FOOD MICROBIOLOGY

Lecturer: Dr. Pham Hoang Nam

pham-hoang.nam@usth.edu.vn
 2 Course description

•Credit points: 3 ECTS

•Time commitment: lecture 30h, practice 12h
•Recommended background knowledge: Fundamental
Microbiology, Food Biochemistry
•Objectives & Outcome
- understand problems from various areas of food microbiology,
- master the fermentation processes and other transformation of
food by microorganisms.
- control the microbial metabolism used in food production as
well as the microbes involved in foodborne intoxication
•Assessment/ Evaluation

Component Attendance Exercises Practice/Reports Midterm Final

Percentage
0 40 60
%
 Some useful references
3

Bibek Ray, Arun Bhunia, 2013. Martin R. Adams, Maurice O.

Fundamental Food Microbiology, Moss, 2008. Food Microbiology.
CRC Press Fifth Edition. RSC Publishing, Third Edition
 4
Some useful references

In Vietnamese
Kieu Huu Anh, 2010. Food
Microbiology, Vietnam National
Education Publisher.
In French
Son Chu Ky, 2011. Microbiologie
Alimentaire (in French), Course,
Hanoi University of Science And
Technology.
Microbiologie alimentaire
Tome 2: Aliments fermentés et
fermentations alimentaires TEC
– DOC 1996 Coordinateurs:
C.M. BOURGEOIS et J.P.
LARPENT
 COURSE CONTENTS

1. History and development of Food

Microbiology
2. Overview on General Microbiology
3. Physicochemical parameters influencing
microbial fermentation.
4. Microbes involved in food fermentation
➢ Beer
➢ Wine
➢ Distilled beverage
➢ Dairy (butter, milk, cheese...)
➢ Vinegar
➢ Meat and fish products
➢ Fermented vegetables
4. Undesirable microbes in food
➢ Microbes in food spoilage
➢ Foodborne diseases
➢ Pathogenic microbes in food
 INTRODUCTION

What is Food Microbiology?

Food microbiology is the study of the
microorganisms that inhibit, create, or
contaminate food, including the study of
➢ microorganisms causing food spoilage
➢ pathogens that may cause disease
especially if food is improperly cooked or
stored
➢ those used to produce fermented foods
such as cheese, yogurt, bread, beer,
and wine...
➢ those with other useful roles such as
producing probiotics
 INTRODUCTION

Brief History....
DID YOU KNOW...
 The first evidence of beer
manufacture has been trace Ninkasi, the ancient Sumerian
to Ancient Babylonia as far tutelary goddess of beer
back as 7000 BC!
 Milk, butter and cheese were
used in Egypt as early as 3000
BC.
 INTRODUCTION

DID YOU KNOW...

 Between 3000 BC and 1200
BC, the Jews used salt from
the Dead Sea for food
preservation
 Salted fishes and meat were
found in the diet of Chinese
and Sumerian people
 Wines was made from 3500
BC by the Assyrians.
 INTRODUCTION

DID YOU KNOW...

 Fermented sausages were
made by the Babylonians
and the Chineses since 1500
BC
 Olive and other oils were
used for food preservation
from 1500 BC
 Smoking meat was
prepared around 1000 BC
 INTRODUCTION

BUT... It is DOUBTFUL whether

the people understood
 Nature of these food
making process
 Nature of these food
preservations techniques
 Nature of food poisoning From 7000 BC to Middle-Ages
and food spoilage
 Relationship between food
and microbes?
 INTRODUCTION

The Early Years

 1658, Athanasius Kircher,
German monk, first
description: presence of
“worms” invisible to the
naked eyes in decaying
bodies, meat, milk and other
subtances
=> Lacked of precision and did
not receive wide acceptance
Athanasius Kircher
(1602-1680)
 INTRODUCTION

1660‐ Robert Hooke observes plants under simple

Microscope, he observed small compartments and
called them “cells” => he was observing cell walls

Robert Hooke (1635-1703)

 INTRODUCTION
Antonie van Leeuwenhoek
was a linen merchant who
needed to analyze his
cloth fibers => curious
about other specimens like
rain water and tooth
plaque. To his surprise he
found “moving
animalcules”!
1676‐ the first to observe
microbes (up to 300X)

Antonie van Leeuwenhoek (1632-1723)

 INTRODUCTION

1688 Francesco Redi‐ published experiments to disprove

spontaneous generation

Francesco Redi
(1626-1697)

=> Life only comes from living

matters!
 INTRODUCTION

▪ Nicolas Appert, French

inventor & businessman
▪ 1778, etablished a brewery
house
▪ 1789-1793: discovered food
preservation process using
thermal canning Nicolas Appert
(1749-1841)
▪ 1802: export canned
products, tested by French
navy
▪ 1810: published his methods,
awarded 12.000 euros!
Appert’s first can
 INTRODUCTION

 1857-Fermentation is
caused by microbes
 1861-Completes S-shaped
flask experiments that
disprove spontaneous
generation
 1862-Introduces
pasteurization to prevent
spoilage
 1881-Develops anthrax
vaccine for animals
Louis Pasteur (1822-1895)
 INTRODUCTION

Golden Age of Food Microbiology: Louis Pasteur

▪ 1861-Completes S-
shaped flask
experiments (swan-
neck flask)

=> Microbes are

everywhere (air)
and will grow in
broth unless killed
by boiling and
prevented from
entering broth

17
 INTRODUCTION

Golden Age of Food Microbiology: Louis Pasteur

▪ 1857- Series of experiments in

Fermentation:
➢ Proved that yeasts are
facultative anaerobes – can live
with or without oxygen
➢ Explained why wine turned sour
=> fermentation of lactic acid
by bacteria
➢ Proved that the alcoolic
fermentation is caused by yeasts
➢ Discovered the method to avoid
juice spoilage: pasteurization
(62.7°C, 30 min) => introduced
commercially
18
 INTRODUCTION

Microbiology after Pasteur era: Robert Koch

 Studied anthrax and

hypothesized that it may be
caused by bacteria
 Invented agar plates for culture
of microbes
 Developed Koch’s postulates
 Germ theory -diseases are
caused by germs and not
because you have sinned or
done something wrong!

Robert Koch (1834-1910)

 INTRODUCTION
Microbiology after Pasteur era
Other significant findings

• 1876, Car Weigert used methylene blue (a

synthetic dye) to stain bacteria in aqueous
suspensions.
• 1878, Joseph Lister isolated Streptococcus
(now Lactococcus) lactis in pure culture by
serial dilution from sour milk.
➢ 1883 Emil Christian Hansen used pure
cultures of yeasts to ferment beer => Hansen
medium
➢ 1884, Hans Christian Gram developed Gram
staining of bacterial cells
➢ 1887, Julius Richard Petri, German
microbiologist designed Petri plate
➢ 1887, Forster demonstrated the ablity of pure
cultures of bacteria grow at 0°C
 INTRODUCTION

Microbiology after Pasteur era

Other significant findings
➢ 1894 , J. Denys associated pyogenic
Staphylococcus with death of a
person who ate meat prepared from
a diseased cow.
➢ 1895 Marie von Ermengem isolated
Bacillus botulinus (Clostridium
botulinum) from contaminated meat
and proved that it caused botulism.
➢ 1896, Marie von Ermengem proved
that Salmonella enteritidis caused a
fatal disease in humans who
consumed contaminated sausage.
➢ 1915, Bacillus coagulant, first isolated
from coagulated milk by Hammer
 INTRODUCTION

Microbiology after Pasteur era

Almost modern
 1910- Paul Ehrlich
introduces the first
medication to treat
bacterial infection,
Salvarsan, an arsenic
derivative to treat syphilis
(giang mai)
 1928-1940s- Alexander
Fleming discovers penicillin
from mold Penicillium
notatum
=> Development of antibiotics
 INTRODUCTION
CURRENT STATUS

Food Food
Spoilage/Food Fermentation
Preservation /Probiotics

FOOD
MICROBIOLOGY

Foodborne Food
Diseases management
 24

Food microorganisms: Classification,

diversity, and characteristics
Bacteria, yeasts, molds, and viruses are
important in food for their ability to:
➢ cause foodborne diseases.
➢ cause food spoilage.
➢ produce food and food ingredients
 Microorganisms
classification

With nucleus Without nucleus

Molds Yeasts Bacteria Archaea

Gram stain
Morphology Protein profile
Reproduction Base composition C+G %
Metabolic patterns (10%)
DNA RNA hybridization (90%)
Nucleotide sequence.
 INTRODUCTION

Remind: General Microbiology

Some elements and tools for succinct

classification of microorganisms
bacteria based on

➢ Phenotypic characters

➢ Metabolic characteristics

➢ Microbial growth (yeast, fungi,

bacteria and phages (viruses)
differentiation and kinetics

➢ Microorganisms in foods
 27 INTRODUCTION
Struture of procaryote
 28 INTRODUCTION

Some elements and tools for succinct classification of

microorganisms bacteria based on phenotypic characters

➢ Form, association, GRAM staining Trophic / energy type

➢ Metabolism of substrates in fermentation respiration (sugars,
etc...)
➢ Presence of certain enzymatic activities (catalase etc.)
➢ Production of various metabolites (H2S, Indole, etc.)
➢ Optimum growth temperature (psychrotrophic, mesophilic,
thermophilic)
➢ Optimal growth pH (acidophilus, neutrophile, basophil)
➢ Tolerance to certain compounds (deoxycholate, salt, Tipol
etc.)
=> Manual tests or through automation
 INTRODUCTION
Classification - Bacteria 29

1. By Shape

Most of bacteria: 0.5–1.5 X 2.0–10 µm in size

2. By diameter
Ex: E.coli 1.5 x 2 µm
 30 INTRODUCTION

Taxonomy : level of classification

31 INTRODUCTION
32 INTRODUCTION
 INTRODUCTION

Scientific nomenclature
Given each microbes 2 names:
• Genus: capitalized
• Species: lowercalse
Both italicized or underlined
Ex: Saccharomyces cerevisiae (S.cerevisiae)
Inspiration for names is varied and imaginative!
(find a new species => new name S. hanoimensis is
possible!!
Escherichia coli: Named by Dr. Escherich, found in colon

Staphylococcus aureus: Looked like a cluster of grapes

(staphylococcus) and was golden yellow (aureus)

Saccharomyces cerevisiae: Myc-fungi who eat sugar

(saccharo) and make cerveza (beer)
 INTRODUCTION
Factors affecting food microorganisms growth
(a). Intrinsic factors
 35 INTRODUCTION

Factors affecting food microorganisms

growth
(a). Intrinsic factors - pH
▪ Important in determining which organisms
can survive and thrive on specific foods
▪ Most bacteria grow best at neutral or weakly
alkaline pH usually between 7 and 8.
▪ Some bacteria can grow within a narrow pH
range of 4.5 and 9.0, e.g. Salmonella
▪ molds grow between 1.5 and 11.0
yeasts grow between 1.5 and 8.5.
 36 INTRODUCTION

(a). Intrinsic factors - pH

 INTRODUCTION

(a). Intrinsic factors - pH

pH 1 2 3 4 5 6 7 8 9 10
Food Molds
microorg Yeast
anisms
lactic bacteria
Staphyloccocus aureus
Acetobacer sp.
E. Coli
Clostridium botulinum
Bacillus cereus
Vibrio sp.
 INTRODUCTION

 Water Activity – Aw: Amount of Water available for

chemical reactions and microbial growth

 0 1
No water Pure water
 Most Fresh Foods - Aw > 0.99 !
 39 INTRODUCTION
Water activity
AW: Molds< Yeasts < Bacteria

• Spoilage Bacteria 0.91

• Spoilage Yeasts 0.88
• Spoilage Molds 0.80
Water activity lowered by:
➢ drying
➢ addition of salt or sugar
➢ freezing
 40 INTRODUCTION
Water activity
 INTRODUCTION

(b). Extrinsic factors

Factors external to the food that affect

microbial growth.
1. Temperature
2. Oxygen
3. Relative humidity of storage
 INTRODUCTION
Temperature Sterilisation

 The most important

factor affecting Pasteurisation
microbial growth
 Optimum: 37°C

 Lower temperatures
retard microbial growth !!!

 Higher temperatures
eliminate microbes

 Refrigeration at 5°C Cooling

retards but does not stop
microbial growth Freezing
 Microorganisms can still
cause spoilage with Frozen food
extended spoilage
 Growth at temperatures
below -10°C has been
observed
 INTRODUCTION

Factors affecting food microorganisms growth

Temperature
 INTRODUCTION
Temperature
Hyperthermophiles grow at > 80oC to 130oC (ex.
Thermotoga)
Thermophiles grow at > 45oC. Often their optimum growth
temperatures is 50oC-70oC.
Bacteria in this group are mainly spore formers and are of
importance in the food industry especially in processed
foods.
Mesophiles grow between 25°C and 40°C, with an
optimum growth temperature close to 37 °C
Some such as Pseudomonas aeroginosa may grow at even
lower temperatures, between 5-43 ° C
None of the mesophilic bacteria are able to grow < 5°C or
> 45°C.
Most pathogenic and beneficial bacteria belong to this
group.
Psychrotrophs grow and reproduce between -10 to 25oC,
cause contamination maily in some dairy products
Psychrophiles or cryophiles capable of growth and
reproduction, in low temperatures, ranging from −20 °C[1]
to +10 °C.
Cause food spoilage at very low temperatures
Present in polar ice,glaciers, snowfields and deep
ocean waters
 INTRODUCTION
Factors affecting food microorganisms growth
Temperature
To prevent growth of unwanted microbes

To eliminate them.....
 INTRODUCTION
Oxygen

 Aerobic– must have oxygen Pseudomonas aeruginosa

 Anaerobic - no oxygen Clostridium botulinum
 Micro-aerophiles- small amounts of oxygen
Helicobacter pylori

 Facultative anaerobes – both aerobic and

anaerobic Escherichia coli, Listeria, Staphylococcus
 47
INTRODUCTION
Oxygen

▪ Anaerobic or facultatively anaerobic

sporeformers are most likely to grow in canned
foods .
▪ Microaerophilic bacteria are most likely to grow in
vacuum packed foods since they have low
oxygen tension
▪ Aerobic bacteria are likely to grow on the surface
of raw meat.
▪ Aerobic molds will grow in insufficiently dried or
salted products
 48 INTRODUCTION
Oxygen

Preservation: Changes with

packaging – canning, MAP
(modified atmosphere
packaging), vacuum
packaging
 49 INTRODUCTION

Relative humidity

• Relative humidiy is the amount

of moisture in the atmosphere
or food environment.
• Foods with low water activity
placed at high humidity
environment take up water,
increase their water activity
and get spoiled easily.
• For example, dry grains stored
in a environment with high
humidity will take up water and
undergo mold spoilage.