FT56-FOOD MICROBIOLOGY

History of Food Microbiology

• 7000 BC – Evidence that the Babylonians manufactured beer
(fermentation). Wine appeared in about 3500 BC.
• 6000 BC – The first apparent reference to food spoilage in
recorded history.
• 3000 BC – Egyptians manufactured cheese (fermentation) and
butter (fermentation, low aw). Several cultures also learned to use
salt (low aw) to preserve meat and other foods around this time.

• 1000 BC – Romans used snow to preserve shrimp (low temp),

records of smoked and fermented meats also appear.
• 1658- Athanasius Kircher reported that,
using a microscope, he had seen minute
living worms in putrid meat and milk. The
magnification power of his microscope was
so low that he could not have seen
bacteria.

• 1664- Robert Hooke described the

structure of molds.
• 1665 – An Italian physician by the
name of Francesco Redi
demonstrated that maggots on
putrefying meat did not arise
spontaneously but were instead the
larval stages of flies. This was the
first step away from the doctrine of
spontaneous generation.
• 1683 – Anton van Leeuwenhoek from the
Netherlands examined and described bacteria
through a microscope.
(person who discovered the microbial world).

• He observed bacteria in saliva, rainwater, vinegar, and

other materials; sketched the three morphological
groups (spheroids or cocci, cylindrical rods or bacilli,
and spiral or spirilla); and also described some to be
motile.
• He called them animalcules, and between 1676 and
1683 he reported his observations to the newly
formed leading scientific organization, The Royal
Society of London.
• 1765 – Italian named Spallanzani tried to
disprove the theory of spontaneous
generation of life by demonstrating that
beef broth which was boiled and then
sealed remained sterile. Supporters of
the theory discounted his work because
they believed his treatment excluded O2,
which they thought was vital to
spontaneous generation.
• 1795 – The French government offered
12,000 francs to anyone who could
develop a practical way to preserve
food. A French confectioner named
Nicholas Appert was issued the patent
after showing that meat could be
preserved when it was placed in glass
bottles and boiled. This was the
beginning of food preservation by
canning.
• 1837 – Schwann demonstrates that
healed infusions remain sterile in
the presence of air (which he
passed in through heated coils),
again to disprove spontaneous
generation.
• 1838- Ehrenberg (who introduced
the term bacteria) had proposed at
least 16 species in four genera

• 1875- Ferdinand Cohn had developed

the preliminary classification system
of bacteria.
• Cohn also was the first to discover
that some bacteria produced spores.
• The first person to really appreciate and understand the causal
relationship between microorganisms in infusions and the
chemical changes that took place in those infusions was Louis
Pasteur. Through his experiments, Pasteur convinced the
scientific world that all fermentative processes were caused by
microorganisms and that specific types of fermentations (e.g.
alcoholic, lactic or butyric) were the result of specific types of
microorganisms.

• In 1857 he showed that souring milk was caused by microbes and

in 1860 he demonstrated that heat destroyed undesirable
microbes in wine and beer. The latter process is now used for a
variety of foods and is called pasteurization. Because of the
importance of his work, Pasteur is known as the founder of food
microbiology and microbiological science. He demonstrated that
air doesn’t have to be heated to remain sterile using his famous
swan-necked flasks that finally disproved spontaneous generation.
 Microorganisms in the Food System
1. Helpful microorganisms 2. Spoilage microorganisms 3. Pathogenic
 FOOD MICROBIOLOGY: Microorganisms in Food
Current Status
Bacteria Viruses
A. Food Fermentation/
Probiotics

B. Food Spoilage
Fungi (Molds & Yeasts) Parasites

C. Food borne Illnesses

1. Food poisoning
2. Food-borne disease
 Important Bacterial Groups in Foods
Bacterial Groups Characteristics Example
1. Lactic Acid Bacteria Produce relatively large quantities of lactic acid Lactococcus, Leuconostoc, Pediococcus,
from carbohydrates . Lactobacillus, and Streptococcus thermophilus

2. Acetic Acid Bacteria Produce acetic acid. Acetobacter aceti

3. Propionic Acid Produce propionic acid and are used in dairy Propionibacterium freudenreichii
Bacteria fermentation.

4. Butyric Acid Bacteria Produce butyric acid. Clostridium butyricum

5. Proteolytic Bacteria Hydrolyze proteins because they produce Micrococcus, Staphylococcus, Bacillus, Clostridium,
extracellular proteinases. Pseudomonas, Alteromonas, Flavobacterium,
Alcaligenes, some in Enterobacteriaceae, and
Brevibacterium

6. Lipolytic Bacteria Able to hydrolyze triglycerides because they Micrococcus, Staphylococcus, Pseudomonas,
produce extracellular lipases. Alteromonas, and Flavobacterium
 Bacterial Groups Characteristics Example
7. Saccharolytic Able to hydrolyze complex carbohydrates Bacillus, Clostridium, Aeromonas, Pseudomonas,
Bacteria and Enterobacter

8. Thermophilic Able to grow at 50°C and above Bacillus, Clostridium, Pediococcus, Streptococcus,
Bacteria and Lactobacillus

9. Psychrotrophic They are bacteria that are able to grow at Pseudomonas, Alteromonas, Alcaligenes,
Bacteria refrigerated temperature (≤5°C). Flavobacterium, Serratia, Bacillus, Clostridium,
Lactobacillus, Leuconostoc, Carnobacterium,
Brochothrix, Listeria, Yersinia, and Aeromonas

10. Thermoduric Able to survive pasteurization temperature Micrococcus, Enterococcus, Lactobacillus,

Bacteria treatment Pediococcus, Bacillus (spores), and Clostridium
(spores).
11. Halotolerant They are bacteria that are able to survive high Bacillus, Micrococcus, Staphylococcus,
Bacteria salt concentrations (≥10%) Pediococcus, Vibrio, and Corynebacterium

12. Aciduric Bacteria Able to survive at low pH (<4.0) Lactobacillus, Pediococcus, Lactococcus,
Enterococcus, and Streptococcus
 Bacterial Groups Characteristics Example
13. Osmophilic Bacteria Can grow at a relatively higher osmotic Staphylococcus, Leuconostoc, and Lactobacillus
environment than that needed for other
bacteria.
14. Gas-Producing Produce gas (CO2, H2, H2S) during Leuconostoc, Lactobacillus, Propionibacterium,
Bacteria metabolism of nutrients. Escherichia, Enterobacter, Clostridium, and
Desulfotomaculum

15. Slime Producers Produce slime because they synthesise Xanthomonas, Leuconostoc, Alcaligenes,
polysaccharides. Enterobacter, Lactococcus, and Lactobacillus

16. Spore Formers Have the ability to produce spores . Bacillus, Clostridium, and Desulfotomaculum

They are further divided into aerobic

sporeformers, anaerobic sporeformers, flat
sour sporeformers, thermophilic
sporeformers, and sulfide-producing
sporeformers.
 Bacterial Groups Characteristics Example

17. Aerobes Require oxygen for growth and Pseudomonas, Bacillus, and Flavobacterium
multiplication.

18. Anaerobes Cannot grow in the presence of oxygen . Clostridium

19. Facultative Able to grow in both the presence and Lactobacillus, Pediococcus, Leuconostoc, enteric
Anaerobes absence of oxygen. pathogens, and some species of
Bacillus, Serratia, and coliforms

20. Coliforms Used as an index of sanitation. Escherichia, Enterobacter, Citrobacter, and

Klebsiella

21. Fecal Coliforms Also used as an index of sanitation. Escherichia coli

22. Enteric Pathogens Cause gastrointestinal infection. Pathogenic Salmonella, Shigella, Campylobacter,
Yersinia, Escherichia, Vibrio, Listeria, hepatitis A
 Important Fungi, Viruses and Parasites in Foods

Specific Group Examples of Microorganisms

Molds - produce mycotoxins (food-borne intoxication)
(Aspergillus, Alternaria, Fusarium, Geotrichum, Mucor, Penicillium, Rhizopus)

Yeast - important in food because of their ability to cause spoilage.

- used in food bioprocessing and to produce food additives.
(Saccharomyces, Pichia, Rhodotorula, Torulopsis, Candida, Zygosaccharomyces)

Virus - Hepatitis A and Norwalk-like viruses

-Enteric viruses (poliovirus, echo virus, and Coxsackie virus)

Parasites Trichinella spiralis, Angiostrongylus cantonensis, Trichuris trichiura, etc.

 Natural Sources
Foods of plant origin: Foods of animal origin:
• surfaces of fruits, vegetables and grains • skin, hair, feathers, gastrointestinal tract,
• pores in some tubers (e.g., radish and onion). urinogenital tract, respiratory tract, and milk
ducts (teat canal) in udders of milk animals.

- harbor pathogens such as Salmonella serovars, pathogenic Escherichia

coli, Campylobacter jejuni, Yersinia enterocolitica, and Listeria
monocytogenes without showing symptoms.
- Molds, yeasts, lactic acid bacteria, and bacteria from genera
-Laying birds have been suspected of asymptomatically carrying
Pseudomonas, Alcaligenes, Micrococcus, Erwinia, Bacillus,
Salmonella Enteritidis in the ovaries and contaminating the yolk during
Clostridium, and Enterobacter can be expected from this
ovulation.
source.
- Fish and shellfish also carry normal microflora in the scales, skin, and
- Pathogens, especially of enteric types, can be present if the
digestive tracts (Vibrio parahaemolyticus, Vib. vulnificus, and Vib.
soil is contaminated with untreated sewage.
cholerae are of major concern from these sources).
An understanding of the sources of microorganisms in food is important
to:
• Develop methods to control access of some microorganisms in the
food.
• Develop processing methods to kill them in food.
• Determine the microbiological quality of food, as well as set up
microbiological standards and specifications of foods and food
ingredients.
Air Water
• Microorganisms are present in dust • Although potable water does not contain
and moisture droplets in the air. coliforms and pathogens (mainly enteric
• Generally, dry air with low dust content types), it can contain other bacteria capable of
and higher temperature has a low causing food spoilage, such as Pseudomonas,
microbial level. Alcaligenes, and Flavobacterium.
• Spores of Bacillus spp., Clostridium
spp., and molds, and cells of some • Can also be a source for viral and parasitic
Grampositive bacteria (e.g., infection.
Micrococcus spp. and Sarcina spp.), as
well as yeasts, can be predominantly
present in air.
Soil Equipment
• Because microorganisms can multiply in • Small equipment, such as cutting boards,
soil, their numbers can be very high knives, spoons, and similar articles, because
(billions/g). of improper cleaning, can be sources of cross
contamination.
• Many types of molds, yeasts, and
bacterial genera (e.g., Enterobacter,
• Salmonella, Listeria, Escherichia,
Pseudomonas, Proteus, Micrococcus,
Enterococcus, Micrococcus, Pseudomonas,
Enterococcus, Bacillus, and Clostridium)
Lactobacillus, Leuconostoc, Clostridium,
can enter foods from the soil.
Bacillus spp., and yeasts and molds can get in
• Source of enteric pathogenic bacteria, food from equipment.
parasites and viruses in food.
Food Handlers Food Ingredients
• The presence of minor cuts and infection in • Can be a source of both spoilage and
hands and face and mild generalized diseases pathogenic microorganisms.
(e.g., flu, strep throat, or hepatitis A in an early
stage) can amplify the situation. • Various spices generally have very high
• In addition to spoilage bacteria, pathogens populations of mold and bacterial spores.
such as Staph. aureus, Salmonella serovars,
Shigella spp., pathogenic E. coli, and hepatitis A
can be introduced into foods from human
sources.
• Foods might be contaminated
with microorganisms from
several other sources, namely:
• packaging and wrapping
materials,
• containers,
• flies, vermins, birds, house
pets, and rodents.
 INTRINSIC FACTORS (FOOD ENVIRONMENT)

Intrinsic factors of a food include:

1. pH
2. Moisture content
3. Oxidation–reduction potential (Eh)
4. Nutrient content
5. Antimicrobial constituents
6. Biological structures
 pH • In general, molds and yeasts are able to grow
at lower pH than do bacteria, and Gram-
• most microorganisms grow best negative bacteria are more sensitive to low pH
at pH values around 7.0 (6.6– than are Gram-positive bacteria.
7.5)
• The pH range of growth for molds is 1.5 to 9.0;
for yeasts, 2.0 to 8.5; for Gram-positive bacteria,
Classification of foods based on pH 4.0 to 8.5; and for Gram-negative bacteria, 4.5
requirement: to 9.0.
• high-acid foods (pH below 4.6)
• low-acid foods (pH 4.6 and • When the pH in a food is reduced below the
lower limit for growth of a microbial species, the
above). cells not only stop growing but also lose
viability, the rate of which depends on the
extent of pH reduction.
 Water Activity (Aw)
• In general, the minimum Aw values for growth of microbial
• The free water in a food is groups are as follows:
-most molds, 0.8, with xerophilic molds as low as 0.6;
necessary for microbial growth. -most yeasts, 0.85, with osmophilic yeasts, 0.6 to 0.7;
-most Gram-positive bacteria, 0.90; and Gram-negative
bacteria, 0.93. (exceptions are growth of
• It is necessary to transport Staphlococcus aureus at 0.85 and halophilic bacteria
at 0.75).
nutrients and remove waste
materials, carry out enzymatic • When the Aw is reduced below the minimum level required
reactions, synthesize cellular for growth of a microorganism, the cells remain viable for a
while.
materials, and take part in other
biochemical reactions, such as • But if the Aw is reduced drastically, microbial cells in a
hydrolysis of a polymer to population lose viability, generally rapidly at first and then
more slowly.
monomers.
 Oxygen Requirement
• Aerobes need free oxygen for energy • Anaerobes (obligate or strict anaerobes)
generation, as the free oxygen acts as cannot grow in the presence of even small
the final electron acceptor through amounts of free oxygen as they lack the
aerobic respiration.
superoxide dismutase necessary to
• Facultative anaerobes can generate scavenge the toxic oxygen free radicals.
energy if free oxygen is available, or
they can use bound oxygen in • Microaerophiles grow better in the
compounds such as NO3 or SO4 as presence of less oxygen.
final electron acceptors through
anaerobic respiration.
• If oxygen is not available, then other
compounds are used to accept the
electron (or hydrogen) through
(anaerobic) fermentation.
 Nutrient Content
Carbohydrates Proteins
• All microorganisms normally found in • Microorganisms differ greatly in their
food metabolize glucose, but their ability to metabolize food proteins.
ability to utilize other carbohydrates • Most transport amino acids and small
differs considerably. peptides in the cells; small peptides are
• This is because of the inability of some then hydrolyzed to amino acids inside
microorganisms to transport the
specific monosaccharides and the cells.
disaccharides inside the cells and the • Microorganisms also produce
inability to hydrolyze polysaccharides extracellular proteinases and peptidases
outside the cells. to hydrolyze large proteins and peptides
• Molds are the most capable of using to small peptides and amino acids before
polysaccharides. they can be transported inside the cells.
 Nutrient Content
Lipids Minerals and Vitamins
• Lipids are, in general, less • Microorganisms need several
preferred substrates for the elements in small amounts,
microbial synthesis of energy and such as phosphorous, calcium,
cellular materials. magnesium, iron, sulfur,
• Some beneficial intestinal manganese, and potassium.
microorganisms, such as
Lactobacillus acidophilus strains, • Some microorganisms can
can metabolize cholesterol and are synthesize vitamins in foods .
believed to be capable of reducing
serum cholesterol levels in
humans.
Antimicrobial Constituents • Milk casein as well as some free fatty acids have been shown to
be antimicrobial under certain conditions.

• Eggs contain lysozyme, as does milk, and this enzyme, along

• The inhibitors, depending on their with conalbumin, provides fresh eggs with a fairly efficient
mode of action, can prevent or reduce antimicrobial system.
growth of and kill microorganisms.
• The hydroxycinnamic acid derivatives (p-coumaric, ferulic,
caffeic, and chlorogenic acids) found in fruits, vegetables, tea,
molasses, and other plant sources all show antibacterial and
• eugenol in cloves, allicin in garlic, cinnamic
some antifungal activity.
aldehyde and eugenol in cinnamon, allyl
isothiocyanate in mustard, eugenol and • Lactoferrin is an iron-binding glycoprotein that is inhibitory to a
thymol in sage, and carvacrol (isothymol) number of foodborne bacteria and its use as a microbial blocking
and thymol in oregano. agent on beef carcasses.

• Ovotransferrin appears to be the inhibitory substance in raw

• Cow’s milk contains several antimicrobial egg white that inhibits Salmonella enteritidis.
substances, including lactoferrin ,
• Cell vacuoles of cruciferous plants (cabbage, Brussels sprouts,
conglutinin, and the lactoperoxidase broccoli, turnips, etc.) contain glucosinolates, which upon injury
system. or mechanical disruption, yield isothiocyanates.
 Biological Structures
• The natural covering of some foods
provides excellent protection against the
entry and subsequent damage by
spoilage organisms.

• structures as the testa of seeds, the

outer covering of fruits, the shell of nuts,
the hide of animals, and the shells of
eggs.

• The skin covering of fish and meats such

as beef and pork prevents the
contamination and spoilage of these
foods, partly because it tends to dry out
faster than freshly cut surfaces.
 EXTRINSIC PARAMETERS
• The extrinsic parameters of foods are not substrate dependent. They
are those properties of the storage environment that affect both the
foods and their microorganisms. Those of greatest importance to the
welfare of foodborne organisms are as follows:

1. temperature of storage
2. relative humidity of environment
3. presence and concentration of gases
4. presence and activities of other microorganisms
 Temperature of Storage
• The lowest temperature at which a Psychrotrophs - microorganisms that
microorganism has been reported grow well at or below 7◦C and have their
to grow is −34◦C; the highest is optimum between 20◦C and 30◦C.
somewhere in excess of 100◦C.
Mesophiles - those that grow well
between 20◦C and 45◦C with optima
between 30◦C and 40◦C.

Thermophiles - those that grow well at

and above 45◦C with optima between
55◦C and 65◦C.
 Relative Humidity of
Environment
• The RH of the storage environment
is important both from the
standpoint of Aw within foods and
the growth of microorganisms at the
surfaces.

• Foods that undergo surface spoilage

from molds, yeasts, and certain
bacteria should be stored under
conditions of low RH.
Presence and Concentration of Gases in the Environment
• Carbon dioxide (CO2) is the single most important atmospheric gas that
is used to control microorganisms in foods.
• Carbon dioxide (CO2) and Oxygen (O2) are the two most important
gases in modified atmosphere packaged (MAP) foods.
• Ozone (O3) is the other atmospheric gas that has antimicrobial
properties, and it has been tried over a number of decades as an agent
to extend the shelf life of certain foods.
 Presence and Activities of other Microorganisms
• Some foodborne organisms produce substances that are either
inhibitory or lethal to others.

• These include antibiotics, bacteriocins, hydrogen peroxide, and organic

acids.
END.
Microbial Growth Characteristics
MICROBIAL REPRODUCTION OR GROWTH

- An increase in number or mass of • The process includes several steps, such as DNA
vegetative cells of bacteria, yeasts, replication and separation, partition between the DNA,
invagination and septum formation of wall and cell
and molds is customarily used to separation.
reflect growth for microorganisms. • In this process, one cell asexually divides into two
BACTERIA cells, each an essentially true replica of the original cell.
In bacteria, a form of sexual recombination can occur
Binary Fission that involves transfer of genetic materials from a donor
to a recipient cell (e.g., conjugation).
- Bacteria reproduce by a process
• However, this is quite different from sexual
called transverse binary fission, or, reproduction, which is facilitated through the union of
two cells (gametes) of opposite mating type.
simply, binary fission.
 FUNGI VIRUSES
• Yeasts and molds can also reproduce
asexually. • Viruses do not reproduce by themselves.
• A yeast cell produces a bud that
initially is much smaller in size and • Instead, they attach on the surface and
remains attached to the surface of inject their nucleic acid inside specific
the original cell. host cells, which then replicate the viral
• Molds can grow in size by cell division nucleic acid and produce viral particles.
or elongation at the tip of a hypha.
• They can also form large numbers of
asexual spores on specialized
structure.
• Both yeasts and molds can also
reproduce sexually.
Generation Time (or Doubling Time)
Generation time - the time that a single cell
takes to divide into two.

• However, in practice, generation time is

referred to as the doubling time for the Where:
entire population. G - is generation time (minutes)
0.3 - constant (value of log102, indicates
• In a population of a microbial species, not doubling)
all cells divide at the same time or at the t - duration of study (min)
same rate. log10x - initial
log10z - final cell numbers per milliliter or
• In general, under optimum conditions of colony-forming units (CFUs) per
growth, bacteria have the shortest milliliter.
generation time, followed by yeasts and
molds.
 Optimum Growth

• When the cells of a microbial • The growth range and optimum

species are exposed to a factor growth of a microorganism under a
(e.g., temperature) beyond the specific parameter provide valuable
growth range, the cells not only information for its inhibition,
stop growing, but, depending on reduction, or stimulation of growth
the situation, may be injured or in a food.
lose viability.
Growth Curve
 NATURE OF MICROBIAL GROWTH IN FOOD
• Mixed Population • Growth in Succession or Diauxic Growth
- in a mixed population, the intrinsic - Microorganisms that can
and extrinsic environments dictate metabolize two or more nutrients in a
which one, two, or a few in the initial food, one preferred over the other and
mixed population will become
predominant and produce specific present in limiting concentrations, show
changes in a food. growth in stages separated by a short lag
phase.
• Sequence of Growth - Initially a bacterial strain grows by
- depending on the environment, one utilizing the preferred nutrient and after a
or two types may grow optimally and short lag of adaptation grows by utilizing
create an environment in which they
can no longer grow rapidly. the other nutrient.
• Symbiotic Growth Synergistic Growth
- Symbiosis, or helping one another, during - This is observed during symbiotic
growth often occurs in food containing two growth of two or more microbial types in a
or more types of microorganisms. food.
- One type may produce metabolic - In synergistic growth, each type is
products that the second type needs for capable of growing independently and
proper growth, but cannot produce by producing some metabolites at lower rates.
itself.
Antagonistic Growth
- In turn, the second species produces a - Two or more types of microorganisms
nutrient that stimulates the first one to present in a food can adversely affect the
grow better. growth of each other, or one can interfere with
the growth of one or more types; sometimes
one can kill the other.
END….