A COMPARATIVE ANALYSIS OF BREAD PRESERVATION TECHNIQUES TO

IMPROVE SHELF LIFE IN YOLA NORTH, ADAMAWA STATE

BY

IBRAHIM, kabiru

AEE/18U/1411

OCTOBER, 2024
 A COMPARATIVE ANALYSIS OF BREAD PRESERVATION TECHNIQUES TO
IMPROVE SHELF LIFE IN YOLA NORTH, ADAMAWA STATE

BY

IBRAHIM, kabiru

AEE/18U/1411

A PROJECT WORK SUBMITTED TO THE DEPARTMENT OF AGRICULTURAL

AND ENVIRONMENTAL ENGINEERING, FACULTY OF ENGINEERING, MODIBBO
ADAMA UNIVERSITY, YOLA IN PARTIAL FULFILMENT OF THE REQUIREMENT
FOR THE AWARD OF BARCHELOR OF ENGINEERING (B.ENG.) IN
AGRICULTURAL AND ENVIRONMENTAL ENGINEERINGMODIBBO ADAMA
UNIVERSITY, YOLA.

OCTOBER, 2024
 DECLARATION
I hereby declare that, this project was written by me and it's a record of my own research work
supervised by Prof. A. U. BABAYE of the Department of Agricultural and Environmental
Engineering, Modibbo Adama University, Yola. It has not either in part or in full being submitted
for the award of any degree or certification. Reference made to published literatures has been dully
acknowledged.

------------------------------------- --------------------------------
IBRAHIM, kabiru Date
(AEE/18U/1411)

iii
 CERTIFICATION
This is to certify that, this project work titled “Bread Preservation Technique: A Review of Natural
And Modern Methods”, Adamawa State. By Ibrahim kabir (AEE/18U/1411) under the supervision
of Prof. A. U. Babaye of the Department of Agricultural and Environmental Engineering, Modibbo
Adama University Yola.

-------------------------- --------------------------
Prof. A. U. Babaye Date
(Supervisor)

-------------------------------- ---------------------------
Engr. Prof. A. B Ankidawa Date
(Head of Department)

------------------------------- ---------------------------
Name………………………… Date
External Examiner

iv
 DEDICATION
This project is dedicated to Allah, the Most Merciful and Compassionate. I am grateful for the
guidance and strength that has been bestowed upon me throughout this journey. May this work be
a source of benefit and blessings to all who encounter it, and may it be a means of earning Your
pleasure and forgiveness. Ameen

v
 ACKNOWLEDGMENT
I would like to express my heartfelt gratitude and acknowledgement to the following individuals
and groups who have supported me throughout my journey: First and foremost, my honorable
parents, Engr Mohd Kabiru Dikko and Mallam Fadimatu Kabiru, for their untiring love, guidance,
and encouragement. I also extend my appreciation to the entire Bancharas family and Mallam
Bulama family for their support and kindness. To my colleagues at Alkama Bread Company Nig
Ltd, I thank you all, particularly the management and staff, for your cooperation and camaraderie.
My gratitude also goes to my course mates, lecturers, and project supervisor, Prof A.U Babayi, for
their valuable guidance, expertise, and mentorship. Thank you all for your contributions to my
success. May Allah reward you abundantly.

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 ABSTRACT

Bread spoilage in Nigeria poses significant challenges due to the hot and humid climate, which
accelerates the growth of harmful microorganisms, leading to economic losses and potential
health risks. The primary aim of this study was to assess and compare the effectiveness of Natural
preservation method and modern preservation methods in extending the shelf life of bread. The
methods involved monitoring the weight loss of bread loaves preserved using a garlic honey
mixture (Natural), refrigeration (modern), and room temperature storage (non-natural/modern
method) over a 20-day period. The results indicated that bread stored under refrigeration
exhibited the least spoilage, with only 1.0% weight loss by day 20, compared to 2.0 % in the garlic
honey mixture and 6.2% at room temperature. The study concludes that while Natural preservation
methods provide some level of preservation, modern techniques like refrigeration are significantly
more effective in reducing spoilage and extending bread shelf life, though their widespread
application may be limited by economic and technological barriers in certain regions of Nigeria.
The study recommended that to improve the effectiveness of bread preservation, it is suggested to
enhance natural methods by exploring combinations of natural preservatives and optimizing their
usage. Increasing the accessibility of modern preservation techniques, particularly through
affordable and energy-efficient refrigeration units, is crucial. Simple experimental methods, such
as monitoring weight loss, should be encouraged for easy and cost-effective spoilage assessment.
Finally, integrating natural and modern methods could provide a more sustainable and adaptable
approach to extending bread shelf life, especially in regions with limited resources.

vii
 Table of Contents
CHAPTER ONE ............................................................................................................................. 1
INTRODUCTION....................................................................................................................... 1
1.1 Background of Study ......................................................................................................... 1
1.2 Statement of Problem ........................................................................................................ 2
1.3 Aim and Objectives ........................................................................................................... 2
1.3.1 Aim ................................................................................................................................. 2
1.3.2 Objectives ....................................................................................................................... 3
1.4 Significance of the study ................................................................................................... 3
1.5 Scope and Limitations ....................................................................................................... 4
1.5.1 Scope of the study........................................................................................................... 4
1.5.2 Limitations ...................................................................................................................... 4
CHAPTER TWO ............................................................................................................................ 5
LITERATURE REVIEW............................................................................................................ 5
2.1 Natural preservation methodand Modern Techniques for Food Preservation ................... 5
2.1.1 Modern techniques ......................................................................................................... 5
a. Pasteurization ....................................................................................................................... 5
b. Edible Coatings.................................................................................................................... 6
c. Active Packaging ................................................................................................................. 6
2.1.2 Natural Techniques ......................................................................................................... 8
a. Canning ................................................................................................................................ 8
b. Freezing ............................................................................................................................... 9
c. Refrigeration ...................................................................................................................... 10
d. Fermentation ...................................................................................................................... 13
e. Addition of Sugar: Jellies and Jams ................................................................................... 13
2.2 Plant Extracts as Bio‐Preservative ................................................................................... 13
2.3 Microbial Fermentation ................................................................................................... 15
2.3.1 Lactic acid bacteria: ...................................................................................................... 15
2.3.2 Yeast ............................................................................................................................. 16
CHAPTER THREE ...................................................................................................................... 18
MATERIALS AND METHOD ................................................................................................ 18
3.1 Methodology.................................................................................................................... 18
3.1.1 Weight loss Method ...................................................................................................... 18

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 3.1.2 Procedure ...................................................................................................................... 18
3.1.2.1 Initial Preparation: ..................................................................................................... 18
3.1.2.2 Application of Preservation Methods: ....................................................................... 18
3.1.2.3 Monitoring and Data Collection: ............................................................................... 19
3.2 Materials used .................................................................................................................. 20
3.3 Data Analysis:.................................................................................................................. 20
CHAPTER FOUR ......................................................................................................................... 21
RESULTS AND DISCUSSION ............................................................................................... 21
4.1 Spoilage Rate and Weight Loss (Natural preservation methodMethod) ......................... 21
4.2 Spoilage Rate and Weight Loss (Modern Method) ......................................................... 21
4.3 Spoilage Rate and Weight Loss (Non- Natural/Modern Method) ................................... 22
4.4 ANOVA Data Interpretation ........................................................................................... 34
CHAPTER FIVE .......................................................................................................................... 35
CONCLUSSION AND RECOMMENDATION ...................................................................... 35
5.1 CONCLUSSION ............................................................................................................. 35
5.2 RECOMMENDATION ................................................................................................... 36
REFERENCES ............................................................................................................................. 38

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 LIST OF TABLES

Table 4.1: Natural preservation methodmethod of preservation (garlic-honey mixture) ............. 23

Table 4.2: Modern method of preservation (Refrigeration).......................................................... 26
Table 4.3: Stored Bread at Room Temperature (Non-Modern/Natural preservation
methodMethod) ............................................................................................................................. 28
Table.4.4 Comparison Table of Preservation Methods Based on Effectiveness .......................... 31
Table 4.4: Percentage Loss Over 20 Days (2-Day Intervals) ....................................................... 32
Table 4.5 ANOVA Results ........................................................................................................... 33

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 LIST OF FIGURES
Fig 2.1 Coating of Bread with Honey (Williams & Oladipo, 2018)............................................... 7
Fig 2.2 Bread Refrigeration (Lechowich, 1988) ........................................................................... 12
Fig 2.3 Mold growth on bread (Axel et al., 2017) ........................................................................ 14

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 CHAPTER ONE

INTRODUCTION

1.1 Background of Study

Food preservation is the process of handling and treating food to control its spoilage by stopping
the attack and growth of food-borne disease-causing microbes; avoiding oxidation of fats
(rancidity); and maintaining the nutritional value, texture, and flavour of the food. It is commonly
recognized that chemicals, bacteria from the surrounding area, and enzymes included in the food
itself can cause food products to decay if they are exposed to them. In addition, food and food
products must be carried from one location to another to be consumed. During transit, there are
chances to deteriorate the food, loss or decrease in morphological attraction, and reduction in the
nutritional value of the food. According to the World Health Organization (WHO), around 600
million people roughly one out of every ten people on the earth – become unwell after eating
contaminated food, and 420 000 people die each year, resulting in the loss of 33 million healthy
lifestyles.

For thousands of years, bread is still one of the dominant food sources of the human diet, with the
manufacturing of yeast‐based and sourdough bread being one of the earliest biotechnological
mechanisms (El, 2015). Amidst its medium growth rate (122,000 t in 2007 to 129 t in 2016), it
earned approximately $358 billion in global revenue in 2016 (Garcia et al., 2019). It is also a
magnificent energy source, including protein, iron, calcium, and various vitamins (Rosell, 2011).
Commercially available bread and biscuits contain nearly 7.5 and 7.8% protein (Saranraj &
Geetha, 2012). Bakery products are ideal for fiber addition, as fiber intake has declined in the
European diet partially due to cereal adjustments. Due to the easily spoiled nature of this food, its
quality and palatability degrade during preservation, resulting in changes in physiological,
biochemical, sensorial, and microbial properties (Cauvain & Young, 2007). Mold and fungal
deterioration are the primary causes of significant financial detriments in packed bread items.

Bread, a staple food across many cultures, holds significant importance in the diet of many
Nigerians. Consumed daily by a substantial portion of the population, bread's shelf life and quality
are critical factors that influence its consumption and economic value. In Nigeria, bread

1
preservation has always been a concern due to the country's hot and humid climate, which
accelerates the spoilage process (Necidová et al., 2019).

Historically, bread preservation in Nigeria has relied on several Natural preservation methods
aimed at extending shelf life and maintaining quality. These methods include: Sun Drying, Use of
Natural Preservatives, and Storage Techniques which involve wrapping bread in leaves or cloth
and storing it in cool, dry places (Jimoh & Agboola, 2020). This helps to reduce exposure to
moisture and airborne contaminants. With technological advancements, modern methods have
emerged to address the limitations of Natural techniques, providing more effective and efficient
solutions. Some of these methods include: Chemical Preservatives, Modified Atmosphere
Packaging (MAP), Refrigeration and Freezing, and Irradiation.

1.2 Statement of Problem

Bread spoilage in Nigeria, driven by the hot and humid climate, leads to significant economic
losses and health risks due to the rapid growth of harmful microorganisms. Natural preservation
methods, while accessible and cost-effective, often fail to provide sufficient protection against
spoilage. In contrast, modern techniques, though more effective, are not widely feasible due to
economic constraints and limited technological accessibility, especially in rural areas (Adebayo,
2019). Additionally, the use of chemical preservatives raises health concerns among health-
conscious consumers. This study addresses the inadequacy of existing preservation methods in
Nigeria by critically reviewing Natural preservation method and modern techniques, evaluating
their effectiveness, feasibility, and acceptability within the Nigerian context. The goal is to identify
gaps and propose practical solutions to enhance bread quality, extend shelf life, and reduce
economic losses while addressing consumer health concerns (Eze & Onwuka, 2018).

1.3 Aim and Objectives

1.3.1 Aim

This study aims to enhance bread quality and extend it shelf life by comparing the effectiveness
and feasibility of Natural versus modern bread preservation techniques within the Nigerian
context.

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1.3.2 Objectives

1. Assess the effectiveness of Natural preservation method in extending bread shelf life
2. Assess the effectiveness of modern methods in extending bread shelf life
3. Use simple experiments to measure spoilage rates under different preservation conditions.
4. Compare the effectiveness of Natural preservation method and modern preservation
methods in preventing bread spoilage

1.4 Significance of the study

This study on bread preservation techniques in Jimeta, Adamawa State, Nigeria, has significant
importance for many reasons. These reasons include:

a. Economic Impact: By identifying and promoting effective bread preservation methods,

the study can help reduce economic losses for local bakers and consumers caused by rapid
spoilage. This can lead to increased profitability for bakers and more affordable bread for
consumers (Norman & Potter, 2006).
b. Sustainable Practices: By comparing Natural and modern preservation techniques, the
study can highlight sustainable and cost-effective methods that are accessible to both urban
and rural populations in Jimeta. This is particularly important for areas with limited access
to advanced technologies.
c. Health Benefits: The study aims to identify preservation methods that minimize the
growth of harmful microorganisms, thereby reducing health risks associated with
consuming spoiled bread (Williams & Oladipo, 2018). This is crucial for public health in
Jimeta, where high temperatures and humidity accelerate spoilage.
d. Consumer Awareness: The research will provide valuable insights into consumer
preferences and concerns regarding bread preservatives (Norman & Potter, 2006). This can
help bakers and producers align their practices with consumer expectations, potentially
increasing customer satisfaction and trust.
e. Cultural Preservation: By documenting and evaluating Natural bread preservation
methods, the study helps preserve cultural heritage and indigenous knowledge. It ensures
that valuable Natural practices are not lost in the face of modern technological
advancements (Adebayo, 2019).

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 f. Academic Contribution: The study adds to the body of knowledge in food science and
preservation, providing a localized case study that can be referenced in future research. It
contributes to the academic discourse on balancing Natural and modern preservation
methods in developing regions (Williams & Oladipo, 2018).

1.5 Scope and Limitations

1.5.1 Scope of the study

This study focuses on the bread preservation methods available in Jimeta, Adamawa State, Nigeria,
in other to investigate and compare the effectiveness of Natural and modern techniques employed
by local bakers and households. It includes identifying Natural preservation method s like sun
drying, natural preservatives, and local storage techniques, as well as modern methods such as
chemical preservatives and refrigeration. The study will evaluate the effectiveness of these
methods through experiments measuring spoilage rates while in storage.

1.5.2 Limitations

a. Time Constraints: The duration of the study may be too short to observe long-term effects
of certain preservation methods on bread quality and spoilage rates.
b. Environmental Variability: Temperature and Humidity are the two most important
factors in food processing hence, Variations in temperature, humidity, and other
environmental factors during the study period can impact the spoilage rates and
effectiveness of preservation methods, leading to variability in results.
c. Limited number of Sample: The study may involve a limited number of bread samples
and preservation methods due to time and resource constraints, which could affect the
generalizability of the results.
d. Geographical Scope: The study is limited to Jimeta, Adamawa State, Nigeria. Findings
and recommendations may not be directly applicable to other regions with different
climatic, economic, or cultural conditions.
e. Resource Constraints: Limited access to advanced laboratory equipment and facilities
may restrict the depth and accuracy of experimental analyses.

4
 CHAPTER TWO

LITERATURE REVIEW

2.1 Natural and Modern Techniques for Food Preservation

2.1.1 Modern techniques

a. Pasteurization

Pasteurization is a thermal processing technique used to extend the shelf life of food products,
including bread, by killing or inactivating most spoilage-causing microorganisms and enzymes.
Unlike sterilization, which aims to eliminate all microorganisms, pasteurization targets those that
are most likely to cause spoilage or disease (Lianou et al., 2016). This method involves heating
the bread to a specific temperature for a defined period and then cooling it rapidly to prevent
overcooking.

The process of pasteurization for bread typically involves heating the product to temperatures
between 70°C and 85°C for several minutes. This range is sufficient to destroy pathogenic bacteria,
yeasts, and molds that could spoil the bread or pose health risks (Garcia et al., 2019). The exact
temperature and time combination can vary depending on the type of bread and the desired shelf
life extension. After heating, the bread is cooled quickly to halt the cooking process and preserve
its sensory qualities such as taste, texture, and appearance (Lianou et al., 2016).

One of the primary advantages of pasteurization is its ability to significantly reduce microbial load
without compromising the bread's quality. This method maintains the bread's nutritional content
and organoleptic properties, making it nearly indistinguishable from fresh bread to consumers.

However, the pasteurization process requires specialized equipment, which can be costly and
might not be accessible to small-scale or Natural bakers. The process also consumes a significant
amount of energy, contributing to operational costs (Rosell, 2011). Moreover, while pasteurization
is effective against many microorganisms, it may not eliminate all pathogens, particularly spore-
forming bacteria, necessitating the use of complementary preservation methods to ensure complete
safety.

5
b. Edible Coatings

Edible coatings are thin layers of edible materials applied to the surface of food products, including
bread, to provide a barrier against environmental factors that cause spoilage. These coatings can
be made from a variety of natural substances such as chitosan, beeswax, alginate, pectin, and plant-
based gums. The primary function of edible coatings is to protect the bread from moisture loss,
oxygen exposure, and microbial contamination, thereby extending its shelf life (Williams &
Oladipo, 2018).

The application of edible coatings to bread involves dissolving the coating material in a solvent,
typically water, and then applying it to the bread surface by dipping, spraying, or brushing. The
coated bread is then allowed to dry, forming a protective film (Jimoh & Agboola, 2020). Chitosan,
derived from chitin found in crustacean shells, is a popular choice for edible coatings due to its
antimicrobial properties. Beeswax and plant-based gums like guar and xanthan gum are also
commonly used for their ability to form a durable and moisture-resistant barrier.

One of the key advantages of edible coatings is their natural composition, making them safe and
appealing to health-conscious consumers who prefer products free from synthetic preservatives.
Edible coatings help maintain the moisture content of bread, preventing it from drying out and
staling. They also inhibit the growth of spoilage-causing microorganisms on the bread's surface,
further extending its shelf life. Additionally, some coatings can enhance the bread's appearance by
providing a glossy finish (Eze & Onwuka, 2018).

However, the process of applying edible coatings can be complex and labor-intensive, requiring
uniform application to ensure effectiveness. Some coatings may alter the taste or texture of the
bread, which could affect consumer acceptance. Furthermore, the durability of edible coatings can
diminish over time, necessitating careful formulation and application to maintain their protective
properties (Williams & Oladipo, 2018).

c. Active Packaging

Active packaging is an innovative food preservation technique that involves the use of packaging
materials designed to interact actively with the food product to extend its shelf life (Adebayo,
2019). Unlike Natural passive packaging, which serves only as a barrier, active packaging can

6
absorb, release, or otherwise regulate substances within the package to maintain the quality and
safety of the food.

Fig 2.1 Coating of Bread with Honey (Williams & Oladipo, 2018)

7
For bread, active packaging technologies include oxygen scavengers, antimicrobial agents, and
moisture regulators.

Oxygen scavengers are materials that absorb residual oxygen within the package, thereby reducing
the oxygen level to prevent aerobic microbial growth and oxidative spoilage (Eze & Onwuka,
2018). These scavengers are often incorporated into sachets or integrated into the packaging film
itself. By removing oxygen, these scavengers help maintain the bread's freshness and prevent the
growth of molds and other spoilage organisms (Cauvain & Young, 2007).

Antimicrobial packaging involves the incorporation of antimicrobial substances into the packaging
material. These substances, which can include natural extracts like essential oils or synthetic
antimicrobials, are gradually released into the bread's environment to inhibit the growth of spoilage
microorganisms (Dave & Ghaly, 2011). This method can significantly extend the shelf life of bread
while reducing the need for chemical preservatives directly added to the dough.

Moisture regulators in active packaging help control the moisture level within the package. This
can include desiccants that absorb excess moisture to prevent mold growth or humidity control
films that maintain an optimal moisture balance to keep the bread from drying out or becoming
too moist (Axel et al., 2017). Maintaining the correct moisture level is crucial for preventing both
microbial spoilage and staling.

Active packaging provides several benefits, including enhanced protection against spoilage and
contamination, improved shelf life, and potentially reduced need for chemical additives. However,
active packaging materials can be more expensive than Natural packaging, increasing the overall
cost of the final product. The technology involved is also more complex, requiring careful
integration into existing packaging processes. Additionally, consumer perception of active
packaging can vary, with some consumers expressing concerns about the safety and naturalness of
packaging that interacts with their food (Jimoh & Agboola, 2020).

2.1.2 Natural Techniques

a. Canning

Canning In undeveloped countries, canned goods are rarely consumed by the population. This may
be due to a high cost out of reach for the average customer. The high expense of canning results

8
from the significant demand for energy and freshwater during the canning process. In contrast,
various varieties' canned food is shipped from underdeveloped countries to their more developed
counterparts in rich countries. Developing countries, for example, account for over 25% of the
fruits and vegetables preserved in cans imported by European countries. A large quantity of foreign
currency is generated by various varieties of canned fish, vegetables, and fruits for the benefit of
low-income countries. Canning is a type of food preservation that is accomplished with a
combination of processes, including heating and cooling. Canning inhibits the growth of
microorganisms and inhibits the activity of enzymes. First, the raw materials must be properly
treated because some foods, particularly fish, contain harmful microorganisms such as Clostridium
botulinum, which can be fatal. In canning, all the foodstuffs must not be heated in the same manner
(Van et al., 2004). On the other hand, the best quality can be achieved by ensuring proper heating
conditions and employing fresh, healthy ingredients. The number of primary microorganisms
present and the amount of internal water present are high in fish and meat. The pH of those foods,
on the other hand, is virtually neutral. Once all present microbes have been eliminated, the
challenge of creating a product that is safe to consume becomes extremely difficult. Permanent
heating in a pressure sterilizer at temperatures that can reach more than 100 degrees Celsius, on
the other hand, can be a risk-free choice. The pH of those foods, on the other hand, is virtually
neutral. Once all present microbes have been eliminated, the challenge of creating a product that
is safe to consume becomes extremely difficult. Permanent heating in a pressure sterilizer at
temperatures that can reach more than 100 degrees Celsius, on the other hand, can be a risk-free
choice. (Van et al., 2004). Protein-rich foods are sealed hermetically in cans or jars after being
heated. Plant-based foods do not need to be heated before being packed in cans. Vinegar, acetic
acid, or even oil is put in the can to limit microbial growth; finally, canned products can be stored
for a long time without refrigeration.

b. Freezing

Freezing is an ancient and widely used method of preserving food. Unlike other methods, freezing
preserves food's taste, texture, and nutritional content. Freezing is a low-temperature technique in
which microorganisms cannot reproduce, chemical reactions are reduced, and cellular metabolic
responses are hindered (Delgado & Sun, 2001). The preservation of food goods through freezing
ensures that they retain their quality over an extended time. Canning and dehydration are also long-

9
term preservation methods for food materials; however, freezing is generally considered preferable
to these methods. (Fennema, 1977). Freezing has been efficaciously engaged for the long-standing
preservation of many foods, providing an expressively prolonged shelf life. The process includes
dropping the product temperature usually to −18 °C or lower than this (Fennema et al., 1973). The
physical condition of food material is modified when energy is detached by cooling below freezing
temperatures. However, if the temperature is extremely chilly, it slows down the chemical
modifications that impact the qualities for which food spoils and slows down the increase of
microorganisms. (George, 1993). Despite its outstanding preservation and quality assurance
capabilities, frozen food has not been widely adopted in developing nations. On the other hand,
domestic-level freezers are widespread among the wealthy and upper-middle classes in emerging
countries. People in underdeveloped countries generally dislike frozen foods. Finally, it is one of
the least industrialized food preservation strategies in developing countries. Freezing has been
increasingly popular in recent years for preserving various foods, including fruits and vegetables.
In both rich and emerging countries, deploying freezing techniques is comparable. (Barbosa et al.,
2005). Freezing has been increasingly popular in recent years for preserving various foods,
including fruits and vegetables. In both rich and emerging countries, deploying freezing techniques
is comparable. (Dave & Ghaly, 2011). Depending on the species, meat contains 50–75 percent
water, and freezing turns the greatest amount of water into ice. The phenomenon of meat freezing
is very quick, and nearly 75% of tissue fluid freezes at −5 °C. While the temperature is decreased,
the freezing rate is increased, and around 98% of water freezes at −20 °C. However, widespread
crystal formation occurs at −65 °C (Dave & Ghaly, 2011).

c. Refrigeration

Refrigeration is a method for short-term preservation of food quality as it may delay spoilage and
growth of pathogenic microorganisms. Recent concern for several pathogenic or suspected
organisms that will grow at refrigerator temperatures (< 6°C) has surfaced. These include C.
botulinum type E, Vibrio parahaemolyticus, Yersinia enterocolitica, and Listeria monocytogenes
(Lechowich, 1988). The refrigerated food industry increases the availability of fresh, refrigerated
foods to consumers. The presence of delicatessens in supermarkets has increased this market.
Products range from fresh salads of all types to entrees with 21 to 29 days (El-Hag, 1989). Two
processes may be used for some of these new refrigerated products. The "sous-vide" process

10
involves packaging in a vacuum pouch, cooking in the package, cooling rapidly, and refrigerating.
Cooking may be in a pressure cooker, moist steam oven, or water bath. The "nouvelle carte"
process differs concerning the type of package used. A plate is included in the pouch, and there is
a sleeve with an overwrap. The rapid cooking methods used for these products are designed to
minimize quality changes. As this industry grows, more problems associated with pathogenic
organisms could occur. Problems can be prevented if handlers of these products in the food
distribution chain, as well as in institutional settings and at the consumer level, do not allow
temperature abuse to take place and if additional methods for ensuring inhibition of the growth of
these microorganisms are employed in the production of the products. As precooked vacuum-
packaged meat products become popular with consumers, concern for potential outbreaks of food-
borne illnesses also will increase (Lechowich, 1988). Examined precooked beef and pork packages
obtained from the retail market and a meat processor. Pathogenic and indicator organisms were
not found in the samples. A storage study showed that storage at one °C resulted in a longer shelf
life than at 5 or 10°C, emphasizing the need for careful control. The cook/chill system used in
foodservice operations is a system that involves the use of refrigerator storage of precooked foods
before reheating for consumption. Shelf life is not expected to be as long in this system as for the
techniques previously described because vacuum packaging and cooking in the packaging are not
employed. Many studies have reported the microbiological and sensory quality of foods in a
cook/chill system. Reported that turkey slices taken from rolls cooked conventionally, held in the
refrigerator for not more than 2.5 hr, and then reheated for 30 or 40 sec in a microwave did not
have higher coliform or aerobic counts than freshly prepared slices. The investigators stressed the
need to follow guidelines (HEW, 1978) for reheating to an internal temperature of at least 74°C.
A critical point in the cook/chill system is cooling the product linger-Snyder and Matthews (1988)
(Ollinger-Snyder & Matthews, 1988). round that 5 hr was required for cooling of 900-g loaves to
7°C. The use of slower cooling methods in a food service operation than in commercial production
of refrigerated entrees (Lechowich, 1988) increases the need for careful handling to avoid post-
cooking, prechilling contamination of products.

11
Fig 2.2 Bread Refrigeration (Lechowich, 1988)

12
d. Fermentation

The fermentation method uses microorganisms to preserve food. This method involves the
decomposition of carbohydrates with microorganisms and enzymes (Amit et al., 2017). Bacteria,
yeasts, and moulds are the most common groups of microorganisms involved in the fermentation
of a wide range of food items, such as dairy products, cereal-based foods, and meat products.
Fermentation enhances the nutritional value, healthfulness, and digestibility of foods. This is a
healthy alternative to many toxic chemical preservatives (Lewin, 2012).

e. Addition of Sugar: Jellies and Jams

Jellies and jams are preserved by their sugar content, which is high enough to prevent the growth
of microorganisms, except for mould growth on the surface. Commercially, products are protected
by a vacuum cap and maybe pasteurized after the containers are filled. Recommends that home-
prepared jellies and jams be processed in a boiling water bath (Holmes, 1989). Because they are
formed of fruit juice or a fruit extract in water, jellies are transparent things that may be seen
through. On the other hand, Jams contain all or almost all of the insoluble solids of the fruit because
they are made from whole, crushed, macerated, or pureed fruit, which is employed in their
production. Technically, jams and preserves are the same things, with the exception that the term
preserves is reserved for products that contain entire fruit. The gelation of pectin is caused by the
addition of sugar in the presence of an acidic environment. Hydrogen bonding between hydroxyl
groups and between hydroxyl and carboxyl groups is responsible, at least in part, for the rigidity
of fruit jellies (Whistler & Dabiel, 1985). The interrelationships between the three key ingredients,
pectin, sugar, and acid, are critical to the overall quality of the fruit preserve. When pectin or acid
levels are inadequate, gel formation is prevented; a stiff jelly is produced when sugar levels are
insufficient.

2.2 Plant Extracts as Bio‐Preservative

Plant extracts have been extensively studied as bio‐preservatives, as plants contain many essential
antifungal compounds, for example, phenolic compounds, glucosinolates, cyanogenic glycosides,
oxylipins, and alkaloids (Axel et al., 2017); there is a thriving interest in natural ingredients with
multifunctional properties in food as well. Most edible plant parts include trace amounts of plant
defense substances (phenolic acids), categorized as hydroxybenzoic and hydroxycinnamic acids.

13
Hydroxybenzoic acids are frequently found in larger phenolic compounds, such as hydrolyzable
tannins. Hydroxycinnamic acids occur as esters of glycerol, tartaric, shikimic, and quinic acids, as
well as glycosylated derivatives (Maldanado & Schieber, 2015). Plant extracts can inhibit harmful

Fig 2.3 Mold growth on bread (Axel et al., 2017)

14
bacteria from adhering to the host cell membrane. As a result, it reduces bacterial attachment to
host cell surface membranes, and thus sometimes it becomes a potential anti‐adhesive agent.

Study (Wei et al., 2009) investigated the antifungal efficacy of different raisin extracts and by‐
products in Natural bread. Compared to a sample containing no preservatives, the bread produced
with raisin paste and raisin water separate (7.5%) exhibited the best mold‐ reducing abilities. Leaf
extracts of cherry laurel (Prunus laurocerasus L.) were recommended as potential bio‐preservatives
after demonstrating a very low MIC (mg/mL) against a variety of bread spoilage fungi (Sahan,
2012). Further, showed that bread produced with a mix of sourdough and pea flour hydrolysate
fermented by the antifungal strain Lb. plantarum 1A7 had the most extended shelf life. It is also
effective against P. roqueforti. Moreover, bread formulations containing free or liposome‐
encapsulated garlic extract (0.65 mL/100 g of dough) were found to be more microbiologically
stable than controls, inhibiting molds such as P. herquei, F. graminearum, and A. flavus for five
days (Panila et al., 2019).

2.3 Microbial Fermentation

Naturally, organic and flavorful bread with a long shelf life was achieved naturally through an
expanded fermenting operation: sourdough (Melini & Melini, 2018). The word “sourdough”
describes a particular bread recipe in which flour, water, LAB, and yeast organ‐ isms are fermented
together. Because of their remarkable antifungal activity, lactic acid bacteria (LAB) and
antagonistic yeasts have received particular attention among natural agents (Axel et al., 2016) and
are herein discussed below.

2.3.1 Lactic acid bacteria:

LAB metabolic products enhance bread′s organoleptic and technological aspects, as well as its
textural characteristics, along with its shelf life, nutritional value, and beneficial aspect
(anticarcinogenic and cholesterol reduction abilities), during the fermentation of the dough ().
They can also be adopted to replace chemical preservatives, ensuring a clean label and increased
consumer acceptance (Pawlowska et al., 2012)

Lactic acid bacteria (LAB) have been utilized in fermented foods for over about 4000 years. It is
naturally found in foods or introduced as pure cultures (Schnurer & Magnusson, 2005). It is also
GRAS‐certified (generally recognized as safe) and has an extended application history in various

15
cereal fermentations, particularly in the baking industry. LAB′s adaptability is re‐ markable, not
just in terms of catabolic and anabolic pathways but also in changing environmental conditions
(Buck et al., 2009).

It has also been employed as a starter culture in the food business for centuries, which may produce
several bioactive compounds, along with fatty acids, bacteriocins, organic acids, hydrogen
peroxide, indole lactic acid, and phenyl lactic acid. They also have an anti‐aflatoxigenic effect
(Guimaraes et al., 2018). Particular lab strains that have gripping bio‐preservation action on bread
when adopted as starter cultures include Lactobacillus amylovorous DSM 19,280, Lactobacillus
fermentum Te007, Lactobacillus acidophilus ATCC 20079, Lactobacillus paralimentarius PB127,
Lactobacillus brevis R2D, Lactobacillus rossiae LD108, Lactobacillus hammesii, Lactobacillus
paracasi D5, Pediococcus pentosaceus KTU 05‐8 and KTU 05‐10, Lactobacillus pentosus G004,
Lactobacillus plantarum, Lactobacillus reuteri R29, Lactobacillus rhamnosus, Lactococcus BSN,
Pediococcus acidilactici KTU05‐7, as well as Leuconostoc citreum C5 and HO12 (Axel et al.,
2016). Additionally, adding 15–20% sourdough significantly (p = 0.0001) increased bread volume
and crumb porosity, based on the LAB strain, and reduced acrylamide for‐ mation by an average
of 23% (for LUHS51 and LUHS206) and 54% (for LUHS71 and LUHS225), respectively, in
comparison with regular bread. Also, the most dominating species of the conventional sourdough
microbiota, Lactobacillus sanfranciscensis, has been found to have a favorable impact on several
important quality features of sourdough, notably dough rheological qualities, bread texture and
aroma, and shelf‐life conservation (Zhang et al., 2019).

2.3.2 Yeast

Numerous authors have experimented with using incompatible yeasts as bio‐ control agents. They
can be used as bio‐preservatives as they retain some of the essential features that enhance their
acceptability. They compete for nutrients with fungal pathogens and their higher rate of nutrient
utilization significantly contributes to a bio‐preservative nature. Yeast produces killer toxins, also
called mycotoxins, which showed bioprotective attributes against food spoiling microorganisms
and pathogens (Rima & Steve, 2012). Some yeast genera produce extra‐ and intracellular
compounds which possess antibacterial properties. Production of ethanol of high concentration
and organic acids which results in the change of pH of the medium also responsible for the
effectiveness of yeasts as bio‐preservatives (Muccilli & Restuccia, 2015). Many of them can

16
sustain residence on dry surfaces due to their low requirements for water and nutrients. With Lb.
plantarum 1A7 as a starter, the yeast Wickerhamomyces anomalus LCF1695 (previously
recognized as Pichia anomala) was occupied for sourdough fermentation. It was found that when
P. roqueforti DPPMAF1 was artificially inoculated (102 conidia ml1) with this combination and
stored at room temperature, the microbiological shelf life was elongated to at least 14 days. As
well as Penicillium paneum KACC 44834, out‐ growth on white pan bread leavened with
Penicillium anomala SKM‐T was significantly reduced compared to standard baker′s yeast, and
improved the shelf life. In contrast, bread composed of propionic acid from cultured yeast extract
contained less ethanol and had a better shelf life against mold growth than bread formulated with
non‐fermented yeast extract (Gardner et al., 2002).

17
 CHAPTER THREE

MATERIALS AND METHOD

3.1 Methodology

3.1.1 Weight loss Method

The weight loss method is a straightforward and effective technique used to measure the rate of
spoilage in bread. It involves monitoring the weight of bread loaves over a specified period to
quantify the loss of moisture and other components due to microbial activity and environmental
factors. By regularly weighing the bread and calculating the percentage of weight lost, researchers
can assess the effectiveness of various preservation methods. This method provides a clear
indication of spoilage, as increased weight loss typically correlates with higher rates of microbial
growth and spoilage. The simplicity and quantitative nature of the weight loss method make it a
valuable tool for comparing Natural and modern bread preservation techniques (Adebayo, 2019).

3.1.2 Procedure

3.1.2.1 Initial Preparation:

I obtained and labeled the bread loaves (total of Nine bread loaves with labels A1-A3 for un
preserved, Natural technique and for modern technique.) for each preservation method, making
sure that all the loaves were from the same batch to maintain consistency throughout the
experiment. I carefully recorded the initial weight of each bread loaf using a digital weighing scale,
which provided precise measurements. This initial weight was documented as the baseline for all
future comparisons, allowing for accurate tracking of any changes in weight over time due to the
preservation methods applied. (Jimoh & Agboola, 2020)

3.1.2.2 Application of Preservation Methods:

a. Natural preservation method:

i. Natural Preservatives: I selected fresh garlic bulbs, peeled and chopped the cloves, then ground
them into a smooth paste using a food processor (blender). I measured equal parts (100 ml) of
honey and garlic paste, mixed them thoroughly until well blended, and obtained a thick, spreadable
consistency. After choosing three fresh bread loaves, I lightly moistened their surfaces to help the

18
mixture adhere better. I used a basting brush to apply the honey-garlic mixture evenly over the top,
sides, and bottom of each loaf, ensuring uniform coverage. I allowed the loaves to sit at room
temperature for a 15minutes to let the mixture dry and adhere. Finally, I stored the bread loaves in
parchment paper in a cool, dry place and observed them over the next few days to monitor the
preservation effect. (Adebayo, 2019)

b. Modern Method

i. Refrigeration: I stored the three loaves of bread in a refrigerator set to a temperature of 4°C.
Before placing them in the refrigerator, I ensured that each loaf was securely wrapped in plastic
wrap to minimize exposure to air and moisture, which could accelerate spoilage (Pawlowska et
al., 2012). I placed the loaves on a shelf in the middle of the refrigerator, away from the back wall
where temperatures might fluctuate due to the cooling mechanism (Axel et al., 2017). By
maintaining a consistent temperature of 4°C, I aimed to slow down the growth of mold and
bacteria, thereby extending the shelf life of the bread. Over the storage period, I periodically
checked the loaves for any signs of spoilage, such as weight loss, mold growth and changes in
texture.

c. Storage: I stored the preserved bread loaves in a controlled environment where the temperature
was kept stable between 20°C and 30°C to prevent any condensation or spoilage. The storage area,
selected for its cool and dry conditions, was monitored closely for both temperature and humidity
levels. To maintain low humidity, which is crucial for preventing mold growth, I used desiccators
and silica gel packs, keeping the humidity between 50% and 60%. Each loaf was wrapped in
breathable materials (parchment paper) to protect them while allowing some air circulation.
Regular checks with digital hygrometers and thermometers were used to ensure the environment
remained optimal, effectively extending the bread's shelf life while preserving its freshness and
quality. (Pawlowska et al., 2012)

3.1.2.3 Monitoring and Data Collection:

I weighed each bread loaf at regular intervals, daily, using a digital scale and recorded the weight
on a sheet. I observed and noted any visible signs of spoilage, such as mold growth, off-odors, or
texture changes. Additionally, I monitored and recorded the environmental conditions, including
temperature and humidity, using a hygrometer and thermometer. (Pawlowska et al., 2012)

19
3.2 Materials used

1. Freshly baked bread loaves (same type and batch)

2. Digital weighing scale (accurate to 0.01 grams)
3. Preservation materials for Natural preservation method (garlic and honey mixture)
4. Preservation materials for modern method (Refrigerator)
5. packaging materials (plastic wrap and parchment paper)
6. Desiccators (for controlled humidity environment)
7. Basting brush (silicone)
8. Measuring cup (ml)
9. Labels and markers
10. Data recording sheet (notebook)
11. Food processor (blender)
12. Thermometer (for monitoring environmental conditions)
3.3 Data Analysis:

I calculated the percentage weight loss for each bread loaf at each interval using the
formula:

Percentage Weight Loss = x 100

Then compared the weight loss and spoilage signs across different preservation methods.
ANOVA was used to analyze the data obtained from different bread preservation methods
because it allows for a comparison of means across multiple groups simultaneously. This
statistical method is ideal for determining if there are significant differences in weight loss
among the various preservation techniques. By using ANOVA, we assessed the overall
effectiveness of each method and identify which techniques significantly reduce spoilage,
providing a robust and comprehensive analysis of the preservation methods' impacts on
bread shelf life (Moore et al., 2021).

20
 CHAPTER FOUR

RESULTS AND DISCUSSION

4.1 Spoilage Rate and Weight Loss (Natural preservation method)

The data demonstrates a gradual increase in weight loss across all three bread loaves over the 20-
day observation period. Initially, weight loss was minimal, with only a 0.2% decrease observed by
the second day, and this trend continued steadily. By the 10th day, the loaves had lost
approximately 1.0% of their initial weight, and spoilage signs were minimal, limited to minor
texture changes. However, the preservation effects of the garlic honey mixture began to diminish
after the 12th day, as evidenced by the appearance of minor off-odors and a 1.2% weight loss. This
trend continued, with mold growth becoming visible by the 16th day, and the weight loss reaching
1.6%. By the end of the 20-day period, all samples had experienced a 2.0% weight loss,
accompanied by extensive mold growth and strong off-odors, indicating significant spoilage. The
steady weight loss can be attributed to moisture evaporation, while the spoilage rate increased
significantly in the latter half of the observation period, showing the limits of the natural
preservative's effectiveness (Axel et al., 2017).

4.2 Spoilage Rate and Weight Loss (Modern Method)

The results of the bread preservation experiment using refrigeration at 4°C reveal the effectiveness
of this modern preservation method in slowing down spoilage (Dave & Ghaly, 2011). Over the
20-day observation period, the weight loss in the bread loaves was minimal, with a steady but slow
reduction in weight. The percentage loss in weight was 0% for the first two days, indicating no
moisture loss or spoilage (Lechowich, 1988). By the 4th day, a slight 0.2% weight loss was
observed, which gradually increased to 1.0% by the end of the 20 days.

Throughout the experiment, there were no significant signs of spoilage, such as mold growth or
off-odors, until the final day, when only slight texture changes were noted. This is in contrast to
the natural preservation method (using a garlic honey mixture), where spoilage indicators like
mold growth and off-odors began to appear by the 16th day. The refrigerated bread loaves
remained visually unchanged for a longer period, highlighting the ability of low temperatures to
inhibit microbial growth and slow down the biochemical processes that lead to spoilage
(Lechowich, 1988).

21
According to Cauvain & Young, (2007) The minimal weight loss observed is primarily due to the
reduction in evaporation and dehydration that refrigeration provides. The stable temperature of
4°C and controlled humidity levels around 60-61% created an environment that significantly
reduced the rate of moisture loss compared to ambient conditions. This consistent preservation
environment resulted in only a 1.0% weight loss by the 20th day, suggesting that the loaves were
well-protected against the typical spoilage factors such as mold, bacteria, and yeast, which thrive
in warmer, more humid environments.

4.3 Spoilage Rate and Weight Loss (Non- Natural/Modern Method)

The results clearly show a significant rate of spoilage when bread loaves are stored at room
temperature without any preservation method. The initial slight weight loss over the first few days
is mainly due to moisture evaporation, leading to dryness and surface hardening (Jimoh &
Agboola, 2020). As the days progress, the bread loaves exhibit more severe spoilage signs,
including mold growth and off-odors, starting as early as day 10. By day 20, all samples show
extensive spoilage, with a weight loss of over 6%, making the bread inedible. Eze & Onwuka,
(2018) This data emphasizes the rapid deterioration of bread stored at room temperature and
highlights the necessity of preservation methods to maintain bread quality over time.

22
Table 4.1: Natural preservation method of preservation (garlic-honey mixture)
Day Sample Weight Percentage Observations Temperature Humidity
(g) Loss (%) (°C)
0 A1 500 0% Fresh, no 23 55
spoilage
0 A2 498 0% Fresh, no 23 55
spoilage
0 A3 502 0% Fresh, no 23 55
spoilage
2 A1 499 0.2% No visible 23 54
spoilage
2 A2 497 0.2% No visible 23 54
spoilage
2 A3 501 0.2% No visible 23 54
spoilage
4 A1 498 0.4% No visible 23 54
spoilage
4 A2 496 0.4% No visible 23 54
spoilage
4 A3 500 0.4% No visible 23 54
spoilage
6 A1 497 0.6% No visible 24 55
spoilage
6 A2 495 0.6% No visible 24 55
spoilage
6 A3 499 0.6% No visible 24 55
spoilage
8 A1 496 0.8% Slight texture 24 55
change
8 A2 494 0.8% Slight texture 24 55
change

23
8 A3 498 0.8% Slight texture 24 55
change
10 A1 495 1.0% Minor texture 24 55
change, no
mold
10 A2 493 1.0% Minor texture 24 55
change, no
mold
10 A3 497 1.0% Minor texture 24 55
change, no
mold
12 A1 494 1.2% Minor off- 25 56
odor, no mold
12 A2 492 1.2% Minor off- 25 56
odor, no mold
12 A3 496 1.2% Minor off- 25 56
odor, no mold
14 A1 493 1.4% Minor off- 25 56
odor, no mold
14 A2 491 1.4% Minor off- 25 56
odor, no mold
14 A3 495 1.4% Minor off- 25 56
odor, no mold
16 A1 492 1.6% Visible mold 25 57
growth, off-
odor
16 A2 490 1.6% Visible mold 25 57
growth, off-
odor

24
16 A3 494 1.6% Visible mold 25 57
growth, off-
odor
18 A1 491 1.8% Increased 25 57
mold growth,
off-odor
18 A2 489 1.8% Increased 25 57
mold growth,
off-odor
18 A3 493 1.8% Increased 25 57
mold growth,
off-odor
20 A1 490 2.0% Extensive 25 57
mold growth,
off-odor
20 A2 488 2.0% Extensive 25 57
mold growth,
off-odor
20 A3 492 2.0% Extensive 25 57
mold growth,
off-odor

25
Table 4.2: Modern method of preservation (Refrigeration)
Day Sample Weight Percentage Observations Temperature Humidity
(g) Loss (%) (°C) (%)
0 A1 500 0% Fresh, no 4 60
spoilage
0 A2 498 0% Fresh, no 4 60
spoilage
0 A3 502 0% Fresh, no 4 60
spoilage
2 A1 500 0% No visible 4 60
spoilage
2 A2 498 0% No visible 4 60
spoilage
2 A3 502 0% No visible 4 60
spoilage
4 A1 499 0.2% No visible 4 60
spoilage
4 A2 497 0.2% No visible 4 60
spoilage
4 A3 501 0.2% No visible 4 60
spoilage
6 A1 499 0.2% No visible 4 60
spoilage
6 A2 497 0.2% No visible 4 60
spoilage
6 A3 501 0.2% No visible 4 60
spoilage
8 A1 498 0.4% No visible 4 60
spoilage
8 A2 496 0.4% No visible 4 60
spoilage
8 A3 500 0.4% No visible 4 60
spoilage
10 A1 498 0.4% No visible 4 60
spoilage
10 A2 496 0.4% No visible 4 60
spoilage
10 A3 500 0.4% No visible 4 60
spoilage
12 A1 497 0.6% No visible 4 61
spoilage
12 A2 495 0.6% No visible 4 61
spoilage
12 A3 499 0.6% No visible 4 61
spoilage

26
14 A1 497 0.6% No visible 4 61
spoilage
14 A2 495 0.6% No visible 4 61
spoilage
14 A3 499 0.6% No visible 4 61
spoilage
16 A1 496 0.8% No visible 4 61
spoilage
16 A2 494 0.8% No visible 4 61
spoilage
16 A3 498 0.8% No visible 4 61
spoilage
18 A1 496 0.8% No visible 4 61
spoilage
18 A1 496 0.8% No visible 4 61
spoilage
18 A3 498 0.8% No visible 4 61
spoilage
20 A1 495 1.0% Slight texture 4 61
change, no
mold
20 A2 493 1.0% Slight texture 4 61
change, no
mold
20 A3 497 1.0% Slight texture 4 61
change, no
mold

27
Table 4.3: Stored Bread at Room Temperature (Non-Modern/Natural preservation method
)
Day Sample Weight Percentage Observations Temperature Humidity
(g) Loss (%) (°C) (%)
0 A1 500 0% Fresh, no 25 65
spoilage
0 A2 498 0% Fresh, no 25 65
spoilage
0 A3 502 0% Fresh, no 25 65
spoilage
2 A1 496 0.8% Slight 25 65
dryness, no
visible
spoilage
2 A2 494 0.8% Slight 25 65
dryness, no
visible
spoilage
2 A3 498 0.8% Slight 25 65
dryness, no
visible
spoilage
4 A1 493 1.4% Surface 25 65
hardening, no
visible
spoilage
4 A2 491 1.4% Surface 25 65
hardening, no
visible
spoilage
4 A3 495 1.4% Surface 25 65
hardening, no
visible
spoilage
6 A1 490 2.0% Noticeable 25 65
hardness,
slight
staleness
6 A2 488 2.0% Noticeable 25 65
hardness,
slight
staleness
6 A3 492 2.0% Noticeable 25 65
hardness,
slight
staleness

28
8 A1 487 2.6% Hardened 25 65
crust, slight
staleness
8 A2 485 2.6% Hardened 25 65
crust, slight
staleness
8 A3 489 2.6% Hardened 25 65
crust, slight
staleness
10 A1 484 3.2% Visible 25 65
dryness,
minor mold
spots
10 A2 482 3.2% Visible 25 65
dryness,
minor mold
spots
10 A3 486 3.2% Visible 25 65
dryness,
minor mold
spots
12 A1 481 3.8% Surface mold 25 66
growth,
noticeable
staleness
12 A2 479 3.8% Surface mold 25 66
growth,
noticeable
staleness
12 A3 483 3.8% Surface mold 25 66
growth,
noticeable
staleness
14 A1 478 4.4% Significant 25 66
mold growth,
stale
14 A2 476 4.4% Significant 25 66
mold growth,
stale
14 A3 480 4.4% Significant 25 66
mold growth,
stale
16 A1 475 5.0% Extensive 25 66
mold growth,
off-odor

29
16 A2 473 5.0% Extensive 25 66
mold growth,
off-odor
16 A3 477 5.0% Extensive 25 66
mold growth,
off-odor
18 A1 472 5.6% Severe mold, 25 66
strong off-
odor
18 A2 470 5.6% Severe mold, 25 66
strong SSoff-
odor
18 A3 474 5.6% Severe mold, 25 66
strong off-
odor
20 A1 469 6.2% Extensive 25 66
spoilage,
inedible
20 A2 467 6.2% Extensive 25 66
spoilage,
inedible
20 A3 471 6.2% Extensive 25 66
spoilage,
inedible

30
Table.4.4 Comparison Table of Preservation Methods Based on Effectiveness

Preservation Initial Final Overall Sensory Effectiveness

Method Weight Weight Spoilage Quality
Loss Loss Rate
(Day 2) (Day 20)
Modern Method 0.0% 1.0% Very Low Maintained Most
(Refrigeration) freshness, Effective
texture, and
aroma
Natural 0.2% 2.0% Moderate Some loss in Moderately
preservation texture and Effective
method (Garlic taste over time
Honey Mixture)
Non- 0.8% 6.2% High Rapid Least
Natural/Modern deterioration in Effective
Method (Room sensory
Temperature) attributes

31
Table 4.4: Percentage Loss Over 20 Days (2-Day Intervals)
Day Natural Modern Method Non-
preservation (Refrigeration) Natural/Modern
method Method
A1 A2 A3
0 0.0% 0.0% 0.0%
2 0.2% 0.0% 0.6%
4 0.4% 0.2% 1.2%
6 0.6% 0.2% 2.0%
8 0.8% 0.4% 2.6%
10 1.0% 0.4% 3.2%
12 1.2% 0.6% 3.8%
14 1.4% 0.6% 4.6%
16 1.6% 0.8% 5.2%
18 1.8% 0.8% 5.8%
20 2.0% 1.0% 6.4%

32
Table 4.5 ANOVA Results
Source of Sum of Degrees of Mean F-statistic p-value
Variation Squares (SS) Freedom Square (MS)
(df)
Between 131.36 2 65.68 30.13 7.00e-11
Groups
Within 42.89 30 1.43
Groups
Total 174.25 32

ANOVA Results:

F-statistic: 30.13
p-value: 7.00e-11

33
4.4 ANOVA Data Interpretation

The ANOVA analysis conducted on the spoilage (percentage loss) of bread loaves across three
preservation methods (Natural preservation method using garlic honey mixture, modern method
using refrigeration, and non-Natural preservation method at room temperature) yielded the
following data’s:

F-statistic: 30.13

p-value: 7.00e-11

The F-statistic of 30.13 indicates a comparison between the variance of the means of different
groups (i.e., the preservation methods) and the variance within each group. The F-statistic
quantifies the ratio of the variance between the group means (how different the preservation
methods are from each other) to the variance within the groups (how consistent the results are
within each method). A higher F-statistic value suggests that the variance between the groups is
much larger than the variance within each group, implying that the groups (preservation methods)
are significantly different from each other in terms of spoilage rate (Montgomery,2017; Jimoh &
Agboola, 2020).

A. Significance of the p-Value:

The p-value obtained from the ANOVA is extremely small 7.00e -11. This value is crucial in
determining whether the observed differences between the means of the three preservation
methods are statistically significant (Moore et al., 2021).

Moore et al., (2021) explained that in most scientific research, a p-value threshold (alpha level) of
0.05 is used to determine statistical significance. If the p-value is less than 0.05, Then, we conclude
that there is a statistically significant difference between the groups.

In this case, the p-value is 7.00e-11, which is far below the standard threshold of 0.05. This
extremely low p-value strongly suggests that the differences in spoilage rates between the
preservation methods are not due to random chance, but are indeed statistically significant.

34
 CHAPTER FIVE

CONCLUSSION AND RECOMMENDATION

5.1 CONCLUSSION

The Natural preservation method (garlic honey mixture) assessment, in extending the shelf life of
bread showed that this method provides a moderate level of preservation. Over the 20-day period,
the bread loaves preserved with the garlic honey mixture exhibited a gradual increase in percentage
loss, starting from 0.6% on day 2 and reaching 2.0% by day 20. This indicates that while Natural
preservation methods can delay spoilage, their effectiveness diminishes over time. Although the
garlic honey mixture slowed down the spoilage process relative to non-preserved bread, it could
not prevent significant spoilage over the 20-day period.

The modern method of refrigeration at 4°C demonstrated a significantly higher effectiveness in

extending bread shelf life. The percentage loss for the refrigerated bread was minimal, beginning
at 0% and increasing only to 1.0% by day 20. This contrast with the Natural preservation method
highlights the superior preservation capabilities of refrigeration, which maintained the bread’s
freshness and texture with negligible spoilage. The controlled low-temperature environment
effectively inhibited the growth of spoilage organisms, making refrigeration the most effective
method tested for preserving bread over an extended period.

Simple experiments measuring spoilage rates under different preservation conditions revealed
clear differences in the effectiveness of these methods. By weighing the bread loaves at regular
intervals and noting signs of spoilage, the experiments provided concrete data showing that bread
stored at room temperature had the highest spoilage rate, with percentage losses as high as 6.2%
by day 20. In contrast, the garlic honey mixture slightly reduced the spoilage rate, while
refrigeration was the most effective, with a percentage loss of only 1.0% by day 20. These
emphasizes the varying degrees of effectiveness in preservation methods, with room temperature
storage proving inadequate, Natural preservation method s offering moderate protection, and
modern refrigeration showing superior results.

Comparing the effectiveness of Natural and modern preservation methods in preventing bread
spoilage, it is evident from the ANOVA analysis and the observed data that modern refrigeration
is far more effective. The ANOVA results showed a significant difference in spoilage rates

35
between the methods, with a p-value of 7.0e-11 indicating that the variations were not due to chance.
Refrigeration maintained the bread's quality with minimal spoilage, while the Natural garlic honey
mixture, although better than room temperature storage, was less effective over time. These results
clearly demonstrate that modern preservation methods like refrigeration are essential for extending
the shelf life of bread, offering far greater protection against spoilage than Natural preservation
method s.

5.2 RECOMMENDATIONS

1. Enhance the Use of Natural Preservation Methods: The study showed that natural
preservation methods, such as the use of a garlic honey mixture, offer some level of
protection against bread spoilage. However, their effectiveness can be improved through
further research. It is recommended to explore combinations of different natural
preservatives, optimize their concentration, and test different application methods. This
could potentially enhance their antimicrobial properties, making them more viable for
extending bread shelf life, especially in areas with limited access to modern technology.
2. Promote and Increase Accessibility to Modern Preservation Techniques: Refrigeration
proved to be the most effective method in extending bread shelf life, minimizing spoilage
over the 20-day period. To capitalize on this, efforts should be made to increase the
accessibility of refrigeration, particularly in rural and economically disadvantaged regions.
Initiatives could include promoting affordable, energy-efficient refrigeration units, as well
as exploring the use of solar-powered or off-grid cooling systems to overcome
infrastructure challenges.
3. Encourage Simple Experimental Methods for Spoilage Monitoring: The study utilized
a straightforward method of measuring spoilage rates through regular weight loss
monitoring, which provided accurate and useful data. It is recommended that similar simple
and cost-effective experimental setups be used for monitoring food spoilage, especially for
small-scale bakers and food processors. This would enable early detection of spoilage,
ensuring timely interventions and reducing economic losses.
4. Integrate Natural and Modern Preservation Methods for Optimal Results: The
comparison between natural and modern preservation methods indicates that while natural
methods can offer some benefits, modern techniques like refrigeration are significantly

36
more effective. Therefore, a combined approach might be beneficial. For example, using
natural preservatives in conjunction with refrigeration could further extend shelf life while
reducing dependency on refrigeration alone. This hybrid method could also be more
sustainable and adaptable for various settings, including those with limited resources.

37
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