas. 2013. Microbiological aspects and shelf life of
processed seafood products. Journal of the Science of Food and Agriculture 93 (5): 1184–1190.
Brackett, R.E. 1987. Microbiological consequences of minimally processed fruits and vegetables.
Journal of Food Quality 10 (3): 195–206.
Brooks, F.T. and C.G. Hansford. 1923. Mould growths upon cold-store meat. Transactions of the
British Mycological Society 8(3): 113–142.
Canadian Food Inspection Agency. 1999. Sampling and testing procedures. In In Meat Hygiene
Manual, edited by Meat and Animal Product Division, 18. Canada: Government of Canada.
Chandler, R.E., and T.A. McMeekin. 1989. Temperature function integration as the basis
of an accelerated method to predict the shelf life of pasteurized, homogenized milk. Food
Microbiology 6 (2): 105–111.
Condurso, C., A. Verzera, V. Romeo, M. Ziino, and F. Conte. 2008. Solid-phase microextraction
and gas chromatography mass spectrometry analysis of dairy product volatiles for the determi-
nation of shelf-life. International Dairy Journal 18 (8): 819–825.
Corradini, M.G., and M. Peleg. 2007. Shelf-life estimation from accelerated storage data. Trends
in Food Science & Technology 18: 37–47.
D’Aoust, J.Y., 1991. Psychrotrophy and foodborne Salmonella. International Journal of Food
Microbiology 13(3): 207–215.
Dabadé, D.S., H.M. den Besten, P. Azokpota, M.R. Nout, D.J. Hounhouigan, and M.H. Zwietering.
2015. Spoilage evaluation, shelf-life prediction, and potential spoilage organisms of tropical
brackish water shrimp (Penaeus notialis) at different storage temperatures. Food Microbiology
48: 8–16.
Dalgaard, P., L. Gram, and H.H. Huss. 1993. Spoilage and shelf-life of cod fillets packed in vac-
uum or modified atmospheres. International Journal of Food Microbiology 19 (4): 283–294.
De Vuyst, L., and B. Degeest. 1999. Heteropolysaccharides from lactic acid bacteria. FEMS
Microbiology Reviews 23 (2): 153–177.
Derossi, A., L. Mastrandrea, M.L. Amodio, M.L.V. de Chiara, and G. Colelli. 2016. Application
of multivariate accelerated test for the shelf life estimation of fresh-cut lettuce. Journal of Food
Engineering 169: 122–130.
Domsch, K.H., Gams, W. and Anderson, T.H. 1980. Compendium of soil fungi. Volume 1.
Academic Press (London) Ltd.
Duncan, D. W., Jr. , and J. T. R. Nickerson. 1961. Effect or environmental and physiological con-
ditions on the exponential phase growth of Pseudomonas fragi (ATCC 4973). In Proc. Low
Temp. Microbiol. Symp., p. 253-262. Campbell Soup Co., Camden , N.J. Panasenko, V.T.,
1967. Ecology of microfungi. The Botanical Review 33(3): 189–215.
El-Gendy, S.M., H. Abdel-Galil, Y. Shahin, and F.Z. Hegazi. 1983. Acetoin and diacetyl produc-
tion by homo-and heterofermentative lactic acid bacteria. Journal of Food Protection 46 (5):
420–425.
Eneroth, Å., S. Ahrné, and G. Molin. 2000. Contamination routes of Gram-negative spoilage bac-
teria in the production of pasteurised milk, evaluated by randomly amplified polymorphic DNA
(RAPD). International Dairy Journal 10 (5–6): 325–331.
Farber, J.M. 1991. Microbiological aspects of modified-atmosphere packaging technology—A
review. Journal of Food Protection 54: 58–70.
Gougouli, M., and K.P. Koutsoumanis. 2010. Modelling growth of Penicillium expansum and
Aspergillus niger at constant and fluctuating temperature conditions. International Journal of
Food Microbiology 140 (2–3): 254–262.
Gram, L., L. Ravn, M. Rasch, J.B. Bruhn, A.B. Christensen, and M. Givskov. 2002. Food spoil-
age—Interactions between food spoilage bacteria. International Journal of Food Microbiology
78 (1–2): 79–97.
Griffiths, M.W., J.D. Phillips, and D.D. Muir. 1984. Methods for rapid detection of post-
pasteurization contamination in cream. International Journal of Dairy Technology 37 (1):
22–26.
Guthertz, Linda S., John T. Fruin, Delano Spicer, and James L. Fowler. 1976. Microbiology of
fresh comminuted Turkey meat. Journal of Milk and Food Technology 39 (12): 823–829.
Guy, R.A., A. Kapoor, J. Holicka, D. Shepherd, and P.A. Horgen. 2006. A rapid molecular-based
assay for direct quantification of viable bacteria in slaughterhouses. Journal of Food Protection
69 (6): 1265–1272.
66 P. J. Taormina
Henyon, D.K. 1999. Extended shelf-life milks in North America: A perspective. International
Journal of Dairy Technology 52 (3): 95–101.
Holley, R.A. 1999. Brochothrix. In Encyclopedia of food microbiology, ed. R.K. Robinson,
C.A. Batt, and P. Patel, 314–318. New York: Academic.
Holley, R.A., M.D. Peirson, J. Lam, and K.B. Tan. 2004. Microbial profiles of commercial,
vacuum-packaged, fresh pork of normal or short storage life. International Journal of Food
Hug, D.H., and J.K. Hunter. 1974. Effect of temperature on histidine ammonia-lyase from a psy-
chrophile, Pseudomonas putida. Journal of Bacteriology 119 (1): 92–97.
in’t Veld, J.H.H. 1996. Microbial and biochemical spoilage of foods: An overview. International
Journal of Food Microbiology 33 (1): 1–18.
Ingram, M., and R.H. Dainty. 1971. Changes caused by microbes in spoilage of meats. Journal of
Applied Bacteriology 34 (1): 21–39.
International Commission on Microbiological Specifications for Foods. 1986. Microorganisms in
foods 2. Sampling for microbiological analysis: Principles and specific applications. 2nd ed.
Toronto: University of Toronto Press.
———. 1996. Microorganisms in Foods 5: Characteristics of Microbial Pathogens. Chapman &
Hall: Blackie Academic & Professional.
———. 2011. Microorganisms in Foods: 8. https://doi.org/10.1007/978-1-4419-9374-8.
Ioannidis, A.G., F.M. Kerckhof, Y.R. Drif, M. Vanderroost, N. Boon, P. Ragaert, B. De Meulenaer,
and F. Devlieghere. 2018. Characterization of spoilage markers in modified atmosphere pack-
aged iceberg lettuce. International Journal of Food Microbiology 279: 1–13.
Jacxsens, L., F. Devlieghere, and J. Debevere. 2002. Temperature dependence of shelf-life as
affected by microbial proliferation and sensory quality of equilibrium modified atmosphere
packaged fresh produce. Postharvest Biology and Technology 26 (1): 59–73.
Janssen, M.M.T., H.M.C. Put, and M.J.R. Nout. 1996. Natural toxins. In Food Safety and Toxicity,
ed. J. de Vries, 7–37. Open University of the Netherlands: Publication Robert B. Stern.
Karpíšková, R., M. Dušková, and J. Kameník. 2013. Weissella viridescens in meat products—a
review. Acta Veterinaria Brno [serial online]. 82 (3): 237–241.
King, A.D., and H.R. Bolin. 1989. Physiological and microbiological storage stability of mini-
mally processed fruits and vegetables. Food Technology 43 (2): 132–135.
Korkeala, H.J., P.M. Makela, and H.L. Suominen. 1990. Growth temperatures of ropy slime-
producing lactic acid bacteria. Journal of Food Protection 53 (9): 793–794.
Koutsoumanis, K. 2001. Predictive modeling of the shelf life of fish under nonisothermal condi-
tions. Applied and Environmental Microbiology 67 (4): 1821–1829. https://doi.org/10.1128/
AEM.67.4.1821-1829.2001.
Kraft, A. 1992. Psychrotrophic Bacteria in Foods: Disease and Spoilage. Boca Raton, FL: CRC
Press Inc.
Labuza, T.P. 1984. Application of chemical kinetics to deterioration of foods. Journal of Chemical
Education 61: 348.
Leroi, F., P.A. Fall, M.F. Pilet, F. Chevalier, and R. Baron. 2012. Influence of temperature, pH and
NaCl concentration on the maximal growth rate of Brochothrix thermosphacta and a bioprotec-
tive bacteria Lactococcus piscium CNCM I-4031. Food Microbiology 31: 222–228.
Lin, M., M. Al-Holy, M. Mousavi-Hesary, H. Al-Qadiri, A.G. Cavinato, and B.A. Rasco. 2004.
Rapid and quantitative detection of the microbial spoilage in chicken meat by diffuse reflec-
tance spectroscopy (600–1100 nm). Letters in Applied Microbiology 39 (2): 148–155.
Longhi, D.A., A. Tremarin, B.A.M. Carciofi, J.B. Laurindo, and G.M.F. de Aragão. 2014. Modeling
the growth of Byssochlamys fulva on solidified apple juice at different temperatures. Brazilian
Archives of Biology and Technology 57 (6): 971–978.
Longhi, D.A., W.F. Martins, N.B. da Silva, B.A.M. Carciofi, G.M.F. de Aragão, and J.B. Laurindo.
2016. Estimation of the temperature dependent growth parameters of Lactobacillus virides-
cens in culture medium with two-step modelling and optimal experimental design approaches.
Procedia Food Science 7: 25–28.
Lund, B.M., T.C. Baird-Parker, and G.W. Gould. 2000. The Microbiological Safety and Quality of
Food. Vol. 1. Gaithersburg, MD: Aspen Publishers, Inc.
Lyhs, U., J.M. Koort, H.S. Lundström, and K.J. Björkroth. 2004. Leuconostoc gelidum and
Leuconostoc gasicomitatum strains dominated the lactic acid bacterium population associated
with strong slime formation in an acetic-acid herring preserve. International Journal of Food
Mackelprang, R., A. Burkert, M. Haw, T. Mahendrarajah, C.H. Conaway, T.A. Douglas, and
M.P. Waldrop. 2017. Microbial survival strategies in ancient permafrost: Insights from metage-
nomics. The ISME Journal 11 (10): 2305.
Man, D. 2002. Shelf Life. London: Blackwell Science Ltd.
Mano, S.B., G.D. Garcia de Fernando, D. Lopez-Galvez, M.D. Selgas, M.L. Garcia, M.I. Cambero,
and J.A. Ordonez. 1995. Growth/survival of natural flora and Listeria monocytogenes on refrig-
erated uncooked pork and Turkey packaged under modified atmospheres. Journal of Food
Safety 15: 305–319.
Martínez, N., M.C. Martín, A. Herrero, M. Fernández, M.A. Alvarez, and V. Ladero. 2011. qPCR
as a powerful tool for microbial food spoilage quantification: Significance for food quality.
Trends in Food Science & Technology 22 (7): 367–376.
Mataragas, M., V. Dimitriou, P.N. Skandamis, and E.H. Drosinos. 2011. Quantifying the spoilage
and shelf-life of yoghurt with fruits. Food Microbiology 28 (3): 611–616.
Mikš-Krajnik, M., Y.J. Yoon, D.O. Ukuku, and H.G. Yuk. 2016. Volatile chemical spoilage indexes
of raw Atlantic salmon (Salmo salar) stored under aerobic condition in relation to microbio-
logical and sensory shelf lives. Food Microbiology 53: 182–191.
Mizrahi, S. 2004. Accelerated shelf life tests. In Understanding and Measuring the Shelf-Life Of
Food, ed. R. Steele. Cambridge, UK: Woodhead Publishing.
Mislivec, P.B. and Tuite, J., 1970. Temperature and relative humidity requirements of species of
Penicillium isolated from yellow dent corn kernels. Mycologia 62(1): 75–88.
Mossel, D.A.A., and M. Ingram. 1955. The physiology of the microbial spoilage of foods. Journal
of Applied Bacteriology 18 (2): 232–268.
Nieminen, T.T., P. Dalgaard, and J. Björkroth. 2016. Volatile organic compounds and
Photobacterium phosphoreum associated with spoilage of modified-atmosphere-packaged raw
pork. International Journal of Food Microbiology 218: 86–95.
O’Brien, J.K., and Robert T. Marshall. 1996. Microbiological quality of raw ground chicken pro-
cessed at high isostatic pressure. Journal of Food Protection 59 (2): 146–150.
O’Connor-Shaw, R.E., R. Roberts, A.L. Ford, and S.M. Nottingham. 1994. Shelf life of minimally
processed honeydew, kiwifruit, papaya, pineapple and cantaloupe. Journal of Food Science 59
(6): 1202–1206.
Pablo, B.D., M.A. Asensio, B. Sanz, and J.A. Ordonez. 1989. The D (−) lactic acid and acetoin/
diacetyl as potential indicators of the microbial quality of vacuum-packed pork and meat prod-
ucts. Journal of Applied Bacteriology 66 (3): 185–190.
Panagou, E.Z., P.N. Skandamis, and G.-J.E. Nychas. 2003. Modelling the combined effect of tem-
perature, pH and aw on the growth rate of Monascus ruber, a heat-resistant fungus isolated from
green table olives. Journal of Applied Microbiology 94 (1): 146–156.
Panagou, E.Z., S. Chelonas, I. Chatzipavlidis, and G.-J.E. Nychas. 2010. Modelling the effect
of temperature and water activity on the growth rate and growth/no growth interface of
Byssochlamys fulva and Byssochlamys nivea. Food Microbiology 27 (5): 618–627.
Park, J.-M., J.-H. Koh, and J.-M. Kim. 2018. Predicting Shelf-life of Ice Cream by Accelerated
Conditions. Korean Journal for Food Science of Animal Sesources 38 (6): 1216–1225.
Peleg, M., M.D. Normand, and M.G. Corradini. 2012. The Arrhenius equation revisited. Critical
Reviews in Food Science and Nutrition 52 (9): 830–851.
Pereira, J.A., L. Dionísio, L. Patarata, and T.J. Matos. 2019. Multivariate nature of a cooked blood
sausage spoilage along aerobic and vacuum package storage. Food Packaging and Shelf Life
20: 100304.
68 P. J. Taormina
Pitt, J.I., and A.D. Hocking. 1997. Fungi and Food Spoilage. 2nd ed. Chapman & Hall: Blackie
Academic & Professional.
Pitt, John I., and Ailsa D. Hocking. 2009. Fungi and Food Spoilage. New York: Springer Science
& Business Media.
Pothakos, V., S. Samapundo, and F. Devlieghere. 2012. Total mesophilic counts underestimate in
many cases the contamination levels of psychrotrophic lactic acid bacteria (LAB) in chilled-
stored food products at the end of their shelf-life. Food Microbiology 32 (2): 437–443.
Pothakos, V., C. Snauwaert, P. De Vos, G. Huys, and F. Devlieghere. 2014. Psychrotrophic mem-
bers of Leuconostoc gasicomitatum, Leuconostoc gelidum and Lactococcus piscium domi-
nate at the end of shelf-life in packaged and chilled-stored food products in Belgium. Food
Price, P.B., and T. Sowers. 2004. Temperature dependence of metabolic rates for microbial
growth, maintenance, and survival. Proceedings of the National Academy of Sciences 101 (13):
4631–4636.
Ricciardi, A., E. Parente, and T. Zotta. 2009. Modelling the growth of Weissella cibaria as a func-
tion of fermentation conditions. Journal of Applied Microbiology 107 (5): 1528–1535.
Riva, M., D. Fessas, and A. Schiraldi. 2001. Isothermal calorimetry approach to evaluate shelf life
of foods. Thermochimica Acta 370 (1–2): 73–81.
Ronsivalli, L.J., and S.E. Charm. 1975. Spoilage and shelf life prediction of refrigerated fish.
Marine Fisheries Review 37 (4): 32–34.
Rouf, M.A., and M.M. Rigney. 1971. Growth temperatures and temperature characteristics of
Aeromonas. Applied Microbiology 22 (4): 503–506.
Rysstad, G., and J. Kolstad. 2006. Extended shelf life milk—Advances in technology. International
Journal of Dairy Technology 59 (2): 85–96.
Samson, R.A., J. Houbraken, U. Thrane, J.C. Frisvad, and B. Andersen. 2010. Food and Indoor
Fungi. Utrecht, The Netherlands: CBS-KNAW Fungal Biodiversity Centre.
Saraiva, C., I. Oliveira, J.A. Silva, C. Martins, J. Ventanas, and C. García. 2015. Implementation of
multivariate techniques for the selection of volatile compounds as indicators of sensory quality
of raw beef. Journal of Food Science and Technology 52 (6): 3887–3898.
Sørheim, O., T. Aune, and T. Nesbakken. 1997. Technological, hygienic and toxicological aspects
of carbon monoxide used in modified-atmosphere packaging of meat. Trends in Food Science
& Technology 8 (9): 307–312.
Sotelo, C.G., and H. Rehbein. 2000. TMAO-Degrading Enzymes. Food Science and Technology,
167–190. New York: Marcel Dekker.
Spencer, R. and C. R. Baines. 1964. The effect or temperature on the spoilage of wet white fish.
Food Technology 18:769–773.
Stannard, C.J., A.P. Williams, and P.A. Gibbs. 1985. Temperature/growth relationships for psy-
chrotrophic food-spoilage bacteria. Food Microbiology 2 (2): 115–122.
Suman, S.P., R.A. Mancini, and C. Faustman. 2006. Lipid-oxidation-induced carboxymyoglobin
oxidation. Journal of Agricultural and Food Chemistry 54 (24): 9248–9253.
Szabo, E.A., K.J. Scurrah, and J.M. Burrows. 2000. Survey for psychrotrophic bacterial pathogens
in minimally processed lettuce. Letters in Applied Microbiology 30(6), 456–460.
Taiti, C., C. Costa, P. Menesatti, S. Caparrotta, N. Bazihizina, E. Azzarello, W.A. Petrucci, E. Masi,
and E. Giordani. 2015. Use of volatile organic compounds and physicochemical parameters for
monitoring the post-harvest ripening of imported tropical fruits. European Food Research and
Technology 241 (1): 91–102.
Taormina, P.J. 2014. Meat and poultry: Curing of meat. Encyclopedia of Food Microbiology
(Second Edition) 2: 501–507.
Ternström, A., A.M. Lindberg, and G. Molin. 1993. Classification of the spoilage flora of raw
and pasteurized bovine milk, with special reference to Pseudomonas and Bacillus. Journal of
Applied Bacteriology 75 (1): 25–34.
Tománková, J., J. Bořilová, I. Steinhauserová, and L. Gallas. 2012. Volatile organic compounds as
biomarkers of the freshness of poultry meat packaged in a modified atmosphere. Czech Journal
of Food Sciences 30 (5): 395–403.
Vaikousi, H., C.G. Biliaderis, and K.P. Koutsoumanis. 2008. Development of a microbial time/
temperature indicator prototype for monitoring the microbiological quality of chilled foods.
Applied and Environmental Microbiology 74 (10): 3242–3250.
Van Zijl, M.M., and P.M. Klapwijk. 2000. Yellow fat products (butter, margarine, dairy and
non-dairy spreads), Chp 29. In The Microbiological Safety and Quality of Food, ed. B.M. Lund,
T.C. Baird-Parker, and G.W. Gould, vol. I. Gaithersburg, MD: Aspen.
Viljoen, B.C., I. Geornaras, A. Lamprecht, and A. Von Holy. 1998. Yeast populations associated
with processed poultry. Food Microbiology 15 (1): 113–117.
Wijtzes, T., J.C. De Wit, J.H.J. in’t Veld, K. Van’t Reit, and M.H. Zwietering. 1995. Modelling
bacterial growth of Lactobacillus curvatus as a function of acidity and temperature. Applied
and Environmental Microbiology 61 (7): 2533–2539.
Wilson, P.D.G., T.F. Brocklehurst, S. Arino, D. Thuault, M. Jakobsen, M. Lange, J. Farkas,
J.W.T. Wimpenny, and J.F. Van Impe. 2002. Modelling microbial growth in structured foods:
Towards a unified approach. International Journal of Food Microbiology 73 (2–3): 275–289.
Xiao, Z., and J.R. Lu. 2014. Strategies for enhancing fermentative production of acetoin: A review.
Biotechnology Advances 32 (2): 492–503.
Yamasato, K., D. Okuno, and T. Ohtomo. 1973. Preservation of bacteria by freezing at moderately
low temperatures. Cryobiology 10: 453–463.
Zeppa, G., L. Conterno, and V. Gerbi. 2001. Determination of organic acids, sugars, diacetyl, and
acetoin in cheese by high-performance liquid chromatography. Journal of Agricultural and
Food Chemistry 49 (6): 2722–2726.
Zhang, Y., Y. Mao, K. Li, P. Dong, R. Liang, and X. Luo. 2011. Models of Pseudomonas growth
kinetics and shelf life in chilled Longissimus dorsi muscles of beef. Asian-Australasian Journal
of Animal Sciences 24 (5): 713–722. https://doi.org/10.5713/ajas.2011.10404.
Zwietering, M.H., J.C. De Wit, H.G.A.M. Cuppers, and K. Vann’t Reit. 1994. Modeling of bacterial
growth with shifts in temperature. Applied and Environmental Microbiology 60 (1): 204–213.
Chapter 4
Impact of Sanitation on Product Shelf Life
Steven Tsuyuki and Margaret D. Hardin
4.1 Introduction
Maximizing and maintaining consistent product shelf life continues to challenge the
food industry. With the health implications (Doyle and Glass 2010; Taormina 2010)
of sodium and the consumers’ desire for products with a “clean” label, food product
susceptibility to microbial spoilage and food safety risks has increased. At the same
time, the economics of food manufacturing has shifted production toward mechani-
zation and automation. Longer production runs utilizing complex equipment with
shorter, more intense sanitation cycles have become the norm. Within the healthy
tension that exists between running the plant to meet customer orders and cleaning
the plant, production will always be “king.” However, the consequence of not hav-
ing enough importance placed on sanitation can lead to the disruption of production
activities due to regulatory enforcement and/or food safety and quality product
issues, and result in product recalls and loss of customer and consumer confidence.
A new paradigm is proposed: one that sees sanitation as supporting production and
providing the foundation for the manufacture of safe finished goods that consis-
tently meet the stated shelf life. In this paradigm, the new production day starts with
a clean plant.
Under practical conditions, the manufacturing facility cannot precisely control all
types of microorganisms found in finished goods, but the plant can control the number
of microbes and the risks associated with product contamination. If the microbial
S. Tsuyuki (*)
Sanitary Design and Corporate Sanitation, Maple Leaf Foods, Mississauga, ON, Canada
e-mail: steven.tsuyuki@mapleleaf.com
M. D. Hardin
Vice President of Technical Services, IEH Laboratories and Consulting Group,
Lake Forest Park, WA, USA
e-mail: mh@iehinc.com

https://doi.org/10.1007/978-3-030-54375-4_4
72 S. Tsuyuki and M. D. Hardin
numbers on or in products are excessive, hurdles such as formulation, packaging con-

ditions, and refrigeration may not prevent the proliferation of pathogens or spoilage
organisms to sufficiently assure product safety or quality attributes at the stated code
date on the product package. Plant sanitation, therefore, cannot be overemphasized
(Allen and Foster 1960). Since the 1980s, the meat industry has developed tactics to
control the persistence and spread of Listeria in the processing environment (Malley
et al. 2015). There is common ground between controlling premature spoilage by
spoilage organisms such as lactic acid bacteria (LAB) and controlling bacterial patho-
gens that are capable of growth at refrigeration temperatures such as Listeria. Both of
these groups of microorganisms are ubiquitous, environmental microbes that often
persist in food-manufacturing environments. The factors that control Listeria also
form the foundation required to control spoilage organisms.
Traffic Control + GMP + Dry, Uncracked Floors + Sanitation + Sanitarry Design = Control
Managing the environment in which post-lethality exposed products interact

with people and equipment is crucial to control both Listeria and lactics. While
cooking can eliminate spoilage organisms such as LAB that may be present in the
raw meats, cooked products may become re-contaminated during handling, peeling,
slicing, portioning, and packaging and recommendations published in the scientific
literature include more frequent cleaning of the packaging equipment (Kempton
and Bobier 1970).
This chapter focuses on the sanitation program utilized by successful food
processors and provides industry best practices on the execution of the seven-step
daily cleaning and sanitation process. In addition, the role of non-daily sanitation
tasks is discussed. The importance of equipment sanitary design and installation
that promotes accessibility and enables sanitation performance to meet levels of
microorganisms necessary to maximize shelf life as well as for pathogen control is
discussed. Finally, performance metrics beyond costs are described including envi-
ronmental swabbing, which often drives continuous improvement in both sanitation
effectiveness and efficiency. A maturity model across these attributes is provided as
a tool for benchmarking and self-assessment.
4.2 Sanitation Program
An effective sanitation program is key to controlling food safety issues and maintain-
ing product shelf life. The type and frequency of cleaning depends on the complexity
of the process, equipment design and accessibility, infrastructure, and types of soils
involved. A complete and effective sanitation program consists of both daily sanitation
task as well as those tasks that are performed less than daily. Together, daily and non-
daily sanitation tasks form the basis of a complete sanitation program and should be
captured in the facility’s Master Sanitation Schedule (MSS). Specialized tasks may be
developed to address a specific piece of equipment and surrounding area or for an
4 Impact of Sanitation on Product Shelf Life 73
infrastructure area such as a product storage cooler. Non-daily tasks such as deep
cleaning are intensified cleaning procedures that often involve the extensive, and more
time-consuming, disassembly of equipment areas that cannot be completed daily.
Production activities may allow food residues and fines from foods, liquids from mari-
nades, pinfeathers, fish scales, seeds, sprouts, and vegetable peelings, to become
trapped inside equipment well beyond what is visible on the surface. The extensive
disassembly of equipment increases access to surfaces such as cracks, crevices, and
pores, which are difficult to reach during daily sanitation, and enables these areas to be
manually cleaned and sanitized. Thorough disassembly of equipment can be time-
consuming and labor-intensive and requires the coordinated efforts of personnel in
production, maintenance, engineering, sanitation, quality assurance (QA), and food
safety departments. Sanitors need support from maintenance personnel to break down
equipment or access cabinets that house electrical components. Some outside expertise
such as a sanitation specialist, equipment manufacturer, refrigeration expert, or the
sanitation chemical representative may also be of value. Due to the extensive nature of
the disassembly process, deep cleaning often occurs on weekends or on planned pro-
longed shutdowns. Extensive disassembly of equipment may lead to opening up previ-
ously unexplored areas of equipment, and therefore, participation on the part of
maintenance personnel to assist in the disassembly process is invaluable. The process
can also lead to opportunities to redesign equipment and/or parts to be more easily
removed and cleaned in the future. During deep cleaning and disassembly, it is impera-
tive to keep track of parts such as screws, bolts, hoses, wear strips, and guides for
equipment such as conveyors. Small parts can be placed in tubs and labeled with the
equipment or conveyor number in order to keep track of conveyor belts and associated
guides, parts, and pieces. A map of the room and lines with a number or color code can
also assist in keeping track of parts and pieces. Following removal, conveyor belts can
be put into vats for scrubbing or laid out on tables in order to access all areas for scrub-
bing and cleaning. Finally, operational sanitation tasks that occur within the production
window should be included in the facility’s Good Manufacturing Practices (GMPs).
Operational sanitation tasks are typically executed by front-line employees or desig-
nees to ensure that the last hour of production is just as clean as the first hour.
4.2.1 Daily Sanitation: Seven-Step Process
Most food-processing plants operate one or two shifts of production, followed by a

third shift for cleaning and sanitizing. Structuring sanitation as a step-by-step
process drives organization, order, and method into what appears to be a chaotic
whirlwind of food debris and water, very much like being inside a dishwasher. If
sanitors follow a defined process, understand their role, are trained on the perfor-
mance expectations for each step, are supported for success, are supervised to work
as a team, and are held accountable for their performance, then consistent outcomes
will be achieved that balance the efficiency of cleaning with consistent sanitation
effectiveness.
Support from plant management and supervision are required for the successful
implementation of a consistent process. Sanitation too often receives the least
amount of support of any function in most plants. It is a tough statement to make,
but the reality is that sanitation happens when most people are sleeping. It is often a
challenge to have people available to verify the sanitation process, particularly when
it occurs in the third shift. It is advisable for the individuals responsible for verifica-
tion of sanitation performance to be present on Thursday night so that when Friday
morning rolls around, the weekend can be used to recover. Sleep deprivation is a
small price to pay to show the sanitation team that they are relevant. This is often a
teachable moment for the members of QA and operations who are verifying the
process, as the challenges the sanitation team may be facing on a day-to-day basis
become evident, and hopefully results in all parties working together to facilitate the
effectiveness of the sanitation process. In some plants, the sanitation shift is moved
to the afternoon shift in order to improve sanitation support. Setting up sanitors for
success is pretty simple. A plant that works production to a defined end time, transi-
tions the room for cleaning by removing all production materials from the area, and
completes specific tasks assigned to production will enable an efficient start to the
sanitation process. Preserving the sanitation window is critical. If production runs
late and transition of the room to sanitation is delayed, too often the expectation is
that the sanitation window is compressed in order for the next production shift to
start up on time. Nothing is more demotivating and stressful than starting sanitation
late, receiving the area in an unacceptable state, and rushing to complete the work
to be on time for production startup. Things do go wrong on occasion, and equip-
ment breaks and goes down, forcing production to run later than originally planned.
In these instances, the amount of time allotted for sanitation must be maintained
even if production needs to start later the next day. This must be non-negotiable as
the sanitation process is too important to maintain the quality of the work and the
safety and the morale of the sanitors. The sanitation shift is not the time to play
catch up for production delays and downtime. Strong supervision is the key to con-
sistent process execution. This is even more important for sanitation. First, absen-
teeism and turnover are often much higher for sanitation than for production. This
places a tremendous burden on recruitment and training. Often, short staffing leads
to “working” supervisors that are unable to monitor and coach their front-line
sanitors to work as a team, and are not available to hold them accountable for their
performance. Many companies are realizing the importance of sanitation and are
investing in the sanitation process by ensuring that they have proper staffing, includ-
ing supervisors, adequate tools to perform their jobs, and offering higher salaries
than are paid to production employees.
The objectives of the seven-step wet sanitation process are to:
1. Remove loose soils and fat films using pressurized hot water
2. Remove protein and residue build-up/films, and any associated microorganisms
using detergent and scrubbing
3. Sanitize surfaces using low-pressure, high-volume chemical application to
further reduce microorganisms to acceptable levels
It is very important to complete each process step satisfactorily, in the correct

order, and to stay in sequence with other sanitors working in the same area. The
seven-step sanitation process and best practice elements are described as follows:
1. Area Preparation/Dry Pickup
During this stage, the area is in transition from production to sanitation. Production
and sanitation employees must work together to complete room transfer tasks. No
water is used at this point. Removing gross soils using squeegees, scrapers, and
shovels is far more efficient and effective than using water to knock down soils.
The amount of dry soils can be captured as a performance metric.
(a) Stage sanitation equipment, tools, and reusable Personal Protective Equipment
(PPE) required for each area/equipment, inspect items for wear/damage, and
repair/replace as required.
(b) Switch air-handling equipment to sanitation mode, if applicable.
(c) Perform a documented room transfer audit with production and sanitation
supervisors to ensure that all production-related tasks are completed. Tasks
include the following:
• Collect and remove all edible and inedible product from the area to be
cleaned.
• Collect and remove all packaging materials and production supplies from
the area to be cleaned.
• Collect and remove all garbage and scrape floors and equipment. Remove
all debris into designated disposal areas.
• Clean, sanitize, and secure all water-sensitive components in the area with
plastic to prevent water damage. Some companies clean and sanitize
water-sensitive components both before covering (prior to sanitation) and
after removal of the cover. The additional step of cleaning after removal of
the covering (plastic) also reduces the risk from cross-contamination that
may occur during the removal of the cover (plastic) and any food debris,
overspray, water, and chemicals that may accumulate on the cover during
sanitation.
(d) Disassemble equipment to the level required for daily sanitation. These tasks
are performed by production, maintenance, or sanitors. Sanitors must ensure
that all equipment has been de-energized and proper Lock-Out Tag- Out
(LOTO) procedures have been performed prior to interacting with equipment.
(e) Remove remaining visible soils from equipment and surrounding areas
using squeegees, scrapers, and shovels.
(f) Inspect the condition of the equipment for missing parts or pieces, improp-
erly secured or covered equipment, damaged parts, or foreign material resi-
due. During this process, the sanitor is able to evaluate the condition of
equipment and see any damaged or worn surfaces and parts that may not be
apparent during production until equipment failure occurs. Reporting these
conditions is invaluable to the preventive maintenance of equipment and
ultimately reduces food safety risks.
2. Pre-Rinse
I have never met a sanitor that upon the first meeting and asking them, “how can
I make your job easier?” did not tell me that they needed more water pressure or
more consistent temperature for hot water. However, the use of excessive pres-
surized water increases food safety risk and will degrade equipment assets by
compromising seals and forcing debris and water deeper into equipment and
enclosures. If water pressure alone is not high enough, sanitors will too often
modify the nozzle tips to spray in a “bullet” style manner, which results in maxi-
mizing point pressure while minimizing dispersion. In effect, sanitors are clean-
ing away soils using something akin to a “waterpik.” In addition to the inefficiency
in using point pressure, the effectiveness of the sanitation process is negatively
impacted due to the potential for water and soil to penetrate deeper into complex
equipment beyond what is accessible and cleaned daily. Growth niches exposed
to water and soils, if inhabited by Listeria or spoilage bacteria, can lead to har-
borage sites that allow bacteria to grow despite daily sanitation. Shedding of the
bacteria occurs through machinery motion, food interaction with equipment, and
the interaction of the exposed process line with the surrounding infrastructure.
Shedding and spreading though transfer vectors result in environmental posi-
tives for Listeria or high microbial counts on food contact and non-food-contact
surfaces that can lead to product contamination during operations. Proper train-
ing, supervision, and coaching will help sanitors maintain a balance between
effective rinsing and water usage.
During pre-rinse, the amount of soils being pushed in all directions by hot water
hoses can be overwhelming. If the room air movement is not adequate, within
minutes, the sanitor can barely see 6 ft. in front of them. The saying goes: “If you
can’t see it or reach it, you can’t clean it.” The pre-rinsing step is a labor-intensive
step and, if well executed, will positively influence the outcome of all remaining
steps in the process. What does good look like? At the end of pre-rinse, there
should be no visible soils on either the equipment or infrastructure. Some indi-
viduals may be fooled into believing the room is ready for production at this stage
but visible cleanliness at this stage of the process does not necessarily mean that
the surfaces and equipment are chemically, physically, and microbiologically
clean. A few companies have a supervisor or third-shift QA technician perform a
“pre-op” type inspection of the equipment and surrounding area at this point to
verify the removal of visible soils before applying detergent. Teamwork is crucial
for success. That is, sanitors must be aware of how their actions impact adjacent
areas. They cannot work from the center of their area and spray soils indiscrimi-
nately and with no regard to adjacent areas outside their immediate area. When
sanitors work as a team, everyone sprays soils to common collection zones and
never sprays in the direction of cleaned equipment causing recleaning (rework).
(a) Pre-rinse in the direction from top to bottom (i.e., from the top of the equip-
ment down to the floor), including the low ceiling and overhead structures
impacted daily by production, and surrounding walls and floors. Use a lad-
der when required to access the top of equipment. Sanitors must pay as
much attention to cleaning the environment surrounding the equipment as

they do to the equipment itself. This includes ceiling, walls, floors, drains,
and any overhead infrastructure surfaces that can be cleaned, such as pipes
and drop cords.
(b) Hot water rinsing must utilize temperatures sufficient to melt the fat soils
without baking on proteins (i.e., 54.4°–71.1°/130–160 °F; Cramer 2013),
and at a sufficient pressure (low boosted to a maximum of 280 psi at 10 gpm)
to remove visible soils, but not at excessive levels that may force soils deeper
into equipment or through enclosure seals. The availability of hot water
must be sufficient to maintain the target temperature and pressure through-
out the facility during peak use. It may be necessary to customize water
pressure for specific applications. A multifunctional team composed of
individuals of varying expertise and responsibility, such as maintenance,
engineering, operations, sanitation, and QA, should review the facility and
equipment to determine the optimum cleaning parameters required to maxi-
mize cleaning effectiveness without degrading assets.
(c) All hoses used to deliver water, foaming cleaner, and sanitizer must have a
proper nozzle and tip. Open-ended hoses will waste water and chemicals.
Nozzle tips must be in place to disperse water/chemical appropriately.
“Bullet” tips must never be used.
(d) Sanitors should be aware of the direction of spray, taking into account
sanitor(s) working in adjacent areas. A well-thought-out plan and training
encourage sanitors to work as a team to direct soils to “soil zones” for col-
lection and minimize the need for recleaning areas and equipment due to
recontamination from overspray.
(e) Sanitors should avoid using the water hose as a “floor sweeper” and should
never use the water hose to “force” soils down the drain or to unblock a
plugged drain. Sweep up floor debris periodically using shovels. Never
return to cleaning food contact equipment after handling drains without
proper hand washing or a change of Personnel Protective Equipment (PPE).
In order to reduce the risk of cross-contamination from drains, many
companies assign a dedicated individual to clean drains, using a separate,
color-coded (generally black) brush and bucket designated and identified for
drain use only.
(f) Sanitors must never place parts directly on the floor or on operations floor
stands. Dirty and clean parts should not be stored on the same table or cart.
Cleaned parts should be positioned on a cart or rack that allows parts to
self-drain. Sanitors must never allow sanitation tools that will touch product
contact surfaces to rest on the floor when not in use.
(g) All sanitors must self-inspect (with a flashlight) their work when the pre-
rinse step is complete. They must inspect from all angles and use all the
senses (vision, touch, and smell) to ensure that there are no visible soils
present on ANY surface. Focusing on the known “hard to clean” areas will
either reveal the need for “spot” rinsing or provide assurance to move on to
the next steps in the process. Some companies include detailed photos of
equipment in the written Sanitation Standard Operating Procedures (SSOPs)

in order to emphasize the cleaning and inspection of these “hard to clean”
areas. The target outcome at the end of the pre-rinse step is to have the
equipment and area free of visible soil and fat films.
(h) Detergent application must not begin until all sanitors in an area have com-
pleted the pre-rinse step and performed a self-inspection.
3. Soap and Scrub
Scrubbing has become a lost “art” and unfortunately, for many companies, this
step is only a “soap and rinse.” While the chemistry of soap has improved, there
is no commercial cleaning product available that has proven claims of “scrub-
bing bubbles,” although many try. Mechanical action is an essential part of the
soaping step. While hot water is effective in removing loose soils and melting
fats, surface films, including biofilms and allergens, can only be removed through
scrubbing. Over time, the lack of scrubbing becomes apparent when stainless
steel equipment surfaces “bead” water, appear dull, or have a “blue” hue.
(a) Sanitors should apply the foam in a side-to-side manner to the equipment
and surrounding area, including the ceiling, walls, floors, and infrastructure
surfaces that can be washed, from the bottom up in the reverse order to pre-
rinsing. The consistency of the foam should be adjusted to a shaving or
whipped cream consistency so that the soap does not run off a vertical sur-
face when applied, thereby allowing for adequate contact time with these
surfaces. Never foam more area than can be scrubbed before the foam dries
on the surface (about 15–20 min). Consider 360° access when foaming. This
will ensure that ALL surfaces are treated to the ability that the sanitor has
access to them.
(b) Ensure that the detergent is compatible with the component composition of
the equipment being cleaned and ensure that sanitors are handling chemicals
safely and wearing protective equipment such protective eyewear (i.e., gog-
gles or face shields), chemical resistant gloves, sleeves, and/or when han-
dling and applying chemicals.
(c) Scrub all foamed surfaces using proper tools and abrasive action. Applying
foam alone will not remove surface organic residues and inorganic stains.
Use extension poles and brushes to scrub surfaces beyond your reach.
(d) Always clean drains as part of the “detergent” step and complete the task
using dedicated personnel and tools such as a color-coded brush, bucket, and
pads.
4. Post-Rinse
Excessive post-rinsing must be avoided. Rinse to remove foam, not debris.
Spraying off debris at this stage using pre-rinse water pressure will result in
overspray and the transfer of soil particles from one equipment surface to
another. It is a true sign that pre-rinse was NOT completed effectively. Switching
to the sanitizer hose for “touch ups” will also reduce excessive humidity that
results in condensation.
(a) Rinse using the same spraying best practices as described for pre-rinsing
above. If possible, reduce water temperature and pressure in order to mini-
mize condensation formation and overspray. Pressure is the easiest to reduce
and can be regulated through a valve or reduced through a nozzle tip that
disperses the water stream. Many companies have discontinued using any
pressurized water for the final rinse, opting instead for a flood rinse, particu-
larly in ready-to-eat or exposed finished product areas. This is due to the
increased risk for cross-contamination from non-product surfaces, such as
floors and drains, to food contact surfaces through the use of boosted water.
Pressurized water causes aerosolization of water-containing soil and bacte-
ria that can then settle on equipment, and facilitates moisture penetration
(and contaminants) into sealed areas that can later seep back out onto prod-
uct contact surfaces and the product.
(b) Self-inspect equipment and surrounding area. Deviations must be addressed
by Sanitation BEFORE preoperation inspection.
(c) Sanitation effectiveness testing is performed using a variety of tests includ-
ing ATP, aerobic plate count (APC), and/or environmental swabbing for tar-
get organisms such as Listeria. Testing should be performed immediately
after the equipment and infrastructure have been rinsed. NEVER “swab” a
surface that is NOT visibly clean. ATP swabs are best taken BEFORE sani-
tizer application. Most companies take all swabs before the application of
sanitizer as a measure of the cleaning process and to limit any potential
interference of the analytical method by the sanitizer.
(d) Switch air-handling equipment to production mode, if applicable.
5. Pre-Operation (Pre-op) Inspection
Pre-operation inspection (pre-op) is the verification step to ensure that the equip-
ment and infrastructure are visibly clean. The main objective of pre-op inspec-
tion is NOT to find the sanitor’s mistakes but rather to confirm that the cleaning
phase of the sanitation process was completed in a satisfactory manner so that the
transition phase to production can proceed. Sanitors must be accountable for
their work through self-inspection and verification by their supervisor prior to
pre-op inspection. When fully executed, an effective pre-op inspection is a mul-
tilayered audit that provides invaluable information and feedback. Differentiate
between a true sanitation “miss” and overspray. Sanitors must be trained to
understand where the sanitation misses are, adapt by taking more time and effort
to tackle the “hard to clean” areas on equipment, and by prioritizing self-
inspection to these areas. Overspray is a consequence of not realizing the impact
of the direction of water spray during rinsing. In fact, sanitors must never use a
pressurized water hose to remediate findings. Water hoses should be removed
from the area after the post-rinse step is completed. Findings can be addressed
using mechanical means such as a bucket and paper towel, pad or brush, followed
by a low-pressure (flood) sanitizer hose. If a significant sanitation miss is found,
and a rewash is needed, this results in a delay in proceeding to the next step in the
sanitation process. Everyone performing either self-inspection or pre-op must be
encouraged to use all senses including visual, touch, and smell. A flashlight and
mirror are critical for visual inspection along with using gloved hands to touch
equipment surfaces and being aware of off-odors emanating from equipment or
infrastructure.
(a) Pre-op inspection must be conducted by an independent, non-sanitation per-
son and not by the sanitor.
(b) When deviations are found, the sanitor must be present to remediate findings
using manual means such as a bucket and a pad or brush and a low-pressure
sanitizer spray. Pressurized hot water should not be used unless a complete
reclean of the area and equipment is required.
(c) Track and trend deviations. Investigate repeat deviations to determine the
root cause. Common root causes include poor equipment sanitary design,
poor accessibility to surfaces that need to be cleaned, and subpar perfor-
mance by inadequately trained and supervised sanitors. Unaddressed repeat
findings are an indication of program failure.
6. Condensation Control
(a) Sanitors assigned to tasks involving condensation control must change out of
their sanitation apparel and wear Production PPE consistent with the area.
Condensation tools dedicated to the task must be handled in a sanitary man-
ner to avoid contaminating overhead surfaces when removing condensation.
7. Sanitization
(a) All equipment and environmental surfaces must be sanitized through the use
of a low-pressure, high-volume sanitizer application (flood) at the “no rinse”
concentration level. Avoid significant pooling on food contact surfaces.
Many companies prefer using a foaming sanitizer to allow employees to eas-
ily see where they have and have not applied the sanitizer. The use of foam
also allows for the appropriate contact time of the sanitizer on equipment
and surfaces (particularly on vertical surfaces such as walls and equipment
frames) prior to inspection and equipment setup. Some companies apply the
sanitizer in a single step while others use a two-step application method. In
the two-step method, which may be used on a daily or weekly basis, or in
case of an event or issue, a higher concentration of sanitizer is applied to all
equipment and environmental surfaces followed by a low-pressure flood
rinse of water to food contact surfaces only, and a reapplication of the sani-
tizer at the approved “no rinse” level to those same food contact surfaces.
(b) Make sure to separate parts when sanitizing them to ensure complete cover-
age. Never sanitize parts that are stacked together or piled in a bin or
barrel.
(c) The sanitation step should be repeated if significant assembly of equipment
has occurred or if a significant amount of time has passed (i.e., maximum of
4 h) prior to production startup.
4.2.2 Non-Daily Sanitation: Deep Cleaning and Intervention
The simplest way to describe the merits of non-daily cleaning is to use the analogy
of dental care. In spite of the best efforts to perform dental hygiene daily, trips to the
dentist routinely involve the physical removal of plaque and tartar by the dental
hygienist. This procedure is preventive in nature since if plaque and tartar are
allowed to accumulate, gums will begin to recede and bleed. Untreated, this could
result in periodontal disease that can lead to other health complications. In much the
same way, daily cleaning of equipment is restricted to surfaces that are accessible.
However, equipment can be complex and consist of multiple layers that often over-
lap each other. Sanitors can only clean those surfaces that they have access to and
that time allows. Recognizing this, non-daily tasks are created to further dismantle
equipment for deep cleaning. Usually, tasks are localized to multicomponent parts
or cabinet enclosures. These tasks are preventive in nature and are scheduled events
and not driven by an event or finding. However, an event or finding can trigger an
additional or previously unscheduled deep cleaning. Deep cleaning involves the
further disassembly of equipment to expose surfaces that are not accessible without
the use of tools and maintenance expertise. Deep cleaning tasks are performed after
the seven-step sanitation process (typically started after the post-rinse step) and
performed without the use of pressurized hot water. Tasks may include, but are not
limited to, the following:
• Hand wiping and sanitizing the inner surfaces of enclosures, guards, and covers
• Removal, soaking, and scrubbing of equipment and conveyor belts
• Removal, soaking, and scrubbing ultra-high-molecular-weight polyethylene
(UHMW) conveyor guides and supports
• Removal, dismantling, soaking, and scrubbing conveyor drive sprockets, static
rollers, conveyor support structures, and equipment components that are com-
posed of multiple pieces that are overlapped or bolted or pressed together
(sandwiched)
An intervention for an event or finding often involves a non-daily task and treat-
ment of the entire piece of equipment as a whole. The two most common forms are
a total teardown followed by intensified deep cleaning of the equipment and/or the
use of a heat intervention such as steam treating or baking equipment. The key
objective is to expose and treat ALL surfaces. In a teardown procedure, all mated or
sandwiched points on equipment are opened up to expose surfaces and allow them
to be physically cleaned and sanitized. In using heat, a conduction is used to heat all
surfaces to temperatures that are lethal to pathogens and spoilage organism. A tem-
perature of 160 °F (71 °C) with a hold time of 20–30 min is recommended to treat
equipment for pathogens such as Listeria (Tompkin 2002). However, other microbes
such a lactics may require higher temperatures or longer hold times, as some of
these organisms tend to be more heat-tolerant. Validation and verification of these
processes will help establish the most effective parameters.
As noted at the annual North American Meat Institute (NAMI) Advanced Listeria
course, the use of heat intervention for equipment is still not a common practice
amongst participants and its use is sometimes a subject of debate in the industry.
However, for those companies that have successfully implemented heat interven-
tion, there are a several key points to note:
1. The process used must be thoroughly assessed by a multifunction team prior to
implementation. There are human safety and equipment safety concerns that
must be considered, particularly when using saturated steam. The process must
be both effective and protect assets, such as electrical components and non-metal
pieces from damage.
2. Temperature probe placement is key. Probes must be placed on surfaces that
represent the cold (hardest to heat up) and hot (easiest to heat up) spots. Do not
hang probes in the air, as air temperature does not accurately represent equip-
ment surface temperature. The duration of time for the procedure to be effective
must be validated.
3. Only equipment components that can withstand the heat treatment are included
in the physical “tent.” Specifically, if ultra-high-molecular-weight (UHMW)
pieces can be removed, then do so since the softening point of this material will
be reached during steaming. If there is a chance that steam will penetrate adja-
cent cabinets, block access, and keep adjacent cabinets under positive pressure
using compressed air.
4. Introduce steam through a manifold that provides even dispersion throughout the
tented area.
5. An efficient steaming process will quickly even out the cold and hot spot tem-
peratures and there will be no evidence of steam leakage.
6. The balance between efficacy and effectiveness is based on a right combination
of time and temperature.
7. Moist heat is far more efficient than dry heat.
8. Tenting is a great option when large equipment is stationary. For equipment that
can be moved, such as slicers, tables, etc., an oven (e.g., smokehouse) and heat-
ing with moist heat is another good option. Some companies have a routine rota-
tion and an assigned and validated oven schedule for heating “problem” pieces
of equipment.
9. For small parts, a hot water COP tank or boil tank is a good option for heat
treatment.
All equipment and surrounding infrastructure must have a non-daily sanitation
task that can be accomplished during non-production time that allows either further
disassembly or accessibility of equipment requiring maintenance support. This may
require operations to temporarily move stored product or equipment from areas to
be treated. These tasks are preventive in nature and are captured as part of an MSS
that schedules non-daily sanitation tasks for the entire year. Weekly planning and
schedule compliance are discussed as an agenda item at the regular sanitation meet-
ing. Ideally, these tasks are coordinated with routine preventative maintenance tasks.
4.2.3 Operational Sanitation
Good manufacturing practices (GMP) are well established in the food industry.
Operational sanitation tasks are a specific subset of sanitation best practices that ensure
that exposed products are handled and processed on equipment and in surrounding areas
that are clean and sanitary throughout the entire production “window.” This becomes a
challenge as the production environment is in a constant state of movement of product,
supplies, equipment, and people. Products are most vulnerable during exposure to the
environment, prior to primary packaging. In the case of ready-to-eat (RTE) cooked
products, the risk extends from the point immediately after thermal lethality step until
the time the product is packaged. For some products such as fresh produce, this point
may begin following the application of an antimicrobial spray or flume. Procedures
must be in place to mitigate the risk of contamination associated with the following:
• Direct and indirect contamination by front-line employees
• Product accumulation in equipment leading to product contamination
• Condensation leading to food contact surface or product contamination
The best practice for operational cleaning of equipment within the production
window is to replace “dirty” equipment parts with “clean” ones, if available, and to
clean dirty parts in a wash area segregated from the production area. Plants should
never use pressurized hot water to clean within the production area, especially when
production is running in the area, such as on an adjacent line or piece of equipment.
The risk associated with overspray to adjacent equipment, from cross-contamination
from floors and drains, and the risk of forcing soils deeper into equipment is too
high. The use of hot water will also increase the risk of room condensation. Dry
cleaning with compressed air is also not recommended. If a mid-shift cleanup can-
not be avoided, the best option would be dry cleaning followed by the use of a low-
pressure/high-volume sanitizer at the “no rinse” concentration.
The routine use of a hand sanitizer to sanitize front-line employees’ hands and
an approved sanitizer to sanitize food contact surfaces are also good options for
maintaining the sanitary condition of product handlers and surfaces. Several com-
panies have made this practice routine by activating an audible buzzer, or flashing
light, every 15 min to remind employees to sanitize their hands, tools, and work
station. Alternately, some companies assign an employee to walk around and apply
sanitizer to employees’ hands and to surfaces. Sanitizers require sufficient contact
time to be effective. “Spray, wait, and wipe” or “spray, wipe, and spray again”
should be practiced without allowing the sanitizer to pool on food contact surfaces.
4.3 Sanitary Design and Access
While no piece of equipment is perfect, equipment engineers and manufacturers

have improved equipment design in recent years. There are many standards to refer
to for equipment and facility design including the 3-A sanitary designs used for
dairy equipment, the European Hygiene Engineering and Design group (EHEDG)
guidelines for equipment and infrastructure, and the NAMI sanitary design princi-
ples and checklists for equipment and facility design (NAMI 2016). The NAMI
design principles offer a practical, simple, and effective approach and include easy-
to-use assessment tools and checklists. These checklists were originally designed to
address pathogenic microorganisms; however, the same rules and best practices are
universal and effective when addressing shelf life concerns as well. One point to
keep in mind is that the “score” is irrelevant. It is much more important is to deter-
mine the marginal and unacceptable attributes of the equipment and decide during
fabrication what issues can be addressed through a redesign. If the sanitary design
issue cannot be “designed out,” the plant must develop procedures to mitigate the
risk associated with the condition such as more frequent disassembly and deep
cleaning. Progressive companies will bring the supplier to the facility after installa-
tion and following several cycles of production/sanitation, to determine whether the
equipment is cleanable in the long term and under actual conditions of use.
One consideration that is often overlooked is equipment access for sanitation.
Equipment access for operators comes in the form of stairs and platforms with the
assumption that this level of access meets the needs of sanitation. In most cases, the
level of access required for sanitation is significantly higher. Operating a piece of
equipment is often performed in a fixed operator/equipment interface; however, the
sanitor requires access from all sides of the equipment. For example, if sanitors only
have access to the operator side of the equipment, how does the sanitor clean the
opposite side? Sanitors must be able to clean from the top of the equipment down to
the floor as well as the undersides of equipment. If adequate access to overhead and
elevated areas is not provided with sufficient height, sanitors must use ladders or,
worse, stand on handrails or not clean from an elevated position. Equipment clear-
ance from the floor must be sufficient to allow sanitors to access the undersides and
surrounding floor. This level of access is also important from an inspection standpoint.
Clean In Place (CIP) systems in equipment such as ovens and freezers should
never rely only on automated systems for cleaning. The individual responsible to
operate the CIP must be trained to verify the CIP cleaning task and remediate manu-
ally as required.
4.4 Verification and Performance Swabbing
First, let us agree that “clean is clean.” Regardless of type of facility or area within
the facility, the definition of “clean” must be universal. That applies to all aspects of
performance measurements. What can vary are the conditions that ultimately trigger
a corrective action. What is most important is to not allow repeat findings with no
corrective actions. There is no better way to demonstrate the ineffectiveness of the
plant’s program to auditors/regulators and to send a message to the sanitation team
that nobody cares than a failure to implement effective corrective actions. A “Seek
and Destroy” approach (Malley et al. 2015) used for addressing Listeria findings can
also be applied for determining the root cause for shelf life issues and implementing
corrective actions that are both effective and sustainable. This may include but is not
limited to an evaluation of sanitary design issues within the facility or equipment,
poor equipment access, and/or sanitation team performance. Performance is often
regarded as the last option however, depending on the level of training, and results of
key performance indicators, performance should not be ruled out completely.
There are three types of performance measurements with each one serving a
specific role:
1. Visual Inspection (Pre-op): The pre-op inspection is performed by someone who
is independent from sanitation, such as a QA technician. Each piece of equip-
ment and the surrounding area and infrastructure is inspected after post-rinse for
the presence of visible debris missed by sanitation or for overspray debris.
Successful completion of a pre-op inspection allows the completion of tasks
required to transition the room from sanitation to production. Pre-op perfor-
mance must be a measurement of what the sanitor has failed to effectively clean
and what the inspector has failed to find.
2. Residue Monitoring (ATP bioluminescence): In this case, a sanitation supervisor
or designate swabs a selected number of equipment sites (usually zone 1) after
post-rinsing but prior to sanitization. The results are obtained within minutes and
measure the presence of residue that includes both organic residue (food soils)
and in some cases, microbes. Frankly, there are mixed views on the merit of this
testing. ATP is generally viewed as a qualitative tool used for training since there
are no data to demonstrate that results are comparable to microbial testing, par-
ticularly at lower levels. To be quantitative, the tool must be able to perform well
in a measurement system analysis (MSA) that measures both repeatability and
reproducibility. This means that the same sample site will obtain consistent
results tested a number of times and that multiple units testing the same sample
site will yield comparable results.
3. Microbial Surveillance (APC, Lactics, Listeria spp., etc.): The type of microbio-
logical testing used to monitor and verify the sanitary conditions of the equipment
and environment and the tests performed may vary by product, process, and
objective for testing. Those tests often used as indicators of sanitary conditions
that may affect shelf life include but are not limited to aerobic plate counts (APC),
lactics, or Enterobacteriaceae (EB). The QA technician or designate swabs a
selected number of equipment and area sites (all zones) immediately after the
equipment and area have completed the final sanitization step. Depending on the
method used for analysis and target organism, microbial testing is often viewed as
a lagging indicator, or historical data, since the results may be received anywhere
from 24 to 48 h or as much as a week from the time the samples are taken.
Post-rinse sampling is an investigative approach to finding potential areas of
equipment or infrastructure harborage within the sanitation process. While APC
swabs can be used during this investigation, many companies prefer swabbing spe-
cifically for organisms such as Listeria for food safety issues or for lactics when
spoilage is a concern. Swabs sites are typically on equipment support legs near the
floor juncture or the floor surface underneath equipment either after the pre-rinse or
post-rinse. The process of rinsing enables water to penetrate equipment surfaces far
beyond what is dismantled or accessible on a daily basis. Close proximity to floor
drains in freezers/ovens or persistent wet spots along floor wall junctures are other
good examples of sample sites. While a positive result will not indicate precisely
where the point of harborage is, it will notify the team that there is an active shed-
ding of a harborage site that requires immediate attention. The advantage of this
approach to sampling is that while sanitation sampling at time zero or : Environmental
Monitoring Program (EMP) sampling during production are transfer point detection
(you found something on a surface but it is most likely not where the contamination
originated from), post-rinse sampling is harborage site detection (a growth niche is
occupied and is shedding). Post-rinse sampling detects issues at the area where
“shedding” occurs and these sites are distinctly different from typical EMP and
sanitation effectiveness sites since harborage sites are not readily available for
sampling.
4.5 Maturity Model
Many companies use maturity models to help them see how their programs compare
in maturity to industry best practices. In this case, the model outlines the progress
and current state of sanitation in the food industry. A manufacturer can use the
model to see where their sanitation programs currently stand in relationship to top
performers in the industry and shows them how they can move through the different
levels of maturity toward continuous improvement and optimization of sanitation,
leading to improvements in product food safety and quality (Table 4.1).
Table 4.1 Sanitation maturity model

Low state Medium state High state
Sanitation process—Sanitation is an organized process that is respected and supported
1. Sanitation documentation 1. SSOPs provide detailed 1. SSOPs are routinely
meets regulatory and audit cleaning instructions and describe verified, updated, and audit
expectations but sanitors are performance expectations. ready. Sanitor and supervisor
not properly trained or held Sanitors are actively supervised training is ongoing and
accountable for their work. for their individual workmanship documented through training
Supervisors cannot oversee and teamwork records. Sanitors use
their team since they are flashlights to self-inspect their
often working themselves work after each rinse step
2. Sanitors work as 2. Sanitors work together to stay 2. Sanitors work in a
individuals, do not stay in in sequence and minimize the predetermined direction that
sequence, and are not aware recontamination of adjacent push soils to collection areas
or care about the equipment or area caused by for ongoing pickup instead of
consequence of overspray overspray directing soils to drains
(continued)

3. Rooms are expected to be 3. Operations, maintenance, and 3. Cross-functional room
released back to production QA support sanitation to transition transfer tasks are verified by
on time regardless of the areas from production to signed check sheets. The
delays in starting sanitation sanitation and vice versa duration allotted to sanitation
caused by extended is consistent regardless of
production or room transfer sanitation startup delays
tasks that are poorly
completed by production
Non-daily tasks—Tasks are deployed beyond daily procedures to address the risk associated
with soil type and equipment complexity
1. There is no formal 1. Every piece of RTE equipment 1. RAW and RTE equipment
requirement for non-daily has a non-daily task that is have non-daily tasks. Heat
tasks other than to scheduled based on the prevention intervention or full equipment
coordinate equipment of findings. There is an awareness teardown is fully deployed.
cleaning with preventative of the benefits of heat treatment Plants modify existing
maintenance tasks as but this practice has not been fully equipment based on findings
specified by the Original deployed and design new equipment to
Equipment Manufacturer eliminate known growth
(OEM) niches and to simplify
non-daily tasks
2. If equipment requires 2. Deep cleaning and teardown 2. Non-daily tasks are
intervention due to issues tasks are fully developed. The effective without degrading
related to food safety or schedule of non-daily tasks is assets. The frequency of deep
quality, the SSOP is altered captured in the MSS cleaning and intervention
by increasing the frequency tasks is optimized by using
of the procedure, using data to determine when tasks
higher chemical are required
concentrations or
introducing new chemicals
3. The MSS is not overtly 3. Non-daily task planning is 3. Non-daily task completion
visible and does not have reviewed at the weekly sanitation and planning is reviewed at
expectations or oversight meeting. Production requirements the weekly sanitation
may postpone tasks requiring meeting. Task completion is a
them to be rescheduled Key Performance Indicator
(KPI) and if tasks are
delayed, there is an escalation
process for resolution
Sanitary design—Sanitary design and access are built into equipment design and installation
1. Design is a consideration 1. Sanitary design is a factor in the 1. Sanitary design is
for the procurement of procurement of equipment by embedded as a requirement
equipment engineering and the cross- for the procurement of
functional plant team equipment. Suppliers and
contractors receive preferred
status based on their ability to
execute against these
principles
(continued)

2. Engineering relies on the 2. Tools such as the NAMI 2. Sanitary design is reviewed
OEM to provide equipment sanitary design checklist are used by key internal stakeholders
that meets sanitary design by key internal stakeholders to at every key stage between
principles. Tools such as the determine the marginal and procurement and installation.
NAMI sanitary design unacceptable conditions of the Deficiencies are designed out
checklist are used to equipment design. There is a bias where possible
generate a compliance toward managing deficiencies
score. Equipment installed through procedures rather than
is “off the shelf” redesign deficiencies that would
add cost or delay delivery
3. Access to all equipment 3. Equipment access for sanitation 3. Platforms and access to
surfaces and access to is a consideration. Issues arising clean equipment from all
internal structures for after installation are managed. The sides is designed in to avoid
sanitation is not considered use of ladders and tools to provide the need for ladders during
prior to installation access during sanitation is an sanitation. Guard removal to
acceptable outcome access internal surfaces do
not require tools
Performance—Driving continuous improvement
1. Sanitation costs are the 1. Performance metrics beyond 1. Sanitation KPIs are
only performance metric costs are tracked, trended, and regularly reviewed at the
that has routine tracking reviewed at the scheduled executive level. Independent
and oversight sanitation meeting. Issue verification and technical
resolution drives continuous support drive program
improvement robustness and value
2. Swabbing is biased 2. Swabbing is biased to finding 2. Swabbing is biased to
toward achieving results problem areas and verifying that verifying control of
that meet corporate targets corrective actions implemented intervention “hurdles.” Swab
are both effective and sustainable data are valued and used as a
predictive tool to ensure that
control measures are working
3. Pre-op inspection is used 3. Pre-op inspection is a coaching 3. Pre-op is a multilayered
to point out what the sanitor moment between the sanitor and verification process where
has missed. When the auditor. Findings are addressed findings (sanitor) and missed
deviations are found, they by manual means or through the findings (auditor) are tracked.
are addressed using the use of the sanitizer hose Repeat findings are addressed
pressurized hot water hose through changes in equipment
design/access or through
performance accountability
4.6 Summary
Sanitation is a key enabler for manufacturing plants to produce safe products that
will consistently meet shelf life. It is a defined process that requires the same level
of supervision and support that production processes receive. Sanitation times must
be respected and the non-daily tasks must be deployed at a frequency that is appro-
priate, given the complexity of equipment, the type of soils present, and without
degrading the assets affected. Sanitation metrics must go beyond costs to measure
effectiveness and efficiency of cleaning. Equipment design must permit cleaning

and provide access so that sanitors can do their work safely and effectively. Finally,
satisfactory performance must be driven through personal accountability and
verified following a multilayered audit approach. Repeat deviations in effectiveness
metrics must drive root cause analysis, leading to effective and sustainable correc-
tive actions.
One last point is that there is a talent gap today in the sanitation industry. The
lack of talented floor supervisors and sanitation managers is an indication that as
leaders, we have failed to invest the effort to attract and retain sanitation talent.
While many plants have chosen instead to use third-party sanitation contractors,
plants must retain control over and manage the sanitation program, holding those
accountable to perform sanitation tasks regardless of whether the labor is internal or
contracted. Food plants that recognize these human resource challenges, and devise
strategies to recruit, train, and retain sanitation personnel are more likely to succeed
in operating an effective sanitation program.
References
Allen, J.R., and E.M. Foster. 1960. Spoilage of vacuum packed sliced processed meats during
refrigerated storage. Journal of Food Science 25: 19–25.
Cramer, Michael M. 2013. Food Plant Sanitation second edition. Boca Raton, FL: Taylor &
Francis.
Doyle, Marjorie Ellin, and Kathleen E. Glass. 2010. Sodium reduction and it’s effect on food
safety, food quality and human health. Comprehensive Reviews in Food Science and Food
Safety 9: 44–55.
Kempton, A.G., and S.R. Bobier. 1970. Bacterial growth in refrigerated, vacuum-packaged lun-
cheon meats. Canadian Journal of Microbiology 16: 287–297.
Malley, T.J.V., J. Butts, and M. Wiedmann. 2015. Seek and destroy process: Listeria monocyto-
genes Process controls in the ready-to-eat and poultry industry. Journal of Food Protection 78:
436–445.
North American Meat Institute (NAMI). 2016. Sanitary equipment design principles: checklist &
glossary. Sanitary Equipment Design Taskforce. Washington, DC: AMI Foundation. https://
www.meatinstitute.org/ht/a/GetDocumentAction/i/97261. Accessed 16 February 2018.
Taormina, Peter J. 2010. Implications of salt and sodium seduction on microbial food safety.
Critical Reviews in Food Science and Nutrition 50: 209–227.
Tompkin, R.B. 2002. Control of Listeria monocytogenes in the food-processing environment.
Journal of Food Protection 65: 709–725.
Chapter 5
Advanced Processing Techniques
for Extending the Shelf Life of Foods
5.1 I ntroduction: What Is Food Spoilage and How

Is it Prevented?
Spoilage is the development of unwanted characteristics of color, texture, odors,

flavors, or other organoleptic or nutritive qualities that make food less desirable
(Montville et al. 2012). Spoilage can be the loss of attractive attributes or the devel-
opment of objectionable attributes. These characteristics can develop through abi-
otic chemical or physical changes in the food (e.g., oxidation and drying), through
metabolic activity of the commodity itself (e.g., over-ripening and respiration), or
through microbial activity from native or contaminating bacteria, fungi, or viruses.
There are many stages within the food production process in which spoilage can
occur, including processing, packaging, distribution, retail display, transport, stor-
age, and during handling by the consumer (Gould 1996). One of the many goals of
the food industry is to delay the onset of food spoilage by extending the amount of
time food can remain in storage before it will become spoiled. Current research
continues across the globe to develop cost-effective food processing techniques that
will help create food products of high quality that are safe and readily storable.
This chapter will focus primarily on food preservation techniques that prevent or
inhibit microbial growth, and the unwanted resulting changes in the food product.
One of the most familiar types of preservation is refrigeration. Refrigeration allows
for low-temperature storage of food products including meat and dairy, which keeps
present microorganisms “at bay” and preventing current populations from multiply-
ing. Other bacterial inhibition techniques include freezing, drying, curing, vacuum
packaging, modified atmosphere packaging, acidifying, fermenting, and addition of
S. M. Hertrich · B. A. Niemira (*)

Food Safety and Intervention Technologies Research Unit, Eastern Regional Research Center,
USDA-ARS, Wyndmoor, PA, USA
e-mail: shertrich@naccme.com; Brendan.Niemira@ARS.USDA.GOV

https://doi.org/10.1007/978-3-030-54375-4_5
92 S. M. Hertrich and B. A. Niemira
preservatives (Gould 1996). Other preservation techniques aim at inactivation or

elimination of microorganisms in food products such as pasteurization, sterilization,
and irradiation (Gould 1996). There are also techniques that prevent the entry of
microorganisms into foods such as aseptic packaging techniques (Gould 1996).
Consumer demands for less heavily preserved, unprocessed foods of high quality
have led to the need for processing technologies that will (1) provide less damage to
the product; (2) preserve the organoleptic and nutritional value of the produce with-
out the use of heat (i.e., nonthermal processing); (3) allow the use of more natural
antimicrobials (i.e., essential oils and spices); and (4) provide different packaging
methods that will extend shelf life (i.e., modified atmosphere packaging) (Gould
1996). This chapter will examine novel food processing technologies, and place
them in context with conventional techniques used to preserve a variety of com-
modities. The emphasis will be on spoilage and food quality, with additional discus-
sion of relevant food safety applications.
5.2 Processing Techniques for Extending Shelf Life
5.2.1 Basic Preservation Techniques

5.2.1.1 Cold Storage/Cooling/Freezing
Refrigeration, also known as cool or cold storage/chilling, refers to the storage of

foods at temperatures from 16 to –2 °C (Montville et al. 2012). The mechanism by
which refrigeration is able to delay food spoilage is by decreasing chemical reaction
rates and subsequently delaying most microbial growth (Montville et al. 2012). The
shelf life of meat, fish, and poultry products can be extended from 1 day to up to
2 weeks with refrigeration (Farkas 2001). The shelf life of fruits and vegetables can
be extended from 1–50 days to up to 300 days with refrigeration, depending on the
product (Farkas 2001).
It is generally recommended that foods are kept out of the “danger zone”—the
temperature range which favors microbial growth in foods which is between 40 and
140 °F (USDA 2013). To keep food safe from the growth of pathogens or spoilage
microbes, hot foods should be kept hot (at or above 140 °F) and cold foods should be
kept cold (below 40 °F). If a food reaches temperatures in the danger zone, bacterial
populations could double in only 20 min. At colder temperatures, some concerns
may remain regarding psychrotrophic bacteria, also known as “cold-tolerant” bacte-
ria, which include Yersinia, Vibrio, Pseudomonas, Listeria, and Aeromonas species.
These types of organisms are made up of more unsaturated fatty acid residues and
more branched-chain fatty acids in their lipid molecules compared to their meso-
philic counterparts, allowing their membrane proteins to be able to function at lower
temperatures (Montville et al. 2012). While these species are able to survive at lower
temperatures, growth is typically slowed which helps to extend the shelf life. The
ability of psychrotrophic bacteria to alter their fatty acid composition to maintain
membrane fluidity is known as homeoviscous adaptation (Montville et al. 2012).
5 Advanced Processing Techniques for Extending the Shelf Life of Foods 93
Another well-known basic form of food preservation is freezing. In general,

foods are initially frozen by contact with cold air, contact with a cold surface, or
submersion into a refrigerant liquid such as liquid nitrogen then stored at –20 °C
(Montville et al. 2012). Depending on the amount of water that is present within a
food, the eutectic temperature (the temperature where the solutes within the food
reach their solubility limit and the available water freezes) will vary. This is the state
which the food reaches the totally frozen state and occurs at −15 to –20 °C for fruits
and vegetables and –40 °C for meats (Montville et al. 2012). Some components of
foods, such as sugars and transmembrane proteins, can protect cells from mechani-
cal injury during freezing. During freezing, many microbes go into osmotic shock
where ice formation within the cells of the organisms causes mechanical injury,
resulting in reduction of populations. However, some pathogens are able to with-
stand extended frozen storage without suffering complete kill (Niemira et al. 2002;
Niemira et al. 2003). Microbial growth can occur during thawing when the contents
of freeze-ruptured food tissues become available for metabolism by associated
microorganisms. In a situation of cycles of freeze–thaw, microbial populations take
advantage of the available nutrients and present a greater risk. Refrozen products
that have been thawed are therefore especially susceptible to microbial spoilage.
Freezing can also alter the sensory qualities of the food. Development of ice crystals
on the surface of foods can occur during temperature fluctuation, or freeze–thawing,
which can also lead to microbial spoilage.
5.2.2 Thermal/Heating
Thermal processing for shelf life extension should be regarded as distinct from
cooking. Cooking transforms raw ingredients, enhancing digestibility, improving
flavor, and eliminating potentially harmful contaminating organism. However, once
cooked, either in the home or at a commercial at point of service, the issue of shelf
life is not of primary concern, as the intention is for near-immediate consumption.
Thermal processing which is intended to extend the shelf life of food will be com-
bined with specialized packaging. Historically, these have been metal cans or glass
bottles; more modern packaging for thermally processed foods include formed plas-
tic bottles or jugs, polylaminate plastic bowls, bags, or pouches, or compound con-
tainers incorporating foils, laminated paper, or other advanced materials. These
containers can readily achieve the familiar shelf-stable “canned” state, or, as with
more sensitive commodities and lower thermal processing regimes, be intended for
subsequent refrigerated storage.
Heating is the most widely utilized method for killing microbes in food. Louis
Pasteur, also referred to as the first food microbiologist, was the first to show that
spoilage of milk, wine, and beer could be prevented by heating for a short time at a
relatively low temperature. This process is now known as pasteurization and is
defined as a mild heat treatment that aims to kill nonspore-forming pathogenic bac-
teria as well as inactivate enzymes and spoilage bacteria. For example, milk is
pasteurized at 161 °F (72 °C) for 15 s. Exposing the milk to high temperatures for
longer periods of time will lead to the development of off-flavors. This is true for
most foods as many of us have experienced; when a food is heated for too long (or
burned), it becomes unpalatable. This is why temperature and time are important
variables to consider when utilizing thermal processing.
Another common form of thermal processing is sterilization which aims to kill
all microbes present within a food product. This type of thermal processing is typi-
cally used in products where the presence of bacterial sporeformers is of concern.
Sporeforming bacteria, including some Clostridium and Bacillus species, can cause
severe disease in humans and are highly resistant to heat when in the dormant sporu-
lated state (Setlow 2003). Sporeforming pathogens are only a threat when they ger-
minate and are able to produce endotoxin (Markland et al. 2013a). The goal of
commercial sterilization in food products is to eliminate any vegetative or germinat-
ing bacteria present and to prevent present spores from germinating in a food prod-
uct as well as extend the product’s shelf life by eliminating spoilage bacteria. Note
that commercial sterility is not the same as absolute sterility, which is a process
aimed at eliminating all microorganisms within a product, not just pathogens.
5.3 Advanced Processing Technologies
5.3.1 High Hydrostatic Pressure
The use of pressure in food processing was first investigated in 1899 by Bert Hite at
the Agricultural Experimental Station of West Virginia University where he discov-
ered that pressure treatment of milk and fruit-based foods could increase their shelf
life (San Martin et al. 2002). HHP-processed foods were first introduced to the
Japanese market in 1990 and have slowly since been introduced in other countries
(San Martin et al. 2002). The technology works by placing food inside of a flexible
sealed pouch that is then placed inside of a sealed vessel filled with water that is
subjected to high pressure (Montville et al. 2012). The water inside of the vessel
acts as the pressure-inducing medium. Not all food products are good candidates for
HHP due to the macromolecular changes that may occur during HHP. For example,
a food product that is porous or contains internal voids, such as peppers, melons, or
raspberries, will become compressed, altering the mouthfeel and sensory qualities
of the product. Foods with a higher moisture content tend to be better candidates for
HHP because they can withstand the compression (Montville et al. 2012). Currently,
HHP is used to commercially process products such as guacamole, pre-sliced deli
meats, juices, and oysters. Researchers and producers are interested in expanding
the application of HHP to a wider variety of foods in order to increase the safety and
shelf life of these foods (Markland 2011; Lou et al. 2012).
According to the research that has been done over the years, microorganisms are
inactivated by HHP through a variety of mechanisms. One hypothesis is that a
pressure-induced decrease in cell volume can lead to cell leakage and death (San
Martin et al. 2002; Chilton et al. 1996); however, the mechanism of inactivation of
cells by HHP can be dependent on the type of pressure applied (cyclic or continu-
ous), temperature, treatment time, strain, cell shape, Gram stain type, growth stage,
and treatment medium (San Martin et al. 2002). HHP of vegetative bacterial cells is
generally more effective at higher temperatures unless the bacterial species contains
the heat shock protein (Hsp), in which case heat would cause a baroresistant effect
(Iwahashi et al. 1996; Li and Gänzle 2016). The resistance of microorganisms to
HHP is largely dependent upon the species and strain of the microorganism and is
extremely variable (Raso et al. 1998), but most vegetative cells of bacteria and yeast
are generally inactivated at pressures around 300–400 MPa at ambient temperature
(Knorr 1995).
One of the current disadvantages of HHP is its inability to inactivate bacterial
spores by pressure alone without altering the sensory qualities of the product (San
Martin et al. 2002; Black et al. 2007). Studies show that HHP in combination with
heat can inactivate spores within a food product; however, the nutritional and organ-
oleptic qualities of some foods cannot withstand the thermal treatment (Paidhungat
et al. 2002; Wuytack et al. 2000). Complete inactivation of spores remains a top
priority for high-pressure food processors. It is important to understand the physiol-
ogy of spores, especially those that pertain to spore inactivation by HHP (Black
et al. 2007).
5.3.2 Ultraviolet and Pulsed Light
Application of ultraviolet light (UV) was first used in France for disinfection of
drinking water (Masschelein 2002). UV is currently used for the preservation of
solid and liquid foods. Because of its limited wave penetration, it is most effective
for the inactivation of microorganisms on the surfaces of foods or clear liquids. It
has been often used as an alternative to pasteurization of liquids and juices because
the lack of heat helps to preserve the fresh flavor as well as extend the shelf life.
Pulsed light is another method of food preservation that uses intense and short
pulses of broad-spectrum white light. One of the advantages of pulsed light over UV
light is that it has a greater penetration depth (Bialka et al. 2008).
UV uses an electromagnetic spectrum from 100 to 400 nm and can be classified
into four spectrum regions including UV-A (315–400 nm), UV-B (280–315 nm),
UV-C (200–280 nm), and vacuum UV (100–200 nm) (Keklik et al. 2012). The
UV-C class is considered to be the most effective regarding microbial contamina-
tion of food products (Keklik et al. 2012). The wavelength of pulsed white light
ranges from ultraviolet to infrared (Li and Farid 2016). The mechanism by which
UV and pulsed light inactivate microorganisms is by damaging the organism’s DNA
(Li and Farid 2016).
Advantages of using UV and pulsed light technology for food preservation
include the relatively low cost and the lack of the generation of chemical residues
within the product (Hijnen et al. 2006). Like other nonthermal technologies, one of
the limitations of UV light technology is that it has little effect on the killing of
bacterial spores; however, it has been shown to help improve the lethal effects of
other subsequent treatments (Li and Farid 2016; Gayán et al. 2013). Pulsed light, on
the other hand, has been found to effectively inactivate spores in some products (Li
and Farid 2016). This is most likely due to the thermal stress that is induced upon
the spore coat leading to structural damage of the cell (Hijnen et al. 2006). Another
one of the disadvantages of UV light technology is the limited amount of transmit-
tance through products (Gomez-Lopez et al. 2012; Keklik et al. 2012). To help
overcome this, UV food treatment chambers have been designed so that fluids flow
through it in a thin layer (Gomez-Lopez et al. 2012). Some of the major applications
of UV in the food industry include treatment of liquid foods specifically juices,
milk, and liquid egg products (Li and Farid 2016). Pulsed light is less widely uti-
lized in the industry, although promising results have been observed for inactivation
of bacterial spores in cornmeal (McDonald et al. 2000) and sucrose syrup (Chaine
et al. 2012). More studies need to be performed for the optimization of pulsed light
technology for the preservation of foods.
5.3.3 Ultrasonication
Ultrasonication is a more recently developed technology that preserves foods

through a phenomenon known as cavitation that is created by ultrasound waves
(Delmas and Barthe 2015). Cavitation occurs when vapor cavities, or bubbles, are
formed within a liquid and the ultrasonic energy continues to increase until the
vapor cavities begin to rapidly collapse (implode), creating a shock wave that leads
to short periods of high temperature and pressure throughout the liquid (Butz and
Tauscher 2002; Montville et al. 2012; Delmas and Barthe 2015). The tiny bubbles
which are filled with heat and reactive oxygen species including hydrogen peroxide
are released through the breakage of chemical bonds and are able to kill microbial
cells (Montville et al. 2012). There are two types of ultrasound including high fre-
quency (megahertz range) and low frequency (kilohertz range) (Montville et al.
2012). The frequency necessary to inactivate microorganisms through ultrasonica-
tion is 20–100 kHz (Chandrapala et al. 2012).
While this technology was first described to have the potential to kill microor-
ganisms in 1933 (Szent-Gyorgyi 1933), its potential as a technology to preserve
foods was not recognized until the last few decades (Li and Farid 2016). This tech-
nology is not effective for the inactivation of microorganisms when used alone;
however, it can be effective when used synergistically with other technologies, spe-
cifically thermal technologies (Li and Farid 2016). It has also been shown to increase
the efficacy of HHP (Raso et al. 1998). The use of ultrasonication along with ther-
mal treatments (thermosonication) has been shown to help reduce the time needed
to sterilize a product, which helps to maintain the nutritional and organoleptic quali-
ties of the product (Chandrapala et al. 2012; Mason et al. 2015). Thermosonication
studies demonstrated that ultrasonication combined with thermal treatment at 70 °C
significantly enhanced the inactivation of Bacillus cereus spores in skim milk, liq-
uid beef, cheese products, and rice porridge compared to heating the products alone
(Evelyn and Silva 2015).
Because the effects of cavitation are stronger than the adhesion forces (van der
Waals attraction) on surfaces, ultrasonication may be most effective at surface dis-
infection of foods specifically fresh produce or processing equipment (Montville
et al. 2012). It could potentially also serve as a good preservation process for liquid
products including liquid egg products and juices. The use of the technology is cur-
rently expanding in the wine and beverage industries as well (Montville et al. 2012).
While ultrasonication shows promise for use as a preservation technology in foods,
more studies are needed to address its limitations.
5.3.4 Irradiation
Irradiation uses electrons or photons of sufficient energy that ionizes the molecules
they contact. This ionization is usually enacted in water molecules which are the
single greatest component of foods, leading to the creation of reactive hydrogen and
hydroxyl radicals (Niemira 2014). Unlike UV, which is not powerful enough to ion-
ize and has limited penetrability, ionizing radiation can penetrate solid foods to a
depth of 4–6 cm in the case of electrons, but penetrating as much as 40–50 cm in the
case of photons (either gamma rays or X-rays). The three types of ionizing radiation
are generated by different methods. Gamma rays are produced by certain radioiso-
topes, with cesium-137 and cobalt-60 being the most commonly used for food pro-
cessing. Ionizing electrons (known as electron beams or “e-beams”) are produced
by using magnetic fields to accelerate electrons to high energies, up to 10 MeV. X-rays
are created by directing accelerated electrons into a metal target, usually a dense
alloy of tungsten. The interaction of the electrons with the metal releases a shower
of high-energy X-rays.
All three types of irradiation have been shown to extend the shelf life and improve
the safety of fruits, vegetables, meats, poultry, seafood, eggs, and processed foods
(Niemira et al. 2002; Alvarez et al. 2006; Niemira and Cooke 2010). Irradiation may
be used at doses up to 1 kGy to delay sprouting, slow ripening and/or maturation,
and extend shelf life of stored produce. Other commodities have different statutory
dose limitations, depending on the purpose of irradiation. For example, iceberg let-
tuce and spinach can be treated with up to 4.0 kGy to improve safety (CFR 2017).
5.3.5 Cold Plasma
An emerging technology for food processing is cold plasma, which for practical
purposes may be regarded as a form of ionized gas. Cold plasma is generated by
ionizing gases or gas mixtures with high-voltage electricity or microwaves; the
resulting plasma serves as a nonthermal antimicrobial intervention for foods

(Niemira et al. 2014). The antimicrobial modes of action are primarily related to (1)
chemical reactions with cellular structures; (2) UV damage of cellular components;
and (3) UV-mediated DNA strand breakage. A wide array of equipment designs to
generate cold plasma have been reported, and are an area of active research and
development. In some cases, this research seeks to adapt mature thermal plasma
systems used for surface treatment applications in industries such as textiles, print-
ing, polymer processing, or electronics (McHugh and Niemira 2016). Cold plasma-
mediated inactivation of human pathogens is a primary goal of these food safety
research efforts, with associated extension of shelf life as a concomitant goal
(Lacombe et al. 2015; Min et al. 2016).
Key areas for further development of cold plasma include the determination of
the precise modes of action for various operating conditions; optimization of feed
gas mixtures mated with optimized equipment designs for antimicrobial efficacy,
cost and efficiency; and improving compatibility with existing food handling and
packaging systems. Another body of emerging body of work is the use of cold
plasma to sanitize packaging materials, food contact surfaces, and areas where
sanitizer-resistant pathogen biofilms form (Niemira et al. 2014).
5.3.6 Pulsed Electric Fields
The first interest in the use of electric fields for food processing began in the 1930s,
before the advent of energy-efficient electronic control systems (Montville et al.
2012). Modern pulsed electric field (PEF) systems use about 80% less energy than
thermal treatments (Kempkes 2010), but are overall more expensive (Montville
et al. 2012). PEF technology can be described by high-intensity electric fields which
are varied between 20 and 80 kilovolts (kV) per centimeter for a very short time
(1–100 ms) (Amiali and Ngadi 2012; Raso et al. 2014). PEFs are able to kill vegeta-
tive bacteria through a process known as electropermeabilization (Montville et al.
2012). This process is defined as an electric sock which momentarily opens pores
within a bacterial cell’s plasma membrane allowing the entry of other macromole-
cules into the cell, which then leads to cell death. While the mechanism by which
PEF kills bacterial spores is not fully elucidated, it is suspected that the mechanism
is similar to that of vegetative bacteria (Li and Farid 2016).
In the food industry, a solid food product is passed through PEFs in a continuous
system such as on a conveyor belt. The efficacy of the PEF treatment, and its ability
to kill microorganisms will depend on the type of food product being tested and
depend largely on the type of organism present. It is believed that, in general, Gram-
negative bacteria are most susceptible to PEF treatment and bacterial and yeast
spores are most resistant (Barbosa-Canovas et al. 1999; Yonemoto et al. 1993).
Other factors that affect microbial reductions by PEF include process conditions
(field strength, pulse width, pulse frequency, total treatment time, input energy),
production conditions (flow rate, holding time, temperature), and food properties
(pH, conductivity, particulate) and it is recommended that scale-up and validation

studies in a specific PEF system for specific products be performed (Jin et al. 2015).
Some argue that high cost is a limitation for commercial use of PEF technology
(Kempkes 2010); however, this issue remains controversial.
Initial commercial interests for use of PEF technology mostly surrounded juice
and beverage products including tomato juice (Min et al. 2003). In 2016, testing for
a pilot-scale batch PEF unit was released for the processing of French fries, potato
chips, and other specialty potato products and raw materials including sweet pota-
toes, cassava, beetroot, and carrots (PotatoPro 2016). With the use of this system,
potato cells are electroporated in order to release intracellular compounds, such as
reducing sugars involved in the Maillard browning reaction, which reduces the ten-
dency of browning during frying (PotatoPro 2016). This undesirable browning
gives the potatoes a burned, or overcooked, appearance. This new innovation could
open up the possibilities for the processing of solid plant-based products. Other
applications of PEF include mild preservation of beverages and semi-liquid food
products, extraction processes such as extraction of antioxidants, extraction of oil
and protein from algae, extraction of sugar from sugar beets, and extraction of nutri-
ents or fibers from peels and stems (Pulsemaster 2017). PEF can also be applied for
the removal of acrylamide, concentration of protein from potatoes, and enhance-
ment of production processes for cooked ham and dry sausage (Pulsemaster 2017).
5.3.7 Ozone
The antimicrobial effectiveness of ozone has been shown to be much higher than
that of chlorine and to affect a broader spectrum of microorganisms than chlorine
and other disinfectants (Hirneisen et al. 2010). Bacterial spores have also been
shown to be inactivated by ozone (Markland et al. 2013b; Young and Setlow 2004).
Studies involving the inactivation of spores by oxidizing agents suggest that inacti-
vation is a result of oxidative damage to the spore’s inner membrane (Markland
2011). Ozone can also degrade mycotoxins and pesticides present in foods (Karaca
and Velioglu 2007). In addition, there is little concern of residual ozone in treated
food products due to the rapid decomposition of ozone into oxygen (Graham 1997)
and ozone is currently certified for use on organic foods.
Ozone is applicable in the food industry to treat process water, as a fruit and
vegetable wash, in fruit and vegetable storage, and in recycled water (Hirneisen
et al. 2010). Aqueous ozone has been used to increase the shelf life of apples, straw-
berries, and in juices such as apple cider and orange juice (Hirneisen et al. 2010);
however, the efficacy of aqueous ozone is largely dependent upon the presence of
organic residues, pH, and temperature of the aqueous medium (Hoigné and Bader
1975; Karaca and Velioglu 2007). In general, ozone is more effective at lower tem-
peratures, below pH 5.0, and higher humidity (Karaca and Velioglu 2007).
There are some limitations regarding the use of ozone for food preservation.
Ozone is a very reactive molecule that has the ability to inactivate a broad range of
microorganisms; however, it also reacts with nearly all organic and inorganic com-
pounds (Karaca and Velioglu 2007). Therefore, the higher the amounts of organic
matter present in a food product, the lower the effectiveness of ozone. Ozone may
also cause slight deleterious effects on the quality and physiology of food products
such as losses in sensory quality including enzymatic browning, antioxidants, vita-
mins, and minerals (Karaca and Velioglu 2007). Exposure of humans and animals to
high levels of ozone can also have detrimental effects on health, which causes con-
cern for workers in processing plants. In the United States, OSHA (Federal
Occupational Safety and Health Administration) limits exposure to ozone to a 0.1-
ppm threshold for continuous exposure for an 8-h period and 0.3 ppm for a 15-min
period (Suslow 2004).
5.4 Conclusions
There are many advantages to the use of advanced processing techniques, yet there
are limitations that prevent wider utilization by the food industry. The majority of
these technologies are only able to successfully treat food products of a certain
moisture content, composition, or surface area. Bacterial sporeformers have also
proven to be a challenge for nonthermal technologies as heat is necessary for com-
plete inactivation. More attention should be given to technologies that may prevent
the germination of spores. For example, research has shown that when used syner-
gistically, or in various combinations with one another, these technologies as well as
the addition of mild heat can effectively inactivate spores. Another limitation regard-
ing these technologies is the lack of consumer acceptance. Consumer education and
marketing will be a crucial component as these technologies continue to be devel-
oped. Overall, further research should be performed in regard to the advanced pro-
cessing techniques discussed in this chapter in order to expand their applications.
This will allow food companies to be able to continue to provide their consumers
with a high variety of products that are safe and shelf-stable.
References
Alvarez, I., B.A. Niemira, X. Fan, and C.H. Sommers. 2006. Inactivation of Salmonella serovars
in liquid whole egg by heat following irradiation treatments. Journal of Food Protection 69
(9): 2066–2074.
Amiali, M., and M.O. Ngadi. 2012. Microbial decontamination of food by pulsed electric fields. In
Microbial Decontamination in the Food Industry, Novel Methods and Applications, 407–449.
Washington, DC: Woodhead Publishing.
Barbosa-Canovas, G.V., M.M. Gongora-Nieto, U.R. Pothakamury, and B.G. Swanson. 1999.
Preservation of Foods with Pulsed Electric Fields. San Diego, CA: Academic.
Bialka, Katherine L., Ali Demirci, Paul N. Walker, and Virendra M. Puri. 2008. Pulsed UV-light
penetration of characterization and the inactivation of Escherichia Coli K12 in solid model
systems. Transactions of the ASABE 51.
Black, E.P., P. Setlow, A.D. Hocking, C.M. Stewart, A.L. Kelly, and D.G. Hoover. 2007. Response
of spores to high-pressure processing. Comprehensive Reviews in Food Science and Food
Safety 6: 103–119.
Butz, P., and B. Tauscher. 2002. Emerging technologies: Chemical aspects. Food Research
International 35 (2–3): 279–284.
Chaine, A., C. Levy, B. Lacour, C. Riedel, and F. Carlin. 2012. Decontamination of sugar syrup by
pulsed light. Journal of Food Protection 75 (5): 913–917.
Chandrapala, Jayani, Christine Oliver, Sandra Kentish, and Ashokkumar Muthupandian. 2012.
Ultrasonics in food processing. Ultrasonics Sonochemistry 19 (5): 975–983.
Chilton, P., N.S. Isaacs, B. Mackey, and R. Stenning. 1996. The effects of high hydrostatic
pressure on Bacteria. In High Pressure Research in the Biosciences and Biotechnology, ed.
K. Heremans. Belgium: Leuven University Press.
Code of Federal Regulations. 2017. 21 CFR.179: Irradiation in the production, processing,
and handling of food. http://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfcfr/CFRSearch.
cfm?fr=179.26. Accessed 30 January 2017.
Delmas, H., and L. Barthe. 2015. Ultrasonic mixing, homogenization, and emulsification in
food processing and other applications. In Power Ultrasonics Applications of High-Intensity
Ultrasound, 757–791. Washington, DC: Woodhead Publishing.
Evelyn, E., and Filipa V. Silva. 2015. Thermosonication versus thermal processing of skim Milk
and beef slurry: Modeling the inactivation kinetics of Psychrotrophic Bacillus Cereus spores.
Food Research International 67: 67–74.
Farkas, J. 2001. In Food Microbiology: Fundamentals and Frontiers, ed. M.P. Doyle, L.R. Beuchat,
and T.J. Montville, 2nd ed. Washington, DC: Woodhead Publishing.
Gayán, E., I. Álvarez, and S. Condón. 2013. Inactivation for bacterial spores by UV-C light.
Innovative Food Science and Emerging Technologies 19: 140–145.
Gomez-Lopez, V.M., T. Koutchma, and K. Linden. 2012. Ultraviolet and pulsed light processing
of fluid foods. In Novel Thermal and Non-Thermal Technologies for Fluid Foods. Cambridge,
MA: Academic.
Gould, G.W. 1996. Methods for preservation and extension of shelf life. International Journal of
Food Microbiology 33 (1): 51–64.
Graham, D.M. 1997. Use of ozone for food processing. Food Technology 51 (6): 72–75.
Hijnen, W.A., E.F. Beerendonk, and G.J. Medema. 2006. Inactivation credit of UV radiation for
viruses, Bacteria and protozoan OoCysts in water: A review. Water Research 40 (1): 3–22.
Hirneisen, K.A., E.P. Black, J.L. Cascarino, V.R. Fino, D.G. Hoover, and K.E. Kniel. 2010.
Comprehensive. Reviews in Food Science and Food Safety 9 (1): 3–20.
Hoigné, J., and H. Bader. 1975. Ozonation of water: Role of hydroxyl radicals as oxidizing inter-
mediates. Science 190 (4216): 782–784.
Iwahashi, H., K. Obuchi, S. Fuji, K. Fujita, and Y. Komatsu. 1996. The reason why Trehalose
is more important for Barotolerance than Hasp104 in Saccharomyces Cerevisiae. In High
Pressure Research in the Biosciences and Biotechnology, ed. K. Heremans. Belgium: Leuven
University Press.
Jin, T.Z., M. Guo, and H.Q. Zhang. 2015. Upscaling from benchtop processing to industrial scale
production: More factors to be considered for pulsed electric field food processing. Journal of
Food Engineering 146: 72–80.
Karaca, H., and Y. Velioglu. 2007. Ozone applications in fruit and vegetable processing. Food
Reviews International 23: 91–106.
Keklik, N.M., K. Krishnamurthy, and A. Demirco. 2012. Microbial decontamination of food by
ultraviolet (UV) and pulsed UV light. In Microbial Decontamination in the Food Industry,
Novel Methods and Applications, Woodhead Publishing Series in Food Science, Technology
and Nutrition, ed. A. Demirci and M.O. Ngadi, 344–369. Washington, DC: Woodhead
Publishing.
Kempkes, M.A. 2010. Pulsed Electric Field (Pef) systems for commercial food and juice pro-
cessing. In Case Studies in Novel Food Processing Technologies, Innovations in Processing,
Packaging and Predictive Modeling, Woodhead Publishing Series in Food Science, Technology
and Nutrition, ed. C. Doona, K. Kustin, and F.E. Feeherry, 73–102. Washington, DC: Woodhead
Publishing.
Knorr, D. 1995. High pressure effects on plant derived foods. In High Pressure Processing of
Foods, ed. D.A. Ledward, D.E. Johnston, R.G. Earnshaw, and M. Hasting. Nottingham:
Nottingham University Press.
Lacombe, A., B.A. Niemira, J.B. Gurtler, X. Fan, J. Sites, G. Boyd, and H. Chen. 2015.
Atmospheric cold plasma inactivation of aerobic microorganisms on blueberries and
effects on quality attributes. Food Microbiology 46 (2015): 479–484.
Li, X., and M. Farid. 2016. A review on recent development in non-conventional food sterilization
technologies. Journal of Food Engineering 182: 33–45.
Li, H., and M. Gänzle. 2016. Some like it hot: Heat resistance of Escherichia coli in food. Frontiers
in Microbiology 7: 1763.
Lou, F., P. Huang, H. Neetoo, J. Gurtler, B.A. Niemira, H. Chen, X. Jiang, and J. Li. 2012. High
pressure inactivation of human norovirus virus-like particles: Evidence that the capsid of
human norovirus is highly pressure resistant. Applied and Environmental Microbiology 78:
5320–5327.
Markland, S.M. 2011. Characterization of Superdormant Spores of Bacillus Cereus and Bacillus
Weihenstephanensis. Masters Thesis. Department of Animal and Food Sciences. University
of Delaware, Newark DE. http://udspace.udel.edu/handle/19716/11705. Accessed 4
November 2016.
Markland, S.M., D.F. Farkas, K.E. Kniel, and D.G. Hoover. 2013a. Pathogenic Psychrotolerant
Sporeformers: An emerging challenge for low-temperature storage of minimally processed
foods. Foodborne Pathogens and Disease 10 (5): 413–419.
Markland, S.M., K.E. Kniel, P. Setlow, and D.G. Hoover. 2013b. Nonthermal inactivation of het-
erogeneous and Superdormant spore populations of Bacillus Cereus using ozone and high pres-
sure processing. Innovative Food Science and Emerging Technologies 19: 44–49.
Mason, T.J., F. Chemat, and M. Ashokkumar. 2015. Power ultrasonics for food processing. In
Power Ultrasonics, 815–843. Washington, DC: Woodhead Publishing.
Masschelein, Willy J. 2002. In Ultraviolet Light in Water and Wastewater Sanitation, ed. Rip
G. Rice. Washington, DC: Lewis Publishers.
McDonald, K.F., R.D. Curry, T.E. Clevenger, K. Unklesbay, A. Eisenstrack, J. Golden, and
R.D. Morgan. 2000. A comparison of pulsed and continuous ultraviolet light sources for the
decontamination of surfaces. IEEE Transactions on Plasma Science 28 (5): 1581–1587.
McHugh, T., and B.A. Niemira. 2016. Cold plasma processing of foods. Food Technology
Magazine 3 (16): 68–72.
Min, S., Z.T. Jin, and Q.H. Zhang. 2003. Commercial scale pulsed electric field processing of
tomato juice. Journal of Agricultural and Food Chemistry 51 (11): 3338–3344.
Min, S., S.H. Roh, B.A. Niemira, J.E. Sites, G. Boyd, and A. Lacombe. 2016. Dielectric bar-
rier discharge atmospheric cold plasma inhibits Escherichia coli O157:H7, Salmonella,
Listeria monocytogenes, and Tulane virus in Romaine lettuce. International Journal of Food
Microbiology 237 (2016): 114–120.
Montville, Thomas J., Karl R. Matthews, and Kalmia E. Kniel. 2012. Food Microbiology: An
Introduction. Washington D.C.: ASM Press.
Niemira, B.A. 2014. Irradiation, microwave and alternative energy-based treatments for low
water activity foods. In Microbiological Safety of Low aw Foods and Spices, ed. M. Doyle,
J. Kornacki, and J. Gurtler, 389–401. New York, NY: Springer.
Niemira, B.A., and P. Cooke. 2010. Escherichia coli O157:H7 biofilm formation on lettuce and
spinach leaf surfaces reduces efficacy of irradiation and sodium hypochlorite washes. Journal
of Food Science 75 (5): M270–M277.
Niemira, B.A., X. Fan, and C.H. Sommers. 2002. Irradiation temperature influences product qual-
ity factors of frozen vegetables and radiation sensitivity of inoculated Listeria monocytogenes.
Journal of Food Protection 65 (9): 1406–1410.
Niemira, B.A., C.H. Sommers, and G. Boyd. 2003. Effect of freezing, irradiation and frozen stor-
age on survival of Salmonella in concentrated Orange juice. Journal of Food Protection 66:
1916–1919.
Niemira, B.A., G. Boyd, and J. Sites. 2014. Cold plasma rapid decontamination of food contact
surfaces contaminated with Salmonella biofilms. Journal of Food Science 79 (5): M917–M922.
Paidhungat, M., B. Setlow, W.B. Daniels, D. Hoover, E. Papafragkou, and P. Setlow. 2002.
Mechanisms of induction of germination of Bacillus subtilis spores by high pressure. Applied
PotatoPro. 2016. Pulsed Electric Field for French Fries and Chips: Quantify your benefits with
Solidus. http://www.potatopro.com/news/2016/pulsed-electric-field-french-fries-and-chips-
quantify-your-benefits-solidus. Accessed 7 November 2016.
Pulsemaster. 2017. FAQ about pulsed electric field processing: What are typical applications of
PEF processing? https://www.pulsemaster.us/pef-pulsemaster/faq. Accessed 13 February 2017.
Raso, J., M.M. Gongora-Neito, G.V. Barbosa-Canovas, and B.G. Swanson. 1998. Influence of sev-
eral environmental factors on the initiation of germination and inactivation of Bacillus Cereus
by high hydrostatic pressure. International Journal of Food Microbiology 44 (1–2): 125–132.
Raso, J., S. Condon, and I. Alvarez. 2014. Non-thermal processing: Pulsed electric field. In
Encyclopedia of Food Microbiology, ed. C.A. Bratt, 966–973. New York: Academic.
San Martin, M.F., G.V. Barbosa-Canovas, and B.G. Swanson. 2002. Food processing by high
hydrostatic pressure. Critical Reviews in Food Science and Nutrition 46 (6): 627–645.
Setlow, P. 2003. Spore germination. Current Opinion in Microbiology 6 (6): 550–556.
Suslow, Trevor V. 2004. Ozone applications for postharvest disinfection of edible horticultural
crops. University of California ANR, 1–8.
Szent-Gyorgyi, A. 1933. Chemical and biological effects of ultra-sonic radiation. Nature 131
(3304): 278.
United States Department of Agriculture. 2013. “Danger Zone”. http://www.fsis.usda.gov/wps/
portal/fsis/topics/food-safety-education/get-answers/food-safety-fact-sheets/safe-food-han-
dling/danger-zone-40-f-140-f/CT_Index. Accessed 4 November 2016.
Wuytack, E.Y., J. Soons, F. Poschet, and C.W. Michiels. 2000. Comparative study of pressure-
and nutrient-induced germination of Bacillus subtilis spores. Applied and Environmental
Yonemoto, Yoshimasa, Tetsuo Yamashita, Masafumi Muraji, Wataru Tatebe, Hiroshi Ooshima,
Jyoji Kato, and Akira Kimura. 1993. Resistant of yeast and bacterial spores to high voltage
electric pulses. Journal of Fermentation and Bioengineering 75 (1): 99–102.
Young, S.B., and P. Setlow. 2004. Mechanisms of Bacillus Subtilis spore resistance to and killing
by aqueous ozone. Journal of Applied Microbiology 96 (5): 1133–1142.
Chapter 6
Packaging of Perishable Food Products
Cynthia Ebner, Angela Morgan, and Clyde Manuel
6.1 Introduction
While often overlooked or simply disregarded as waste, packaging is an integral

component of the modern-day food production system, allowing for the benefits of
food processing to be maintained long after production. Without food packaging,
modern food products would have limited shelf life, be susceptible to potentially
hazardous and deleterious contamination, and would lack numerous features
designed for convenience that many consumers have grown accustomed to. With
these considerations in mind, it is easy to appreciate the important role of packaging
in delivering food of acceptable quality to the consumer.
In this chapter, the reader is presented with a broad overview of the world of food
packaging, with a specific emphasis on content relevant to the packaging of perishable
food products, such as meats, cheeses, and fresh produce. This chapter is not intended
to be a comprehensive study of all aspects of food packaging, as there are numerous
excellent reviews that fit this role (Del Nobile and Conte 2013; Robertson 2012).
C. Ebner
Sealed Air Corporation, Charlotte, NC, USA
A. Morgan
Aptar Group, Crystal Lake, IL, USA
C. Manuel (*)
GOJO Industries, Akron, OH, USA
e-mail: manuelc@GOJO.com

https://doi.org/10.1007/978-3-030-54375-4_6
106 C. Ebner et al.
6.2 Packaging Functions
6.2.1 Containment
Quite possibly, the most basic function of a package, containment, allows for the trans-
port and storage of food and other goods. At first glance, it may seem easy to dismiss
this function of a package as blatantly obvious, but its importance is nevertheless
acknowledged; simply put, a package will not function properly if it does not contain
its contents. Some of the earliest forms of packaging, such as pottery and bags, were
originally intended for the containment of food. In one instance, chemical analysis of
the lipid content on ancient pottery from Japan suggests that ceramic pots were used to
store, transport, and process fish and seafood as far back as 15,000 years ago, many
years before the advent of farming practices (Lucquin et al. 2016). The time frame of
this discovery suggests that the widespread adoption of pottery may not have coin-
cided with the settling of mankind due to farming, but in fact may have arisen much
earlier during the age of hunter gatherers.
6.2.2 Protection
Perhaps the most important function of a food package is to protect its contents
from becoming unfit for consumption. A product may become unfit for consump-
tion if its perceived quality falls below a level of consumer acceptance, or if it is no
longer safe to consume. In general, factors that contribute to the loss of product
acceptability fall into one of three categories: physical, chemical, and biological.
Packaging can play a major role in preventing product deterioration from each of
these three factors.
Food products are subjected to physical abuse throughout their entire life cycle,
including production, transit, and even during storage and display on store shelves.
A damaged package will lead to loss of quality and reduced shelf life of a product,
as the damaged package is no longer able to protect the food product from the ele-
ments. Because of this, great care is often taken to design a packaging system that
minimizes the effects of physical abuse. On the production floor, a packaged prod-
uct will have to withstand the abuse of both manual and automatic handling. For
example, it is not uncommon for vacuum packaged meat products to puncture while
being transported on conveyor systems, especially if the system is in a state of dis-
repair or if processing and packaging line speeds are exceedingly fast. To combat
this, packaging materials used for vacuum packaged meat products are sometimes
designed with heavier gauge materials that are more resistant to abuse due to their
increased thickness. During transportation, products typically experience mechani-
cal shock in the forms of drops and vibrations. Materials such as corrugated paper-
board are typically used for building pallets for shipment purposes, as it offers
exceptional crush protection.
6 Packaging of Perishable Food Products 107
Many food products can become unacceptable as the result of degradative chem-
ical processes. Oxygen, moisture, and light are the primary extrinsic factors that can
initiate and accelerate these processes, and the material and type of food packaging
can be tailored to minimize these chemical processes. Meat is often vacuum pack-
aged in materials with oxygen barrier properties in order to slow the progression of
rancidification, which occurs when oxygen accelerates the breakdown of unsatu-
rated fatty acids into volatile aldehydes and ketones, which emit an unpleasant and
rancid odor. Moisture gain or loss is another factor that contributes to the loss of
quality of food products. Products with extremely low water moisture contents,
such as crackers or potato chips, are often packaged in materials that have moisture
barrier properties in order to prevent staling caused by an influx of moisture into the
package. Finally, excessive light exposure can reduce the shelf life of many foods,
especially those with a high fatty acid content. Beer is a product familiar to consum-
ers that is particularly susceptible to the deleterious effects of light. Packaging of
beer in metal kegs, cans, or brown bottles, has been shown to significantly increase
the shelf life of beer beyond that of clear packages, since the rate of photolysis of
hop alpha acids is reduced in these packages (Heyerick et al. 2003). Beer that has
undergone photolysis of hop alpha acids is easily recognizable by the consumer as
having a cardboard or skunky taste.
Packaging can also protect food from biological spoilage. Pests such as insects
and rodents can eat through packaging materials and compromise the contents of
the package. This can be prevented by packaging food in materials that are pest-
resistant (often by increasing material thickness), storing palletized products prop-
erly in warehouses, and utilizing pest control solutions within the storage
environment. Spoilage and pathogenic microorganisms can render a food inedible
and even unsafe if allowed to grow to high levels, and packaging can serve as a bar-
rier to entry for these microorganisms. The foodborne pathogen, Listeria monocyto-
genes, for example, is a contaminant in the environment of some food processing
plants, especially those that produce ready-to-eat (RTE) foods including meat prod-
ucts. As an additional L. monocytogenes control measure, many RTE meat proces-
sors apply a post-packaging lethality step on the finished product by hot water
immersion. This involves the use of a higher heat-resistant type of packaging film
and results in both a reduction in pathogen numbers and extended shelf life for the
product.
6.2.3 Convenience
Many design aspects of food packages have arisen from the need for added conve-
nience throughout the life cycle of a product. For example, a product’s distribution
chain can be optimized and streamlined when the product is distributed by organiz-
ing finished product into units at varying steps. At the primary level, products are
individually packaged for display purposes and for consumer purchase. At the sec-
ondary level, multiple units destined for product display are organized into a large
108 C. Ebner et al.
package (typically made of corrugated paperboard case) that adds convenience

when transporting products at the store level. Finally, at the tertiary level, secondary
units of products are organized together for the purposes of shipment, usually on a
stretch-wrapped pallet. Organizing products into these units simplifies the logistics
involved in transporting many products over long distances.
In recent years, numerous commercial examples of innovative convenience fea-
tures have emerged with direct ties to the shelf life of a food product. In many
cases, these features fall under the category of intelligent packaging, which is
defined as packaging that can track, sense, and/or measure some aspect of the
contained product and then communicate this information to the consumer or user.
While still in their commercial infancy, these intelligent packages can communi-
cate the shelf life status to the consumer. Intelligent packaging is discussed in
more detail later in this chapter.
6.2.4 Communication
The package that encloses food also provides a platform for communicating impor-
tant information to the consumer. Food companies can differentiate themselves
from their competitors by incorporating marketing or branding materials into their
package designs. In the United States, the Food and Drug Administration (FDA)
regulates much of the information that appears on the outside of a primary food
package intended for consumer purchase. Information such as statements of iden-
tity, nutritional facts, ingredient declarations, package contents, net weight, and
serving sizes are examples of information required by law. These regulations are
designed to assist consumers in making informed decisions on their food purchases.
It should be noted that messages tied to product shelf life, such as “use by,” “best
before,” “sell by,” or “expired by” dates, are not regulated and thus not required by
law (the sole exception to this being infant formula).
6.3 Materials Used in Food Packaging
Food can be packaged in a wide variety of materials. The ideal choice of material
for a particular food product is influenced by a variety of factors, including cost,
appearance, flexibility, durability, ease of implementation with production process,
and compatibility with food. For perishable products such as packaged meat, sea-
food, and fruits and vegetables, one of the most important variables to consider is
permeability of the material to moisture, oxygen, carbon dioxide, and also light, as
these factors are highly influential on various processes that dictate the shelf life of
a food product. These factors are not discussed at length in this chapter but are dis-
cussed in general terms.
6.3.1 Metal
Metals have a variety of advantages as a packaging material. They offer superior

physical durability, barrier protection from gas transmission and light penetration,
and are highly recyclable. Steel and aluminum are the two most frequently used
metals for food packaging. Most cans produced in the United States each year are
made of steel, which is typically coated with a thin layer of electrically deposited tin
to enhance corrosion resistance (hence the name “tin can”). Aluminum, while more
expensive than steel, offers advantages in the form of lighter weight and enhanced
corrosion resistance. Most metal containers used for food products are lined with an
inert protective enamel coating that prevents contact between the product and the
metal itself. This is especially important for high acid foods such as soda, juices,
and tomato-based products, as contact with the metal itself will rapidly leach metal-
lic ions into the food itself. The leaching of metallic ions into the food leads to a
reduction in product quality due to both flavor loss and nutrient loss. A well-known
example of this is the rapid loss of ascorbic acid (vitamin C) in the presence of
metallic ions. In addition to rigid containers such as cans and trays, metallic layers
can also be incorporated into flexible packages by lamination or by metalizing plas-
tic film in a vacuum chamber, thus incorporating a high-quality barrier into a flexi-
ble package. Metal packaging is most often used for shelf stable products, as its
properties lend itself well to the harsh processing conditions these products undergo.
6.3.2 Glass
Glass is a non-crystalline amorphous solid derived from the heating of silica oxides
in the presence of various additives. A number of additives in the form of carbonates
are typically used in the glass-making process depending on the desired effect on
the finished product. For example, sodium and potassium carbonate are often added
to the glass formula in order to lower the melt temperature, making the glass easier
to work with during the manufacturing process. During the manufacturing process,
furnaces heat the raw material mixture to approximately 1500 °C, a temperature at
which the raw materials melt into a viscous liquid and can then be molded into
shapes. Upon cooling, the molds harden into a non-crystalline amorphous solid.
Glass is a highly recyclable material, with recycled broken glass (cullet) constitut-
ing a large proportion (15–50%) of the ingredients used in a formulation for new
glassware. The advantages of glass include its ability to be formed into a variety of
shapes, its impermeability to moisture and gases, and its non-reactivity with food.
These factors make glass an ideal packaging material for long-term storage of prod-
ucts that might be susceptible to flavor loss. Some disadvantages of glass include
heavy weight (which increases shipping costs) and breakability from physical dam-
age or rapid temperature fluctuations. Consumers often interpret glass packaging as
110 C. Ebner et al.
an indicator of higher product quality, especially for dressings, sodas, and juices
(Risvik 2001).
6.3.3 Plastics
The word plastic describes any material that can be molded into various shapes and
forms while soft and then set into a rigid or flexible form. Plastics can be catego-
rized as thermoset or thermoplastic materials. Thermoset materials solidify into an
irreversible rigid state, usually offering exceptional durability at the expense of
reduced recyclability. On the other hand, thermoplastic materials soften when heat
is applied. Nearly all plastics used in food packaging are considered thermoplastic.
Within the context of food packaging, the term plastic typically describes synthetic
materials derived from petroleum by-products such as ethylene and methane,
although natural materials such as cellulose and lactic acid are sometimes used.
Plastic materials are created through a process called polymerization, which links
together individual monomeric units together to create higher molecular weight
polymeric materials. This is the reason the term plastic and polymer are sometimes
used interchangeably, although this can be a source of confusion.
Plastics have numerous advantages as a material for food packaging. They are
relatively inexpensive, lightweight, and can be molded into a variety of shapes,
which can help to reduce costs associated with transportation of the material. Owing
to its flexibility, plastics are also highly resistant to denting and shattering (though
they are highly susceptible to puncturing). Plastics are also extremely versatile
materials, and the polymerization process allows for the creation of a wide variety
of materials with various functions and properties depending on the end goal. The
major disadvantage of plastics is their relatively poor recyclability when compared
to other materials, such as glass or metals. This is especially true for more compli-
cated plastic food packaging materials, such as multilayered laminates or co-
extruded materials.
6.4 Specific Plastic Polymers Used in Food Packaging
6.4.1 Polyethylene
Polyethylene is created from the polymerization of ethylene, a gaseous by-product

from the petroleum industry. It is the most abundant plastic material produced, with
an average annual global production of ethylene resin around 80–90 million metric
tons per year (Strom and Rasmussen 2011). It has a variety of advantages that make
it useful for food packaging, including ease of production, low cost, excellent form-
ability, strength, good resistance to moisture and chemicals, and highly recyclable.
Polyethylene materials can be classified into functional groups based on their

polymer chain branching and density. Low-density polyethylene (LDPE) is a highly
branched polymer, which interferes with the ability of the polymer to stack tightly
into itself, thus decreasing its density (Fig. 6.1). The low density of LDPE results in
the material having a low melting temperature, which makes it a good heat sealable
material. LDPE is also relatively transparent with good optical properties and good
moisture resistance. These favorable properties make LDPE frequently used in
packaging applications that require a film structure.
Linear low-density polyethylene (LLDPE) has less branching than LDPE, result-
ing in a relatively linear polymer (Fig. 6.1). This linearity results from its unique
manufacturing process, which uses butene, hexene, and octene in the copolymeriza-
tion process. The resultant polymer has a much narrower melt temperature range
than typical LDPE, and also higher tensile strength and impact resistance. This
allows for a thinner gauge film to be used in many applications. Disadvantages of
LLDPE include poorer optical properties than LDPE and difficulty of manufacture
and processing.
High-density polyethylene (HDPE) is a linear polymer with a higher density
than LDPE or LLDPE. The primary advantage of HDPE over LDPE is enhanced
durability due to its high tensile strength. It also has a higher melting temperature
and increased resistance to cracking at lower temperatures. HDPE has poor optical
properties, and so is rarely used in applications requiring package clarity. Plastic
milk containers are one of the best-known examples of a food package based on
HDPE. Additional examples of objects comprised of HDPE include bottle caps,
industrial piping, and in some instances, even fuel cells for automobiles.
Fig. 6.1 Depiction of the

general linearity properties
of HDPE, LLDPE, and
LDPE
112 C. Ebner et al.
6.4.2 Polypropylene
Polypropylene (PP) is a plastic material made from the catalytic addition of propylene
monomers into a polymer (Fig. 6.2). Depending on the catalyst conditions, the resul-
tant polypropylene polymer can take one of three configurations (Fig. 6.3): isotactic
(methyl groups are on one side of the carbon chain), syndiotactic (methyl groups are
evenly dispersed on both sides of the carbon chain), and atactic (methyl groups are
randomly dispersed on both sides of the carbon chain). Atactic PP is a low-quality and
low-value by-product of the PP polymerization process, as it is a soft, rubbery mate-
rial with a lower melting temperature than that of isotactic or syndiotactic
PP. Commercial PP is primarily in the isotactic and syndiotactic forms, as these are
more crystalline and predictable than atactic. PP is an extremely tough and dense
polymer, with enhanced resistance to chemicals and heat. Its high melting temperature
makes it a good polymer for use in such applications as hot filled products, retort
pouches, and microwavable containers. Unlike the polyethylenes, PP has poor trans-
parency, and appears as a hazy material when compared to other polymers. Common
examples of PP in food packaging include rigid trays and food storage containers.
Fig. 6.2 Polymerization of H3C R

propylene into R
CH2
polypropylene
CH3 CH3
propylene monomer polypropylene polymer
Fig. 6.3 Various configurations of polypropylene and their structures

Fig. 6.4 Chemical

structure of polystyrene
6.4.3 Polystyrene
Polystyrene (PS) is a material made from the addition of styrene monomers. The
resulting polymeric chain has a benzene ring attached at every other carbon
(Fig. 6.4). PS is rather inexpensive but suffers from several properties that make it
impractical for use in the packaging of perishable foods. PS is a very brittle mate-
rial, making it a poor choice when mechanical stability is required. PS is also a
poor barrier to moisture and oxygen, meaning it is not practical for use with tech-
nologies such as modified atmosphere packaging. PS can be stretched during the
extrusion process, resulting in a product called oriented PS film. Oriented PS film
has improved optical properties but remains extremely brittle. This form of PS is
sometimes used as an inexpensive alternative to PP trays, especially for packaging
of meats. Expanded PS (EPS) is an extremely common material with insulative
properties. EPS is produced by the addition of a blowing agent, usually pentane,
into EPS resin. When heat is applied to the EPS resin beads, the beads expand to
several times their original size. The beads are then molded together into a closed-
cell foam with excellent insulative properties. Typical food applications for EPS
are egg trays, meat trays, and coffee cups. In North America, consumers generally
refer to EPS as “Styrofoam.” This is technically incorrect, as the name “Styrofoam”
is owned and trademarked by the Dow Chemical Company and is used as a build-
ing insulation material.
6.4.4 Polyvinyl Chloride
Polyvinyl chloride (PVC) is a polymer produced from the addition of vinyl chloride
monomers (Fig. 6.5). This plastic material offers good optical and strength qualities
while being resistant to oils and other hydrophobic compounds. PVC is prone to
degradation at high temperatures, and therefore most PVC is produced using plasti-
cizers in the production process in order to lower the melting temperature of the
product. PVC films produced using these plasticizers often have very good stretch
properties, making them excellent materials for overwrapping some food products,
such as meat.
114 C. Ebner et al.
H2C
Cl
R
Cl
Cl R
vinyl chloride monomer polyvinyl chloride polymer
Fig. 6.5 Polymerization of vinyl chloride into polyvinyl chloride
The use of plasticizers in the production of PVC (and other plastic materials) has
been highly controversial. Recent studies have demonstrated detectable levels of
bisphenol-A (BPA) in the urine of a reference population of adults in the United
States (Calafat et al. 2005). This is alarming as BPA is a known carcinogen with
endocrine-disrupting properties and has been associated with an increased risk of
developing breast and prostate cancers in mammals (Seachrist et al. 2016). While
there has yet to be a scientific consensus on the risks of BPA exposure in food pack-
aging materials, many states in the United States have already banned the use of
BPA in products intended for use in infants and children. At the federal level, The
National Institute of Environmental Health Sciences (NIEHS) and National
Toxicology Program (NTP) have recently funded over $30 million in grants for
research to further investigate the risks of BPA exposure (Schug et al. 2013).
6.4.5 Polyvinylidene Chloride
Polyvinylidene chloride (PVDC) is similar in structure to PVC, except that it has

two chlorine atoms per monomeric unit. Commercial polymerizations of PVDC
typically include a co-monomer to lower the melt temperature of the material. The
major advantage over PVDC over PVC is its excellent moisture and gas barrier
properties. Due to high costs, PVDC is often used as an individual component in
more complicated multilayered packaging materials, for example, in some barrier
shrink bags for vacuum packaging beef. This material is frequently sold to house-
holds as a film wrap for food. Saran® Wrap is an example of a popular brand of
PVDC film wrap used for food. Originally created by the Dow Chemical Company,
this PVDC-based film wrap was first marketed to households in 1953. The SC
Johnson Company purchased the rights to Saran® Wrap in 1998, and shortly there-
after reformulated the product to be based on polyethylene.
6.4.6 Ethylene Vinyl Alcohol and Polyvinyl Alcohol
Ethylene vinyl alcohol (EVOH) and Polyvinyl alcohol (PVOH) are two polymers
frequently used in multilayered packaging applications for their excellent gas bar-
rier properties. This barrier function is due to the OH group contained in their
monomeric units. Two main disadvantages of these materials are their high cost and
high solubility in water. For this reason, these two polymers are usually used in
combination with other polymers in a multilayered package, in order to protect the
EVOH or PVOH from hydrolysis. EVOH is the most frequently used polymer for
oxygen barrier properties in multilayered packaging systems.
6.4.7 Polyethylene Terephthalate
Polyethylene terephthalate (PET) is one of the most widely used polymers for food
packaging materials. This material is commonly produced as a product of a trans-
esterification reaction with ethylene glycol and dimethyl terephthalate, or by an
esterification reaction with ethylene glycol and terephthalic acid. PET is a member
of the “polyester” family of materials, which means that it includes an ester func-
tional group in its polymer chain. The term polyester is commonly used to describe
PET materials. The material has relatively good optical and barrier properties, is
stable over a wide temperature range, and has excellent mechanical and chemical
resistance. This makes PET an excellent material for a variety of food packaging
applications, especially those that are exposed to extreme temperature ranges.
6.4.8 Polyamides
Polyamides are a wide group of polymers that incorporate an amide group into its
polymeric backbone. In the United States, these materials are commonly referred to
as nylon materials, although this term is technically a former DuPont trademark.
Polyamide materials are resistant to high temperatures and mechanical stress. They
are relatively good gas and odor barriers, but often have poor water barrier proper-
ties. An advantage of polyamide as a barrier material over EVOH is that its barrier
performance is not typically impacted by moisture content. Polyamide materials are
extensively used in the packaging of cheese and dairy products and cured meat
products.
6.4.9 Polycarbonates
Polycarbonates (PC) are polymers containing carbonate groups in its backbone.

They are formed by the polymerization of Bisphenol-A (BPA) with phosgene. The
resultant material is clear, extremely durable, and very resistant to heat. This mate-
rial is commonly used in the manufacture of refillable water containers, frozen food
trays designed for oven reheating, and some carbonated beverages. Despite the
advantages of polycarbonates as a packaging material, the use of BPA in the polym-
erization process is highly controversial (see section above on polyvinyl chloride).
116 C. Ebner et al.
6.4.10 Ionomers
Ionomers are copolymers of ethylene and methacrylic or acrylic acid that is partially
neutralized by metal cations, typically zinc or sodium. Ionomers often have a highly
desirable balance of properties, including excellent optical clarity, high puncture
resistance, enhanced durability and toughness (relatively to PE films), low heat seal
initiation temperatures, broad seal strength over a wide variety of temperatures, and
excellent resistance to oils and lipids. Ionomers are often used in multilayered coex-
truded or laminated packaging materials as tie layers, facilitating adhesion of two
relatively incompatible materials.
6.5 Packaging of Specific Perishable Food Products
6.5.1 Overview
In the following section, specific examples of packaging technologies and their

application to extending the shelf life of perishable food products are considered.
Prior to their discussion, it is first important to understand the main packaging tech-
nologies used to extend the shelf life of these products: vacuum packaging and
modified atmosphere packaging (MAP).
6.5.2 Vacuum Packaging
Initially developed by Henri de Poix of the Dewey Almy Chemical Company prior to
World War II, the process was finally patented in 1945, and involved storing frozen
meat quarters in a latex rubber bag that was subjected to a vacuum (De Poix 1945).
While today’s version of vacuum packaging relies on plastic polymer materials instead
of latex rubber, the fundamental process remains the same: (a) the food product is first
placed into a flexible and impermeable, plastic bag or container; (b) the atmosphere in
the package is removed by evacuation; and (c) the plastic bag or container is hermeti-
cally sealed to prevent transmission of moisture or gases into the sealed package. In
vacuum packaging, the objective is to remove as much oxygen as possible within the
package. In general, modern vacuum packaging technologies are able to achieve oxy-
gen levels of around 0.5–1.5% at the point of packaging (Kelly et al. 2018).
6.5.3 Modified Atmosphere Packaging
Modified atmosphere packaging (MAP) is a process where a mixture of gases, typi-

cally purified nitrogen, oxygen, or carbon dioxide, is flushed into a package just
prior to the sealing step. Packages used in a MAP process typically consist of a rigid
tray with an impermeable or semi-permeable multi-layered barrier film on top. On

packages with an impermeable barrier, the gas mixture chosen remains constant
throughout the life of the product. Semi-permeable layers allow for gas exchange
between the package and the environment, which is critical for fruits and vegetables
that respire long after harvest.
The mixture of gases chosen for MAP depends upon the desired effect. Oxygen
is often added to prevent creating a strict anaerobic environment, which may favor
the proliferation of spoilage and/or pathogenic bacteria in some foodstuffs (Farber
2016). Carbon dioxide is utilized for its antimicrobial effect, as it is able to easily
penetrate into bacterial cells, causing cell death via cell wall collapse and subse-
quent leaking of cellular contents (Oulé et al. 2006). Carbon dioxide is also is more
soluble than oxygen in water, fats, and oils, and thus has a limited ability to pene-
trate into the surface of foods. Nitrogen is solely utilized as inert filler, as it dis-
places other reactive gases. Less common gases utilized in MAP technology include
carbon monoxide and sulfur dioxide. In meat products, carbon monoxide can be
used to permanently “fix” myoglobin into the carboxymyoglobin state, which then
appears as a bright red pigment (Djenane and Roncalés 2018). However, this prac-
tice is highly controversial due to issues with consumer acceptance. Carbon monox-
ide can also be used to prevent the browning of fruits and vegetables. Sulfur dioxide
is sometimes used as an antimicrobial, especially in preventing mold and bacteria
from growing on fruits; however, it can cause allergy-like reactions in some sensi-
tive consumers and therefore is rarely used (Thompson 2010).
6.5.4 Packaging of Fresh Red Meat
Research on consumer preference of fresh red meat has demonstrated color to be the
primary factor influencing consumer purchasing decisions, with consumers having a
preference for red meat with a bright cherry red color (Font-i-Furnols and Guerrero
2014). Myoglobin, a meat pigment found in the muscle tissue of animals, is responsible
for the familiar red color of beef. Myoglobin is a protein that stores oxygen in the
muscle tissue of animals. This function is attributed to a heme group within myoglobin,
which has a strong affinity for iron and oxygen. In meat, the color is largely determined
by the oxidation state of the myoglobin molecule, which can be myoglobin (purple),
oxymyoglobin (bright red), or metmyoglobin (brown). In the absence of oxygen, the
myoglobin (purple) state predominates with the heme molecule containing an iron
atom in the ferrous (+2) oxidation state bound to a water molecule (H2O). When
exposed to oxygen for a short period of time, the myoglobin state changes to oxymyo-
globin (bright red) where the heme molecule contains an iron atom in the ferrous (+2)
oxidation state bound to an oxygen molecule, resulting in the familiar “bloom” to con-
sumers. When fresh meat has been exposed to oxygen for a long period of time, or
when it has been cooked, it will convert to the metmyoglobin state (brown). This is
when the iron atom in the heme molecule is in the ferric (+3) state, as it has lost an
electron. Meat color can also be impacted by age of the animal at slaughter, animal diet,
and level of exercise. Since the metmyoglobin (brown) state of meat is seen as a sign of
118 C. Ebner et al.
poor quality (even though it is not associated with reduced quality from a microbial
standpoint), most efforts to extend the shelf life of red meat products seek to delay or
prevent the formation of metmyoglobin.
Microbial spoilage is also a major factor in influencing the shelf life of fresh red
meat. Given that essentially all packaged meat products are produced, shipped, and
stored under refrigeration conditions, the predominant spoilage microflora are psy-
chrotrophs, most notably Pseudomonas species in aerobic packaged meat, and
Lactobacillus and Brochothrix species in meat packaged under reduced oxygen
conditions. When allowed to flourish on meat, Pseudomonas species produce pro-
teolytic enzymes that break down amino acids into foul smelling volatile com-
pounds, appearing as highly offensive off odors to the consumer. Pseudomonas
species can also cause slime to develop on meat as a result of tissue proteolysis. This
slime often appears on spoiled meat as a green discoloration.
Given the severity of proteolytic spoilage and reduced shelf life associated with
more aerobic type packaging systems, the vast majority of packaging solutions that
seek to extend the shelf life of fresh red meat are focused on reducing the oxygen
content of the package itself. Vacuum packaging is by far the most common method of
choice for shipment and delivery of fresh red meat primals and subprimals, and can
achieve a shelf life of approximately 7–12 weeks (Voges et al. 2007). This shelf life
can vary depending upon temperature conditions, beef source, and initial microbial
load. At the store, the vacuum packages are then opened and broken down into indi-
vidual steaks by the butcher. The individual cuts are placed onto a foam tray with a
moisture absorbent pad wrapped with a clear oxygen and moisture permeable plastic
film. The high permeability of the plastic overwrap material allows for oxygen to come
into contact with the meat, producing the bright cherry red “bloom” that consumers
prefer. While this approach for retail packaging of beef is very inexpensive, it also
results in a short shelf life of approximately 4–7 days, depending on storage conditions
and initial beef quality. Vacuum packaging of individual steaks and cuts is possible and
is sometimes seen; however, consumers tend to find the purple color of the native
myoglobin found in vacuum packaged beef unacceptable (Voges et al. 2007).
Innovations in packaging technology have resulted in fresh red meat products
being packaged under MAP conditions. These packages typically consist of meat
placed inside a semi-rigid barrier tray and sealed with a barrier top web. An advan-
tage of this approach is that the product is not physically compressed, as can happen
at the corners of vacuum bags. This compression can result in exudation of cellular
components from the muscle tissue itself, also known as “purge,” which many con-
sumers find undesirable. Gas mixtures for red meat packaged using MAP technol-
ogy usually contain high amounts of oxygen (50–80%), with carbon dioxide as the
remainder. The high oxygen content helps to ensure that the muscle tissue remains
a bright red color, while the carbon dioxide serves to help control spoilage microor-
ganisms. An unintended consequence of the high oxygen content in these packages
is the tendency for the meat to spoil due to lipid oxidation, usually after a period of
around 2 weeks. Low-oxygen MAP packaging generally uses gas mixtures of
around 30% carbon dioxide and 70% nitrogen. The shelf life of refrigerated red
meat packaged under these conditions can range anywhere from 3 to 5 weeks. A
disadvantage is that, depending on the gaseous mixture of the MAP system, these
conditions favor the purple color of the native myoglobin form in the muscle tissue,
which some consumers may not prefer. Another MAP system commonly used for
case-ready fresh meats is the master pack system that consists of several air perme-
able overwrapped packaged placed in a large pouch commonly referred to as a
master pack or mother bag. The master pack is impermeable to oxygen and mois-
ture. The retailer then removes the oxygen permeable case-ready packages from the
high barrier master package and the product blooms within several minutes after
exposure to air.
6.5.5 Packaging of Poultry Products
Unlike beef, the shelf life of poultry products is primarily tied to the microbial pro-
duction of off flavors and odors, and not to color. The microorganisms primarily
responsible for these off flavors and odors are Pseudomonas, Achromobacter,
Enterobacter, and Shewanella species (Russell et al. 1995). In a sensory panel on
expired poultry products, consumers described off flavors from these microorgan-
isms as “sulfur,” “dishrag,” “ammonia,” “wet dog,” “skunk,” “dirty socks,” and
“rancid fish” (Russell et al. 1995). Poultry products are typically packaged under
MAP conditions more frequently than vacuum conditions. Gas mixtures for poultry
products packaged under MAP conditions can vary greatly, though in general,
researchers have identified better shelf life performance when packaged at carbon
dioxide levels of 20% or higher in order to prevent proliferation of aerobic spoilage
bacteria (Rossaint et al. 2014). Under these conditions, a shelf life of 23 weeks is
typically achievable. The majority of poultry retail packages consist of a rigid or
foam tray sealed by a barrier overwrap film. This allows for the package to be gas
flushed to extend shelf life, all while maintaining the traditional look of a retail
poultry overwrap package.
A major safety consideration of MAP packaged poultry concerns the growth of
Campylobacter jejuni, a mesophilic pathogenic organism that thrives in low oxygen
environments. Compounding the issue is the widespread distribution of Campylobacter
jejuni in retail poultry products within the United States. In one survey, 70.7% of
chicken samples tested at retail locations tested positive for Campylobacter (Zhao
et al. 2001). Thus, temperature control is critical for poultry products packaged using
MAP gas mixtures that may favor growth of this pathogenic organism.
6.5.6 Packaging of Seafood Products
Seafood products such as freshwater fish, saltwater fish, and shellfish are extremely
perishable products with spoilage resulting from both autolytic changes involving
proteolytic enzymes as well as from microbial degradation. Immediately following
120 C. Ebner et al.
the death of a fish, endogenous enzymes digest muscle tissue into compounds with
off-odor properties (Ahmed et al. 2013). These endogenous enzymes are able to
hydrolyze protein tissue even at refrigeration temperatures. After the autolysis pro-
cess has continued for 3–5 days, these compounds begin to serve as nutrients for a
variety of spoilage bacteria, which then proliferate and produce their own undesirable
odors and off flavors. Typical spoilage organisms for seafood include Pseudomonas
spp., Shewanella spp., Photobacterium spp., Brochothrix spp., and various lactic acid
bacteria (Boziaris and Parlapani 2017). Most seafood is also highly susceptible to
chemical spoilage, primarily in the form of lipid oxidation. This problem is com-
pounded by the fact that these oxidation reactions can occur at freezing temperatures.
Also, most seafood products are comprised of a high percent of unsaturated fatty
acids, which are highly susceptible to oxidation.
Vacuum and MAP packaging can extend the shelf life of seafood products by
shifting the microflora towards non-spoilage organisms. Early research has shown
that high carbon dioxide environments approaching 100% could vastly improve on
the shelf life of seafood products, albeit at the expense of package collapse, a phe-
nomenon that results from the dissolution of atmospheric carbon dioxide into the
muscle tissue of the seafood (DeWitt and Oliveira 2016). The gas compositions
used in MAP packaging of fish and seafood generally fall into one of two catego-
ries: mixtures for fatty fish (a mixture of carbon dioxide and nitrogen) and mixtures
for non-fatty fish (an even mixture of oxygen, carbon dioxide, and nitrogen). The
shelf life of packaged fish products is generally between 10 and 20 days, depending
on the storage temperature and incoming quality of the initial product.
A major risk associated with vacuum and MAP packaged fish is the potential
growth of Clostridium botulinum type E, an anaerobic organism that produces the
potentially deadly botulinum toxin. This risk stems from observations that many fish
may become naturally contaminated with type E strains of Clostridium botulinum
that reside in seabeds (Gram 2001). Originally, oxygen was added to fresh fish pack-
ages in order to prevent the growth of Clostridium botulinum, but it has now been
shown that this organism can grow and produce toxin in packages with relatively
high levels of oxygen added. Exacerbating this issue is the observation that several
strains of type E Clostridium botulinum can produce botulinum toxin at refrigeration
temperatures that normally suppress spoilage organisms (Gram 2001). Thus, a pack-
aged fish product that undergoes temperature abuse could readily develop toxic lev-
els of botulinum toxin without any glaring effects of spoilage organisms. The only
effective way to prevent growth of Clostridium botulinum in vacuum or MAP pack-
aged fish is to keep the product at or below 3 °C at all times.
6.5.7 Packaging of Fresh Produce
After harvest, the tissues of fruits and vegetables continue to undergo metabolic pro-
cesses that affect their sensory attributes and ultimately, quality and shelf life.
Respiration is the primary metabolic process that contributes to the postharvest physi-
ological changes in the fruit or vegetable tissue. In this process, energy in the form of
ATP is generated by the oxidation of glucose-rich compounds (e.g., starches) within
the cells of the plants. Respiration in plants is typically an aerobic process, where
oxygen is consumed and converted into carbon dioxide, water, and energy (in the form
of heat). Plants can also undergo anaerobic respiration in certain situations, although
this process tends to rapidly deteriorate the quality of the fruit or vegetable. In general,
the respiration rate of a fruit or vegetable is proportional to the shelf life of the product,
since this metabolic process is responsible for the senescence of the plant.
Many factors can influence the respiration rate of fruits and vegetables. In fruits,
ethylene gas serves as a plant hormone that regulates the ripening and thus respiration
process. Fruits are generally classified as either climacteric or non-climacteric.
Climacteric fruits are fruits whose ripening is associated with an increase in both eth-
ylene production and respiration, which results in an increase in carbon dioxide pro-
duction. In climacteric fruits, ethylene acts as a trigger for the ripening process.
Examples of climacteric fruits include apples, melons, bananas, avocadoes, and toma-
toes. Non-climacteric fruits, on the other hand, do not increase ethylene production
during the ripening process, and so, the presence of ethylene does not accelerate this
process. Examples of non-climacteric fruits include citrus fruits and strawberries.
Many produce items are packaged using modified atmosphere packaging (MAP)
technologies. MAP technologies generally fall under two separate categories: pas-
sive and active MAP technologies. In passive MAP, an atmospheric barrier is formed
around a perishable fresh produce product. This barrier passively results in a modi-
fication of the interior atmosphere of the package. As the produce undergoes senes-
cence, O2 is consumed and CO2 is released, and the barrier properties of the film
allow the CO2 to build thereby passively modifying the atmosphere, slowing down
respiration and extending shelf life.
Active MAP is the utilization of scavenging materials (either within the packag-
ing film or in the form of sachets) or the infusion of a specific composition of
gasses into the package during sealing. These gaseous mixtures can be custom tai-
lored to desired levels based on the respiration requirements of the produce item.
Another form of active MAP involves one-way gas valves that allowed certain
respiration gases to exit the package while preventing outside gases from entering
the package. Roasted coffee beans are a great example of an active MAP package
that utilizes a one-way valve. As roasted coffee beans age, they emit carbon
dioxide. If carbon dioxide is allowed to build up to high levels within the package,
staling will occur. This is why one-way gas valves are often incorporated into fresh
coffee bean packages, to allow off gassing of carbon dioxide without preventing
any influx of external gases.
Selection of passive or active MAP will depend upon how the fresh produce
ripens and what quality attributes are impacted by that process versus the need to
quickly halt ripening and respiration early in the shelf life. When oriented polypro-
pylene of different thickness (20, 40, and 80 μm) was used to package ready-to-eat
table grapes, it was determined that passive MAP of the highest thickness film
attained 70 days of shelf when stored at 5 °C, which was more than active MAP was
able achieve (Costa et al. 2011). Fresh endive stored at 20 °C under active MAP
122 C. Ebner et al.
Table 6.1 Examples of optimum MAP storage conditions for various fruits and their expected
shelf lives
MAP conditions
Fruit Storage temperature (°C) % CO2 % O2 Shelf life (Days)
Apple 3 3 3 200–300
Avocado 7 5 10 12–56
Banana 15 2 5 21–60
Grapes 2 5 3 40–90
Lemon 15 5 5 130–220
Orange 10 10 5 42–84
Papaya 13 5 8 14–35
Pineapple 15 5 10 12–15
Strawberry 15 10 20 7–15
Adapted from Mangaraj and Goswami (2009)
created by the use of an O2 scavenging sachet, did not change O2 and CO2 partial
pressure during the steady-state period, compared to passive MAP, but induced a
50% reduction of the transient period and delayed greening and browning (Charles
et al. 2008). Active MAP of romaine lettuce with a gas mixture of 10% O2, 10%
CO2, and 80% N2 delayed growth of autochthonous lettuce microflora, but not
Salmonella and even favored the survival of the pathogen, possibly due to the elimi-
nation of its natural antagonists (Horev et al. 2012). The effects of the passive MAP
on lettuce were less pronounced. These varying results highlight the importance of
selecting MAP conditions that are specific to the respiration requirements of the
produce item in question, while considering the intended outcome of the MAP
package itself (Table 6.1).
6.5.8 Packaging of Dairy and Cheese Products
Fresh pasteurized milk, once bottled in glass, has been regularly packaged in blow
molded polymers since the 1960s. The vast majority of fluid milk is packaged in
HDPE containers, although some polyethylene terephthalate (PETE) and low-
density polyethylene (LDPE) examples can be found. Half gallons are also sold in
a three-layered carton format (polyethylene, paperboard, polyethylene) with a shelf
life of 15–21 days at refrigeration temperatures (Fromm and Boor 2004).
Fermented dairy products such as block cheeses are often packaged in multi-layered
co-polymer materials that are designed to exhibit a variety of properties to enhance the
shelf life of these foods. Cheeses are often packaged in reduced oxygen environments
in order to prevent unwanted growth of microbial spoilage organisms, such as mold.
High carbon dioxide flushing of MAP packaged cheese is also common, as it displaces
oxygen and assists in shelf life extension by inhibiting growth of mold and other spoil-
age organisms. In these conditions, CO2 levels can range from 50% to 100%. The typi-
cal shelf life of high CO2 MAP packaged hard cheeses (such as cheddar) is 4–6 weeks
depending on storage conditions. High CO2 conditions are also highly effective at
extending the shelf life of cottage cheeses, which typically have a shelf life of
2–4 weeks. When cottage cheese packages are flushed with high CO2, shelf life
extended to approximately 8–12 weeks, depending on storage conditions. However, it
has been reported that cottage cheese can take on a “soda-like” flavor, as the CO2
absorbs into the product itself. Cheese packages also usually contain a moisture barrier
to prevent the product from drying out. Nylon is becoming more popular in many co-
polymeric cheese packages due to its enhanced durability and strength. This is very
important for packaging of cheese blocks, as the sharp corners can puncture packages
during transportation and handling.
6.6 Active Packaging Technologies
6.6.1 Overview
Packaging materials have evolved from providing a simple inert, passive containment
for food products into packaging that can provide “active functions” such as extend-
ing shelf life and maintaining or improving desirable conditions of the packaged
food. Active packaging is designed to deliberately release components into the pack-
aged food or the surrounding environment. Two popular active packaging concepts
include materials or packages that scavenge unwanted substrates or gases from the
packaging environment, or materials or packages that emit gases or antimicrobials
into the food or package environment (Suppakul et al. 2003). As mentioned previ-
ously in this chapter, the spoilage of many meat and food products is attributed to the
production of unpleasant off odors. These off odors are generated either by bacterial
growth and proliferation, endogenous enzyme activity, or by degradative chemical
reactions (e.g., oxidation of lipids). Regardless of the mechanism, these off odors
contribute to a reduction in product shelf life. Thus, it is evident that active packaging
technologies such as odor-scavenging materials and antimicrobial packaging have
the potential to positively influence and extend the shelf life of these products.
For an active packaging technology to be commercially viable, it must have sev-
eral desirable attributes. First and foremost, the active components must be approved
for use in direct food contact applications, meaning it will be non-toxic, non-
allergenic, and harmless to human health. The active components must be easily
incorporated into the package, by incorporation into the film, tray, sachet, label, and
must be able to function under conditions of use for the package (e.g., shipment and
storage temperatures, and lighting conditions). The active components also must not
adversely impact the organoleptic profile of the product, nor mask any types of
spoilage. Finally, the active components should be invisible to the customer. This is
particularly a challenge for active components that are directly incorporated into a
film, as this can cause a film become opaque, or otherwise degrade the optics of the
film to such an extent it becomes unfit for packaging applications. This is especially
124 C. Ebner et al.
the case where the film customer or final user desires a clear film in which the con-
tents of the package can be visually inspected from outside the package. A few
examples of active packaging technologies are discussed below.
6.6.2 Oxygen Scavenging Technologies
The shelf life of oxygen-sensitive food (e.g., fatty fish, fatty cuts of beef, and pack-
aged nuts) can be extended by reducing the amount of oxygen within a package.
Vacuum and MAP packaging technologies often extend shelf life based on this con-
cept. Additionally, a variety of active packaging technologies have been developed
that scavenge the available oxygen within a package. One approach is through the
inclusion of a material (other than the package itself) capable of consuming oxygen.
These materials usually consist of sachets with oxygen scavenging properties.
Modern oxygen scavenging sachets usually contain a mixture of iron and sodium
chloride. Moisture within a package activates the oxygen scavenging material by
reacting with iron and oxygen to form iron oxide, also known as rust. This process
is accelerated by the presence of sodium chloride, which acts as a catalyst for the
rusting process. Using this process, the oxygen level within a package can be
reduced to 0.01% or lower (Brandon et al. 2009). An advantage of this iron-based
oxygen scavenging system is that it does not produce any offensive off odors or
unwanted flavors. A disadvantage of this system is that a moisture content of greater
than 50% is required for the sachets to work, rendering them inappropriate for use
in packaged dry products (Brandon et al. 2009).
More recently, sachet free oxygen scavenging technologies have been introduced
into the market. These sachet-free technologies rely on oxygen scavenging poly-
mers within the film itself to extend product shelf life. A major advantage of this
approach is the prevention of accidental ingestion of oxygen scavenging sachets.
One patented approach to creating an oxygen scavenging film is to incorporate a
resin with a large number of unsaturated bonds into the barrier layer. The film is
then irradiated, which “triggers” the oxygen scavenging effect of the film by increas-
ing the reactivity of the unsaturated bonds with ground state oxygen (Beckwith
et al. 2016). A frequent application of oxygen scavenging films is in packaging of
high-value bread, cake, and cereal products, especially those using a preservative-
free formulation. In these products, oxygen scavenging films can extend shelf life
by preventing the growth of several fungal spoilage organisms, including Penicillium
spp. (Nielsen and Rios 2000). Additional examples of oxygen scavenging technolo-
gies are provided below in Table 6.2.
Table 6.2 Select examples of commercially available oxygen scavenging technologies

Oxygen scavenging technology Delivery method Commercial product examples
Metal powders (Fe, cu, etc.) Sachet, labels, tray, Ageless® series—Mitsubishi gas and
extrusion films chemical company
Sorbent system—Impak corporation
O-busters®—Sachets and strips -
Dessicare Inc.
O2Block®—Nanobiomatters
Oxy-guard®—Clariant
FreshMax® labels and sachets—
Multisorb technologies
ShelfPlus O2®—Albis
Ascorbic acid and ascorbate Sachets and gaskets Celox® series—GCP applied
salts technologies
Darex® MB2003—GCP applied
technologies
Catechol Sachet Tamotsu—OhE chemicals, Inc.
Photosensitive dyes Film Zero2—CSIRO Australia
Enzymes, e.g., glucose oxidase, Sachet Bioka—Bioka Ltd.
alcohol oxidase
MXD-6 (nylon) Film, bottles Oxbar®—Plastipack
Unsaturated polymers Extrusion films, trays, Amosorb®—PolyOne
bottles Cryovac® OS films—Sealed Air
Corporation
Pd catalysts/H2 Sachets, films, labels HyGuard™—Polyone
6.6.3 Carbon Dioxide Scavenging Technologies
While CO2 is often incorporated into modified atmosphere packages for its bacterio-
static properties, sometimes the buildup of this gas within a package is a highly
undesirable property. In some foods, for example, cottage cheeses, buildup of CO2
can have a negative impact on some of the flavor attributes of the product which
negatively impact shelf life. In other foods, especially fermented foods, the buildup
of CO2 over time can compromise package integrity. In each case, CO2 scavenging
technologies can serve as a solution to the problem.
Broadly speaking, CO2 scavenging technologies fall into one of two categories by
mode of action: physical and chemical absorption. In chemical absorption, calcium
salts and alkaline solutions can be utilized to react with residual CO2 in a package.
The most frequently used salt for this reaction is calcium hydroxide, which has favor-
able properties that make it suitable for food contact use. Sodium carbonate is also
used, although this compound is only suitable for scavenging CO2 from moist pack-
age environments, since water is a requirement for the chemical reaction responsible
for the consumption of CO2. In physical absorption, CO2 gas molecules adsorb onto
a physical substrate and become trapped in the physical network of the substrate. The
two most common substrates used for this purpose are activated carbon and zeolites.
126 C. Ebner et al.
Activated carbon is characterized by an amorphous porous structure with an extremely

large surface area. An advantage of activated carbon is that its non-polar structure
means that moisture inside a package has relatively little impact on the rate of CO2
adsorption. Zeolites, which are microporous, aluminosilicate minerals with a com-
plex three-dimensional structure, are also used for physical adsorption of CO2 gas.
The surface area and structure of zeolites can be customized by varying the cation
(e.g., sodium and calcium) used for formation of the zeolite. Many zeolites have a
higher affinity for water vapor than CO2 gas, which can be a disadvantage for the use
of zeolites as CO2 scavengers in high moisture food products.
There are numerous examples of the successful use of CO2 scavenging technolo-
gies to extend shelf life of foods. Some fruits, such as pears, can undergo acceler-
ated browning reactions when CO2 levels build up to high levels within a package.
In one study, researchers found pears packaged in CO2 scavenging film and sub-
jected to long-term cold storage had enhanced shelf life versus pears store in non-
scavenging film (Nugraha et al. 2015).
6.6.4 Odor Scavenging Packaging
Odor scavenging technologies are designed specifically to control undesirable odors

within a package (de Abreu et al. 2011). Plastic polymer-based packaging has long
been known to absorb volatile compounds from food, which is commonly referred to
as “scalping” (Van Willige et al. 2002). Conversely, plastic packaging materials can
also impart undesirable volatile compounds or odors into food. Although generally
regarded as a negative attribute, it is possible to construct a package where the absorp-
tion of compounds by food becomes a desired feature in controlling odor profiles. For
some foods, the oxidation of lipids can result in the development of “rancid” odors.
These products, such as nuts and oils, benefit from using a combination of oxygen
scavengers and odor absorbers in their packaging. Other foods such as bone-in-meats,
sausage, ham, salami, pepperoni, poultry, and processed poultry meats such as turkey
pepperoni can generate sulfur type off odors during distribution and storage. Hydrogen
sulfide and other sulfur containing compounds, such as thiols or mercaptans, are
generated during the normal shelf life of these products as the by-products of enzy-
matic or microbial degradation of sulfur containing amino acids such as cysteine.
These odors are extremely unpleasant, especially hydrogen sulfide (rotten egg) and
ethyl mercaptan (skunk). These off odors are particularly a problem in high oxygen
barrier packaging, which effectively traps the off odor inside the package. Although
the product may still be perfectly safe for consumption, the odors are released upon
opening the package, causing consumers to regard the product as spoiled.
Other off odors may be composed of additional chemical classes, such as alde-
hydes, acids, ketones, and amines. For example, fish and seafood generate amine
odors while dairy products may generate sour or “buttery” odors. Thus, it becomes
important to know the profile for undesirable volatile compounds for the products
being packaged, so that an appropriate scavenger system can be used. Two impor-
tant caveats to designing an active odor-scavenging package are that the scavenger
materials not “scalp” the desirable odors from the packaged product and that the use
of the scavenger material does not mask product spoilage.
Being volatile compounds, odors may be removed within a packaging system by
physical means through adsorption or absorption. Adsorption is a physical and/or
chemical process by which one substance becomes attached to another through
physical and/or chemical interactions. More specifically, adsorption is the adher-
ence, binding, or attraction of atoms, molecules, or ions to the surface of another
material. In adsorption, binding to the surface is frequently reversible, but com-
pounds that have taste or odor typically tend to bind strongly. Adsorbents are fre-
quently characterized by having very large surface areas per unit weight. Typical
adsorbents include activated carbon and silica. Absorption is the penetration of one
substance into the inner structure of another. The most common industrial absor-
bents include zeolites, molecular sieves, and cyclodextrins.
Odor compounds may also be removed within a package using chemical means.
This is known as chemisorption. During a spontaneous reaction, molecular bonds
may be broken or created as the volatile odor compounds react with the scavenging
agent. These processes could include synthesis reactions, reduction/oxidation reac-
tions, or acid–base reactions. For example, one method for the removal of hydrogen
sulfide is through a reaction with a metal ion such as iron or copper, resulting in the
formation of a metal sulfide. An overview of typical odor scavenging technologies
and their applications is listed in Table 6.3.
Table 6.3 Frequently used odor scavenging applications

Scavenging agent Mechanism(s) Application
Activated carbon Adsorption Sachet, pad, label, tray
Alumina Adsorption Sachet, pad, label, tray, film
Chemisorption
Baking soda Chemisorption Sachet, pad, label
Cyclodextrins Absorption Sachet, pad, label, tray, film
Layered double hydroxides Adsorption, absorption, Sachet, pad, label, tray, film
(LDH), e.g., hydrotalcite chemisorption
Metals: e.g., cu, Zn, Fe, Ni, Ag Chemisorption Sachet, pad, label, tray, film
Metal oxides/salts: e.g., ZnO, Chemisorption Sachet, pad, label, tray, film
MgO CaO, Ca(OH)2, Fe2O3
Molecular sieves Absorption Sachet, pad, label, tray, film
Adsorption
Organic acids, e.g., citric acid, Chemisorption Sachet, pad, label
ascorbic acid
Polyalkylene imine, e.g., Chemisorption Film, tray, label
polyethylene imine
Silica gel Absorption Sachet, pad, label, tray, film
Adsorption
Zeolites Absorption Sachet, pad, label, tray, film
Zinc Ricinoleate Chemisorption Sachet, pad, label, tray, film
128 C. Ebner et al.
6.6.5 Moisture Control Technologies
For many foods, excess moisture within a package can have a major negative impact
on shelf life. In dry powdered products, excess moisture can cause clumping and
caking to occur and in hard candies, and can soften the product, leading to a highly
undesirable mouth feel. In many packaged meat products, moisture loss in the form
of purge occurs over time and must be controlled. Ultimately, moisture control tech-
nologies help control the water activity within a package, which minimizes both
microbial growth and unwanted sensory changes within a product. Desiccants are
often incorporated into packages of dry food products as sachets. Silica gels,
calcium oxide, and clays are frequently used as desiccants. Absorbent pads are
frequently used in fresh meat packages to absorb excess purge. These are typically
made of silica gel or cellulosic fibers.
6.6.6 Antimicrobial Packaging Technologies
Antimicrobial packaging technologies extend the shelf life of food by killing

unwanted spoilage organisms that contribute to loss of sensory attributes. Additionally,
antimicrobial packaging technologies may be designed to kill or help control growth
of pathogens, such as Listeria monocytogenes, Salmonella spp. and Escherichia coli,
which may be present on the surface or within food. Antimicrobial coatings and films
are some of the more promising antimicrobial packaging technologies and have been
researched extensively for several decades. In this approach, the antimicrobial ingre-
dient is incorporated directly into the coating or film, which is then in direct contact
with the food. Table 6.4 below shows several recent research examples of antimicro-
bial coatings and films and their applications in food packaging.
When the antimicrobial film is in direct contact with a food surface, the antimi-
crobial ingredient diffuses into the food which prevents microbial growth. Because
of this, antimicrobial films must be in direct contact with a food for antimicrobial
activity to take place. A key challenge with designing these antimicrobial films
involves calibrating the rate of diffusion of the antimicrobial ingredient into the
food. If the rate of diffusion is too fast, then the antimicrobial activity is quickly
depleted. If the rate of diffusion is too slow, minimal antimicrobial activity may
occur. Another challenge is that many food matrices may inactivate or reduce the
antimicrobial activity of these active ingredients. For example, fatty foods may
reduce the antimicrobial activity of many bacteriocins, such as nisin, which is effec-
tive against Gram-positive bacteria, such as L. monocytogenes (Franklin et al. 2004).
Many approaches to incorporating antimicrobial ingredients into films and coatings
have been explored. Edible films, typically made of cellulosic materials, have been
formulated with naturally derived bacteriocins, such as nisin. Nisin has extremely
potent antimicrobial activity against Gram-positive bacteria, such as L. monocyto-
genes. When individually packaged hot dogs were coated with cellulose materials con-
Table 6.4 Select references on antimicrobial agents incorporated into films and coatings
Food
Antimicrobial contact
ingredient matrix Foods Targeted microorganisms References
Oregano and LDPE Culture Salmonella Typhimurium, Solano and de
thyme essential only Escherichia coli, Listeria Rojas Gante
oils monocytogenes (2012)
Nisin Cellulose Frankfurters Listeria monocytogenes Franklin et al.
coating (2004)
Lauroyl arginate Chitosan Culture Salmonella Typhimurium, Haghighi et al.
ethyl (LAE) and gelatin only Escherichia coli, Listeria (2019)
monocytogenes, Campylobacter
jejuni
Rosemary and Whey Culture E. coli, S. aureus, and Ribeiro-Santos
cinnamon protein- only Penicillium spp. et al. (2017)
based film
Bacteriocin 729 Poly-lactic Fish filets Listeria monocytogenes, Woraprayote
acid film Staphylococcus aureus, et al. (2018)
Pseudomonas aeruginosa,
Aeromonas hydrophila,
Escherichia coli, Salmonella
Typhimurium
taining nisin, a 2-log10 reduction in Listeria monocytogenes counts was observed over
a 60-day storage window (Franklin et al. 2004). Researchers have also incorporated
nisin into PVC and linear low-density polyethylene (LLDPE) into packaging systems,
and observed that it significantly reduced populations of Salmonella Typhimurium
when used to package fresh poultry (Natrajan and Sheldon 2000). While nisin has
shown good antimicrobial activity across a broad spectrum of bacteria, one concern
involves the risk of bacteria becoming resistant to its activity via mutations. To address
this concern, antimicrobial film approaches using nisin often include a second bacte-
riocin in order to reduce the risk of resistance causing mutations from occurring.
Essential oils such as those derived from oregano, thyme, cinnamon, and pimento,
have long been known to exhibit antimicrobial properties, and are often incorpo-
rated into films in a research setting. In one study, a 4.0% (w/w) combination of
oregano and thyme oils incorporated into extruded LLDPE films was able to inhibit
Salmonella Typhimurium, Listeria monocytogenes, and Escherichia coli O157:H7
using standard culture methods (Solano and de Rojas Gante 2012). One major dis-
advantage of essential oils comes from their impact on the sensory attributes of a
food product. While these compounds are highly active against a wide range of
bacteria, they tend to confer major sensory properties (flavor, aroma) into a food.
While research on antimicrobial films for spoilage organisms and pathogen con-
trol has been promising, few technologies and approaches have successfully transi-
tioned into the real world. One issue is that laboratory evidence rarely translates into
real-world efficacy. For example, many laboratory experiments are reliant upon
food simulants, rather than complex food matrices. In real-world food systems, the
130 C. Ebner et al.
composition of pH, salt, carbohydrate, protein, and fats within a food can drastically
change, which may impact the performance of the antimicrobial film. Storage con-
ditions of the packaged food itself can also have a major impact on the performance
of the antimicrobial films. This is mainly due to temperature and humidity having a
major impact on the rate of diffusion of antimicrobial ingredients. As diffusion
slows, so does the antimicrobial activity. When promising antimicrobial films are
adapted from the laboratory into a real-world setting, what is often seen is an initial
spike in antimicrobial efficacy, which quickly falters due to lack of diffusion of the
antimicrobial into the food, a negative interaction of the antimicrobial with food
components, or a saturation of the food with the antimicrobial ingredient.
Finally, a major challenge with antimicrobial films regards their difficulty in
manufacturing. Most laboratory-scale experiments are performed using a casting
process, rather than extrusion. Extrusion is the production process of choice for
polymeric films due to its high line speeds and ability to create a uniform product.
The high heat and pressures encountered during extrusion often destroy any antimi-
crobial activity of many antimicrobial ingredients. Future research investigating
novel compounds should always incorporate stability testing under various tem-
peratures and pressures to ensure compatibility with the extrusion process.
6.7 Intelligent Packaging
According to the United States Department of Agriculture, each year in the United
States, about 30–40% of the entire food supply is wasted (USDA 2019). This repre-
sents 133 billion pounds of food and is valued at over $162 billion. An emerging
area of packaging, termed “intelligent packaging,” seeks to use innovative technolo-
gies to help reduce food waste.
Intelligent packaging has been defined as “materials and articles that monitor the
condition of packaged food or the environment surrounding the food” (EFSA 2009).
In other words, intelligent packaging technologies seek to communicate informa-
tion about a food package to the consumer so that they may make an informed deci-
sion. In general, intelligent packaging falls under three broad categories: (1) data
storage devices, (2) indicators, and (3) sensors.
Intelligent packaging technologies that monitor data are intended to improve sup-
ply chain efficiency. The most frequently encountered data monitoring approaches
are barcodes and Radio Frequency Identification Devices (RFID) tags. Barcodes uti-
lize patterns to store information about a package. They are widely used to assist in
inventory control and have also experienced increasing adoption for traceability pur-
poses. RFID tags are more expensive but are able to store far more data than bar-
codes. RFID tags are becoming increasingly adopted for use in inventory management,
and for the promotion of safe food by enhancing traceability in the event of a food-
borne illness outbreak. RFID tags consist of a small antenna wired to a small micro-
processor, allowing the tag to communicate to other devices via radio waves.
Indicators communicate the presence or absence of an analyte or a condition to

the consumer. Often, they do this via change in color. Indicators may be placed on
or outside a package. Time and Temperature indicators (TTIs) and freshness indica-
tors are two examples of commercially available intelligent packaging devices.
TTIs monitor the storage of a product along the production chain, specifically, the
temperature and time of holding throughout a process, usually transportation. As
discussed previously, time and temperature are two critical variables that have a
negative impact on the shelf life of a product if they are not controlled. TTIs help to
keep transportation processes in control with respect to time and temperature, thus
leading to food that retains its quality better. TTIs can monitor a critical temperature
or they can monitor the history of time and temperature along an entire process.
Freshness indicators communicate information about the quality of a food to the
consumer. These devices often monitor microbiological metabolites that are corre-
lated with a loss of quality. For example, volatile nitrogen compounds and biogenic
amines are often associated with the loss of quality of a meat product. When the
amount of metabolite exceeds a threshold, a chemical reaction takes place that leads
to a color change of the freshness indicator. An example of this is the SensorQ sen-
sor from Food Quality Sensor International Inc., (Lexington, MA). This sensor is
placed inside of a package and monitors the level of amines generated by microbio-
logical degradation of a meat product. Amine oxidases present in the sensor react to
the presence of amines by initiating a change of color, communicating to the con-
sumer that the product is no longer fresh.
Sensors are different from indicators, in that they use energy to trigger a response.
They are designed to quantify or detect key analytes of interest. A sensor receives a
signal on its receptor from the analyte. This signal is then converted into energy that
controls the communicative ability of the device. Chemical sensors receive a signal
from a chemical analyte, for example, the amount of CO2 within a package.
Biosensors receive the signal from a biological material, such as the presence of an
antigen that may be associated with a pathogen.
6.8 Concluding Thoughts
Packaging has grown to play a major role in everyday consumer life. Significant
advances in food packaging technology over the last century have assisted in
enhancing the shelf life of many products, all while providing an elevated level of
convenience to the consumer. The future should focus on adapting innovative tech-
nologies that may control pathogenic and spoilage organisms to the real world. As
we have seen, many of these technologies are promising in laboratory settings, but
they do not always behave the same when translated into real-world applications.
Additionally, while current packaging technologies assist in the enhancement of
shelf life, the fact that food waste is still a major global concern suggests that
research and innovation in this area need to continue. Future research should also
focus on sustainability and improving the environmental impact of packaging mate-
132 C. Ebner et al.
rials. Unfortunately, many of the materials frequently used in food packaging are
not recyclable, which can have a detrimental impact on the environment. Though
sometimes the benefits of food packaging go unnoticed compared to the above limi-
tations, the importance of these materials in our everyday lives cannot be understated.
References
Ahmed, Z., O.N. Donkor, W.A. Street, and T. Vasiljevic. 2013. Activity of endogenous mus-
cle proteases from 4 Australian underutilized fish species as affected by ionic strength,
pH, and temperature. Journal of Food Science 78 (12): C1858–C1864. https://doi.
org/10.1111/1750-3841.12303.
Beckwith, S., F. B. Edwards, J. Rivett, C. L. Ebner, T. Kennedy, , R. McDowell, and D. V. Speer.
2016. Multilayer film having an active oxygen barrier layer with radiation enhanced active bar-
rier properties. United States Patent 9452592. Cryovac, Inc. Duncan, SC
Boziaris, I.S., and F.F. Parlapani. 2017. Specific spoilage organisms (SSOs) in fish. In The
Microbiological Quality of Food Foodborne Spoilers, ed. Antonio Bevilacqua, Maria
Rosaria Corbo, and Milena Sinigaglia, 61–98. Amsterdam: Elsevier. https://doi.org/10.1016/
B978-0-08-100502-6.00006-6.
Brandon, K., M. Beggan, P. Allen, and F. Butler. 2009. The performance of several oxygen scaven-
gers in varying oxygen environments at refrigerated temperatures: Implications for low-oxygen
modified atmosphere packaging of meat. International Journal of Food Science & Technology
44 (1): 188–196. https://doi.org/10.1111/j.1365-2621.2008.01727.x.
Calafat, A.M., Z. Kuklenyik, J.A. Reidy, S.P. Caudill, J. Ekong, L.L. Needham, and L. L. 2005.
Urinary concentrations of Bisphenol A and 4-Nonylphenol in a human reference population.
Environmental Health Perspectives 113 (4): 391–395. https://doi.org/10.1289/ehp.7534.
Charles, F., C. Guillaume, and N. Gontard. 2008. Effect of passive and active modified atmosphere
packaging on quality changes of fresh endives. Postharvest Biology and Technology 48 (1):
22–29.
Costa, C., A. Lucera, A. Conte, M. Mastromatteo, B. Speranza, A. Antonacci, and M.A. Del Nobile.
2011. Effects of passive and active modified atmosphere packaging conditions on ready-to-eat
table grape. Journal of Food Engineering 102 (2): 115–121.
de Abreu, D.A.P., J.M. Cruz, and P.P. Losada. 2011. Active and intelligent packaging for the Food
industry. Food Reviews International 28 (2): 146–187. https://doi.org/10.1080/87559129.201
1.595022.
De Poix, H.M.J.T. 1945. Process for Preserving Perishable Foodstuffs. Dewey and Almy Chemical
Company: Cambridge, MA.
Del Nobile, M.A., and A. Conte. 2013. Packaging for Food Preservation. New York, NY: Springer.
https://doi.org/10.1007/978-1-4614-7684-9.
DeWitt, C., and A. Oliveira. 2016. Modified atmosphere systems and shelf life extension of fish
and fishery products. Food 5 (4): 48. https://doi.org/10.3390/foods5030048.
Djenane, D., and P. Roncalés. 2018. Carbon monoxide in meat and fish packaging: Advantages and
limits. Food 7 (2): 12. https://doi.org/10.3390/foods7020012.
European Food Safety Authority (EFSA). 2009. Guidelines on submission of a dossier for safety
evaluation by the EFSA of active or intelligent substances present in active and intelligent
materials and articles intended to come into contact with food. EFSA Journal 7 (8): 1208.
Farber, J.M. 2016. Microbiological aspects of modified-atmosphere packaging technology - A
review. Journal of Food Protection 54 (1): 58–70. https://doi.org/10.4315/0362-028X-54.1.58.
Font-i-Furnols, M., and L. Guerrero. 2014. Consumer preference, behavior and perception about
meat and meat products: An overview. Meat Science 98 (3): 361–371. https://doi.org/10.1016/j.
meatsci.2014.06.025.
Franklin, N., K.D. Cooksey, and K. Getty. 2004. Inhibition of Listeria monocytogenes on the sur-
face of individually packaged hot dogs with a packaging film coating containing Nisin. Journal
of Food Protection 67 (3): 480–485.
Fromm, H.I., and K.J. Boor. 2004. Characterization of pasteurized fluid milk shelf-life attributes.
Journal of Food Science 69 (8): M207–M214.
Gram, L. 2001. Potential hazards in cold-smoked fish: Clostridium botulinum type E. Journal
of Food Science 66 (35): S1082–S1087. https://doi.org/10.1111/j.1365-2621.2001.tb15527.x.
Haghighi, H., R. De Leo, E. Bedin, F. Pfeifer, H.W. Siesler, and A. Pulvirenti. 2019. Comparative
analysis of blend and bilayer films based on chitosan and gelatin enriched with LAE (lauroyl
arginate ethyl) with antimicrobial activity for food packaging applications. Food Packaging
and Shelf Life 19: 31–39.
Heyerick, A., Y. Zhao, P. Sandra, K. Huvaere, F. Roelens, and D. De Keukeleire. 2003. Photolysis
of hop-derived trans-iso-alpha-acids and trans-tetrahydroiso-alpha-acids: Product identifica-
tion in relation to the lightstruck flavour of beer. Photochemical and Photobiological Sciences
2 (3): 306–314.
Horev, B., S. Sela, Y. Vinokur, E. Gorbatsevich, R. Pinto, and V. Rodov. 2012. The effects of
active and passive modified atmosphere packaging on the survival of Salmonella enterica sero-
type Typhimurium on washed romaine lettuce leaves. Food Research International 45 (2):
1129–1132.
Kelly, C.A., M. Cruz-Romero, J.P. Kerry, and D.P. Papkovsky. 2018. Assessment of performance
of the industrial process of bulk vacuum packaging of raw meat with nondestructive optical
oxygen sensing systems. Sensors 18 (5): 1395. https://doi.org/10.3390/s18051395.
Lucquin, A., K. Gibbs, J. Uchiyama, H. Saul, M. Ajimoto, Y. Eley, A. Radini, C.P. Heron, S. Shoda,
Y. Nishida, J. Lundy, P. Jordan, S. Isaksson, and O.E. Craig. 2016. Ancient lipids document
continuity in the use of early hunter-gatherer pottery through 9,000 years of Japanese prehis-
tory. Proceedings of the National Academy of Sciences of the United States of America 113
(15): 3991–3996. https://doi.org/10.1073/pnas.1522908113.
Mangaraj, S., and T.K. Goswami. 2009. Modified atmosphere packaging of fruits and vegetables
for extending shelf-life: A review. Fresh Produce 3 (1): 1–31.
Natrajan, N., and B.W. Sheldon. 2000. Efficacy of nisin-coated polymer films to inactivate
Salmonella Typhimurium on fresh broiler skin. Journal of Food Protection 63 (9): 1189–1196.
Nielsen, P.V., and R. Rios. 2000. Inhibition of fungal growth on bread by volatile components from
spices and herbs, and the possible application in active packaging, with special emphasis on
mustard essential oil. International Journal of Food Microbiology 60 (2–3): 219–229. https://
doi.org/10.1016/S0168-1605(00)00343-3.
Nugraha, B., N. Bintoro, and H. Murayama. 2015. Influence of CO2 and C2H4 adsorbents to
the symptoms of internal browning on the packaged ‘silver bell’ pear (Pyrus communis L.).
Agriculture and Agricultural Science Procedia 3 (1): 127–131.
Oulé, M.K., K. Tano, A.-M. Bernier, and J. Arul. 2006. Escherichia coli inactivation mechanism
by pressurized CO2. Canadian Journal of Microbiology 52 (12): 1208–1217. https://doi.
org/10.1139/w06-078.
Ribeiro-Santos, R., A. Sanches-Silva, J.F.G. Motta, M. Andrade, I. de Araújo Neves, R.F. Teófilo,
M.G. de Carvalho, and N.R. de Melo. 2017. Combined use of essential oils applied to protein
base active food packaging: Study in vitro and in a food simulant. European Polymer Journal
93: 75–86.
Risvik, E. 2001. The Food and I sensory perception as revealed by multivariate methods. In Food,
people and society, ed. L.J. Frewer, E. Risvik, and H. Schifferstein, 23–37. Berlin, Heidelberg:
Springer. https://doi.org/10.1007/978-3-662-04601-2_3.
Robertson, G.L. 2012. Food Packaging: Principles and Practice. 3rd ed. Boca Raton, FL: CRC
Press Taylor & Francis.
Rossaint, S., S. Klausmann, U. Herbert, and J. Kreyenschmidt. 2014. Effect of package perfo-
ration on the spoilage process of poultry stored under different modified atmospheres. Food
Packaging and Shelf Life 1 (1): 68–76. https://doi.org/10.1016/j.fpsl.2014.01.002.
134 C. Ebner et al.
Russell, S.M., D.L. Fletcher, N.A. Cox, and N. A. 1995. Spoilage Bacteria of fresh broiler chicken
carcasses. Poultry Science 74 (12): 2041–2047. https://doi.org/10.3382/ps.0742041.
Schug, T. T., J. J. Heindel, L. Camacho, K. B. Delclos, , P. Howard, A. F. Johnson,, J. Aungst,
D. Keefe, R. Newbold, N. J. Walker, R. Thomas Zoeller, and J. R. Buchen. 2013. A new approach
to synergize academic and guideline-compliant research: The CLARITY-BPA research pro-
gram. Reproductive Toxicology 40:35–40. doi:https://doi.org/10.1016/j.reprotox.2013.05.010.
Seachrist, D.D., K.W. Bonk, S.-M. Ho, G.S. Prins, A.M. Soto, and R.A. Keri. 2016. A review of
the carcinogenic potential of bisphenol A. Reproductive Toxicology 59: 167–182. https://doi.
org/10.1016/j.reprotox.2015.09.006.
Solano, A.C.V., and C. de Rojas Gante. 2012. Two different processes to obtain antimicrobial
packaging containing natural oils. Food and Bioprocess Technology 5 (6): 2522–2528.
Strom, E.T., and S.C. Rasmussen. 2011. 100+ Years Of Plastics. Leo Baekeland and Beyond.
Washington, DC: American Chemical Society. https://doi.org/10.1021/bk-2011-1080.
Suppakul, P., J. Miltz, K. Sonneveld, and S.W. Bigger. 2003. Active packaging technologies with
an emphasis on antimicrobial packaging and its applications. Journal of Food Science 68 (2):
408–420. https://doi.org/10.1111/j.1365-2621.2003.tb05687.x.
Thompson, A.K. 2010. Controlled Atmosphere Storage of Fruits and Vegetables. 2nd ed.
Wallingford, Oxfordshire, UK: CABI.
United States Department of Agriculture. 2019. Estimates of Food Waste. https://www.usda.gov/
foodwaste/faqs. Accessed 31 November 2019.
Van Willige, R., D. Schoolmeester, A. Van Ooij, J. Linssen, and A. Voragen. 2002. Influence
of storage time and temperature on absorption of flavor compounds from solutions by
plastic packaging materials. Journal of Food Science 67 (6): 2023–2031. https://doi.
org/10.1111/j.1365-2621.2002.tb09495.x.
Voges, K.L., C.L. Mason, J.C. Brooks, R.J. Delmore, D.B. Griffin, D.S. Hale, W.R. Henning,
D.D. Johnson, C.L. Lorenzen, R.J. Maddock, R.K. Miller, J.B. Morgan, B.E. Baird,
B.L. Gwartney, and J.W. Savell. 2007. National beef tenderness survey–2006: Assessment of
Warner–Bratzler shear and sensory panel ratings for beef from US retail and foodservice estab-
lishments. Meat Science 77 (3): 357–364. https://doi.org/10.1016/j.meatsci.2007.03.024.
Woraprayote, W., L. Pumpuang, A. Tosukhowong, T. Zendo, K. Sonomoto, S. Benjakul, and
W. Visessanguan. 2018. Antimicrobial biodegradable food packaging impregnated with
Bacteriocin 7293 for control of pathogenic bacteria in pangasius fish fillets. LWT 89: 427–433.
Zhao, C., B. Ge, J. De Villena, R. Sudler, E. Yeh, S. Zhao, D.G. White, D. Wagner, and J. Meng. 2001.
Prevalence of Campylobacter spp., Escherichia coli, and Salmonella serovars in retail chicken,
Turkey, pork, and beef from the greater Washington, D.C., area. Applied and Environmental
Microbiology 67 (12): 5431–5436. https://doi.org/10.1128/AEM.67.12.5431-5436.2001.
Chapter 7
Beyond the Standard Plate Count:
Genomic Views into Microbial Food
Ecology
7.1 Introduction
Genomic approaches to preserving and extending shelf life, as well as improving

functional properties and nutritional quality.
For more than half a century, spoilage-causing microbes have been identified
with some success primarily relying on traditional selective and nonselective cul-
ture techniques (Waite et al. 2009). Advancements in genomics research have
helped scientists to better understand how microbes interact with one another and
their environment, how they infect their host, and how they have evolved over
time. In the field of food science, genomics have helped us to better understand the
microbial ecology of microbial communities on foods and food contact surfaces.
Microbial ecology is of special interest to food microbiologists because it describes
the ability of microbes to function in complex food environments (Floros et al.
2010). While studying individual species is important, traditional cultivation meth-
ods of spoilage microbes can be laborious, time-consuming, and biased by selec-
tive germination and outgrowth conditions on nutrient agar plates (Davey 2011;
Kort et al. 2008).
Microbial spoilage is a complex ecological process involving the presence of
multiple species with specific niches, as well as metabolic dependencies, which can
be easily overlooked using basic culture-dependent techniques which favor the
growth of single microbial species (Gram et al. 2002). Understanding how these
individual species function within a diverse microbial environment can help us to
understand the “bigger picture” of how these species interact in complex biological
systems. These dynamic microbial communities include nonpathogenic spoilage
S. M. Hertrich · B. A. Niemira (*)

Food Safety Intervention Technologies Research Unit, Eastern Regional Research Center,
USDA-ARS, Wyndmoor, PA, USA
e-mail: sarmark@udel.edu; Brendan.Niemira@USDA.GOV

https://doi.org/10.1007/978-3-030-54375-4_7
microbes which affect food qualities including taste, flavor, texture, appearance, and
shelf life (Floros et al. 2010). Genomics can also help scientists to better understand
the process of fermentation, which is the microbial degradation of organic com-
pounds within food products and is one of the oldest and most commonly used
forms of food preservation (Sieuwerts et al. 2008). Because food fermentations are
typically carried out by mixed cultures consisting of multiple microbial species,
understanding the population dynamics of these cultures can help to increase their
industrial performance (Sieuwerts et al. 2008).
Culture-independent genomics tools, such as genome sequencing, have allowed
scientists to more accurately estimate the microbial diversity of foods (Floros et al.
2010). Until recently, scientists were limited to looking at DNA sequences of a small
set of genes among small set of organisms at one given time. Now, scientists are able
to obtain complete information about the organization and genetic composition of
entire genomes (Watson et al. 2008) at a considerably low cost. In addition to micro-
bial genomics, which is the study of the genomes of individual microbes, microbial
metagenomics is the culture-independent functional and sequence-based analysis of
the heterogeneous microbial genomes from a particular environment (Riesenfeld et al.
2004; Handelsman et al. 1998; Casas and Rohwer 2007; Gilbert and Dupont 2011;
Casas and Maloy 2014).
Other “omics” tools including proteomics and metabolomics provide a better
understanding of metabolites, enzyme activity, and metabolic fluxes (Floros et al.
2010), which also play a role in food degradation. Changes to the environment,
including deforestation, irrigation, coastal zone degradation, wetland modification,
expansion of urban areas, global climate change, and other human interventions,
can lead to changes in the structure, composition, and dynamics of microbial com-
munities (Patz et al. 2004) in which our food is grown or produced. When used
appropriately, “omics” tools can be used to help scientists generate models that can
identify changes in microbial ecology and, therefore, prevent spoilage of foods and
food crops (Casas and Maloy 2014).
7.2 Recombinant DNA Technology
Recombinant DNA (rDNA) technology is a nontraditional alternative to conven-

tional breeding practices, which allows for the transfer of a large number of specific
genes into edible plants (Kumar and Kumar 2015). This technology, also referred to
as genetic engineering or transgenic technology, allows for the genetic modification
of edible plants to prevent spoilage, increase crop yields, decrease exposure to her-
bicides and pesticides, decrease production costs, and ultimately the creation of
more nutritious food products at cheaper prices. In addition, rDNA technology has
also allowed for the reduction or elimination of allergens from foods in order to
protect those individuals who suffer from food allergies (IFT 2000). It is believed
that the use of rDNA technology will allow us to feed the world in the future, when
the population will have multiplied considerably, and there will be less land and
7 Beyond the Standard Plate Count: Genomic Views into Microbial Food Ecology 137
resources available to grow crops. Crops that are tolerant to local soil and weather
conditions, particularly in developing countries, will allow for the growth of
nutritious foods at an affordable cost.
Recombinant DNA technology involves the “cloning” of a specific genomic tar-
get into a “transformed” organism of interest. Cloning of bacterial DNA describes
the ability to construct recombinant DNA molecules and maintain them within
cells. This process typically involves the use of a vector which provides all the nec-
essary information that allows the new DNA to be inserted into the cell, allows the
recombinant vector to multiply independently, and carries a selectable marker
which distinguishes transformed cells from non-transformed cells (Watson et al.
2008). The most common types of vectors are plasmids, which are small (~3 kb)
circular DNA molecules found naturally in many bacteria. Restriction enzymes are
used to cut specific segments of the vector DNA where the recombinant DNA is to
be inserted and ligation enzymes join the ends of the inserted DNA sequence to the
new cell.
7.3 Whole Genome Sequencing
In 1953, Watson and Crick solved the structure of DNA by studying the crystallo-
graphic data produced by Rosalind Franklin and Maurice Wilkins (Watson and Crick
1953; Zallen 2003). This discovery allowed future scientists to understand the con-
cepts of DNA replication and encoding proteins in nucleic acids; however, the ability
to “read” DNA sequences did not develop for quite some time (Heather and Chain
2016). The first studies of microbial genomes began in the 1970s with the sequenc-
ing of two bacteriophage genomes (Wooley et al. 2010; Fiers et al. 1976; Sanger
et al. 1978). The development that was considered the “major breakthrough” in DNA
sequencing technology came about in 1977, known as Sanger’s “chain termination”
or dideoxy technique (Sanger and Nicklen 1977). Then, in 1995 the first bacterial
genome, Haemophilus influenza, was sequenced and regarded as a huge step for the
microbiology community (Wooley et al. 2010; Fleischmann et al. 1995).
Advanced molecular techniques, specifically whole genome sequencing, have
helped scientists to better understand gene expression in individual microbial cells
(Floros et al. 2010). Whole genome sequencing arrays have also allowed scientists
to observe the complex regulation of gene expression within the cell that occurs
during infection (Toledo-Arana et al. 2009). For example, scientists now understand
exactly which virulence genes in specific species of bacteria and viruses are upregu-
lated in the host in order for infection to occur.
With information provided by whole genome sequencing, food scientists will be
able to develop cost-effective approaches for detection and control of the strains that
are likely to cause disease (Floros et al. 2010). Sequencing of plant and animal
genomes will allow scientists to study their evolution and domestication and, there-
fore, facilitate establishment of more effective breeding programs (Qin et al. 2014).
Specialized intervention technologies in food processing plants, as well as other
points in the food production chain, can also be developed to prevent contamination
with foodborne pathogens (Floros et al. 2010). Strain monitoring may help identify
dominant microbial strains which drive food spoilage, as well as strains that drive the
fermentation process (Ecrolini 2013), which would benefit future food production
and sustainability strategies. For example, scientists used whole genome sequencing
coupled with traditional culturing techniques to identify the specific spoilage organ-
isms in vacuum packaged ham (Piotrowska-Cyplik et al. 2017). They were also able
to identify the specific spoilage organisms which were responsible for changes in pH
and other organoleptic characteristics during deterioration of the quality of the meat.
7.4 Metagenomics
Over the last two decades, the decrease in the cost of whole genome sequencing has
significantly increased accessibility to a large number of microbial genomes. The
availability of this information has changed the nature of microbiology, as well as
the way we study microbial ecology (Wooley et al. 2010). Scientists are now able to
study and compare microbial genomes, side by side, which has opened up new
fields of study including comparative genomics and systems biology. These areas of
study are highly beneficial because microbes very rarely live in single species com-
munities. Microbes most often interact with each other, as well as their environ-
ment, which could also potentially include a host organism (Wooley et al. 2010).
Metagenomics describes the study of genetic material recovered from environ-
mental samples such as food, soil, and water. Metagenomics allows scientists to
study the variety microbial organisms within a sample, even if the organisms are
unculturable in the laboratory (Coughlan et al. 2015). This type of technology allows
scientists to discover how microbes function in communities. For example, scientists
now better understand the diversity and complexity of the microbial communities
that reside in the human gastrointestinal tract, many of which were previously unable
to be cultured in the laboratory by traditional methods (Ley et al. 2008; Wooley et al.
2010). This type of analyses can reveal the identity of a species present within a
sample, as well as provide insight into the metabolic activities and functional roles
of the microbes present within a sample population (Langille et al. 2013).
A typical metagenomics analysis begins with the isolation of high-quality micro-
bial DNA from an environmental sample which should represent all species present
within the sample, qualitatively and quantitatively (Coughlan et al. 2015). Metagenomic
DNA is then directly sequenced and analyzed using bioinformatics to determine the
functional traits of the microbial organisms which are present in the initial sample
(Coughlan et al. 2015). Data are typically analyzed using computer software to infer
phylogenetic relationships with existing genomic databases (Santiago-Rodriguez
et al. 2016). Operational Taxonomic Units (OTUs) are similar sequences of DNA that
serve as markers of species relatedness. The abundance of these OTUs are calculated
to determine the taxonomy and diversity of the microbial c ommunity present within
the sample (Kuczynski et al. 2012). These techniques can be applied to help identify
specific organisms and enzymes involved in food spoilage, as well as novel enzymes
from natural sources to aid in food processing reactions (Coughlan et al. 2015).
7.5 CRISPR-Cas
Clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-

associated sequences (Cas) make up the adaptive immune system of bacteria, which
protects against invasive genomic elements (Barrangou and Marraffini 2015).
CRISPR arrays confer immunological memory and surveillance mechanisms in
bacteria, and cas genes encode effector proteins when the cell is under attack (Selle
and Barrangou 2015). Immunity mediated by CRISPR-Cas is categorized into three
molecular processes including acquisition, expression, and interference (Barrangou
2013; Barrangou and Marraffini 2015). Acquisition occurs when foreign genetic
elements, such as bacteriophage DNA, enter a host cell. The foreign genetic element
“samples” a novel spacer sequence from the host cell’s chromosome using a copy
and paste process, which leads to the formation of an additional CRISPR repeat-
spacer unit within the host cell. Expression and RNA biogenesis occur in mature
surveillance CRISPR RNAs (crRNAs), which comprise a portion of the “sample”
CRISPR spacer sequence and define the target sequence in which the mobile genetic
element is attempting to invade. Interference against mobile genetic elements is
driven by Cas nucleases, which allow recognition of the target sequence. Cas nucle-
ases perform cleavage of complementary DNA elements to interfere with the inva-
sion of the mobile genetic element and are defined by the crRNA guide sequence
(Selle and Barrangou 2015). Scientists have found a way to reprogram CRISPR-Cas
system to edit and remodel the genomes of a variety of organisms.
It is believed that the development of the CRISPR-Cas system has reinvigorated
the field of functional genomics in a scientific area that has recently been defined by
whole genome sequencing (Selle and Barrangou 2015). Scientists are now using the
CRISPR-Cas system to perform genome editing in a diverse range of organisms
including eukaryotes and prokaryotes. Early application of CRISPR-Cas technol-
ogy arose from food-science-driven research when industrial starter cultures for
milk fermentation processes were characterized (Barrangou et al. 2007). The tech-
nology has also been applied to other organisms of interest across the field of food
science including yeast, corn, rice, and tomatoes (Selle and Barrangou 2015). Other
applications include genome editing for improvement of growth and disease resis-
tance in food crops and animals (Selle and Barrangou 2015).
7.6 Foodomics
The term “foodomics” has been defined as a new discipline, which describes the
study of food and nutrition through the application of advanced “omic” technolo-
gies to improve the confidence, health, and well-being of the consumer (Cifuentes
2009; Herrero et al. 2010; Herrero et al. 2012). Foodomics is regarded as a global
discipline in which food, nutrition, advanced analytical techniques, and bioinfor-
matic tools are brought together (Herrero et al. 2012; Andjelković et al. 2017). The
suffixes “ome” and “omics” are derived from the term “genome,” which was created
by Hans Winkler in 1920 (Capozzi and Bordoni 2013). The suffix is now used to
describe some of the major high-throughput “omics” methodologies including
genomics, transcriptomics, proteomics, metabolomics, allergomics, lipidomics,
nutrigenomics, as well as many others. These disciplines include the study of genes,
RNA transcripts, proteins, metabolites, allergens, fats, and other nutrients that can
be found within the cells of foods and food ingredients. Advanced technologies
including mass spectrometry (MS)-based tools are commonly used to carry out
“omics” based studies (Herrero et al. 2012). Other technologies including nuclear
resonance spectroscopy and liquid chromatography (LC) are commonly used in
metabolomics-based studies (Gibbons et al. 2015).
Foodomics is a systems science that can be used to aid in the development of
transgenic foods, metabolic study for compound profiling, biomarker analysis
related to food quality, the effects of food bioactivity on human health, as well as
address food safety issues (Herrero et al. 2012; Quigley et al. 2012). These tools can
provide a health assessment of plants and animals as food producers (García-Cañas
et al. 2012), as well as the production and monitoring of food quality (Gašo-Sokač
et al. 2011). Foodomics can also help scientists to further examine the rates and
causes of change in foods during storage in order to extend shelf life. For example,
food products including red wine (Arapitsas et al. 2016) and iceberg lettuce (Garcia
et al. 2016) have been investigated using LC and MS platforms to determine the
impact of storage specifications on their metabolic profile (Castro-Puyana et al.
2017; Bayram and Gökırmaklı 2018). Scientists have also used variety of foodo-
mics techniques, including MC, LC, and genomics, to demonstrate that the shelf life
of fruits can be enhanced by suppressing ripening-specific enzymes and slowing the
rate of fruit softening (Meli et al. 2010).
7.7 Current Challenges and Research Needs
Novel approaches to enhance the production and shelf life of foods are warranted in
order to feed the world’s growing population. The emerging fields of science as
described in this chapter can help to further our knowledge regarding the effects,
relevance, and significance of the changes that occur in foods over time. The avail-
ability of advanced genomics and genetics tools will influence the integration of a
mechanistic and evolutionary approach to tackling food quality and food safety
obstacles (West et al. 2006; Sieuwerts et al. 2008). As the field of food science con-
tinues to progress, we are likely to see a movement from the study of the single
components of food toward a systems approach, where we can better understand
how all the components of a product form a complex network associated with spe-
cific biological functions (Santiago-Rodriguez et al. 2016). These approaches may
be used more frequently, as the technology becomes more advanced and the cost
continues to become more and more affordable.
References
Andjelković, U., M.Š. Gajdošik, D. Gašo-Sokač, T. Martinović, and D. Josić. 2017. Foodomics
and food safety: Where we are. Food Technology and Biotechnology 55 (3): 290–307.
Arapitsas, P., A. Della Corte, H. Gika, L. Narduzzi, F. Mattivi, and G. Theodoridis. 2016. Studying
the effect of storage conditions on the metabolite content of red wine using HILIC LC–MS
based metabolomics. Food Chemistry 197: 1331–1340.
Barrangou, R. 2013. CRISPR-Cas systems and RNA-guided interference. Wiley Interdisciplinary
Reviews RNA 4: 267–278.
Barrangou, R., and L.A. Marraffini. 2015. CRISPR-Cas systems: Prokaryotes upgrade to adaptive
immunity. Molecular Cell 54: 234–244.
Barrangou, R., C. Fremaux, H. Deveau, M. Richards, P. Boyaval, S. Moineau, D.A. Romero, and
P. Horvath. 2007. CRISPR provides acquired resistance against viruses in prokaryotes. Science
315: 1709–1712.
Bayram, M., and Ç. Gökırmaklı. 2018. Horizon scanning: How will metabolomics applica-
tions transform food science, bioengineering, and medical innovation in the current era of
Foodomics? OMICS 22 (3): 177–183. https://doi.org/10.1089/omi.2017.0203.
Capozzi, F., and A. Bordoni. 2013. Foodomics: A new comprehensive approach to food and nutri-
tion. Genes & Nutrition 8: 1–4.
Casas, V., and S. Maloy. 2014. Genomic and metagenomic approaches for predicting pathogen
evolution. In One Health: People, Animals, and the Environment, ed. R.M. Atlas and S. Maloy,
227–235. Washington, DC: American Society for Microbiology.
Casas, V., and F. Rohwer. 2007. Phage metagenomics. Methods in Enzymology 421: 259–268.
Castro-Puyana, M., R. Pérez-Míguez, L. Montero, and M. Herrero. 2017. Application of mass
spectrometry-based metabolomics approaches for food safety, quality and traceability. Trends
in Analytical Chemistry 93: 102–118.
Cifuentes, A. 2009. Food analysis and foodomics. Journal of Chromatography. A 1216: 7109–7110.
Coughlan, L.M., P.D. Cotter, C. Hill, and A. Alvarez-Ordóñez. 2015. Biotechnological appli-
cations of functional metagenomics in the food and pharmaceutical industries. Frontiers in
Microbiology 6: 672.
Davey, H.M. 2011. Life, death, and in-between: Meanings and methods in microbiology. Applied
Ecrolini, D. 2013. High-throughput sequencing and metagenomics: Moving forward in the culture-
independent analysis of food microbial ecology. Applied and Environmental Microbiology 79
(10): 3148–3155.
Fiers, W., R. Contreras, F. Duerinck, G. Haegman, D. Iserentant, et al. 1976. Complete nucleotide
sequence of bacteriophage ms2 RNA: Primary and secondary structure of the replicase gene.
Nature 260: 500–507.
Fleischmann, R.D., M.D. Adams, O. White, R.A. Clayton, E.F. Kirkness, et al. 1995. Whole-
genome random sequencing and assembly of haemophilus influenza rd. Science 269: 496–512.
Floros, J., R. Newsome, and W. Fisher. 2010. Feeding the world today and tomorrow: The impor-
tance of food science and technology: An IFT Scientific Review. Comprehensive Reviews in
Food Science and Food Safety 9: 572–599.
Garcia, C.J., R. García-Villalba, Y. Garrido, M.I. Gil, and F.A. Tomás-Barberán. 2016. Untargeted
metabolomics approach using UPLC-ESI-QTOF-MS to explore the metabolome of fresh-cut
iceberg lettuce. Metabolomics 12: 138.
García-Cañas, V., C. Simó, M. Herrero, E. Ibáñez, and A. Cifuentes. 2012. Present and future chal-
lenges in food analysis: Foodomics. Analytical Chemistry 48: 10150–10159.
Gašo-Sokač, D., S. Kovač, and D.J. Josić. 2011. Use of proteomic methodology in optimization and
processing and quality control of food of animal origin. Food Technology and Biotechnology
49: 397–412.
Gibbons, H., A. O’Gorman, and L. Brennan. 2015. Metabolomics as a tool in nutritional research.
Current Opinion in Lipidology 26: 30–34.
Gilbert, J.A., and C.L. Dupont. 2011. Microbial metagenomics: Beyond the genome. Annual
Review of Marine Science 3: 347–371.
Gram, L., L. Ravn, M. Rasch, J.B. Bruhn, A.B. Christensen, and M. Givskov. 2002. Food spoilage-
interactions between food spoilage bacteria. International Journal of Food Microbiology 78
(1–2): 79–97.
Handelsman, J., M.R. Rondon, S.F. Brady, J. Clardy, and R.M. Goodman. 1998. Molecular bio-
logical access to the chemistry of unknown soil microbes: A new frontier for natural products.
Chemistry & Biology 5: R245–R249.
Heather, J.M., and B. Chain. 2016. The sequence of sequencers: The history of sequencing DNA.
Genomics 107 (1): 1–8.
Herrero, M., V. García-Cañas, C. Simo, and A. Cifuentes. 2010. Recent advances in the application
of CE methods for food analysis and foodomics. Electrophoresis 31: 205–228.
Herrero, M., C. Simó, V. García-Cañas, E. Ibáñez, and A. Cifuentes. 2012. Foodomics: MS-based
strategies in modern food science and nutrition. Mass Spectrometry Reviews 31: 49–69.
Institute of Food Technologists (IFT). 2000. Who Benefits from rDNA Biotechnology-Derived
Foods? http://www.ift.org/knowledge-center/read-ift-publications/science-reports/expert-
reports/biotechnology-and-foods/who-benefits-from-rdna-biotechnology-derived-foods.
aspx. Accessed 23 October 2017.
Kort, R., B.J. Keijser, M.P. Caspers, F.H. Schuren, and R. Montijn. 2008. Transcriptional activity
around bacterial cell death reveals molecular biomarkers for cell viability. BMC Genomics 9:
590.
Kuczynski, J., J. Stombaugh, W.A. Walters, A. Gonzalez, J.G. Caporaso, and R. Knight. 2012.
Using QIIME to analyze 16S rRNA gene sequences from microbial communities. Current
Protocols in Microbiology Unitas 1E: 5.
Kumar, S., and A. Kumar. 2015. Role of genetic engineering in agriculture. Plant Archives 15: 1–6.
Langille, M.G.I., J. Zaneveld, J.G. Caporaso, D. McDonald, D. Knights, J.A. Reyes, et al. 2013.
Predictive functional profiling of microbial communities using 16s rRNA marker gene
sequences. Nature Biotechnology 31: 814–821.
Ley, R.E., M. Hamady, C. Lozupone, P.J. Turnbaugh, R.R. Ramey, J.S. Bircher, M.L. Schlegel,
T.A. Tucker, M.D. Schrenzel, R. Knight, and J.I. Gordon. 2008. Evolution of mammals and
their gut microbes. Science 320: 1647–1651.
Meli, V.S., S. Ghosh, T.N. Prabha, N. Chakraborty, S. Chakraborty, and A. Datta. 2010.
Enhancement of fruit shelf life by suppressing N-glycan processing enzymes. PNAS 107 (6):
2413–2418.
Patz, J.A., P. Daszak, G.M. Tabor, A.A. Aguirre, M. Pearl, J. Epstein, N.D. Wolfe, A.M. Kilpatrick,
J. Foufopoulos, D. Molyneux, D.J. Bradley, and Working Group on Land Use Change and
Disease Emergence. 2004. Unhealthy landscapes: Policy recommendations on land use change
and infectious disease emergence. Environmental Health Perspectives 112: 1092–1098.
Piotrowska-Cyplik, A., K. Myszka, J. Czarny, K. Ratajczak, R. Kowalski, R. Biegańska-Marecik,
J. Staninska-Pięta, J. Nowak, and P. Cyplik. 2017. Characterization of specific spoilage
organisms (SSOs) in vacuum-packaged ham by culture-plating techniques and MiSeq next-
generation sequencing technologies. Journal of the Science of Food and Agriculture 97:
659–668.
Qin, C., C. Yu, Y. Shen, X. Fang, L. Chen, J. Min, et al. 2014. Whole-genome sequencing of cul-
tivated and wild peppers provides insights into Capsicum domestication and specialization.
PNAS 111 (14): 5135–5140.
Quigley, L., O. O’Sullivan, T.P. Beresford, R.P. Ross, G.F. Fitzgerald, and P.D. Cotter. 2012. High-
throughput sequencing for detection of subpopulations of bacteria not previously associated
with artisanal cheeses. Applied and Environmental Microbiology 78: 5717–5723.
Riesenfeld, C.S., P.D. Schloss, and J. Handelsman. 2004. Metagenomics: Genomic analysis of
microbial communities. Annual Review of Genetics 38: 525–552.
Sanger, S.F., and A.R.C. Nicklen. 1977. DNA sequencing with chain-terminating. Proceedings of
the National Academy of Sciences 74: 5463–5467.
Sanger, F., A.R. Coulson, T. Friedmann, G.M. Air, B.G. Barrell, et al. 1978. The nucleotide
sequence of bacteriophage phix174. Journal of Molecular Biology 125: 225–246.
Santiago-Rodriguez, T.M., R. Cano, and R. Jiménez-Flores. 2016. Potential applications of
metagenomics to assess the biological effects of food structure and function. Food & Function
7 (10): 4160–4169.
Selle, K., and R. Barrangou. 2015. CRISPR-based technologies and the future of food science.
Journal of Food Science 80 (11): R2367–R2371.
Sieuwerts, S., F.A.M. de Bok, J. Hugenholtz, and J.E.T. van Hylckama Vlieg. 2008. Unraveling
microbial interactions in food fermentations: From classical to genomics approaches. Applied
Toledo-Arana, A., O. Dussurget, G. Nikitas, N. Sesto, H. Gvet-Revillet, D. Balestrino, E. Loh,
J. Gripenland, T. Tiensuu, K. Vaitkevicius, M. Barthelemy, M. Vergassola, M.-A. Nahori,
G. Soubigov, B. Regnault, J.-Y. Coppee, M. Lecvit, J. Johansson, and P. Cossart. 2009. The
Listeria transcriptional landscape from saprophytism to virulence. Nature 459: 950–956.
Waite, J.G., J.M. Jones, and A.E. Yousef. 2009. Isolation and identification of spoilage microor-
ganisms using food-based media combined with rDNA sequencing: Ranch dressing as a model
food. Food Microbiology 26: 235–239.
Watson, J., and F. Crick. 1953. Molecular structure of nucleic acids. Nature 171: 709–756.
Watson, J.D., T.A. Baker, S.P. Bell, A. Gann, M. Levine, and R. Losick. 2008. Molecular Biology
of the Gene. 6th ed. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press.
West, S.A., A.S. Griffin, A. Gardner, and S.P. Diggle. 2006. Social evolution theory for microor-
ganisms. Nature Reviews. Microbiology 4: 597–607.
Wooley, J.C., A. Godzik, and I. Friedberg. 2010. A primer on metagenomics. PLoS Computational
Biology 6: 1–13.
Zallen, D.T. 2003. Despite Franklin’s work, Wilkins earned his Nobel. Nature 425: 15.
Chapter 8
The Changing Shelf Life of Chilled,
Vacuum-Packed Red Meat
John Summer, Paul Vanderlinde, Mandeep Kaur, and Ian Jenson
8.1 Introduction
The Australian red meat industry has a long history of export to a large number of
distant markets. The advent of vacuum packing made a significant change to the
industry and its ability to supply high-quality product to these markets. This chapter
describes the attempts to develop a solid Australian meat export trade prior to the
development of vacuum packing, and then the incremental, but significant, improve-
ments in shelf life since that time. While the attention of the industry, and this chap-
ter, is on beef, comparisons are made to the shelf life of lamb.
In 2016, Australia exported red meat to more than 130 countries. The industry
has come a long way from its origins, which began with seven Zebu cattle, 44
sheep, 19 goats and 32 pigs that landed with wobbly legs in 1788 after months at sea
on the First Fleet which colonised Australia. This motley herd formed the basis of
Australia’s livestock industry, and quickly outgrew the consumption needs of the
local population, leading to the need to export, almost entirely to England, the
mother country.
J. Summer
M&S Food Consultants, Deviot, TAS, Australia
P. Vanderlinde
Vanderlinde Consulting, Carbrook, QLD, Australia
M. Kaur
University of Tasmania, Hobart, TAS, Australia
I. Jenson (*)
Meat & Livestock Australia, North Sydney, NSW, Australia
e-mail: ijenson@mla.com.au

https://doi.org/10.1007/978-3-030-54375-4_8
146 J. Summer et al.
A great deal is known about the early Australian meat industry, thanks to two
definitive texts: A Settlement Amply Supplied by Dr. Keith Farrer (1980) and A
Review of Research since 1900 by Dr. Jim Vickery (1990). This introduction owes
much to their work, which shows how problems with shelf life when shipping across
both hemispheres were solved.
Mechanical refrigeration (‘cold on demand’) revolutionised the food industry,
replacing an existing global trade in natural ice. For Australia, refrigeration offered
commercial possibilities and, in 1873, the SS Norfolk was loaded with a trial ship-
ment of frozen meat to England; the trial was unsuccessful when the circulating
brine system failed.
In 1879, the SS Strathleven, fitted with mechanical refrigeration was loaded with
mutton, beef and butter, which were then frozen on board. After a 64-day voyage the
Strathleven arrived in London with a 34-tonne cargo in excellent condition.
Organoleptic testing using an untrained consumer panel proceeded at a lunch on board
the vessel for 150 tasters, and samples to Queen Victoria received the royal assent.
Problems caused by a lack of knowledge of how to thaw frozen meat meant the
Australian trade soon took second place to chilled meat from Argentina and
Paraguay. Successfully landed in London in 1907 after a 14-day voyage, their prod-
uct was markedly superior to Australian frozen meat because there was no ‘drip’,
and it attracted a price premium.
For the next 60 years, Australian research concentrated on trying to solve the
problem of delivering chilled meat to distant markets. Various processes were
investigated, and scientists from the forerunner of the Commonwealth Scientific
and Industrial Research Organisation (CSIRO) worked on commercialising the
effect of carbon dioxide on extending shelf life of chilled meat (Empey and Vickery
1933). The first trial shipment of chilled beef under carbon dioxide took place in
1933, with forequarters held in gas-tight cargo spaces. There were great problems
due to gas leaks and the consignment was landed in the United Kingdom with
incipient spoilage. Fitting out vessels with gas-tight storage rooms proved difficult,
and World War 2 brought an end both to the export trade and its research and
development.
However, the advent of flexible packaging in the 1960s allowed the chilled
vacuum-packed (VP) meat trade to move towards commercial reality; vacuum
packaging technology progressed quickly thereafter so that, by the 1980s, shelf life
of around 14 weeks at −1 °C was considered the industry norm. More recently, stor-
age trials carried out in Australia on VP beef primals (whole muscles) indicate shelf
life of around 28 weeks (Small et al. 2012) and Canadian researchers have reported
similar extended shelf life (Youssef et al. 2014).
In this chapter, we discuss possible reasons for this significant extension in shelf
life of chilled VP beef primals. We begin the journey by focusing on early R&D in
Australia and New Zealand, countries for which the technology presented obvious
commercial advantages. A review by Nottingham (1982) presents the early state of
the art led by researchers at CSIRO (e.g. Grau 1979) and the New Zealand Meat
Research Institute (e.g. Gill and Newton 1979).
We then focus on the question of whether shelf life extension has resulted from
improvements in hygiene and refrigeration, leading to a shift in microbial commu-
8 The Changing Shelf Life of Chilled, Vacuum-Packed Red Meat 147
nities in the vacuum pack. We also seek to distinguish between a focus solely on
bacterial numbers in evaluating end of shelf life and one of sensory testing as the
primary consideration.
8.2 ensory and Microbiological Quality of Chilled Red

S
Meats
Shelf life of chilled meats is determined when product is considered unacceptable

by any one of the following purchasers in the supply chain:
• Brokers or wholesalers, who move product to the next purchaser
• Further processors, who portion meat for retail or food service use
• Retailers, who display product for sale
• Food service operators, who prepare meat for consumption
• Consumers, who purchase meat for consumption
For the final consumer, there are three occasions when quality is assessed:
• At purchase, when the meat appearance is evaluated, particularly meat colour
and amount of drip (exudate, also known as purge)
• On opening, when odour and whether the meat is slimy/sticky due to excessive
bacterial growth are major criteria
• On consumption, when odour, flavour and texture are important criteria
More detailed criteria by which purchasers assess chilled product packed in vari-
ous ways are presented in Table 8.1.
The relationship between the microbiological profile and sensory changes differs
between meat packed under aerobic conditions and that packed under atmospheres
containing high concentrations of CO2 (20% or more) such as occurs in vacuum
(VP) and modified atmosphere packs (MAP). Inside a vacuum package, residual
oxygen is consumed due to ongoing muscle respiration and bacterial growth and
CO2 is produced. This results in an atmosphere of <1% O2 at 20–40% CO2 (Egan
et al. 1988). Under aerobic storage, shelf life ends quickly (around 12 days at
0–2 °C) because of the growth of psychrotrophic, Gram-negative bacteria which
become biochemically active, such as Pseudomonas and Shewanella. In VP and
MAP meats stored close to 0 °C, shelf life is extended past 20 weeks because of the
inhibition of Gram-negative spoilage bacteria by the high CO2 concentration and the
microbial communities selected for by the storage conditions: lactic acid bacteria
(LAB), which are relatively biochemically innocuous.
Growth of LAB in VP meat was demonstrated by Egan (1983) with a typical sig-
moid growth curve observed in VP primals stored at 0 °C (Fig. 8.1), where the popu-
lation of LAB increased to a maximum of log10 7–8 cfu/cm2 after around 10 weeks.
The LAB were a small proportion of the aerobic plate count (APC) initially, and
came to dominate the APC within the first few weeks. Shelf life was deemed ended
after around 14 weeks due to persistent off-odours when packs were opened.
Table 8.1 Sensory criteria of red meat through shelf life

Retail meat package Positive quality attributes Attributes at end of shelf life
Over-wrapped tray Pink-red bloom Loss of bloom, brown discolouration
(non-aged meat) Odour of fresh meat Off-odours, off-flavours, slime
MAP—High Pink-red bloom Loss of bloom, brown discolouration
oxygen, high CO2 Odour of fresh meat Off-odours, off-flavours, slime
Excessive purge (drip)
Vacuum pack Purple meat colour, tight pack Unacceptable, persistent odour
Short-lived confinement odour Meat discoloured (brown, grey or green)
Minimal purge (drip) in intact pack
Excessive purge (drip)
8
APC
7
bacterial count (log10 /cm2)
LAB
6
5
4
3
2
1
0
0 5 10 15
Storage time (weeks)
Fig. 8.1 Increase in the APC and LAB counts on VP beef primals stored at 0 °C (after Egan 1983),
arrow shows time at which product was considered spoiled
These early studies indicate a complex relationship between the bacterial popu-
lation in vacuum packs and the end of shelf life as evidenced by sensory evaluation;
the attainment of maximum concentration per se was found not to equate with end
of shelf life, one corollary being that it is the composition of the population and the
build-up of metabolic end products which are critical in determining shelf life.
8.3 Then and Now: Sensory Aspects of Shelf Life
In the early 1970s, sensory panels recorded maximum acceptability of VP beef

around 7 weeks, allowing the Japanese market to become a commercial reality. A
total shelf life of 10–12 weeks at 0 °C was possible until ended by flavour changes
described by panellists as sour, acidic and cheesy (Newton and Rigg 1979).
More recently, anecdotal evidence from meat exporters suggested that shelf life
of VP beef has improved greatly (>20 weeks) compared with that found in the
1980s. To evaluate this suggestion, the CSIRO set up a trial on VP beef primals
stored at −0.5 °C. At the expected end of shelf life (20 weeks), all samples were still
of excellent sensory quality; no samples remained to determine the actual shelf life
(Small et al. 2009).
A second, more extensive trial was set up using VP cube rolls and striploins
sourced from six abattoirs located from a broad latitudinal range from Tasmania in
the south (42°S) to far-north Queensland (19°S) to include stock from both temper-
ate and subtropical areas. Vacuum packs were stored at −0.5 °C for up to 30 weeks.
A trained sensory panel rated primals well for appearance and odour up to 28 weeks,
after which there was a marked decline in score, particularly for striploins. Steaks
cooked from striploins and cube rolls stored for 26 weeks scored well for aroma and
flavour with sour flavours only noted at 28 weeks (Small et al. 2012).
In addition to documenting extraordinarily long shelf life, Small et al. (2012)
recorded bacterial growth curves that did not conform with the traditional sigmoid
curve comprising lag, exponential, stationary and decline phases, and the median
counts barely reached 6 log10 cfu/cm2. In addition, there were large variations in
counts on each sampling day, often of the order of 4–5 log10 cfu/cm2, both within
and between processing establishments which had started with similar contamina-
tion levels.
The authors found it difficult to explain their findings, though they noted a New
Zealand study of vacuum-packed beef striploins stored at −1 °C for 16 weeks in
which the APC similarly did not follow the expected sigmoid curve, barely reaching
6 log10 cfu/cm2 by the end of the trial (Penney et al. 1998). These observations are
typical for growth of bacteria near their growth/no-growth boundary (McMeekin
et al. 2002) where the lag phase prior to growth can vary considerably between
samples.
In the trial of Small et al. (2012), the temperature in the storage rooms averaged
−0.5 °C, cycling daily over the 30-week storage period between −2.7 °C and
+1.8 °C. Near the minimum temperature, and at a meat pH of 5.4, a proportion of
the community may be close to their growth: no-growth boundary. By contrast, near
the maximum daily temperature and on meat of higher pH, a proportion of the com-
munity will be capable of growth.
Canadian researchers also report on extended shelf life of VP primals with strip-
loins stored at either −1.5 °C or +2 °C being acceptable to taste panellists after 23
and 17 weeks, respectively (Youssef et al. 2014).
Thus, it can be concluded that shelf life of VP beef primals has increased mark-
edly over recent years and, in the following sections, we follow the developments in
process hygiene, packaging technology and product storage temperatures which
have paralleled these increases.
8.4 Then and Now: Factors That Affect Shelf Life
Early reports of total bacterial levels were determined as the APC at 25 °C incubated
for 96 h. These conditions allowed maximum recovery of slower growing organ-
isms and organisms unable to grow at 30 °C or 35 °C, temperatures which are often
used for bacterial counts. Historically, at packing, the APC of beef primal cuts cen-
tred around 3–4 log10 cfu/cm2 (Egan 1983; Penney et al. 1998), with Egan et al.
(1988) listing three prerequisites for optimal shelf life of VP meats:
• APC at packing no more than 2–3 log10 cfu/cm2
• Packaging film with low oxygen permeability (≤25 cm3/m2/day at 23 °C)
• Good control of temperature throughout the storage period
These prerequisites offer a template against which current processing and stor-
age of VP beef may be assessed.
8.4.1 Low Total Counts at Packing
To a large degree, the hygienic quality of beef cuts (primal and manufacturing meat)
is linked with that of the carcasses from which they are derived. For Australian
meat, it is possible to construct a 70-year comparative profile by utilising APC data
from Grau (1979) and three national baseline studies (Table 8.2).
It should be emphasised that the data quoted by Grau (1979) were gathered at a
single abattoir whereas baseline data reflect national average data. Nonetheless, the
progressive reduction in APC appears to reflect the radical changes which the indus-
try underwent beginning with the introduction of HACCP-based QA systems in the
mid-1990s.
It is a reasonable assumption that improvements in the microbiological condition
of the chilled carcass will be passed to the meat cuts fabricated from them and this
is borne out by the most recent national baseline study of Australian primal cuts at
the time of packing. As indicated in Table 8.3, the mean APC of two high-value
primals, striploins and outsides, was <2 log10 cfu/cm2 (Phillips et al. 2012), thus
Table 8.2 Beef carcass Log10

contamination in APC/cm2 References
Australia, 1937–2004
1937 3.88 Grau (1979)
(Sumner et al. 2011)
1964 3.90 Grau (1979)
1978 2.79 Grau (1979)
1994 3.02 Vanderlinde et al. (1999)
1998 2.43 Phillips et al. (2001)
2004 1.33 Phillips et al. (2006)
Table 8.3 Aerobic plate counts (25 °C for 4 days) on Australian VP striploins and outsides
(Phillips et al. 2012)
Concentration (log10 cfu/cm2)
Primal cut Mean SD Maximum
Striploin (n = 572) 1.3 1.0 5.3
Outside (n = 572) 1.5 1.0 4.2
meeting the first essential criterion for optimal shelf life attainment cited by Egan
et al. (1988) of low total counts at the time of vacuum packing.
It seems clear, therefore, that improvements in process hygiene have occurred
and have contributed to significantly lower levels of bacterial contamination on pri-
mals at the time of vacuum packing. Since signs of bacterial spoilage do not occur
until some time after bacterial counts have reached their maximum, such improve-
ments are likely to have had an impact on increasing shelf life of vacuum-packed
meat over recent decades.
8.4.2 Low-Permeability Packaging Films
Technical information on vacuum packaging films and packaging technologies in

the early days of VP chilled meats (Anon. 1970) reflects a technology in its infancy:
impermeable films are described without any reference to oxygen transmission
rates, and packaging technologies are broadly assigned to either Evacuation and
Sealing Without a Chamber, or Vacuum Sealed in a Chamber.
An early edition of CSIRO’s Meat Research News Letter (Anon 1971) describes
the first steps in the export of chilled beef cuts citing the advantages of refrigerated
containers and concluding: ‘Vacuum packs in air-impermeable films extend the stor-
age life by about two and a half times the period possible in air. They have the advan-
tage of minimising weight loss while allowing ageing during this safe distribution life’.
The transmission rates for films routinely used for vacuum packaging have
improved since the introduction of flexible film vacuum packing technology.
Transmission rates in the range of 30–40 cm3/m2/day at 25 °C were reported in 1985
(Gill and Penney 1985) to 18.6 cm3/m2/day at 23 °C in recent years (Kiermeier et al.
2013). The multilayer structure of packaging materials has become more robust, plus
tailored to fit specific cuts, thus reducing the likelihood of air entrapment or the devel-
opment of leaks. Vacuum machines have also evolved, allowing rapid sealing of prod-
uct (about 25 products/min) and form/fill machines are increasingly giving abattoirs
the ability to manufacture retail-ready products in addition to the traditional primal/
sub-primal in a vacuum bag.
Thus, meat in vacuum packs today is encased in superior packaging films with
lower oxygen transmission, has reduced chance of leakage and is less likely to
entrap oxygen, all contributing to greatly reduce the opportunity for the those
microbes requiring oxygen or lower carbon dioxide concentration to grow and spoil
product.
8.4.3 Temperature Throughout the Supply Chain
The optimum temperature for storage of meat was defined by Gill et al. (1988a) as
−1.5 ± 0.5 °C, a temperature which minimises growth of spoilage organisms while
preventing freezing of the product. The same workers also showed that small rises
in temperature reduced shelf life significantly; at temperatures of 0 °C, 2 °C or 5 °C,
the storage life was reduced by about 30%, 50% or 70%, respectively, compared
with storage at −1.5 °C (Gill et al. 1988b).
Storage temperature is especially important in international trade where sea
freight from Australasia to Europe may occupy up to 50 days, to which must be
added time in the exporting country from packing to despatch, plus time required in
the importing country for further processing, distribution and retailing.
Exporters routinely include data loggers in cartons of chilled VP primal cuts, a
routine precaution in the event that shipments are delayed, or that the container
temperature is higher than the −1 °C set point normally specified. A recent collation
of time and temperature data for 135 sea voyages from Australia to markets in
Europe, USA, Asia and the Middle East demonstrated that the mean temperature
was usually close to 0 °C for the entire voyage, including brief temperature spikes
during loading, unloading and trans-shipping. Occasionally, a higher temperature
occurred which, on a long voyage, for example, to Europe, would prompt the
importer to utilise the consignment as rapidly as possible (Sumner 2016).
Thus, it is likely that temperature control through the supply chain has improved
over recent decades.
8.5 hen and Now: Microbial Communities on VP Beef

T
Primals
Egan et al. (1988) considered three factors (product hygiene, film permeability and
storage temperature) as important in achieving long shelf life but did not discuss
microbial communities.
The selective effect of carbon dioxide against Gram-negative aerobes was estab-
lished in the 1930s (Haines 1933) and applied in the VP meat context by Ingram
(1962). As mentioned earlier, trade in chilled VP beef preceded a deep knowledge
of the microbial communities which developed on the meat surface. However, work
through the 1970s established that provided meat of normal pH (5.4–5.8) was
packed in a film of low oxygen permeability (<100 cm3/m2/day at 25 °C) and an
atmosphere of 20% carbon dioxide and <1% oxygen, lactic acid bacteria (LAB)
would grow to be the dominant flora at the time of spoilage (Egan 1983). However,
if the meat pH were higher than normal and/or the film allowed a less inhibitory
CO2:O2 ratio, spoilage due to bacteria other than LAB could occur with B. thermo-
sphacta causing early spoilage due to the presence of unacceptable dairy-like odours
(Campbell et al. 1979). As well, if film permeability were relatively high (150 cm3/
m2/day), Shewanella putrefaciens could spoil high pH meat due to production of

hydrogen sulphide giving rise to rotten egg-type odours and greening of the meat.
Finally, at higher than optimum storage temperatures, Enterobacteriaceae could
grow on high-pH meat producing hydrogen sulphide and malodourous amines such
as cadaverine and putrescine (Dainty et al. 1979).
Still in the early days of the new VP technology, the knowledge that spoilage was
due to the predominance of lactic acid bacteria, particularly species of Lactobacillus,
was based on the phenotypic and biochemical characteristics of organisms growing
on selective media such as MRS agar. However, there were numerous reports (e.g.
Hitchener et al. 1982) of ‘unusual lactobacilli’ from VP meat which were unable to
grow on acetate agar and, on the basis of biochemical, physiological and chemical
criteria, Collins et al. (1987) ascribed several atypical lactobacilli to a new genus,
Carnobacterium.
More recent studies cite the predominant organisms in VP beef as Carnobacterium
divergens, C. piscicola, Lactobacillus sakei, L. curvatus, Leuconostoc gelidum,
Leuc. carnosum and B. thermosphacta (Ercolini et al. 2006; Fontana et al. 2006;
Jones 2004; Nissen and Sorheim 1996; Sakala et al. 2002).
Modern genetic methods (metagenomics) provide opportunities to investigate
the microbial communities and the possibility that the selection of certain bacteria
may also affect shelf life. In a recent study, molecular techniques were used to fur-
ther investigate the microbial communities extracted as rinsates from VP primals
acquired by Small et al. (2012). Early in the shelf life, the microbial community on
both cube rolls and striploins primal cut was dominated by Gram-negative, aerobic
bacteria (Pseudomonas spp.). By week 16, various species of the lactic acid bacteria
including Carnobacterium were detected and, by week 30, C. maltaromaticum or
C. divergens had become dominant.
Carnobacteria have a number of positive effects on refrigerated meat (see review
by Leisner et al. 2007) because they produce lactic acid, lowering the pH of meat,
and some strains synthesise antibacterial compounds (bacteriocins), which may
inhibit slower growing microorganisms. C. divergens and C. maltaromaticum have
been studied extensively as protective cultures in order to inhibit growth of Listeria
monocytogenes in fish and meat products and C. maltaromaticum has been shown
to inhibit a wide range of bacterial isolates found in vacuum-packed beef (Zhang
et al. 2015).
Recent work on VP beef primals has shown that storage temperature causes a
shift in the profile of microbial communities in the vacuum pack. Low storage tem-
peratures (≤2 °C) favoured the growth of lactic acid bacteria whereas storage at
higher temperatures (4 °C or 8 °C) resulted in increased prevalence of non-LAB
Enterobacter, Serratia and Enterococcus, bacterial species usually associated with
meat spoilage (Ross et al. 2016).
The finding of significant differences in microbial communities during storage at
‘low’ versus ‘high’ temperatures is particularly relevant when exporting to markets
where maintaining the integrity of the cold chain can be difficult (e.g. >40 °C is not
unusual for much of the Middle Eastern year), or where storage and distribution
infrastructure is not able to maintain the cold chain close to 0 °C.
8.6 Shelf Life of Vacuum-Packed Lamb
The shelf life of lamb is accepted as being shorter than beef, with the Australian
industry often claiming about 12 weeks at −0.5 °C though this shelf life may be
exceeded (Kiermeier et al. 2013). The shelf life of lamb has been reviewed by Mills
et al. (2014), and among the reasons for its shorter shelf life are the following:
• Higher bacterial levels of lamb primals at packing (Phillips et al. 2013).
• Increased difficulty packing smaller primals, and bone-in primals.
• Lamb primals typically contain several muscle groups, most of which have a
higher pH than beef primals.
• Most lamb primals contain both fat and lean surfaces, which leads to localised
areas of high pH and less inhibition of microbes.
8.7 Conclusions
At the outset of this chapter, we undertook to pronounce on whether shelf life exten-
sion has resulted from improvements in hygienic processing, packing technology
and the cold chain, leading to a shift in microbial communities in the vacuum pack.
Based on the evidence we have assembled, our conclusions are, firstly, that
microbial communities have probably not changed over the decades, and that
Carnobacterium has always dominated the microbial community on VP meat.
Secondly, we conclude that extended shelf life of VP beef primals has been due to
improvements in initial count, barrier film permeability, vacuum machines, and
temperature control through storage and shipping.
On the other hand, there is much still to adduce about how conditions under
vacuum affect communities. For instance, in the normal commercial setting of low-
permeability packaging and storage below 0 °C, does pH become the key factor in
determining the ultimate microbial community? If so, is this why Small et al. (2012)
found some VP beef replicates with APCs of 7–8 log10 cfu/cm2 while others on the
same sampling day had log10 3–4 cfu/cm2? Is the higher pH of lamb primals, which
contain a number of muscle groups, the reason why lamb shelf life is only about a
half that of beef primals under the same storage conditions? Do variations of breed-
ing lines of livestock impart variations of chemical structures of protein, collagen
and/or fats that impact microbial metabolism and eventual organoleptic attributes of
meat during shelf life? Is the proportion of psychrotrophic, CO2-tolerant bacteria on
cuts at the time of vacuum packing the critical microbial determinant of shelf life?
These are all questions that need to be answered if we are to have a complete under-
standing of the factors that determine shelf life of VP meat.
References
Anonymous. 1970. The use of packaging films for chilled fresh meats. Meat Research News Letter
70/6. Brisbane, Australia: CSIRO Division of Food Research.
———. 1971. Shipment of chilled beef cuts. Meat Research News Letter 71/2. Brisbane, Australia:
CSIRO Division of Food Research.
Campbell, R., A. Egan, F. Grau, and B. Shay. 1979. The growth of Microbacterium thermosphac-
tum on beef. Journal of Applied Bacteriology 47: 505–509.
Collins, M., J. Farrow, B. Phillips, S. Ferusu, and D. Jones. 1987. Classification of Lactobacillus
divergens, Lactobacillus piscicola, and some catalase-negative, asporogenous rod-shaped
bacteria from poultry in a new genus, Carnobacterium. International Journal of Systematic
Bacteriology 37: 310–316.
Dainty, R., B. Shaw, C. Harding, and S. Michanie. 1979. The spoilage of vacuum packed beef
by cold tolerant bacteria. In Cold tolerant microbes in spoilage and the environment, SAB
Technical Series, ed. A. Russell and R. Fuller, vol. 13. London: Academic.
Egan, A. 1983. Lactic acid bacteria of meat and meat products. Antonie Van Leeuwenhoek 49:
327–336.
Egan, A., I. Eustace, and B. Shay. 1988. Meat packaging—maintaining the quality and prolonging
the storage life of chilled beef, pork and lamb. Proc 34th international congress of meat science
and technology.
Empey, W., and J. Vickery. 1933. The use of carbon dioxide in the storage of chilled beef. Journal
of the Council of Scientific and Industrial Research 6: 233–243.
Ercolini, D., F. Russo, E. Torrieri, P. Masi, and F. Villani. 2006. Changes in the spoilage-related
microbiota of beef during refrigerated storage under different packaging conditions. Applied
and Environmental Microbiology 72: 4663–4671.
Farrer, K. 1980. A settlement amply supplied: Food technology in nineteenth century Australia.
Melbourne: Melbourne University Press.
Fontana, C., P. Cocconcelli, and G. Vignolo. 2006. Direct molecular approach to monitoring bac-
terial colonization on vacuum-packaged beef. Applied and Environmental Microbiology 72:
5618–5622.
Gill, C., and K. Newton. 1979. Spoilage of vacuum-packaged dark, firm, dry meat at chill tempera-
tures. Applied and Environmental Microbiology 37: 362–364.
Gill, C., and N. Penney. 1985. Modification of in-pack conditions to extend the storage life of
vacuum packaged lamb. Meat Science 14: 43–60.
Gill, C., D. Phillips, and J. Harrison. 1988a. Product temperature criteria for shipment of chilled
meats to distant markets. In Proc 1st International Refrigeration Conference, 40–47. Brisbane:
Refrigeration for Food and People.
Gill, C., D. Phillips, and M. Loeffen. 1988b. A computer program for assessing the remaining
storage life of chilled red meats from product temperature history. In Proc 1st International
Refrigeration Conference, 35–39. Brisbane: Refrigeration for Food and People.
Grau, F. 1979. Fresh meats: Bacterial association. Archiv für Lebensmittelhygiene 30: 81–116.
Haines, R. 1933. The influence of carbon dioxide on the rate of multiplication of certain bacteria
as judged by viable counts. Journal of Society of Chemical Industry 52: 13T.
Hitchener, B., A. Egan, and P. Rogers. 1982. Characteristics of lactic acid bacteria isolated from
vacuum-packaged beef. Journal of Applied Bacteriology 52: 31–37.
Ingram, M. 1962. Microbiological principles in prepacking meat. Journal of Applied Bacteriology
25: 259–281.
Jones, R. 2004. Observations on the succession dynamics of lactic acid bacteria populations in
chill-stored vacuum-packaged beef. International Journal of Food Microbiology 90: 273–282.
Kiermeier, A., M. Tamplin, D. May, G. Holds, M. Williams, and A. Dann. 2013. Microbial growth,
communities and sensory characteristics of vacuum and modified atmosphere packaged lamb
shoulders. Food Microbiology 36: 305–315.
Leisner, J., B. Laursen, H. Prévost, D. Drider, and P. Dalgaard. 2007. Carnobacterium: Positive and
negative effects in the environment and in foods. FEMS Microbiological Reviews 31: 592–613.
McMeekin, T., J. Olley, D. Ratkowsky, and T. Ross. 2002. Predictive microbiology: Towards the
interface and beyond. International Journal of Food Microbiology 73: 395–407.
Mills, J., A. Donnison, and G. Brightwell. 2014. Factors affecting microbial spoilage and shelf
life of chilled vacuum-packed lamb transported to distant markets: A review. Meat Science 98:
71–80.
Newton, K., and W. Rigg. 1979. The effect of film permeability on the storage life and microbiol-
ogy of vacuum packed meat. Journal of Applied Bacteriology 47: 433–441.
Nissen, H., and O. Sorheim. 1996. Effects of vacuum, modified atmospheres and storage tempera-
ture on the microbial flora of packaged beef. Food Microbiology 13: 183–191.
Nottingham, P. 1982. Microbiology of carcass meats. In Meat Microbiology, ed. M. Brown, 13–65.
London, New York: Applied Science Publishers.
Penney, N., R. Bell, and S. Moorhead. 1998. Performance during retail display of hot and cold
boned beef striploins after chill storage in vacuum or carbon dioxide packaging. Food Research
International 31: 521–527.
Phillips, D., J. Sumner, J. Alexander, and K. Dutton. 2001. Microbiological quality of Australian
beef. Journal of Food Protection 64: 692–696.
Phillips, D., D. Jordan, S. Morris, I. Jenson, and J. Sumner. 2006. A national survey of the micro-
biological quality of beef carcasses and frozen boneless beef in Australia. Journal of Food
Protection 69: 1113–1117.
Phillips, D., K. Bridger, I. Jenson, and J. Sumner. 2012. An Australian national survey of the micro-
biological quality of frozen boneless beef and beef primal cuts. Journal of Food Protection 75:
1862–1866.
Phillips, D., S. Tholath, I. Jenson, and J. Sumner. 2013. Microbiological quality of Australian
sheep meat in 2011. Food Control 31: 291–294.
Ross, T., J. Bowman, M. Kaur, C. Kocharunchit, and M. Tamplin. 2016. Microbial Ecology and
Physiology. University of Tasmania Report (Meat & Livestock Australia project G.MFS.0289).
Sakala, R., H. Hayashidani, Y. Kato, T. Hirata, Y. Makino, A. Fukushima, T. Yamada, C. Kaneuchi,
and M. Ogawa. 2002. Change in the composition of the microflora, on vacuum-packaged beef
during chiller storage. International Journal of Food Microbiology 74: 87–99.
Small, A., A. Sikes, and I. Jenson. 2009. Prolonged storage of chilled vacuum packed beef from
Australian export abattoirs. Proc: International Conference on Meat science and Technology,
Copenhagen, August 2009, pp.1294–1297. http://www.icomst2009.dk/fileadmin/documents/
ICOMST_PS8_Final.pdf.
Small, A., I. Jenson, A. Kiermeier, et al. 2012. Vacuum-packed beef primals with extremely long
shelf life have unusual microbiological profile. Journal of Food Protection 75: 1524–1527.
Sumner, J. 2016. The impact of transport to Australia’s distant markets on the shelf-life of beef
and sheep primals. Australian meat processors corporation (AMPC) report 2106.1075, North
Sydney, Australia.
Sumner, J., A. Kiermeier, and I. Jenson. 2011. Verification of hygiene in Australian manufacturing
beef processing–Focus on Escherichia coli O157. Food Protection Trends 31 (8): 514–520.
Vanderlinde, P., B. Shay, and J. Murray. 1999. Microbiological quality of Australian beef carcass
meat and frozen bulk packed beef. Journal of Food Protection 61: 437–443.
Vickery, J. 1990. Food science and Technology in Australia: A review of research since 1900.
Sydney: CSIRO Publishing.
Youssef, M., C. Gill, and X. Yang. 2014. Storage life at 2°C or −1.5°C of vacuum-packaged bone-
less and bone-in cuts from decontaminated beef carcass. Journal of the Science of Food and
Agriculture 94: 3118–3124.
Zhang, P., J. Baranyi, and M. Tamplin. 2015. Interstrain interactions between bacteria isolated from
vacuum-packaged refrigerated beef. Applied and Environmental Microbiology 81: 2753–2761.
Index
A Clean label products, 6, 27

Active packaging Clostridium, 94
antimicrobial packaging technologies, Clostridium botulinum, 28, 32–34, 120
128, 130 Clustered regularly interspaced short
carbon dioxide scavenging technologies, palindromic repeats (CRISPR), 139
125, 126 Code dates
components, 123 date labelling, 10
food/package environment, 123 expiration, perishable food accounts, 10
moisture control technologies, 128 food products, 9
odor scavenging technologies, 126, 127 foodservice workers, 9
oxygen scavenging technologies, 124, 125 food waste, 10, 11
Aerobic plate count (APC), 17, 53, 147 government entities, 9
Aluminum, 109 JD, 9
Ambient chain, 1 open date coding, 10
Amplified fragment length polymorphism open dating, 9
(AFLP) analysis, 53 perishable foods, 10
Animal food, 12 product box/package, 9
Antimicrobial packaging technologies, quality degradation, 10
128, 130 sell/display by, 10
use by dates, 10
use by label, 10
B WTP, 10, 11
Bacillus cereus, 28, 30–32, 97 Cold chain, 153, 154
Bacillus species, 94 Cold plasma, 97, 98
Bisphenol-A (BPA), 114, 115 Cold storage, 92, 93
Blockchain, 5 Commonwealth Scientific and Industrial
Brochothrix, 118 Research Organisation
(CSIRO), 146
Computational methods, 7
C Consumers, 2, 6, 8, 9, 13
Campylobacter jejuni, 119 acceptance, 19, 20
Carbon monoxide, 117 sensory analysis, 19
Carnobacteria, 153 Cooked ham products, 3
Carnobacterium, 153, 154 Cooling, 92, 93
Chilled food supply chain, 1 CRISPR associated sequences (Cas), 139

https://doi.org/10.1007/978-3-030-54375-4
158
CRISPR RNAs (crRNAs), 139 vendor-managed supply chains, 5

Culture-independent genomics tools, 136 Foodomics, 139, 140
Customer demands, 6 Food packaging
communication, 108
containment, 106
D convenience, 107, 108
Date labelling, 8, 10 dairy and cheese products, 122, 123
Designing food distribution systems, 4 EVOH, 115
Destruction, 13 food products, 105
Direct-to-consumer food distribution fresh produce, 120–122
models, 4 fresh red meat, 117–119
Distressed product, 11, 12 intelligent packaging, 130, 131
Diversion, 12 ionomers, 116
Donation, 12 MAP, 116, 117
materials
food product, 108
E glass, 109
Enterobacteriaceae, 153 metals, 109
Escherichia coli, 7 plastics, 110
Ethylene vinyl alcohol (EVOH), 114 modern-day food production system, 105
Expanded PS (EPS), 113 PC, 115
Expiration date PET, 115
destruction, 13 polyamides, 115
distressed product, 11, 12 polyethylene, 110, 111
diversion, 12 poultry products, 119
donation, 12 PP, 112
scandals, 13, 14 protection, 106, 107
Expired products account, 10 PS, 113
PVC, 113, 114
PVDC, 114
F PVOH, 115
Federal Occupational Safety and Health seafood products, 119, 120
Administration (OSHA), 100 vacuum packaging, 116
Food and Drug Administration (FDA), 108 Food preservation processing
Foodborne pathogens, 7 aseptic packaging, 92
Food businesses, 3 bacterial inhibition, 91
Food logistics characteristics, 91
blockchain, 5 cold plasma, 97, 98
designing food distribution systems, 4 cold storage, 92, 93
direct-to-consumer food distribution consumer demands, 92
models, 4 cooling, 92, 93
distribution structure, 4 food industry, 91
end of shelf life, 3 freezing, 92, 93
food industries, 4 HHP, 94, 95
food processors, 4 irradiation, 97
forecast-based ordering model, 5 microorganisms, 92
fresh food products limits, 3 ozone, 99, 100
MTO, 5 PEF, 98, 99
MTS, 5 pulsed light, 95, 96
OPP, 5 spoilage, 91
perishability, 3, 4 thermal/heating, 93, 94
RFID, 5 ultrasonication, 96, 97
shelf life, perishable food products, 4 ultraviolet light, 95, 96
supply chain structure, 5 Food processors, 4
159
Food products, 12 Ionizing radiation, 97

Food quality, 17, 18 Ionomers, 116
Food safety factors Irradiation, 97
B. cereus (see Bacillus cereus)
C. botulinum (see Clostridium botulinum)
challenge studies, 35–36 J
L. monocytogenes (see Listeria Julian dates (JD), 9
monocytogenes) Just Noticeable Difference (JND), 20
microorganisms, 27
pathogens, 28, 36, 37
RTE products, 28 L
seasonal products, 28 Lactic acid bacteria (LAB), 53, 72, 147
shelf life, 27, 28 Lactics, 17
Y. enterocolitica (see Yersinia Lactobacillus, 118, 153
enterocolitica) Lactobacillus viridescens, 63
Food science techniques, 6 Linear low-density polyethylene (LLDPE),
Foodservice/retail customers, 7 111, 129
Foodservice workers, 9 Lipid content, 106
Food supply chains, 1 Listeria monocytogenes, 28, 34–35, 107, 129
Food waste, 10, 11 Lock-Out Tag-Out (LOTO) procedures, 75
Forecast-based ordering model, 5 Logistics, 3
Freezing, 92, 93 Low-density polyethylene (LDPE),
Fresh cut produce, 3 111, 122
Frozen foods supply chain, 1
M
G Made to order (MTO), 5
Genomics Made to stock production (MTS), 5
CRISPR-Cas, 139 Metagenomics, 138
culture-independent genomics tools, 136 Metagenomics techniques, 18
foodomics, 139, 140 Metals, 109
functional properties, 135 Microbial degradation, 18
metagenomics, 138 Microbial ecology, 135
microbial spoilage, 135 Microbiological profiles, 18, 19
nutritional quality, 135 Microbiological quality, 147, 148
“omics” tools, 136 Microbial spoilage, 7, 118, 135
rDNA technology, 136, 137 Microorganisms, 92, 96
whole genome sequencing, 137, 138 Modified atmosphere packaging (MAP), 116,
Glass, 109 117, 121
Good manufacturing practices (GMPs), Moisture control technologies, 128
35, 73, 83 Molecular techniques, 18, 137
Multilayered audit approach, 89
Multiple-hurdle approach, 35
H Myoglobin, 117
High-density polyethylene (HDPE), 111
High hydrostatic pressure (HHP), 94, 95
Homeoviscous adaptation, 92 N
N-acyl homoserine lactones (AHL), 56
National Institute of Environmental Health
I Sciences (NIEHS), 114
Intelligent packaging, 130, 131 National Toxicology Program (NTP), 114
Inventory management systems, 12 Nonthermal processing, 92
160
O food quality, 17, 18

Open date coding, 10 microbiological profiles, 18, 19
Open dating, 9 shelf-life of food, 1
Operational sanitation, 83 Quality personnel, 6
Operational Taxonomical Units (OTUs), 138 Quantitative PCR (qPCR), 18
Operations personnel manufacture product, 6
Order penetration point (OPP), 5
Oxygen scavenging technologies, 124, 125 R
Ozone, 99, 100 Radio Frequency Identification Devices
(RFID), 5, 130
Ready-to-eat (RTE), 28
P Recombinant DNA (rDNA) technology,
Pathogen growth, 6, 7 136, 137
Penicillium spp., 124 Reduced oxygen packaging (ROP), 33
Perishability, 1, 4 Refrigerated, processed foods of extended
Perishable foods, 1 durability (REPFEDs), 34
Perishable products, 5 Refrigerated storage temperatures, 52, 53
Personal Protective Equipment (PPE), 75 Regulators, 2
Photobacterium phosphoreum, 19 Regulatory, 7, 8
Physical decomposition, 23 Retailers, 7
Plastics, 110 Routine shelf life testing, 20, 22, 23
Polyamides, 115
Polycarbonates (PC), 115
Polyethylene, 110, 111 S
Polyethylene terephthalate (PETE), 115, 122 Sanitation
Polymers, 110–112, 115, 116 area preparation/dry pickup, 75
Polypropylene (PP), 112 condensation control, 80
Polystyrene (PS), 113 controlling bacterial pathogens, 72
Polyvinyl alcohol (PVOH), 114 deep cleaning, 73, 81–82, 84
Polyvinyl chloride (PVC), 113, 114 food safety, 71
Polyvinylidene chloride (PVDC), 114 maturity model, 86–88
Predictive microbiology models, 18 microbial spoilage, 71
Processor, 3 MSS, 72
Pseudomonas species, 118 non-negotiable, 74
Psychrotrophic bacterial pathogens, 41, 62 operational (see Operational sanitation)
Psychrotrophic pathogens, 7 performance, 74
Psychrotrophic strains, 31 post rinse, 78–79
Pulsed electric field (PEF), 98, 99 pre-operation (Pre-op) inspection, 79–80
Pulsed light, 95, 96 pre-rinse, 76–78
sanitary design, 72, 80, 83–85
sanitization, 80
Q soap and scrub, 78
Quality deterioration rates structuring, 73
APC/lactics, 17 swabbing, 72, 79, 84–86
factors, 14 wet sanitation process, 74
first-order reaction, 14–16 Sanitation Standard Operating Procedures
loss, food quality, 15–17 (SSOPs), 78
order of the reaction, 15 Scandals, 13
shelf life of foods, 14 Seafood products, 119, 120
temperature, 14 “Seek and Destroy” approach, 84
zero-order reaction, 15, 16 Sensors, 131
Quality factors Sensory, 147, 148
consumer acceptance, 19, 20 Shelf life determinations
161
code dates (see Code dates) science, 6, 7

organoleptic properties, 2 shelf life days, 2
perishable foods, 2 Standard Plate Count (SPC), 53
quality deterioration rates (see Quality Styrofoam, 113
deterioration rates) Survival analysis, 20
quality factors (see Quality factors) Sustainable companies, 13
sensory properties, foods, 2
stakeholders (see Stakeholders)
Shelf life extension, 6 T
Shelf life measurement methods, 53 Thermal processing, 93, 94
Shelf-life of food Thermosonication, 96
definition, 1 Thiobarbituric reactive substances
perishable foods, 1 (TBARS), 56
quality factors, 1 Third party logistic providers (3PL), 4
types, food supply chains, 1 Time and Temperature indicators
Shelf life testing (TTIs), 131
APC/lactics, 22 Total Plate Count (TPC), 53
beneficial, 21 Trimethylamine oxide (TMAO), 56, 60
deterioration, food, 20
microbial counts, 22
organoleptic, 20 U
routine, 20, 22, 23 Ultrasonication, 96, 97
stages, 20, 21 Ultraviolet light (UV), 95, 96
Shewanella putrefaciens, 153 Undesirable microorganisms, 8
Short-wavelength near-infrared (SW-NIR), 54
Spoilage
accelerated shelf life testing, 54–55 V
fresh produce, 57–58 Vacuum packaging, 116
fresh seafood, 60–61 Australian red meat industry, 145
growth of microorganisms, 41 chilled red meats, 147
microbial growth, 55–56 low-permeability packaging films, 151
milk and dairy products, 55–56 low total counts, 150, 151
plant-based protein products, 59–60 mechanical refrigeration, 146
processed meat, poultry and fish, 55–56 microbial communities, 152, 153
psychrotrophic bacteria, 53, 64 organoleptic testing, 146
raw meat and poultry, 55–56 shelf life, 148–150, 154
shelf life determination, 53–55 supply chain, 152
temperature, 42–53 temperature, 152
Stakeholders Vendor-managed supply chains, 5
business development, 2, 3
commercialization, food products, 2
consumers, 2, 8, 9 W
logistics, 2–5 Whole genome sequencing, 137, 138
operations team, 2 Willingness to pay (WTP), 10, 11
regulators, 2
regulatory, 7, 8
retail/foodservice end users, 2 Y
sales, 2 Yersinia enterocolitica, 28–30

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Food Microbiology and Food Safety

Food Safety and

LinkedIn: Emily Phuong

More information about this series at http://www.springer.com/series/7131

Food Microbiology and Food Safety

© Springer Nature Switzerland AG 2021

In the twenty-first century, a growing global population and development in formerly

Jacksonville, FL, USA Peter J. Taormina

1 Purposes and Principles of Shelf Life Determination�������������������������� 1

Cynthia Ebner Sealed Air Corporation, Charlotte, NC, USA

© Springer Nature Switzerland AG 2021 1

1.1.1.1 Business Development

variability in demand and transportation (Ahumada and Rene Villalobos 2009).

Vendor-­managed supply chains mean perishable foods produced on demand factor-

psychrotrophic or psychrotolerant bacterial pathogens like Listeria monocytogenes,

subjected to class II recalls (United States Department of Agriculture, Food Safety

Consumers form quality expectations of food products based on perceived cues

1.1.2  ssuring Quality and Wholesomeness of Food through

1.1.2.2 Food Waste

Lettuce Carrots Chicken

1.1.3 Expiration = Decision Time

1.1.3.1 Distressed Product

1.2 Principles of Shelf Life Determination

1.2.1 Quality Deterioration Rates

If that equation is integrated, it becomes Eq. (1.3):

Amount left ( A ) = Initial amount ( A0 ) − kθ (1.3)

Integrating Eq. (1.6) gives a logarithmic function of the first-order as shown in

1.2.2 Defining Quality Factors

1.2.2.1 Microbiological Profiles

Fig. 1.5 Growth of

phosphoreum in cod fillets stored at 0 °C (Dalgaard et al. 1997). Growth of the

1.2.2.2 Consumer Acceptance

1.2.3 Shelf Life Testing

This book focuses on perishable foods, as there is little to no microbial degradation

© Springer Nature Switzerland AG 2021 27

avoiding products containing ingredients that are not recognizable or perceived as

2.2 Food Safety Concerns for Extended Shelf Life Foods

2.2.1 Yersinia enterocolitica

2.2.2 Bacillus cereus

2.2.3 Non-Proteolytic Clostridium botulinum

Clostridium botulinum is an anaerobic, spore-forming microorganism capable of

Non-proteolytic Group II strains of C. botulinum have an optimum temperature

2.2.4 Listeria monocytogenes

Listeria is a ubiquitous organism, widely distributed in the environment including

2.3 Challenge Studies and Shelf Life

These precooked, prepackaged convenience foods have increased in popularity

MacDonald, E., M. Einöder-Moreno, K. Borgen, L. Thorstensen Brandal, L. Diab, Ø. Fossli,

contributing to outbreaks and description of outbreak categories. Journal of Food Protection

Spoilage is characterized by any change in a food product that results in unacceptable

© Springer Nature Switzerland AG 2021 41

temperature-dependent rates of spoilage as well as growth temperature ranges of

3.2 Role of Temperature in Shelf Life

is not widespread, opportunities do occasionally arise for industry collaboration with

3.3 Storage Temperatures and Microbial Growth

3.3.1 Freezing Temperatures

Leuconostoc mesenteroides 4, 3c MRS agar – – Isothermal Korkeala et al. (1990)

Table 3.2 Minimum growth temperature (°C) of selected fungi

Table 3.2 (continued)

Table 3.2 (continued)

Jacksonville, FL, USA Peter J. Taormina

1 Purposes and Principles of Shelf Life Determination�� 1

1.1.1.1 Business Development

Vendor-managed supply chains mean perishable foods produced on demand factor-

1.1.2 ssuring Quality and Wholesomeness of Food through

1.1.2.2 Food Waste

1.1.3 Expiration = Decision Time

1.1.3.1 Distressed Product

1.2 Principles of Shelf Life Determination

1.2.1 Quality Deterioration Rates

1.2.2 Defining Quality Factors

1.2.2.1 Microbiological Profiles

1.2.2.2 Consumer Acceptance

1.2.3 Shelf Life Testing

2.2 Food Safety Concerns for Extended Shelf Life Foods

2.2.1 Yersinia enterocolitica

2.2.2 Bacillus cereus

2.2.3 Non-Proteolytic Clostridium botulinum

2.2.4 Listeria monocytogenes

2.3 Challenge Studies and Shelf Life

3.2 Role of Temperature in Shelf Life

3.3 Storage Temperatures and Microbial Growth

3.3.1 Freezing Temperatures

3.3.2 Refrigeration Temperatures

3.4 Shelf-Life Determination

3.5 Accelerated Shelf-Life Testing for Refrigerated Foods

3.6 Chemical Indicators of Microbial Growth

3.7 Shelf Life of Specific Food Types

3.7.1 Fresh Produce

3.7.2 Pasteurized Milk and Dairy Products

3.7.3 Plant-Based Protein Products

3.7.4 Fresh Seafood

3.7.5 Raw Meat and Poultry

3.7.6 Further Processed Meat, Poultry, and Fish

4.2 Sanitation Program

4.2.1 Daily Sanitation: Seven-Step Process

4.2.2 Non-Daily Sanitation: Deep Cleaning and Intervention

4.2.3 Operational Sanitation

4.3 Sanitary Design and Access

4.4 Verification and Performance Swabbing

4.5 Maturity Model

5.1 I ntroduction: What Is Food Spoilage and How

5.2 Processing Techniques for Extending Shelf Life

5.2.1 Basic Preservation Techniques

5.3 Advanced Processing Technologies

5.3.1 High Hydrostatic Pressure

5.3.2 Ultraviolet and Pulsed Light

5.3.5 Cold Plasma

5.3.6 Pulsed Electric Fields

6.2 Packaging Functions

6.3 Materials Used in Food Packaging