Foood Microbiology 2

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MCB 403: Food Microbiology (3 Unit)

The course will examine the relationship between micro –organism and food substances
with respect to food production and nutrition. The distribution, role and significance of
micro-organisms in food. The intrinsic and extrinsic parameters of food that effect
microbial growth. Food spoilage and food borne diseases, indices of food sanitary growth
and food micro-biological standards. History of micro-organisms in food. Spoilage of
foods: meat, fish, cereals, cereal produce, dairy products, canned foods. Micro-organisms
associated with local foods such as Ogi (akamu), Ugba, Palmwine, Bread, Garri, Ogiri,
Kulikuli, Nono, etc. Lactic acid fermentations: butter and cheese production, yoghurt
manufacture, pickles, sauerkraut etc. Foodborne diseases- infections and intoxications.
Food preservation and the growth curve. Indices of food sanitary growth and
microbiological standards. Quality control of food products. Food plant sanitation and
food control acts.

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INTRODUCTION
Food microbiology is the study of the microorganisms that inhabit, create, or contaminate
food, including the study of microorganisms causing food spoilage. Food microbiology
as the name infers is the aspect of microbiology that deals with food. Food
microbiologists must understand microbiology and food systems and be able to integrate
them to solve problems in complex food ecosystems.
Raw plants or animal food materials may be assumed to contain some microorganisms.
These microorganisms may be natural microflora or contaminant microflora. Natural
microflora present in the environment from which the food is obtained. Contaminant
microflora is those present from external sources. The natural microflora of foods vary
with the food and location from which it was obtained e.g. Streptococcus lactis occurring
naturally in milk because the organism is found on the exterior of the udder & adjacent
areas. It then easily gets into the milk as it is been drawn.
Also, Mucor, Rhizopus & Bacillus spp. occurring commonly in foods because their
spores are present in large number in the air settling fast on foods. Pseudomonas and
Alcaligenes spp. are found on the hides, hooves, air and gut of sheep & cattle & are
therefore important sources of contamination on meat products.
Animals also have the normal intestinal floras swallowed with the food & water.
Some of the microorganisms are ejected with faecal material and if fresh meat is
improperly handled; it may be contaminated by such organisms. The frequency of
isolation of such organism from fresh meat may give an erroneous impression that those
particular organisms constitute a natural microflora on meat and water products whereas
they are really contaminant microflora.

In such instances, the plant materials obtained from the field are expected to carry the
spores or vegetative cell of the prevalent microorganism. Most micro-organisms normally

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resident on plants are bacteria except in situation where the soil is heavily infected by a
particular mould.
Fishes and other aquatic food materials have the indigenous microorganism of the waters
in sea and they are found as resident or natural microflora. The population and variety of
these microorganisms varies with changes in their population in water and the prevailing
environmental conditions such as pH, temperature, amount of dissolved O 2, amount of
organic materials etc. e.gs of organisms commonly associated with fishes are
Pseudomonas, Flavobacterium, Achromobacter, Aeromonas, Micrococcuus.
Pseudomonas species and spore forming bacteria are found on wheat and oats in
dry weather whereas the lactic acid bacteria like Leuconostoc and Lactobacillus spp. are
found in wet weather. Also, Salmonella and Shigella spp. are both gastro intestinal
pathogens, they are found on vegetables and fruits harvested from farm irrigated with
sewage water. The same organisms are also found on meat contaminated with faeces
from infected animals and can also be found in milk from infected animals especially if
the milk was not drawn aseptically.
Claviceps purpurea is a fungus and is referred to as ergot fungus, it occasionally
attacks wheat and barley but more commonly attacks age flour make from such infected
grains may be contaminated if improperly treated before milling. These occasionally
micro floras are more commonly referred to as food contaminant. The microorganism
present as natural or occasional microflora on raw food materials may bring about
desirable or undesirable changes in the food of supplied with necessary conditions for
growth e.g. Pseudomonas and Erwinwa spp. cause bacteria soft rot on vegetable
especially on injury sites such as points of excision and breaks in the epidermis.
Lactobacillus spp. produces souring in masses of vegetables packed together in wet,
warm conditions especially in vegetables rich in sugars. This souring is a desirable
change in vegetable preservation by a process referred to as pickling.

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Factors affecting microbial growth in foods
These factors are similar to those that affect the growth of microorganisms in culture.
1. Substrate availability
Microorganisms found associated with foods depends on such foods for nutrient
supply and substrate availability including specific nutrient requirement, availability
of nutrients in utilizable forms etc. As long as a substrate is not a limiting factor,
microbial growth may continue with attendant enzymatic changes that bring about
deterioration or flavor development e.g. some spp. of Staphylococcus responsible for
locust bean fermentation require biotin for growth; when this is lacking, growth does
not take place.
2. The physical state of the food i.e. either dried or frozen or dried, heated, fresh solid,
liquid also affects the types of microorganism that will survive in such foods. This is
because the physical state affects the chemical composition and as such determines
the enzymes that may be able to act.
3. Chemical Properties of the foods: This determines the extent of microbial growth.
All bacteria associated to foods are heterotrophic and they make use of common
proteins, CHOs and fat. Many bacteria ferment CHOs and some ferment amino acids
to produce a variety of organic acids and simple sometimes odorous molecules e.g.
some spp. of Pseudomonas e.g.
a. Syringae is actively protolytic and lipolytic. It produces stale, oxidized, rancid or
bitter flavours on vegetable.
b. Yeasts are also found on sugary foods because t hey derive energy from
converting sugar to CO2 and H2O. Sometimes, if ammonia is available, they
convert it into yeast cell substance.
c. Rhizopus and Mucor spp. are the most commonly occurring fungi occurring on
starchy and sugary foods such as sweet potato, eba, bread, cooked rice etc.
d. Leuconostoc mesenteroides metabolizes the fructose portion of sucrose molecule
to CO2, lactic acid, acetic acid or ethanol in addition to CO2 and lactic acid.

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4. Effect of Temperature: The temperature at which food is eaten have great
influence on the rate and extent of microbial growth e.g. members of the order
Mucorales such as Mucor and Rhizopus spp. occur on starchy foods ate at moderate
room temperature and produce stolons which run along the surface. Penicillium and
Aureobasidium spp. grow commonly on foods under refrigerator. Thammedium
spp. grow readily on cold store, cured, wheat. Streptococcus thermophilus can grow
when the cured derived from fermented milk is heated to a high temperature, the
organisms is therefore useful in cheese making.

Temperature is the most efficient means to control microbial growth. Based on their
tolerance of broad temperature ranges, microorganisms are roughly classified as follows:

1. Psychrophies grow only at refrigeration temperatures.


2. Psychrotrophs grow well at refrigeration temperatures, but better at room
temperature.
3. Mesophiles grow best at or near human body temperature, but grow well at room
temperature.
4. Thermophiles grow only at temperatures about as hot as the human hand can
endure, and usually not at all at or below body temperature. NOTE

a. Some psychrotrophic microorganisms grow very slowly in foods below freezing,


but usually not below 19°F. There are a few reports of growth, usually of molds, at
14°F, but no reliable reports of growth below that temperature. This means that the
standard storage temperature for frozen foods, O°F, does not permit microbial
growth. However, many microorganisms survive freezing.
b. Most psychrotrophs have difficulty growing above 90°F.
c. Most foodborne disease organisms are mesophiles. A good rule of thumb is to
store perishable foods below 40°F or above 140°F.
d. In the temperature range where both mesophilic and psychrotrophic organisms
grow (about 41°F. to about 90°F), the psychrotrophs grow more rapidly, causing

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spoilage and at the same time frequently interfering with the growth of foodborne
disease organisms.

5. Ionic Requirements: Members of the family, Halobacteriaceae are halophilic.


They are found growing and causing spoilage of salted ocean fish that is improperly
refrigerated. E.g. Pediococcus thalophilus grow well in 7% NaCl solution and the
organism is useful in oriental fermentation of soya beans (soya-iru). Lactic acid
bacteria tolerate high salt concentrations. The salt tolerance gives them an advantage
over other less tolerant species and allows the lactic acid fermenters to begin
metabolism, which produces acid, which further inhibits the growth of non-desirable
organisms. Leuconostoc is noted for its high salt tolerance and for this reason,
initiates the majority of lactic acid fermentations.

6.pH: pH is a term used to describe the acidity or alkalinity of a solution. At pH 7,


there is an equal amount of acid (hydrogen ion: H +) and alkali (hydroxyl ion: OH-),
so the solution is “neutral”. pH values below 7 are acidic, while those above 7 are
alkaline. pH expresses the H + concentration logarithmically, that is, in multiples of
10. For example, at pH 5 there are 10 times as many H + as at pH 6; at pH 3 there
are 100 times as many H + as at pH 5, and so on.

pH has a profound effect on the growth of microorganisms. Most bacteria grow best at
about pH 7 and grow poorly or not at all below pH 4. Yeasts and molds, therefore,
predominate in low pH foods where bacteria cannot compete. The lactic acid bacteria are
exceptions; they can grow in high acid foods and actually produce acid to give us sour
milk, pickles, fermented meats, and similar products. Leuconostoc contribute off-flavors
to orange juice.

Mean pH Values of Selected Foods (Lopez, 1987)


pH Value Selected Foods
2.3 Lemon juice (2.3), Cranberry sauce (2.3)
3.0 Rhubarb (3.1)

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Applesauce (3.4), Cherries, RSP (3.4)
Berries (3.0 – 3.9), Sauerkraut (3.5)Peaches (3.7), Orange juice (3.7)
Apricots (3.8)
Cabbage, red (4.2), Pears (4.2)
4.0
Tomatoes (4.3)
Ravioli (4.6)
4.6
Pimientos (4.7)
Spaghetti in tomato sauce (4.9)
Figs (5.0)Onions (5.2)
5.0
Carroes (5.2)
Green Beans (5.3), Beans with pork (5.3)Asparagus (5.5), Potatoes (5.5)
Lima beans (5.9), Tuna (5.9), Tamales (5.9)
Codfish (6.0), Sardines (6.0), Beef (6.0)
Pork (6.1), Evaporated milk (6.1)
6.0
Frankfurters (6.2), Chicken (6.2)
Corn (6.3)
Salmon (6.4)
Crabmeat (6.8), Milk (6.8)
7.0 Ripe olives (6.9)
Hominy (7.0)

Many of the investigators who reported these values also determined that adverse
factors, such as low temperature or low water activity, increased the minimal pH for
growth. But the processor can be sure that these minimal values will prevent growth of
these pathogens under any and all circumstances.

7. Moisture: The requirement for water by microorganisms found on foods could be


in form of free liquid water or water droplets in the atmosphere. Fungi spores can
tolerate dry environment so they are more important on dry foods e.g. bread, grains,
rice, millet, wheat, sorghum & the flours of grain. Spores of Aspergillus are
important on these types of foodstuffs. Bacteria & yeast are found on foods with
free water like in water droplet on foods & vegetable surface & wet foods etc.
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8. Inhibitors: Such as toxins and other metabolites that may slow down or prevent
the growth of other microorganisms in the food e.g. species of Penicillium,
Aspergillus & Claviceps purpurea produce antimicrobial toxins but they are also
poisonous when foods containing such toxins are ingested by man.
9. Associations : In food fermentations there is usually a succession of organism on
the substrates e.g. bacteria starts the digestion & fermentation of starchy foods like
corn; when some acids has been formed and the pH is lowered, yeast then continues
the fermentation because they are more acid tolerant. This type of association is
synergistic. Various other associations between food microorganisms may be
antagonistic e.g. Penicillium produces penicillin to prevent other organism from
colonizing the surface.

10. Oxygen: Oxygen is essential for growth of some microorganisms; these are called
aerobes. Others cannot grow in its presence and are called anaerobes. Still others
can grow either with or without oxygen and are called microaerophilic. Strict
aerobes grow only on food surfaces and cannot grow in foods stored in cans or in
other evacuated, hermetically sealed containers. Anaerobes grow only beneath the
surface of foods or inside containers. Aerobic growth is faster than anaerobic.
Therefore, in products where both conditions exist, such as in fresh meat, the surface
growth is promptly evident, whereas subsurface growth is not.
11. Water Activity: Water activity (aw) is a term describing the availability of water to
microorganisms. It is only roughly related to percent moisture. Pure water has an a w
of 1.00, and the atmosphere above the water in a closed container will have an
equilibrium relative humidity (ERH) of 100%. If we add an ounce of rocks to a
quart of water in such a container, the ERH and the aw will not change. But if we
add an ounce of salt, the ERH will fall to about 98 % and the aw to 0.98. Rocks do
not dissolve in water but salt does, thereby reducing the proportion of water that can
enter the atmosphere. Likewise, the amount of water available to microorganisms
present in the solution is reduced. Yet the percent moisture is the same in the
container with rocks as it is in the container with, salt, namely, 98%.
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The GMP regulations for low-acid canned foods defined water activity as the vapor
pressure of the food product divided by the vapor pressure of pure water under identical
conditions of pressure and temperature. The regulations define low-acid foods as foods,
other than beverages, with a finished equilibrium pH value greater than 4.6 and a water
activity greater than 0.85.

The water activity (aw) limits for growth of principal foodborne disease organisms.*
Microorganism Minimal aw for growth
Salmonella 0.945
Clostridium botulinum 0.95
Clostridium perfringens 0.93
Staphylococcus aureus 0.86**
Vibrio parahaemolyticus 0.94

*These limits are the lowest reported, with all other growth conditions optimal. If other
conditions are less than optimal, the minimal aw will be higher.

Most bacteria fail to grow in a food or other medium where the aw is lower than 0.94.
Bacteria require a higher aw than yeasts, which in turn require a higher aw than molds.
Thus, any condition that lowers the aw first inhibits bacteria, then yeasts, and finally
molds. But each species has its limits which are interrelated with other growth factors.
Table 2 gives the aw limits for growth of principal foodborne disease organisms held
under otherwise optimal conditions. Certain molds and bacteria can grow on fish
immersed in saturated salt solution where the aw is about 0.75. Some molds can grow in
foods with aw 0.62 – 0.65. At these lower limits, growths are very slow. The aw of fully
dried foods, such as crackers or sugar, is about 0.10 and such products are
microbiologically stable because of this factor alone. The stability of intermediate
moisture foods (aw 0.75 – 0.90), such as dried fruits, jams, and soft moist pet foods,
depends on combinations of factors, such as low aw, low pH, pasteurization, chemical
additives, and impervious packaging.

FOOD FERMENTATIONS

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Fermentation is a metabolic process that converts sugar to acids, gases and/or alcohol. It
occurs in yeast and bacteria, but also in oxygen-starved muscle cells, as in the case of
lactic acid fermentation. Fermentation is also used more broadly to refer to the bulk
growth of microorganisms on a growth medium. It involves a change in the chemical
composition of natural foods to being about desired or undesired flavor, textural changes
and improves the keeping quality of foods or it may cause spoilage of foods. These
changes are brought about by various microorganisms.
Fermentation is the anaerobic production of energy from CHOs by microorganisms to
alcohols, acids, gas produced as by products. The byproducts bring about flavor changes
to the food substance & may even be useful in preserving the foods. Fermentation in food
processing is the conversion of carbohydrates to alcohols and carbon dioxide or organic
acids using yeasts, bacteria, or a combination thereof, under anaerobic conditions.
Fermentation usually implies that the action of microorganisms is desirable.

Uses

The primary benefit of fermentation is the conversion of sugars and other carbohydrates
into preservative organic acids, e.g. converting juice into wine, grains into beer,
carbohydrates into carbon dioxide to leaven bread, and sugars in vegetables.

Food fermentation has been said to serve five main purposes:

 Enrichment of the diet through development of a diversity of flavors, aromas, and


textures in food substrates
 Preservation of substantial amounts of food through lactic acid, alcohol, acetic
acid, and alkaline fermentations
 Biological enrichment of food substrates with protein, essential amino acids, and
vitamins
 Elimination of antinutrients
 A decrease in cooking time and fuel requirement

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TYPES OF FERMENTATION

A) Alcoholic fermentations:
These are fermentation process in which ethanol & CO 2 are the major end
products. Yeasts especially of the Genera Saccharomyces and Torulopsis are very
important in this type of fermentation.
Energy obtained from simple sugars is reduced to CO 2 & water with ethanol e.g.
beer, wine, bread & liquors such as spirit, such like gin, whiskey, brandy, rum etc.
other important. Food materials obtained from alcoholic food materials includes
vitamins, simple cell proteins & dried food yeasts.

The first example of alcoholic food fermentation is bread.


1. Science of bread making

Bread is the product of baking a mixture of flour, water, salt, yeast and other ingredients.
The basic process involves mixing of ingredients until the flour is converted into a stiff
paste or dough, followed by baking the dough into a loaf.
To make good bread, dough made by any process must be extensible enough for it to
relax and to expand while it is rising. A good dough is extensible if it will stretch out
when pulled. It also must be elastic, that is, have the strength to hold the gases produced
while rising, and stable enough to hold its shape and cell structure.
Two proteins present in flour (gliadin and glutenin) form gluten when mixed with water.
It is gluten that gives dough these special properties. Gluten is essential for bread making
and influences the mixing, kneading and baking properties of dough.
Bread making involves the following basic steps: mixing ingredients, rising
(fermentation), kneading, second rising, baking and cooling
The moistened flour called dough is mixed with yeast & the yeast used in
Saccharomyces cerevisiae and allowed to stand in a warm place for 2-5 hours. The sugar
present is rapidly fermented by the yeast to ethanol & CO2. The CO2 is entrapped within
the dough & it causes it to rise while the alcohol is lost during baking. The dough must

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have good elasticity, strength and stability to withstand the pressure of its increasing gas
without breaking. α and β – amylases in flour hydrolyse the starch to maltose and the
yeast produce maltase and invertase & also zymase which hydrolyses maltose & sucrose
to simple sugars, CO2 & alcohol.

The salt add flavor and it toughens the gluten & also retards fermentation. Natural flours
are short of α – amylase & this can be supplemented by using fungi amylase.

2. BEER MAKING
Beers are made from grains such as barley, rice, maize, guinea corn, sorhugm etc.
In the case of barley, the grains are cleaned, soaked in water between 15-20 0C and
allowed to germinate, this process is called ‘malting’. The malt produced contains starch,
amylases, proteins and protelyotic enzymes. The malted grains are then usually dried by
kilning and stored for future use. The malt is crushed with other starchy materials refers
to as adjuncts, mix with water and allowed to stand at a control temperature for
hydrolysis of starch & protein to take place, this process is called mashing. After the
starch has been converted to sugar in a process known as saccharification, the mixture is
then boiled to inactive enzymes & filter. The filtrate is referred to as wort.
The filtrate is then treated with hops to give flavor and to act as preservative against
bacteria. It is filter again and the hopwort is pitched with the appropriate trait of
Saccharomyces cerevisiae, the process of which is an act of inoculating the wort
(pitching) with S. cerevisiae and allowed to ferment for about 10 days at low temperature
between 10-150C until it gets to the desired alcohol content usually between 3.6 – 5.2%.
After fermentation, the beer is allowed to mature in storage tanks at temperature near 0 0C
to encourage settling of proteins & other nitrogenous materials and yeast cells. The beer
is then filtered for preservation, carbonated, clarified by filtration again. It is then bottled
or canned. Variation in alcoholic concentration of different beer type is related to the
amount of starch conversion allowed during mashing, length of fermentation and
temperature of fermentation.

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Microbiologist duties include maintaining an active, healthy yeast population and
developing pure cultures for use. At the end of fermentation, the yeast is collected from
the tank & test for viability. This is done by staining method. The yeast cells are stained
with methylene blue, those cells that absorbed the methylene blue are dead cells while
others are viable.
Living cells
% viability= X 100
Total no .of cells
3. WINE MAKING
Wines are made from crushed grains called must. The soluble sugars present in the must
i.e. glucose & fructose are fermented naturally by a succession of organism naturally
present on grapes.
The most important of them are S. cerevisiae variety of ellipsoideus which ferments the
glucose more readily than fructose. However, S. elegans ferments fructose more than
glucose. Control fermentation could be by inoculating malt with starter – cultures of
microorganisms after first treating with SO2 in order to destroy the natural microflora on
grapes. The temperature is controlled in such a way that the rate of fermentation is
controlled and death of yeast cells is prevented. Long slow fermentations low
temperatures produce wines with flavor, bitterer than rapid fermentation at high
temperature. Usually temperatures between 7-180C are used for white wines, 12-180C for
sparkling wines, 21-270C for red wines; in order to allow the pigments and tannins to be
released from the skin. The alcohol content of wine is between 9-14%.
After fermentation, the wine is stored to allow maturing, clarified and bottled.

SPARKLING WINES
These are table – wines in which C0 2 is produced by a secondary fermentation. e.g
Champagne. This wine undergoes a second alcoholic fermentation in the bottle under
pressure. As the C02 thus produced carbonates the wine, is released & gives a typical
popping sound when the bottle is opened. During maturation, many wines undergo a
second fermentation called the malo-lactic fermentation caused by strains of
Lactobacillus, Leuconostoc, Pediococcus. They convert the malic acid which is always
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present in grapes to lactic acid, this give a sour flavor to the wine and this reduces the
alcoholic content. The malo-lactic fermentation is referred to thus as wine-spoilage.

B. LACTIC FERMENTATION
Lactic acid fermentation is caused by some fungi and bacteria. The most important lactic
acid producing bacteria is Lactobacillus. Other bacteria which produce lactic acid
include:

 Leuconostoc mesenteroides
 Pediococcus cerevisiae
 Streptococcus lactis
 Bifidobacterium bifidus.

This type of fermentation is employed in the food industry for the purposes of
preservation and development of flavor. Vegetables & milk products are the foods
most commonly preserved by lactic acid fermentation. Foods may be fermented
singly or in a mixture depending on the culture e.g. among the Orientals vegetables
such as cabbages, peppers, radish soy-beans may be fermented in a mixture.

The lactic acid bacteria belong to two main groups – the homofermenters and the
heterofermenters. The pathways of lactic acid production differ for the two.
Homofermenters produce mainly lactic acid, via the glycolytic (Embden–Meyerhof)
pathway. Heterofermenters produce lactic acid plus appreciable amounts of ethanol,
acetate and carbon dioxide, via the 6-phosphoglucanate/phosphoketolase pathway.
The glycolytic pathway is used by all lactic acid bacteria except Leuconostocs, group
III lactobacilli, oenococci and weissellas. Normal conditions required for this pathway
are excess sugar and limited oxygen. Axelsson (1998) gives an in-depth account of the
biochemical pathways for both homo- and hetero-fermenters.

Homolactic fermentation

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 The fermentation of 1 mole of glucose yields two moles of lactic acid;

C6H12O6      2 CH3CHOHCOOH
Glucose lactic acid

 Heterolactic fermentation
 The fermentation of 1 mole of glucose yields 1 mole each of lactic acid, ethanol
and carbon dioxide;

C6H12O6
CH3CHOHCOOH+ C2H5OH+ CO2
carbon
Glucose lactic acid+ ethanol+
dioxide

Major lactic acid bacteria in fermented plant products.

Homofermenter Facultative Obligate heterofermenter


homofermenter

Enterococcus faecium Lactobacillus bavaricus Lactobacillus brevis

Enterococcus faecalis Lactobacillus casei Lactobacillus buchneri

Lactobacillus acidophilus Lactobacillus coryniformis Lactobacillus cellobiosus

Lactobacillus lactis Lactobacillus curvatus Lactobacillus confuses

Lactobacillus delbrueckii Lactobacillus plantarum Lactobacillus coprophilus

Lactobacillusleichmannii Lactobacillus sake Lactobacillus fermentatum

Lactobacillus salivarius   Lactobacillus sanfrancisco

Streptococcus bovis   Leuconostoc dextranicum

Streptococcus thermophilus   Leuconostoc mesenteroides

Pediococcus acidilactici   Leuconostoc paramesenteroides

Pedicoccus damnosus    

Pediococcus pentocacus    

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 From Beuchat (1995).

Lactic acid fermentation is used throughout the world to produce speciality foods:

 Western world: yoghurt, sourdough breads, sauerkraut, cucumber pickles and


olives
 Middle East: pickled vegetables
 Korea: kimchi (fermented mixture of Chinese cabbage, radishes, red pepper, garlic
and ginger)
 Russia: kefir
 Egypt: laban rayab and laban zeer (fermented milks), kishk (fermented cereal and
milk mixture)
 Nigeria: gari (fermented cassava)
 South Africa : magou (fermented maize porridge)
 Thailand : nham (fermented fresh pork)
 Philippines : balao balao (fermented rice and shrimp mixture)

B1. FERMENTED VEGETABLES


The lactic and fermentation of vegetables originated from the preserving effect on the
product. The growth of lactic and bacteria during vegetable fermentations results in:
1) Restriction of the growth of undesirable organism and delay or prevention of the
normal spoilage.
2) Production of unique flavors because of the accumulation of organic acids or by
products giving a characteristic and distinct finished product.
Lactic acid fermentation also gives the sour taste to fermented vegetables such as
traditionally cultured sauerkraut and pickles. The sugars in the cabbage are converted into
lactic acid and serve as a preservative.
The fermentation of vegetables by natural means is usually initiated by Leuconostoc
mesenteroides. When the pH is lowered to about 5.2, species of Lactobacillus replace
them & further lower the pH. The Leuconostoc mesenteroides ferment the hexose sugars
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to about 50% lactic acid & CO2 & a mixture of ethanol & acetic acid. Pediococcus spp.,
Streptococcus faecalis & Lactobacillus plantarum are homo-fermenters; while
Leuconostoc mesenteroides & Lactobacillus brevis are heterofermenters.

(i) SAUERKRAUT
Sauerkraut is the clean, sound product, of characteristic flavor obtained by full
fermentation (lactic) of properly prepared and shredded cabbage in the presence of not
less than 2% nor more than 3% of salt. It contains not less than 1 1/2% of acid expressed as
lactic acid on completion of fermentation.

Lactic acid bacteria are the primary group of organisms involved in sauerkraut
fermentation. They can be divided into three groups according to their types and end
products:

Leuconostoc mesenteroides                an acid and gas producing coccus

Lactobacillus plantarum and L. cucumeris bacilli that produce acid and a small
amount of gas

Lactobacillus pentoaceticus               acid and gas producing bacilli


(L. brevis)

In addition to the desirable bacteria there are a range of undesirable micro-organisms


present on cabbage (and other vegetable material) which can interfere with the sauerkraut
process if allowed to multiply unchecked. The quality of the final product depends
largely on how well the undesirable organisms are controlled during the fermentation
process. Some of the typical spoilage organisms utilize the protein as an energy source,
producing unpleasant odours and flavours.

The fermentation process

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Shredded cabbage or other suitable vegetables are placed in a jar and salt is added.
Mechanical pressure is applied to the cabbage to expel the juice, which contains
fermentable sugars and other nutrients suitable for microbial activity. The first micro-
organisms to start acting are the gas-producing cocci (L. mesenteroides). These microbes
produce acids. When the acidity reaches 0.25 to 0.3% (calculated as lactic acid), these
bacteria slow down and begin to die off, although their enzymes continue to function.
The activity initiated by the L. mesenteroides is continued by the lactobacilli (L.
plantarum and L. cucumeris) until an acidity level of 1.5 to 2% is attained. The high salt
concentration and low temperature inhibit these bacteria to some extent. Finally, L.
pentoaceticus continues the fermentation, bringing the acidity to 2 to 2.5% thus
completing the fermentation.

The end products of a normal kraut fermentation are lactic acid along with smaller
amounts of acetic and propionic acids, a mixture of gases of which carbon dioxide is the
principal gas, small amounts of alcohol and a mixture of aromatic esters. The acids, in
combination with alcohol form esters, which contribute to the characteristic flavour of
sauerkraut. The acidity helps to control the growth of spoilage and putrefactive organisms
and contributes to the extended shelf life of the product. Changes in the sequence of
desirable bacteria, or indeed the presence of undesirable bacteria, alter the taste and
quality of the product.

Effects of temperature on sauerkraut process

The optimum temperature for sauerkraut fermentation is around 21ºC. A variation of just
a few degrees from this temperature alters the activity of the microbial process and
affects the quality of the final product. Therefore, temperature control is one of the most
important factors in the sauerkraut process. A temperature of 18º to 22º C is most
desirable for initiating fermentation since this is the optimum temperature range for the

18
growth and metabolism of L. mesenteroides. Temperatures above 22ºC favour the growth
of Lactobacillus species.

Effects of salt on the sauerkraut process

Salt plays an important role in initiating the sauerkraut process and affects the quality of
the final product. The addition of too much salt may inhibit the desirable bacteria,
although it may contribute to the firmness of the kraut. The principle function of salt is to
withdraw juice from the cabbage (or other vegetable), thus making a more favourable
environment for development of the desired bacteria.

Generally, salt is added to a final concentration of 2.0 to 2.5%. At this concentration,


lactobacilli are slightly inhibited, but cocci are not affected. Unfortunately, this
concentration of salt has a greater inhibitory effect against the desirable organisms than
against those responsible for spoilage. The spoilage organisms can tolerate salt
concentrations up to between 5 and 7%, therefore it is the acidic environment created by
the lactobacilli that keep the spoilage bacteria at bay, rather than the addition of salt.

In the manufacture of sauerkraut, dry salt is added at the rate if 1 to 1.5 kg per 50kg
cabbage (2 to 3%). The use of salt brines is not recommended in sauerkraut making, but
is common in vegetables that have low water content. It is essential to use pure salt since
salts with added alkali may neutralize the acid.

Use of starter cultures

In order to produce sauerkraut of consistent quality, starter cultures (similar to those used
in the dairy industry) have been recommended. Not only do starter cultures ensure
consistency between batches, they speed up the fermentation process as there is no time
lag while the relevant microflora colonize the sample. Because the starter cultures used
are acidic, they also inhibit the undesirable micro-organisms. It is possible to add starters
19
traditionally used for milk fermentation, such as Streptococcus lactis, without adverse
effect on final quality. Because these organisms only survive for a short time (long
enough to initiate the acidification process) in the kraut medium, they do not disturb the
natural sequence of micro-organisms. On the other hand, if Leuconostoc mesenteroides is
added in the early stages, it gives a good flavour to the final product, but alters the
sequence of subsequent bacterial growth and results in a product that is incompletely
fermented. If gas producing rods (for example L. pentoaceticus) are added to the
sauerkraut, this disturbs the balance between acetic and lactic acids - more acetic acid and
less lactic acid are produced than normal - and the fermentation never reaches
completion. If lactic acid, non-gas producing rods (L. cucumeris) are used as a starter,
again the kraut is not completely fermented and the resulting product is bitter and more
susceptible to spoilage by yeasts.

It is possible to use the juice from previous kraut fermentation as a starter culture for
subsequent fermentations. The efficacy of using old juice depends largely on the types of
organisms present in the juice and its acidity. If the starter juice has an acidity of 0.3% or
more, it results in a poor quality kraut. This is because the cocci which would normally
initiate fermentation are suppressed by the high acidity, leaving the bacilli with sole
responsibility for fermentation. If the starter juice has an acidity of 0.25% or less, the
kraut produced is normal, but there do not appear to be any beneficial effects of adding
this juice. Often, the use of old juice produces sauerkraut which has a softer texture than
normal.

(ii) PICKLES/CUCUMBER PICKLES


Cucumber pickles are prepared from immature cucumbers preserved in vinegar & lactic
acid containing salt i.e. NaCl. The cucumber are washed, packed in about 5% salt
solution & allowed to ferment for about 20 days. Leuconostoc mesenteroides,
Pediococcus spp. & Streptococcus faecalis are important in the process. During the
period, the cucumbers absorb some of the salt & lactic acid is produced while acetic acid
20
may also be present. Fresh cucumber may have a low count of the lactic acid bacteria. So
the outer leaves of cabbage may be introduced into the vats to serve as inoculums.

(iii) OTHER VEGETABLES


Vegetables like carrots, okro, green-beans, soyabeans etc. may also be preserved by lactic
acid fermentation & bringing. They may be soaked in brine solution & allow to ferment
naturally by a succession of organism as in other vegetables or they may first be blanched
& starter cultures introduced – blanching leads to a softening of the tissues, so soy-beans
& beans may be preserved by brining.
BRINING
The term refers to the treatment of food with Nacl for preservative purposes. It may be
done in a salt solution of 2-10% cone with the food immersed to allow the salt with the
food immersed to allow the salt to penetrate. Dried brining may be done by treating foods
with salt crystals i.e. salting. The NaCl results from the tissue fermentable sugars & other
nutrient needed by lactic acid bacteria for growth. It also inhibits or completely prevents
growth of most bacteria residing on the food surfaces. It contributes to flavor & texture of
the food. Generally, fermented vegetables are further preserved by canning or bottling; in
which both process create anaerobiosis for anaerobic conditions c further inhibit
microbial action.

B2. FERMENTED MILKS


Fresh milk left undisturbed get sour within one or 2 days depending on the prevailing
environmental condition i.e. the higher the temperature, the faster the rate of souring.
Milk souring is caused by a number of microorganisms many of which are contaminants
introduced into the milk during the milking process either by use of an unhygienic
condition e.g. dirty utensils or through the skin of the cow. Some of the natural
contaminants of milk include Streptococus lactis and S. kefir.

21
(i) YOUGHURT
Yoghurt is made from non-fat or low-fat milk. The milk is pasteurized, homogenized and
cooled to inoculation temperature. It is then inoculated with an equal number of
Streptococcus thermophilus and Lactobacillus bulgaricus. These bacteria ferment the
milk. They ferment the lactose & form lactic acid together with traces of acetaldehyde,
acetone and diacetatyl. All of which give yoghurts its characteristics flavor. Fruits or fruit
– flavor such as strawberry, raspberry, orange, pineapple etc. may be added but must be
pasteurized separately before adding to the yoghurt. After fermentation between 29-46 0C,
the yoghurt is cooled rapidly to 40C to prevent further acid production.
(ii) BUTTERMILK
Buttermilk is obtained by the fermentative activities of Streptococcus cremoris, S.
diacetylactis & Leuconostoc cremoris. The milk is first pasteurized. Low-fat milk or
skin-milk is used.
The first organism i.e. S. cremoris produces lactic acid while the latter two produce the
aroma. The product should have a pleasant acid flavor, a buttery aroma and a smooth
viscous texture. If the milk is deficient in citric acid 0.1% sodium citrate is added before
pasteurization. The shelf-life of butter milk is between 10-14 days.
(iii) BUTTER
This is made from milk – fat. Salt may be added to improve the flavor & keeping quality
e.g. butter anchor. The cream which is the layer of milk – fat globules, on the surface of
standing milk is first separated from the milk. This is done by agitating the milk in
centrifuges or cream separator.
The cream is then neutralized to pH 6.8 – 7.2 and then pasteurized to destroy pathogenic
organism and also to eliminate spoilage organism that may cause spoilage of the food &
thus the flavor.
The cream is then cool to 100 C, stored overnight at the same temperature to permit fat
crystallization. The cream is then churned until the fat breaks i.e. until discrete fat,
globules are formed and begin to separate, and this is usually in about 30-40 minutes.

22
Churning refers to agitating. Chilled water is then added to make the fat firm. The churn
is revolved a few times more and then drawn up the buttermilk. The fat is washed again
in clean water with the churn revolved again & then drained off the washed water. About
1.5 – 2% salt is added and the churn is revolved for about 15mins to work the butter.
Workings break the moisture into smaller droplets & improve the keeping quality – by
limiting the amount of the available nutrient within the droplets. The residual water
during working becomes incorporated into the fat particles. Streptococcus lactis, S.
cremoris sour the cream while Leuconostoc citrivorum and L. detranicum attack the citric
acid to produce diacetyl which give butter its characteristic aroma.

(iv) CHEESE MAKING


Lactic acid bacteria as starter cultures are central to cheese production. This natural
process is a slow process. So modern cheese making makes use of an enzyme (rennin)
that occurs in the abomasums of calf’s stomach. They begin the process by degrading
proteins in the starting material. The proteases of the starter culture, as well as the rennet
in curd and the plasmin in milk, degrade proteins, namely casein, to produce free amino
acids and peptides. These amino acids not only contribute to the basic taste of cheese,
but are also essential precursors to the volatile aroma compounds that are responsible for
the distinct cheese flavours. The free amino acids are converted to volatile components
by amino-acid-converting enzymes (AACEs). Because of the variance in the presence
and/or regulation of AACEs in different strains of LAB, resulting in differences in levels
and types of amino acids produced, their flavour forming qualities also vary ending on
the strain, contributing to the diversity of cheese types and flavours. Controlling the
processes of amino acid degradation during cheese ripening provides a means of
controlling its flavour development. Additionally, the lactic acid produced by the LAB’s
metabolism helps in curdling, and also lowers the pH of the product to preserve against
other microbes. LAB also produce natural antibiotic-like substances called bacteriocins
that act as natural preservatives, such as the nisin produced by Lactococcus lactis, which
prevent spoilage from yeasts and moulds, as well as other bacteria.

23
Cheese is way of preserving milk for long periods of time. In the process, the milk in
cheese becomes something completely unlike milk, but cheese has its own interesting and
delicious properties. Cheese-making is a long and involved process that makes use of
bacteria, enzymes and naturally formed acids to solidify milk proteins and fat and
preserve them. Once turned into cheese, milk can be stored for months or years.

The main preservatives that give cheese its longevity are salt and acids. The basic steps in
cheese making go something like this (for most common cheeses like cheddar):

First, milk is inoculated with lactic acid bacteria and rennet. The lactic acid bacteria
convert the sugar in milk (lactose) to lactic acid. The rennet contains enzymes that
modify proteins in milk. Specifically, rennet contains rennin, an enzyme that converts a
common protein in milk called caseinogen into casein, which does not dissolve in water.
The casein precipitates out as a gel-like substance that we see it as curd. The casein gel
also captures most of the fat and calcium from the milk. So the lactic acid and the rennet
cause the milk to curdle, separating into curds (the milk solids, fats, proteins, etc.) and
whey (mostly water). The curds and whey are allowed to soak until the lactic acid
bacteria create a lactic acid concentration that is just right. At that point, the whey is
drained off and salt is added.

Now the curds are pressed in a cheese press -- lightly at first to allow the escape of the
remaining whey, then severely (up to a ton of pressure) to solidify the cheese.

Finally, the cheese is allowed to age (ripen) for several months in a cool place to improve
its taste and consistency. A sharp cheddar cheese has been aged a year or more. During
this time, enzymes and bacteria continue to modify proteins, fats and sugars in the
cheese. The holes in Swiss cheese occur during ripening -- Swiss cheese is ripened in a
cool place for several weeks, and then put in a warm place (70 degrees F, 21 degrees C or
so) for four to six weeks, where special bacteria ferment the remaining lactose and
produce carbon dioxide bubbles in the cheese.

24
The enzymes (rennin) when added to pasteurized milk acts on combination with the
starter – culture of bacteria and form curd by coagulating the casein. After the curdling
has set to a uniform gel, it is cut into cubes and then cooked to expel the whey. The
cooking temperature determines the texture of the cheese whether it is hard or soft & it
ranges from 32-500C. If cooked at a higher temperature, much of the whey is expelled,
resulting in hard cheese & vice versa. The texture of this type of cheese impacts the
characteristics on it.
The cooked curd is then separated by drawing off the whey & allowed to form large
blocks in a process referred to as matting. This is common in Cheddar cheese. The cheese
is then washed, milled into the desired shape, salted either by the direct addition or by
immersing into brine. The processes of matting, milling, washing & salting contribute to
the differences in the final product. The cheese is then pressed to expel its remaining
whey. All types of cheese are made in this way.
After pressing, then curing or ripening of the cheese take place & is the most important
step that gives each type of cheese its characteristic flavor. This involves keeping the
cheese in room where temperature, humidity etc are controlled to enhance the growth of
the organism desired to make use of the remaining whey trapped in the cheese blocks to
produce end products that give rise to the characteristic flavors. Note that some
organisms might also give rise to undesirable flavors in the process of ripening. The
ripening process may last from a few days to several months or even years.

(v) WARA (CURD)


Curd is unfermented cow or goat milk. The young leaf of Sodom apple (bom-bom)
Calotropis procera are crushed lightly, the juice squeezed out into fresh milk while
stirring. The juice contains a coagulating substance that curdles the milk. The mixture is
heated gently for about 20mins till a yellowish grey whey shows at the edge of the scum.
The floating dirt is then carefully removed with a calabash scoop and the white curd is
exposed. Heating is continued until the desired texture & flavor is obtained. The mixture

25
is then filtered into a basket molds that gives the curd its characteristics shape. The curd
is dropped drawn back into the whey while still hot.

C. ACETIC FERMENTATION
The main desirable fermentation carried out by acetic acid bacteria is the production of
vinegar. Vinegar, literally translated as sour wine, is one of the oldest products of
fermentation used by man. Vinegar is used extensively to give flavor & acts as
preservative e.g. in vegetable processing in the making of salad creams & rice topping.
Also used in meat, fish & poultry curing. There are various types of vinegar but all
contains acetic acid made from ethanol. Apart from acetic acid is the major constituent of
vinegar, small quality of pyruvic, formic, propionic & butyric acids in addition to
glycerol & diacetyl have been identified in vinegar; so each type of vinegar derived its
flavor from the characteristic combination of all other substances.

It can be made from almost any fermentable carbohydrate source, for example fruits,
vegetables, syrups and wine. Whatever the raw material used, the fermentation process
follows a definite sequence. The basic requirement for vinegar production is a raw
material that will undergo an alcoholic fermentation. Apples, pears, grapes, honey,
syrups, cereals, hydrolyzed starches, beer and wine are all ideal substrates for the
production of vinegar. To produce a high quality product it is essential that the raw
material is mature, clean and in good condition. The vinegar process is essentially a two
stage process, where yeasts convert sugars into alcohol, followed by Acetobacter, which
oxidize alcohol to acetic acid. Acetobacter convert alcohol to acetic acid in the presence
of excess oxygen.

Oxidation of alcohol to acetic acid and water

The oxidation of one mole of ethanol yields one mole each of acetic acid and water;

26
C2H5OH  O2 CH3COOH
H2O
+ +
Alcohol   acetic acid Water

This is usually done as a 2 stage process first stage is alcoholic fermentation and the
second stage is acetification.
Alcoholic fermentation is initiated degraded by yeast while acidification is by bacteria.
Summary of the Reactions:
Yeast 2C 2 H 5OH 2C02
1) C 6 H 12O 6 + +
alcoh ol alco h ol anaerobiacally
Bacteria 2 CH 3 CHO H 2 O
2) 2C2H 5OH +
❑ acetalde h yde ❑
2CH 3COOH H 2O
3) 2 CH 3 C H 0 ❑ +
+O 2 acetic acid ❑

Microbes involved in the vinegar process.

The production of vinegar depends on a mixed fermentation, which involves both yeasts
and bacteria. The fermentation is usually initiated by yeasts which break down glucose
into ethyl alcohol with the liberation of carbon dioxide gas. Following on from the yeasts,
Acetobacter oxidize the alcohol to acetic acid and water.

Yeast reaction

C6H12O6 2C2H5OH + 2CO2


ethyl
Glucose yeast carbon dioxide
alcohol +
Bacterial reaction

27
C2H5OH + O2 CH3COOH + H2O
Alcohol   acetic acid Water

The yeasts and bacteria exist together in a form known as commensalism. The
Acetobacter are dependent upon the yeasts to produce an easily oxidizable substance
(ethyl alcohol). It is not possible to produce vinegar by the action of one type of micro-
organism alone.

For a good fermentation, it is essential to have an alcohol concentration of 10 to 13%. If


the alcohol content is much higher, the alcohol is incompletely oxidized to acetic acid. If
it is lower than 13%, there is a loss of vinegar because the esters and acetic acid are
oxidized. In addition to acetic acid, other organic acids are formed during the
fermentation which become esterifies and contribute to the characteristic odour, flavour
and colour of the vinegar.

Acetaldehyde is an intermediate product in the transformation of the reducing sugar in


fruit juice to acetic acid or vinegar. Oxygen is required for the conversion of acetaldehyde
to acetic acid.

In general, the yield of acetic acid from glucose is approximately 60%. That is three parts
of glucose yield two parts acetic acid.

Micro-organisms involved in the fermentation of vinegar.

The organisms involved in vinegar production usually grow at the top of the substrate,
forming a jelly like mass. This mass is known as 'mother of vinegar'. The mother is
composed of both Acetobacter and yeasts, which work together. The principal bacteria
are Acetobacter acetic, A. xylinum and A. ascendens. The main yeasts are
Saccharomyces ellipsoideus and S. cerevisiae. It is important to maintain an acidic
28
environment to suppress the growth of undesirable organisms and to encourage the
presence of desirable acetic acid producing bacteria. It is common practice to add 10 to
25% by volume of strong vinegar to the alcoholic substrate in order to attain a desirable
fermentation.

The alcoholic fermentation of sugars should be completed before the solution is acidified
because any remaining sugar will not be converted to alcohol after the acetic acid is
added. Incomplete fermentation of the juice results in a "weak" product. The acetic acid
strength of good vinegar should be approximately 6%.

Fermentation methods :

Small scale production

Vinegar can be made at home at the small scale by introducing oxygen into barrels of
wine or cider and allowing fermentation to occur spontaneously. This process is not very
rigorously controlled and often results in a poor quality product.

The Orleans process

The Orleans process is one of the oldest and well known methods for the production of
vinegar. It is a slow, continuous process, which originated in France. High grade vinegar
is used as a starter culture, to which wine is added at weekly intervals. The vinegar is
fermented in large (200 litre) capacity barrels. Approximately 65 to 70 litres of high
grade vinegar is added to the barrel along with 15 litres of wine. After one week, a further
10 to 15 litres of wine are added and this is repeated at weekly intervals. After about four
weeks, vinegar can be withdrawn from the barrel (10 to 15 litres per week) as more wine
is added to replace the vinegar.

One of the problems encountered with this method is that of how to add more liquid to
the barrel without disturbing the floating bacterial mat. This can be overcome by using a
glass tube which reaches to the bottom of the barrel. Additional liquid is poured in
29
through the tube and therefore does not disturb the bacteria. Wood shavings are
sometimes added to the fermenting barrel to help support the bacterial mat.

Traditional vinegar which involves partly filling vats with wines & exposing to the air for
acetic acid bacteria to grow in. the acetic acid bacteria develop as a gelatinous pellicle on
the surface. The conversion of ethanol to acetic acid takes several weeks & when the
wine has been slowly converted to vinegar part of it is drawn off at the base and the
original volume restored by replenished with new wines. This slow process leads to the
production of vinegar of better quality with more aroma & flavor.

Other methods of vinegar production are mechanical to increase production rate & are
referred to as quick processes.

Quick vinegar method

Because the Orleans process is slow, other methods have been adapted to try and speed
up the process. The German method is one such method. It uses a generator, which is an
upright tank filled with beechwood shavings and fitted with devices which allow the
alcoholic solution to trickle down through the shavings in which the acetic acid bacteria
are living. The tank is not allowed to fill as that would exclude oxygen which is
necessary for the fermentation. Near the bottom of the generator are holes which allow
air to be drawn in, the air rises through the generator and is used by the acetic acid
bacteria to oxidize the alcohol. This oxidization also releases considerable amounts of
heat which must be controlled to avoid causing damage to the bacteria.

This trickling process utilizes a tank loosely filled with wood shavings. Alcoholic liquors
are distributed over the shavings from the top of the tank. The liquor is acetified as it
trickles down to the base & is recycled until the process is completed i.e. until enough
acetic acid has been formed. Acetic acid bacteria distributed on the showings therefore
have a large surface area for acetification to take place. Temperature is regulated because

30
acetification yields an exothermic process, so cooling coils are inserted to keep the
temperature between 25-300 C.

The submerged culture fermentation method – this process ensures that oxidation to
acetic acid is bidirectional & not unidirectional as in the previous 2 methods. The system
is constantly aerated to keep the acetic acid bacteria in a rapid phase of acetification as
the air introduced disperses the cells & also supply O 2. Cooling coils are also
incorporated to dissipate heat. The process is also very quick. Then filter finished vinegar
to clarify them & then pasteurized at 740 C or there about.

D. OTHER INDIGENOUS MICROBIAL FOOD FERMENTATIONS


Other fermentation procedures are employed for making palatable and nutritious food.
Also, for preserving various foods such as rye, wheat, sorghum, maize, beans etc. this
type of fermentations are practiced mainly in African South – East Asia and in the Far
East by making use of bacteria, mold or yeast. Many fermentation processes in W. Africa
are done in the natural way whereby the process involves a succession of
microorganisms. However, the technology of this process has greatly improved to ensure
uniformity of products. It is now common to find control fermentation of grains,
vegetables, fruits etc. making use of starter – cultures & incubating at specific
temperature for a specified period.

Examples
1) SOY-SAUCE
This is made from soya beans and wheat or rice. It is a dark – brown liquid use as
seasoning in China, Korea, Indonesia etc.
The inoculum is obtained by culturing Aspergillus oryzae and Aspergillus sojae on wheat
– bran until there is good mycelia growth on the fungi. This inoculum is then added to a

31
combination of cooked soy-beans and roasted, crushed, wheat or rice. Incubate at 25 –
300c for 3-5 days.
During this period A. oryzae grows vegetatively and liberate enzymes unto the substrates
while Lactobacillus delbrueckii which is naturally present in wheat bran also produce
enzymes. The cake formed by the ramification of mycelium into the substrate is then
crumbled and covered with about 20% brine, vegetable growth of the organism then
cease but the enzymes previously produced continue to act to break down proteins in the
soy-bean and wheat to glutamic acid, other peptides and amino acids. The amylase break
down the starch to sugars & these are slowly converted to alcohol by fermentative yeasts
which also contribute to the flavor, color and translucent of the final sauce.

2) SOY – IRU
This is also made from soy-beans. The beans are cooked at atmospheric pressure for
about 30mins, then dehaulled and cooked again for about 3hrs; then allowed to cool.
The beans are then placed in calabash tray lined with banana leaves & also covered with
the same leaves. Fermentation is allowed to continue for about 3-5 days at roan
temperature, after which the beans would have been soften ready for eating.
The fermented beans have a light brown colour and the product is use as condiment by
the West Africa population. Species of Staphylococcus are important in the making of
this product.

3) PAP OR OGI

This is fermented maize, guinea corn or sorghum starch. The substrate is soaked in
water & left to ferment & soften naturally.The inhalation of water by the maize
grain activates the hydrolytic enzymes & the starch to hydrolyze the starch to
sugars. The sugars are then fermented by the natural microflora of yeast &
bacteria. Lactobacillus sp. produces lactic acid, CO 2 etc. Leuconostoc
mesenteroides is heteroferentative & thus acts first, succeeded by Lactobacillus
which is homofermentative. This organism continues producing lactic acid

32
lowering the pH, & making the medium unsuitable for further growth of
Lactobacilli. The result is that residual sugars are not fermented. In consequence,
the fermentation vessel develops a yeast population that utilizes the residual sugars
& converts some of them to ethanol & CO2. The maize is then washed after the
period of fermentation & then milled in mortals or between stones or in a
mechanized mill. The product is made into pap by stirring with hot water.

4. GARI PRODUCTION

The food is obtained from cassava fermentation. The cassava tubers are peeled, washed,
grated and allowed to ferment from 4-6 days in sacks at room temperature i.e. between
28-30oC. A longer period of fermentation is required during harmathan because of the
lower temperature which induces lower rate fermentation. The fermentation occurs in two
stages: -

a. Corynebacterium manihotis produces a mixture of organic acids such as lactic,


butyric, acetic acid etc which lowers the pH.
b. The acids then stimulate the growth of Geotrichum candidum which produces
aldehydes and esters responsible for the Gari flavor; the sack soured state.

The HCN in Cassava is eliminated during the period of fermentation through the
hydrolysis of linamarin which evolves gaseous HCN dissolves in water and is leached out
while still standing and during pressing & frying.

5. FUFU

This is also made from cassava. Peel, wash and submerged completely in water for 2-
3days until it is soft. Length of soaking depends on the temperature & while it is soaking
microorganisms like Lactobacillus ferment it is produce mixture of acids but mainly
lactic acid. Then sieved the soft tubers after fermentation and reduce the water content by
pressing. It is prepared by mixing with hot water & turn into a paste. Alternatively, it can
be pounded, wrapped in leaves cooked & pounded again.
33
CLASSIFI CATION, IDENTIFICATION AND ENUMERATION OF
MICROORGANISMS IMPORTANT IN FOOD

The significance of microorganisms in foods depends upon several conditions:

1) the numbers found


2) the types of microorganisms
3) the type of food
4) the treatments to which the food has been exposed
5) the processing or storage treatments the food receive
6) whether the food is to be eaten as it is or heated
7) the individuals who might consume the food.

Microorganisms may have at least one of four functions in a food.

a) They may have a useful function – useful organisms are those which produce
desirable changes in food such as converting milk to cheese, sugar to alcohol,
and cabbage to sauerkraut. These changes are referred to as fermentations. The
organism does not always have to be present; enzymes can be separated from
the organism and used to produce the desired result.
b) Cause spoilage – spoilage is the result of undesirable changes in the odour,
colour, flavor, texture, or appearance of food. Some organisms that do not
directly cause changes in a food may alter the flora so that spoilage organisms
can grow e.g. bacteriophages.
c) Be inert – the inert microorganisms do not find an environment favourable for
growth.
d) Be a health hazard.

Primary Sources of Microorganisms Found in Foods

34
1. Soil and Water – Soil microorganism may enter the atmosphere by the action of
wind and enter the water bodies when it rains. They may also enter water when
rainwater flows over soils into bodies of water. Aquatic organisms can be
deposited onto soils through the actions of cloud formation and rainfall.
2. Plants and Plant Products - Relatively small number of soil and water organisms
that contaminate plants find the plant environment suitable. Those that persist on
plant product have capacity to adhere to plant surfaces so that they are not easily
washed away and because they are able to obtain their nutritional requirements.
E.gs. Corynebacterium, Curtobacterium, Pseudomonas, Xanthomonas and
moulds.
3. Food Utensils – The cutting block in a meat market along with the cutting knives
are contaminated from initial samples and repeated placing of harvested
vegetables in same containers and utensils leads to a build-up of organisms.
4. Intestinal Tract of Humans and Animals - Enterobacteriaceae are expected in
faecal wastes along with intestinal pathogens. This flora becomes a water source
when polluted water is used to wash raw food products.
5. Food Handlers - The microflora on the hands and outer garments of handlers
generally reflects the environment and habits of individuals, and the organisms
may be from soils, nasal cavities, mouth, gastrointestinal tract, waters, dust, and
other environmental sources.
6. Animal Feeds – The organisms in dry animal feed are spread throughout the
animal environment and may be to occur on animal hides. E.gs Salmonellae in
poultry and Listeria monocytogenes in dairy and meat.
7. Animal Hides – The types of organisms found in raw milk can be a reflection of
the flora of the udder and from both the udder and the hide, organisms can
contaminate milk containers and the hands of handlers.
8. Air and Dust - The Gram-positive organisms, a number of moulds and some
yeast persist more in air and dust. The types of organisms in air and dust are those
that are constantly reseeded to the environment.

35
MICROBIAL TYPES IN FOOD

The microbial analysis of food products yields many diverse types of microorganisms.
The predominant types and those capable of causing food spoilage or be a health hazard
includes:

A) Aerobic/Microaerophilic, Motile, Gram- Negative Bacteria

In this group only one genus, Campylobacter, is important in food.

Campylobacters are non-spore forming, Gram-negative rods, oxidase-positive having


one or more polar or amphitrichous flagella which confer a rapid, darting motility and
may be an important feature in pathogenesis. Cells are pleomorphic and may be 0.5-8µm
in length and 0.2-0.5µm in width. It uses amino acids or tricarboxylic acid intermediates
rather than carbohydrates as a source of energy. They cannot ferment or oxidize sugars
and are oxygen-sensitive microaerophiles, growing best in an atmosphere containing 5-
10% oxygen and 3-5% carbon dioxide. All Campylobacter species grow at 37o C; C.
jejuni and C. coli have optima at 42-45o C but cannot survive cooking or pasteurization
temperatures. They do not grow below 28oC and survive poorly at room temperature with
declined viability during chilling or frozen storage. They may persist under these
conditions for prolonged periods and are particularly sensitive to other adverse condition
such as drying and reduced pH.

The principal reservoir is the alimentary tract of wild and domesticated animals and birds.
It is a common commensal of rodents, dogs, cats, dairy cattle, sheep, pigs, poultry and
wild birds. Asymptomatic human carriage also occurs. Under adverse environmental
conditions campylobacters have been reported to adopt a ‘viable non-culturable’ state
where the organisms cannot be isolated by cultural methods but remains infective.
C.jejuni can revert to a culturable state by passage through an animal host.

36
Pathogenesis: Enteropathogenic campylobacter can cause an acute enterocolitis. The
incubation period is from 1 to 11 days, most commonly 3-5 days with malaise, fever,
severe abdominal pain and diarrhea as the main symptoms. Complications are rare but
reactive arthritis can develop leading to a serious neurological disease, Guillain-Barre
syndrome.

Isolation and Identification: They can be grown on peptone-based media including


nutrient broth in an incubation atmosphere of 5-6% oxygen and about 10% carbon
dioxide. A resuscitation stage of 4 h at 37o C in a non-selective environment is
recommended as a recovery and repair process for cells from food or other environmental
sources sublethally injured due to stress from freezing, drying or heating. Media
containing oxygen scavenging compounds such as blood, pyruvate, ferrous salts, and
charcoal and enrichment media including antibiotics (polymyxin B, trimethoprim) are
used. After selective enrichment for 24 and 48 h under microaerobic conditions at 42-
43oC, samples are streaked on to selective plating media. Colonies are non-haemolytic
with flat, watery appearance and an irregular edge and a grey or light-brown colouration.

Infection can be acquired by a number of routes. Direct transmission person-person or


from contact with infected animals (kittens and puppies) and occasionally waterborne
outbreaks.

B) Gram- Negative, Aerobic Rods, and Cocci

This group include the family Pseudomonadaceae with the genera Pseudomonas and
Xanthomonas; the family Halobacteriaceae with the genera Acetobacter and
Gluconobacter; the family Neisseriaceae with the genus Acinetobacter and four other
genera Alcaligenes, Brucella, Flavobacterium and Alteromonas.

37
i) Brucella: Gram- negative, catalase-positive, short oval rods, non-motile and usually
occur singly, in pairs, or, rarely in short chains. It grows optimally around 37 oC and are
heat-labile (63 o C for 7-10 min).It can survive for many days in infected animal milk
provided the acidity remains low (<0.5% lactate). Each of the four species of human
pathogens is associated with a particular host, B. abortus (cattle), B. melitensis (sheep and
goat), B. suis (pigs), and B. canis (dogs). Brucellosis is principally contracted from close
association with infected animals and is an occupational disease of farmers, herdsman,
veterinarians and slaughterhouse workers, or at low risk by consumption of milk or milk
products from an infected animal.

Pathogenesis: Brucellosis is a protracted illness characterized by incubation period of


one to six weeks followed by chronic, relapsing fever accompanied with sweats,
headache, constipation, anorexia, pains in the limbs and back, and weight loss. Treatment
is with a mixture of Tetracycline and Streptomycin. The causative organism is a
facultative parasite and can live intracellular or in extracellular body fluids.

Isolation and Identification: Liver infusions or calf serum is normally added to media
for their growth because Brucella is fastidious. The organism grows slowly and cultures
are incubated for three weeks before they are considered negative, therefore testing foods
for the organism is not practically feasible. Cattles are tested for the presence of
antibodies to the organism in the ‘Ring test’. Stained antigen is mixed with the test milk;
if antibodies to Brucella are present (indicative of infection) then they will cause the
antigen to clump and rise with the milk fat on standing to form an intense blue-violet ring
at the top of the milk.

ii)Acetobacter: These cells are ellipsoidal to straight or slightly curved rods. Young
cultures are Gram-negative, while older cultures are gram variable. They are motile
[peritrichous flagella] or nonmotile.

38
The Acetobacter are noted for the oxidation of ethanol to acetic acid. They oxidize
acetate to lactate to CO2 and H2O .They are found on fruits juices and alcoholic beverages
[bear and wine].

iii)Alcaligenes: The four species in this genus are motile [four to eight peritrichous
flagella] rods, coccal rods, or cocci. They are strict aerobes and oxidase positive

They are widespread in nature, being found in soil, water, decaying matter, and the
intestinal tract of animals. These organisms are involved with spoilage of protein foods
[eggs and dairy products].

C) Gram- Negative, Facultative Anaerobic Rods

The family Enterobacteriaceae includes the genera; Escherichia, Serratia, Shigella,


Salmonella, Citrobacter, Klebsiella, Enterobacter, Erwinia, Proteus and Yersinia. The
family Vibrionaceae includes Vibrio and Aeromonas.

i)Escherichia coli: E. coli is an inhabitant of the gut of humans and other warm blooded
animals where it is the predominant facultative anaerobe though only a minor component
of the total microflora. It is found in soil and water, and in various foods, especially
animal products and foods handled by people. Generally a harmless commensal, it can be
an opportunistic pathogen causing a number of infections e.gs Gram-negative sepsis,
urinary tract infections, pneumonia in immunosuppressed patients, and meningitis in
neonates. The principal species is E. coli. Strains causing diarrhea are classified into
three groups based on their virulence properties: enteropathogenic E.coli (EPEC),
enteroinvasive E. coli (EIEC), and enterotoxigenic E.coli (ETEC). E. coli serotype
0157:H7 (EHEC) is recognized as the cause of a number of haemorrhagic colitis and
haemolytic uraemic syndrome. Enteroadherent E.coli is identified by their characteristic
pattern of adherence to Hep-2-cells.The organisms are members of the coliform group,
which are indicators of faecal contamination. It is a catalase-positive, oxidase-negative,
fermentative, short, non-sporing rod. Differentiated from other members of the family on
39
the basis of sugar fermentation and other biochemical tests known by the acronym
IMViC. These tested for the ability to produce:

i) Indole from tryptophan (I);


ii) Sufficient acid to reduce the medium pH below 4.4, the break
point of the indicator methyl red (M);
iii) Acetoin ( acetylmethyl carbinol) (V); and
iv) The ability to utilize citrate (C).

Most strains of E. coli are indole and methyl red positive and VP and citrate negative.

Pathogenesis: Illness caused by ETEC usually occurs between 12 and 36 h after


ingestion of the organism with symptom ranging from mild diarrhea to a severe cholera-
like syndrome of watery stools without blood and mucus, stomach pains and vomiting.
The ingested organism resists expulsion from the small intestine by adhering to the
epithelium through attachment or colonization factors in the form of fimbriae on the
bacterial cell surface. Two toxin types are produced: the heat-stable (ST), which can
withstand heating at 100 o C for 15 min and acid resistant, and the heat-labile (LT) which
are inactivated at 60 o C after 30min at low pH.

Infection by EIEC results in symptoms of an invasive bacillary dysentery normally


associated with Shigella. EIEC invades and multiplies within the epithelial cells of the
colon causing ulceration and inflammation with clinical features as fever, severe
abdominal pains, and watery diarrhea preceding watery stools containing blood, mucus
and faecal leucocytes.

Symptoms of EPEC infection include vomiting and diarrhea with stools containing
mucus but rarely blood appearing 12-36 h after ingestion of the organism. In infants, the
illness is more severe than other diarrheal infections persisting for longer than 2 weeks in
some cases. Pathogenesis is related to its ability to adhere closely to the enterocyte
membrane causing cell loss from the villus tips.
40
EHEC sometimes known as Verotoxin-producing E. coli (VTEC) can cause
haemorrhagic colitis (a self-limiting, acute, bloody diarrhea that begins with stomach
cramps and watery diarrhea after an incubation period of 3-8 days); haemolytic uraemic
syndrome, characterized by three features renal failure, haemolytic anaemia and
thrombocytopaenia (a drop in the number of blood platelets) and thrombotic,
thrombocytopaenic purpura (related to haemolytic uraemic but includes fever and
neurological symptons).

Isolation and Identification: Selective techniques for E. coli exploit the organism’s
tolerance of bile, aniline dyes and other surfactive compounds and the ability to grow at
temperatures around 44 o C. MacConkey agar contains bile salts (and sometime the
aniline dye, crystal violet) act as inhibitors of Gram-positive and some fastidious Gram-
negative bacteria. Lactose is included as a fermentable carbohydrate with a pH indicator,
usually neutral red. E. coli will produce red colonies like others strong acid producers.

Eosin/ methylene blue agar is a selective and differential medium containing aniline dyes
eosin and methylene blue, the selective agents but also serving as indicator for lactose
fermentation by forming a precipitate at low pH. Strong lactose fermenters produce
green-black colonies with a metallic sheen.

Β- glucuronidase activity is possessed by 95% of E. coli strains and only a limited


number of other bacteria is a biochemical feature increasingly used.

Colonies suspected from selective and differential media can be confirmed by further
biochemical testing.

ii) Aeromonas : Aeromonads are Gram—negative, rods with rounded ends to coccoid
which ferment glucose. They are motile (polar flagella), and oxidase and catalase
positive. These organisms are similar in growth and biochemical characteristics with the
Enterobacteriaceae except for the positive oxidase test and nitrate reduction. The main
habitat is water. The numbers present depends on factors such as nutrient level and
41
temperature. Some strains cause disease (haemorrhagic septicemia) in fish and frogs and
enteritis in humans. A. hydrophila ia neither salt (≤5%) nor acid (min.pH≈6.0) tolerant
and grows optimally at around 28oC. Its most significant feature with regard to any threat
it may pose in foods is its ability to grow down to chill temperatures as low as -0.1 oC in
some strains.

Pathogenesis: In children under five years, mild and self-limiting gastroenteritis is


common mostly characterized by profuse watery diarrhea and sometimes dysenteric
stools. A. hydrophila and A. sobria produce a range of potential virulence factors
including cytotoxic and cytotonic enterotoxins.

Isolation and Identification: Enrichment media e.g alkaline peptone water is used, but
direct plating is done when the number present is high. A. hydrophila grow on a wide
range of enteric media fermenting lactose like coliforms but not xylose as well as bile
ducts.

iii)Vibrio: This genus is in the family of Vibrionaceae. The cells are short, motile,
oxidase positive, curved, or straight rods. Some strains fail to grow without the presence
of NaCL, the optimum concentration being 3.0%.
Strains of V. costicola can tolerate NaCl, concentrations of 23%. This species has been
found in cured meats and curing brines). V. cholerae and V. parahaemolyticus are
important pathogens, causing gastroenteritis in humans. V. cholerae is found in the
intestinal tract of people and animals, in water, and occasionally in food. V. cholera is
found in the intestinal tract of people and animals, in water, and occasionally in found. V.
parahaemolyticus is found in the ocean, in seafoods, and in the intestinal contents of
infected humans. Various other vibrios have been incriminated in the outbreak of
foodborne illness.

iv)Salmonella: The genus Salmonella divided into five subgenera(I-V). These subgenera
are differentiated on the basis of minor biochemical differences. Subgenus I includes the
majority of serotype of this genus, especially the more important serotypes, and subgenus
42
III includes the organisms previously called the Arizona group. Most of the different
types were differentiated on the basis of serological reactions, so they were called
serotypes or serovars. The Salmonella organisms are catalase- positive, oxidase-
negative, with both respiratory and fermentative metabolism. Most of the serotypes are
motile (peritrichous flagella), do not ferment lactose, and grow well in simple media
containing glucose, inorganic nitrogen and mineral salts. Although salmonellae do not
ferment sucrose, strains of S.arizonae contain a plasmid that gives them the ability to
ferment this sugar.

The Salmonellae are found in animals, humans, processing equipment, feed, and various
products. The natural habitat is the intestinal tract of humans and animals. Some
serotypes seem to be localized in a region or a country, but with national and
international travels and trade, the organisms are easily disseminated. The serotypes of
Salmonella are an important cause of foodborne illness.

D) Gram-Negative, Anaerobic Straight, Curved, and Helical Rods

In this category, the family Bacteroidaceae contains twelve genera, including


Bacteroides. These organisms are nonmotile, straight rods, being nonsporeforming
obligate anaerobes; they probably cause no changes in food. However, they are found in
fecal material in very high numbers. Hence, they might be useful as indicators of fecal
contamination of food as well as water.

Gram-Positive Cocci

The Gram-postive cocci include aerobic or facultative anaerobic bacteria in the family
Micrococcaceae with the genera Micrococcus and Staphylococcus. Other Gram-positive
cocci include the genera Streptococcus, Leuconostoc, Pediococcus, and Aerococcus.

43
i)Micrococcus: These spherical cells are strict aerobes, catalase positive, occur singly or
in pairs, and characteristically ivied in more than one plane to form irregular clusters,
tetrads, or cubical packets. They can grow in the presence of 5% salt.

The Micrococci are found in soil, water, and dust, and on the skin of people and other
animals. They are found in several types of foods especially milk and dairy products, on
animal carcasses, and in meat products. They are important as potential spoilage
organisms.

ii)Staphylococcus: These nonmotile cells occur singly, in pairs or in irregular clusters.


They are facultative anaerobes with respiratory and fermentative metabolism. Most
strains can grow in 7.5 percent to 15 percent salt. The organisms usually are sensitive to
chlorine, chloramine, iodine, and iodophores. Although usually sensitive to heat, they are
moderately resistant to radiation.

Both S. aureus and S. epidermidis are commonly found on the skin and mucous
membranes of humans and warm-blooded animals. They are potential pathogens, being
either the primary pathogen or secondary invader.

The staphylococci are found in many types of food products. In general, they are not able
to compete very well with other organisms. S. aureus produces enterotoxins, which are
one of the main causes of food borne illness. S. aureus is found in pimples, boils, acne,
wound infections, and in the nose. It is easily transferred to foods by careless food
handlers. This species is differentiated by its ability to produce coagulase (which clots
blood plasma) and a heat-stable nuclease.

iii)Streptococcus: These organisms have been divided into groups. The groups of
primary importance in food are the lactic streptococci and enterococci. These spherical
cells occur in pairs or chains. With the exception of some strains, they are not motile.
They are facultative and anaerobes with a fermentative metabolism, fermenting glucose

44
primarily by the hexose diphosphate pathway and producing mainly lactic acid. Thus,
they are called homofermentative.

The fermentation of carbohydrates to lactic acid is desirable in products such as cultured


milk, cheese, and sauerkraut, but it is undesirable in products such as fresh milk. They
can utilize sugar in an alcohol fermentation, resulting in lower alcohol an higher acid than
is wanted. Some strains utilize citric acid and form acetoin and diacetyl. The ability to
produce diacetyl in milk is desirable in manufacture of cultured sour cream, butter milk
and butter.

Various functions such as lactose metabolism, proteolytic activity, and hemolysin and
bacteriocin production are mediated by plasmids.

The streptococci are widely distributed being found in air, water, sewage, soil, and plants,
in the intestinal tracts of people and animals, and in various food products.

Some species and strains are somewhat heat resistant, surviving 60⁰ C for 30 min. The
enterococci can survive freezing and frozen storage in food products.

Although some pathogenic types may be transmitted to foods, There is more interest in
the streptococci as possible indicators of fecal contamination, as useful fermentative
organism, and as potential storage bacteria.

iv)Leconostoc: These spherical to lenticular cells occur in pairs or chains. They often
have complex nutrient requirements such as vitamins, amino acids, and a fermentable
carbohydrate. They ferment glucose to lactic acid, ethanol and CO₂.

The organisms are important in fermentation and spoilage of food. They are not
pathogenic although they have physiological differences.

Endospore-forming Rods and Cocci


45
This group includes the genera Bacillus, Clostridium, Sporolactobacillus and
Desulfotomaculum the genus is included with endospore-forming bacteria. The spores
that these organisms produce are different from the vegetative cells. The spores are an
inactive or dormant state of the organisms. The organisms go through certain stages to
change from a vegetative cell to a spore and back to a vegetative cell; sporulation,
germination, and outgrowth.

Sporulation: The mechanisms that trigger spore formulation are not fully known. Most
strains of clostridia produce spores when incubated in a good medium under anaerobic
conditions, 3⁰C to 8⁰C below their optimum growth temperature.

For all species, not all cells sporulate, regardless of the conditions. Spore formation
beings after the exponential growth phase during the stationary phase. Spores formation
can be arbitrarily divided into seven stages;

 division of axial chromatin filament;


 spore septation
 engulfment of the spores protoplast;
 cortex formation
 coat formation
 maturation and
 the free spore stage.

The spore consists of a core surrounded by several layers of mucopeptides and


proteinaceous outer coats.

An organism can initiate sporulation without completing the process and reverting to a
vegetative cell. However, there is a stage at which the cell becomes irreversibly engaged
in sporulation and is committed to complete the process. The composition of the substrate
and the temperature of sporulation can affect the resistance characteristics of the resultant
spores.
46
The main characteristic that is important to food microbiologist is the resistance of spores
to heat, radiation, chemicals, desiccation, and freezing. Quite commonly, the heat
resistance of spores is about 105 times, and radiation resistance is about 10times more
than that of the corresponding vegetative cells. Spores can be stored for long periods and
retain their ability to geminate and produce vegetative cells.

Germination: Since spores are dormant, they must be converted to vegetative cells to be
important in food spoilage or toxin production. The conversion of the heat resistant spore
to vegetative cells would allow less severe thermal process for food preservation.

Spores may need a conditioning treatment prior to germination. This process, called
activation, may be induced by aging, heating, radiation, altering the pH, or with
chemicals. Low number of viable spores does not always germinate immediately. Thus,
canned foods that apparently pass short storage test to determine potential spoilage can
show spoilage after extensive storage. Activation is a reversible process and if, conditions
do not allow germination, the spore reverts to its dormant state.

Outgrowth: The development of a vegetative cell through the first cell division is called
outgrowth. It occurs in a substrate capable of supporting vegetative growth. Outgrowth
proceeds by the swelling of the spore, emergence from the spore coat, elongation, and
cell division. Germination and outgrowth can be followed by determining the synthesis of
RNA, proteins, and DNA.

FOOD SPOILAGE

Food spoilage can be described as changes in form, and/or organoleptic properties of the
food (i.e. change in taste, odour, flavour that can be measured chemically) that renders it
unacceptable to the consumer

47
The changes could be textural, colour, flavour, aroma etc. examples of food spoilage are:
softening of vegetables (textural & flavour changes); browning of apples (colour
changes); souring of milk, rancidity of butter etc. The spoilage may be due to

i. Microbial action
ii. Mechanical damage such as bruising, cutting and freezing
iii. Physiological process such as enzyme action
iv. It could also be due to insect damage

MICROBIAL FOOD SPOILAGE

Microorganisms may cause a lot of food spoilage when good microbiological standards
are not maintained during the processing of such foods. Some of the microbes are present
as natural microflora & some others from sources such as knife, vessels, containers,
hands may be important in this effect. Canned food products may be spoiled due to
inadequate heat process. Uncontrolled or over fermentation of fermented foods could lead
to formation of sharp acid taste. Generally microbial actions are absent or reduce in dried
food, smoked food like fish, meat & grains (rice, beans & garri). It is also slow in
refrigerated foods or totally absent depending on the hygiene & temperature of storage.
But it is high in fresh foods with high water content such as fresh meat, fish, vegetables,
fruits etc. The water present in such foods is easily made use of by the microorganisms&
their action leads to undesirable changes in the food.

1. SPOILAGE OF CEREAL FOODS

Cereals are dried foods but they may be contaminated from various sources such
as water, air, dust, insect, rodents etc. Rains or animals may carry microorganisms
from the tassels to the grain itself and grains may also get easily contaminated
48
during storage. The low moisture content of grains protects them from microbial
attack.

a. Spoilage of cereal grains

Cereal grains are naturally likely to contain spores of molds, yeast and bacteria.
When the atmospheric humidity of the environment is high even when the grain has been
properly dried, there may be growth of molds at the surface. A wet mash of the grains
will also undergo acid fermentation (this is because the bacteria on the surface now
utilize the water) by bacteria and alcoholic fermentation by yeast. Helminthosporium,
Aspergillus, Penicillium and Fusarium spp. are some of the molds commonly found
associated with grains. The aggregation of their spores on the grain causes black point,
green, blue or pink or red colourations.

b. Flour

Flour is the milled grain and so all the nutrients especially CHO are present in the flour.
Aspergillus repens produces growth balls on corn flour at low moisture level of about
13% whereas A.flavus does the same thing at about 16% moisture level & above.

c. Bread

When bread is stored under fairly humid condition, such as when wrapped before cooling
or by keeping in humid store rooms, then the following types of spoilage may occur:

49
i. Rhizopus nigricans: which is commonly referred to as the bread mold may
produce a white cottony growth. Monilia (or Neurospora sitophila) with its pink
conidia gives a colour change to the bread. The spoilage renders the bread
unacceptable to consumers just for the physical appearance.

ii. Ropy bread : This is soft, sticky and discolored and when the bread is sliced,
long string-like threads are formed giving rise to the term ropy from rope. The
bread emanates unpleasant odour due to the action of proteolytic enzymes of
Bacillus subtilis & B. mesentericus. Bread made from unrefined flour is more
liable to this type of damage because the organisms are common in soil air and
grains.
iii. ‘Red’ bread : Red bread is caused by pigments of Serratia marcescens.
iv. Chalky bread: The bread has chalk-like spots due to the growth of the yeast-
like fungi Endomycopsis fibuligera and Trichosporon variable.

2. SUGARY FOODS

These are foods that have a high % of sugar which should normally render them safe
from microbial attack but osmophilic organisms still cause deterioration.

a. Raw Sugar : This includes cane or the untreated juice extract from them.
Fermentation of this may be caused by yeast such as Zygosaccharomyces
nussbaumii, Clostridium saccharolyticum, Penicillium glaucum, Aspergillus
sydowi etc. They yield a mixture of ethanol, furfural, acetone, butanol and gas.
Bacillus stearothermophilus, C. thermosaccharolyticum and C. nigrificans
are thermophilic bacteria found in hot-pressed sugar juices or in the storage tanks.
They are responsible for fermentation of sugars with lactic, formic and acetic
acids as end – products.
b. Honey : Nearly all honeys contain dormant bacteria, mold spores and yeast but
only the yeast cause spoilage by fermentation. Honey is the nectar of flowers

50
concentrated to about 83% sugar. Saccharomyces bailii var. osmophilus and S.
bisporus var. mellis and S. rouxii can cause spoilage by reducing the viscosity.
c. Candies : These include sweet, chocolates and fruits or coconut centered candies.
Gaseous fermentation by Clostridium sporogenes and C. putrificum may cause
the chocolate coating to burst or push out some of the syrup and disrupt the entire
candy.

3. VEGETABLE AND FRUIT SPOILAGE

a. Microbial spoilage of vegetable and fruit may be aided by physical factors like
bruising, crushing, wounding, shredding or physiological factors like increased
respiration and enzyme action especially during ripening which makes more nutrients
available for the microorganisms.

i. Soft rot may be due to pectinolysis by members of Pseudomonas, Erwinia,


Fusarium, Colletotrichum, Rhizoctonia, Sclerotina etc. Yeast rarely proliferate
(multiply) on the surface of fruits and vegetables because they lack enzymes
which can degrade the available substrates and also because they cannot penetrate
them. But they proliferate in tissues exposed by mechanical injury. Erwinia
carotovora causes soft rot of both leafy and root vegetables while Rhizopus may
cause mushiness of wet packed vegetable.
ii. Geotrichum candidum can grow in cracks on tomato fruits and ferment it and
thereby give rise to the development of an acid flavor and acid odour.
iii. Penicillium digitatum (green mould) causes serious losses in leaves, oranges
while P. italicum (blue-mould) attacks grapes and oranges and P. expansum
causes soft broom rot of apples.

3b. Vegetable and Fruit Juice

51
Juice from grapes, lemons, tomatoes, oranges, pineapple, mangoes, pears etc can be
attacked by various species of yeast such as Saccharomyces, Torulopsis and
Zygosaccharomyces spp. These organisms can ferment the sugar present and produce
alcohol under anaerobic condition. Lactic bacteria such as Lactobacillus brevis and
Leuconostoc mesenteroides can produce lactic acid in apple and pear juices.

4. BEVERAGES

i. Yeast and bacteria are important in the spoilage of many beverages. Species
of Saccharomyces and Torulopsis are commonly associated with fermenting
soft acid drink.
ii. Lactobacillus spp .produces lactic acid and cause souring.
iii. L. mesenteroides develop dextrin which form slime balls in the drink.
iv. Moulds of the genera Penicillium and Aspergillus grow as delicate, fluffy,
cottony masses suspended in the liquid.

5. DAIRY PRODUCTS SPOILAGE

Milk, the major raw material on which the dairy industry is based is an excellent medium
for cultivating many kinds of microorganisms because of its rich nutrient composition,
high water content and its near neutral pH (range of 6.7-6.8).

a. Raw/Fresh Milk

This is liquid milk &bacteria enter from the skin, hands and equipment used during the
milking process.

i) Souring : Streptococcus lactis sours milk by producing lactic acid at ambient


temperature. This souring may be accompanied by curdling or coagulation. S.
thermophilus and S. faecalis caused the same thing at temperature between 37-500 C.

52
ii) Ropiness : Alcaligenes viscolactis and Micrococcus freudenreichii cause
surface ropiness at ambient temperature. Enterobacteraerogenes and S. lactis var.
hollandicus cause ropiness throughout the milk.

iii) Butryric acid formation : This is carried out by Clostridium spp. After heat
treatment especially when the condition is unfavorable for lactic acid bacteria.

iv) Proteolysis : This is the hydrolysis of milk protein by S. faecalis and Pseudomonas
fluorescens. This leads to the development of bitter flavours. P. fluorescens produces a
proteolytic enzyme which survives pasteurization even though the organism itself does
not.

v) Ammonia formation: This is caused out by P. fluorescens & Alcaligenes


viscolactis.

vi) Sweet Curdling of milk : This is due to coagulation of milk without any
decrease in pH. It is caused by B. cereus which produces enzyme that digests the
phospholipids membrane surrounding each fat globule and thereby permits the globules
to coalesce and form a curd which rises to the surface.

vii) Colour changes may occur with the flavor and physical changes already
mentioned.

Blue-milk may occur which is due to Pseudomonas syncyanea and S.lactis.

Red-milk due to Serratia marcescens and Brevibacterium erythrogenes.

Brown-milk due to P.putrefaciens.

Yellow – milk caused by Flavobacterium spp.

viii) Souring due to mixed acid production by the fermentative action of Micrococcus,
Mycobacterium and Bacillus spp. when the conditions are unfavorable for lactic. A
53
mixture of lactic, acetic & formic acids together with alcohol & hydrogen gas is produced
with lowering of the pH.

b. Butter

Microorganisms that cause spoilage of butter are usually psychrotrophic & cause spoilage
between 5-100 C. These microorganisms are usually restricted to the moisture droplets
within butter because the nutrients are present in such droplets.

Types of butter spoilage: -

1) Pseudomonas fragi & P. fluorescens hydrolyze the fat to give rise to short chain
fatty acids with the development of a rancid flavor. i.e. the lipolytic action of the
enzymes of these two organisms causes rancidity by hydrolyzing the long chain
fatty acids to short chain fatty acids.

ii) Putrid flavours : This is due to proteolysis by P. putrefaciens

iii) Surface discoloration :

A black discoloration appearing on the surface of butter due to P. nigrifaciens.

Reddish – pink due to the presence of Fusarium culmorum.

Green due to the presence of Penicillium spp.

iv) Surface taints or cheery flavor defects

This is caused also by P. putrefaciens in unsalted or low – salt butter.

v) Malty – flavor :This is produced by S. lactis var maltigenes.

c) CHEESE

54
i) Geotrichum spp. may grow in cottage cheese digesting the casein & developing a
velveting film on the surface.

ii) P. fragi produces flavor defects and give the odour of overripe strawberry.

6. MEAT AND MEAT PRODUCTS

The microorganisms responsible for meat spoilage may be contaminant from the skin, gut
utensils, personnel and storage containers. Raw meat could be spoiled by its own enzyme
by the conversion of protein into simple nitrogenous substances which could be used by
non-proteolytic microorganisms.

a) Raw Meat Spoilage

i) Surface slime form on the raw meat by the species of Pseudomonas and
Alcaligenes.

ii) Rancidity due to the oxidation of fats by Achromobacter and Pseudomonas or by


yeast, it may be catalyzed by light and copper.

iii) Loss of fresh color by the heterofermentative action of Lactobacillus sp – greening


of sausage.

iv) Under anaerobic condition Clostridium novyii may putrefy raw meat by decomposing
the protein and gives rise to fouls smelling gas like NH3, H2S and amines.

v) Colouring due to pigment bacteria

Red spot caused by Serratia marcescens.

Yellow spot by Flavobacterium or Micrococcus

Green spot by Chromobacterium lividum

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Black spot caused by the presence of Cladosporium herbarum.

This mould grows even at temperature below -50C.

Pseudomonas mephilica converts myoglobin to sulf myoglobin in the presence of H2S in


meat stored at 1-20C leading to green discoloration.

vi) Whisker formation due to the aggregation of the mycelia of membersof Mucorales
such as Mucor, Thamnidium and Rhizopus. These organisms produce extensive whisky,
cottony, grey to black growth on the surface of meat in storage rooms.

b) Cured Meat

Meat treated with various curing agents such as NaCl, NO3, NO2 for preservative
purposes may still be spoiled by microbial action.

i) Species of the genus Clostridium such as C. mucosum, C. parabifermentans, C.


septicum etc are responsible for tainting dry cured hams because the salts do not penetrate
fast and they do not yet evenly distributed. So putrefaction takes place in the interior of
the meat by these organisms.

ii) Debaryomyces membraneofaciens var. hollandicus is highly tolerant to salt and


organic acids & so causes film to form on brines used for curing meat.

c) Sausage

i) Slime developing between the wrapper and the meat or in the interior of the meat
and Micrococcus sp. and yeast are responsible for this slime formation.

ii) Bacteria of the genera Microbacterium, Arthrobacter and Lactobacillus may cause
souring of packed sausage between 2-100 C.

d) Hamburger

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i) Putrefaction by Bacillus, Clostridium, Enterobacter, Proteus, Sarcina, Mucor &
Penicillium.

ii) Souring by Pseudomonas, Alcaligenes, Flavobacterium, Micrococcus at low


temperatures.

e) Refrigerated Packed Meat

Film wrappers that permit good air exchange favor the growth of aerobic bacteria such as
Pseudomonas sp. and this leads to putrefaction, off flavours and slime formation.
Wrappers with poor gas exchange encourage the lactic such as Lactobacillus sp,
Streptococcus sp. especially when combined with vacuum packaging.

7) FISH AND OTHER SEA FOODS

The natural microflora of fish depends on the microflora of the water in which they live.
Freshly cut fish has a shining iridescent surface with bright characteristic colorations.
Spoilage of fish & other sea foods include the following:

i) Autolysis may occur while the fish is still in a state of rigor.


ii) Pseudomonas sp. is an important spoilage organism at chilling temperature &
Pseudomonas fluorescens may give rise to greenish – yellow coloration.
iii) Salted fish may be spoilt by P. salinaria, Sarcina and species of Micrococcus.
All these organisms are obligatory halophiles and grow best at warm
temperature; causing proteolysis of the flesh and appearance of red pigment on
the surface.
iv) Smoked fish could be spoilt by mould of various types.
v) Trichosphoron sp. spoils crab meat by its proteolytic action on crab proteins.
vi) Shrimps are spoilt mainly by Achromobacter.

8. POULTRY FOOD SPOILAGE

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The micro-organisms that spoil poultry foods which include meat or rather the fresh and
eggs of chickens, turkey, geese, duck etc are either present on the feathers, on the skin,
feet, gut, cloaca of the birds at the time of slaughter or during egg-laying or were
introduced during handling.

a. Raw poultry meat

i) Autolysis – caused by the enzyme of the body itself.

b. Species of Peudomonas and Achromobacter may be responsible for the


development of unpleasant odours and slime, on dressed poultry at refrigerated
temperature.
c. Eggs Spoilage

(i) Cracks, blood stains, faecal stains are all undesirable defects in fresh eggs.
(ii) Green rot of eggs caused by P. fluorescens at refrigeration temperature.The
bright green fluorescens is observed by candling under u.v light. When green
eggs are broken, the spoilage is evident by a fruity but acrid odour.
(iii) Fungi such as Penicillium, Cladosporium and Sporotrichum species generally
produce mould colonies on the outside of the shell when the R.H of the
environment is high. Mucor and Thamnidium also produce a loose network of
aerial hyphae (whiskers) when the R.H is still higher.
(iv) Black rot of the yolk of the egg and when the egg is broken, it may have a
muddy brown colour and it is also hard. Black rot is caused by the organism
Proteus melanovogenes and certain species of Pseudomonas.
(v) Souring in cold storage which is caused by species of Pseudomonas
(vi) Off-odour and flavor : Aerobacter cloacae gives the odour of hay and
Achromobacter perolens gives a musty odour.

9. Spices and Condiments

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The fungi are the most important microorganisms in their spoilage. Examples include
leafy ones such as bay-leaves, thyme, oregano and mint, parsley while ginger, nutmeg,
peppers, cloves and the dehydrated powder of onions and garlic are referred to as all
spices. Pepper could be colonized by various species of Aspergillus such as A. glaucus,
A. flavus and A. ochraceus, these organisms are notorious for their mycotoxins
production e.g. A. flavus produces aflatoxins. Penicillium sp. is sometimes also present.

CONTROL OF FOOD SPOILAGE

A proper understanding and manipulation of the factors that caused microorganisms to


grow in foods and hence cause spoilage will lead to control of food spoilage. Foods
spoilage can be controlled by:

1). Fermentation

This is one of the oldest methods of food preservation known. Carbohydrates foods such
as maize, sorghum, cassava and fruit juice etc are fermented such that the acid(s) and
ethanol produced make the food unsuitable for microorganisms to grow. This product of
fermentation i.e. the acid(s) lower the pH and reduces the water activity (a w) of the food
i.e. amount of water in the food. Many foods such as vegetables, dairy products, grains,
roots and other tuber crops are preserved this way.

2. Refrigeration

The application of low temperature on foods slows down the activity of spoilage
microorganisms. Low temperature also reduces the activity of self-contained or innate
enzymes in foods of raw plants or animal origin. This lowered/reduced activity leads to
prolonged life for fresh foods. Heated foods can also be further preserved by low temps.
Refrigeration implies the use of temperature between 5 and 10ºC. It is useful for foods
like leafy vegetables, fruits; milk etc. Chilling involves rapid cooling at temperature
around 0ºC. Freezing of foods take place at subzero temperature and this leads to a

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change in state. The water in the food is frozen, such that it is unable for microbes.
Blanched vegetables can also be kept in fridges.

3.Canning

Canning began in France with Francis Appert who succeeded in preserving meat in
stopper bottles kept in boiling water for various period of time. Canning involves the use
of high temperature which kills the microorganisms in the food. Canning also creates an
anaerobic environment such that only anaerobes can grow in canned foods. Fruits,
vegetables, meat, juices, fish etc are canned. Spoilage of canned foods could be

(i) Leaker spoilages


(ii) Incipent thermophilic
(iii) Insufficient heat processes
(iv) Cross underprocessing

All these types of spoilage except thermophilic spoilage can be controlled by Good
Manufacturing Practices (GMP).

4. Use of Chemicals

The chemicals act by inhibiting microbial growth. Examples of chemicals used include
lactic, citric, acetic acids in drinks and beverages, benzoic acids in carbonated soft drinks,
SO2 (a gaseous chemical) and (sodium sulfite) NaSO3 are used as antibrowning agents in
dried foods. CO2 is also used in many drinks to present microbial growth i.e. in
carbonated drinks. NaCl is also used to salt foods.

5). Radiation

Radiation may be applied on foods to kill the microorganisms. There are 2 types of
radiation namely ionizing and non-ionizing radiation. The U.V and micro waves are e.gs
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of non-ionizing radiation. The problem with U.V light is that it acts only on the surface, it
has poor penetration power. Gamma rays is an e.g. of ionizing radiation. This one leaves
a trail of ions in its path as it penetrates the food. Radiation is use for foods like meat,
fish. These are not very common because they are considered as additives to food and
will involve carrying out of series of test to confirm the effects on foods.

6). Drying

This involves lowering of moisture content which inhibits the growth of microorganisms.
E.gs of food preserved by drying are grains e.g. millet, rice, maize. Drying could be by
freeze-drying in which water is removed by sublimation from frozen foods under vacuum
e.gs of freeze-dried foods are meat, fish and vegetables. Sun drying is also possible and
this is practicable in the tropics. Also, drying by frying e.g gari making. In gari, two
things are involved, the high temperature of drying and the removal of water which
prevents growth of microorganisms.

7). Oiling and Greasing

Surface of foods could be coated with oil to reduce evaporation and retard the escape of
CO2 from some foods. For eggs, light paraffin oil could be used after oiling, the excess is
drained off.

8). Smoking

This involves drying and the application of smoked gas as a preservative. The smoked
gas retards microbial growth directly and also from complexes with some of the chemical
constituents of foods leading to reduced growth of microbes. Smoking causes
considerable dried and formation of a glazed surface crust which acts as barrier against
microbial penetration.

METHODS OR DETECTING MICROORGANISMS IN FOODS

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Various techniques are used to determine the number and kind of microorganisms in food
as part of quality control and to detect organism responsible for spoilage or food borne
diseases. This is done by:

1. Direct microscopic examination of food sample

A sample of food is taken and examined under the microscope for fungal spores and
hyphae. Gram stain method to determine bacteria.

2. Visual observation of food samples for signs of spoilage due to microbial


action.
3. Prolonged incubation of fruit samples at suitable temperature and media is
useful in detecting the presence of spores of members of the Bacteriaceae.
4. Enumeration of indicator type of bacteria e.g. E.coli, Pseudomonas
aeroginosa, Staphylococcus aureus are indicators of faecal or human
contamination of unprocessed foods. E.coli; Enterobacter and Serratia species
may also be indicators of post-heat processing examination in the case of
pasteurized and canned foods.
5. Test for microbial metabolites by chromatographic methods, immunological
and biochemical tests. The presence of microbial metabolites such as toxins
even in the absence of overt spoilage is evidence of microbial presence and
action in food.
6. Membrane filtration

This is very useful in determining microbial presence in beverages and water. It is also
used in detecting microbial present in beer, wine and dairy foods. The advantage is that
large volume of foods can be filtered rapidly through membrane filters.

After filtration, the filters are then removed and placed directly on suitable media and
then incubated. After 24 hours, counting is done. The problem with this method is that
some foods especially the dairy foods clock the pores of the filters during the filtration.
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This is prevailed by carrying out pre-filtration to retain the clocking materials by the
larger diameters.

7. A standard plate count is used for many types of food e.g it is routinely done
for milk.

(a) Specific staining procedures of food samples to make observations of the


microorganism present easy.
(b) Direct isolation by plaiting out food samples in suitable media and
incubating. Make counts after the incubation period. Examples of media
that can be used include: -For direct count, cooked meat media are used.
Liver broth to detect the presence of anaerobes in canned foods.
MacConkey Agar for detection of Salmonella, Shigella in shell fish.

8. Malt-extract agar, Potato dextrose agar, Rose Bengal – Chloramphenicol agar,


Wort agar can be used to detect yeasts and mould. Wort agar is very useful for
detecting osmophilic yeasts in sugary foods.Incubate food samples at mimic
temperatures to encourage growth of microorganisms present in foods
preserved by low temperature application e.g. temperature of 15 - 17ºC is
suitable for the development of psychrotrophic organism that might cause
spoilage during refrigerated distribution and storage.

9. Resuscitation treatment.

This is useful for canned foods. A sample of food is plated on solid recovery medium for
some hours at room temperature to eliminate stresses that the organism might have been
subjected to during heat processing. It is then plated on suitable selection media.

FOOD BORNE ILLNESSES


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These may be microbial or non-microbial in origin. Food borne illnesses are condition of
distress experience as a result of consumption of food or drinks.

Non-microbial types

This may be due to intolerance, or sensitively to food and this result in allergic reactions
of various types e.g. the sudden and rapid swelling of the eyes or the whole face
following the ingestion of particular food e.g.

1. Eggs: This is also referred to as immunological hypersensitivity because peculiar


antigen/antibody reaction occurs after the injection of food or inhalation of food dust.
2. Metal poisoning: The metal toxic to man and animals are common in the environment
and processing e.g. Pb, Ar, Ag which are common in water, if in a food processing
plant, water that is high in lead content is used, there may be accumulation in food
which causes illness in the body. Also, Pb, Ar, Cu which are used as fungicides on
plants, may leave residues on the fruits or leaves and when ingested they may be
injurious when accumulated to the threshold level.
3. Phyllatoxins: There are of plant origin. Many plants produce toxins that have adverse
effect on man and animal when accumulated to the threshold level e.g. CN is present
in the starchy root of cassava, also caffeine which is a stimulant is present in coffee
and tea and this could cause excitement.

Microbial foodborne illness (Food Poisoning)

Human illnesses caused by foodborne microorganisms are popularly referred to as food


poisoning. The common use of a single classification is due primarily to similarities of
symptoms of various food-related diseases. Food poisoning may be divided into two
major classes, food infection and food intoxication. Food infection results when foods
contaminated with pathogenic, invasive, food poisoning bacteria are eaten. These bacteria
then proliferate in the human body and eventually cause illness. Food intoxication
follows the ingestion of preformed toxic substances which accumulate during the growth
of certain bacterial types in foods.
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The period of time between the consumption of contaminated foods and the appearance
of illness is called the incubation period. The incubation period can range anywhere from
less than one hour to more than three days, depending on the causative organisms or the
toxic product.

Characteristics of the important bacterial food intoxications and foodborne


infections.

Incubation
Disease Etiologic Agent Symptons
Period

Difficulty in swallowing,
double vision, difficulty
in speech. Occasionally
nausea, vomiting, and
Usually 1 to 2
diarrhea in early stages.
Clostridium days; range
Constipation and
1. Botulism botulinum A.B.E.F 12 hours to
subnormal temperature.
toxin more than 1
Respiration becomes
week
difficult, often followed
by death from paralysis
of muscles of
respiration.

2. Staphylococcal food Staphyloccal 1 to 6 hours; Nausea, vomiting,


poisoning enterotoxin average 3 abdominal cramps,
hours diarrhea, and acute
prostration. Temperature
subnormal during acute
attack, may be elevated
later. Rapid recovery-

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usually within 1 day.

Abdominal pains,
Average
diarrhea, chills, fever,
Specific infection about 18
3. Salmonellosis frequent vomiting,
by Salmonella spp. hours; range
prostration. Duration of
7 to 72 hours
illness: 1 day to 1 week.

Abdominal cramps,
fever, chills, diarrhea,
Shigella sonnei, S. Usually 24 to watery stool (frequently
4. Shigellosis (bacillary flexneri, S. 48 hours; containing blood,
dysentery) dysenteriae, S. range 7 to 48 mucus, or pus), spasm,
boydii hours headache, nausea,
dehydration, prostration.
Duration: a few days.

Headache, malaise,
Escherichia coli Usually 10 to
5. Enteropathogenic fever, chills, diarrhea,
serotypes associated 12 hours;
Escherichia coli vomiting, abdominal
with infant and range 5 to 48
infection pain. Duration: a few
adult infections hours
days.

Abdominal cramps and


diarrhea, nausea, and
Usually 10 to
6. Clostridium malaise, vomiting very
Clostridium 12 hours;
perfringens food rare. Meat and poultry
perfringens range 8 to 22
poisoning products usually
hours
involved. Rapid
Recovery.

7. Bacillus cereus food Bacillus cereus Usually about Similar to Clostridium


poisoning 12 hours; perfringens poisoning

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range about 8
to 16 hours

Abdominal pain, server


Usually 12 to watery diarrhea, usually
8. Vibrio
Vibrio 14 hours; nausea and vomiting,
Parahaemolyticus
parahaemolyticus range 2 to 48 mild fever, chills and
food poisoning
hours headache. Duration: 2 to
5 days.

Many of these illnesses occur in the gastrointestinal tracts while some of the pathogens of
their toxins can also enter into the blood stream and get distributed all over the body
where they can cause localized diseases of organs or tissues.

9. Traveler’s diarrhea: This is caused by the notorious E. coli and some spp. of
Salmonella and Shigella. Symptoms include abdominal pains, diarrhea, nausea,
dehydration, chills; symptoms usually appear within 2-5days of consuming
infected food and water. The above two illness are common and the main source
of infection is water.
10. Typhoid fever: This is caused by Salmonella typhi. Symptoms include abdominal
distention, constipation, fever, headache, loss of appetite; nausea, vomiting,
diarrhea and complications may include intestinal bleeding and pneumonia.
11. Asiatic cholera: Common in Asia, caused by Vibro cholerae. Symptoms include
large amount of mucus in stool, loss of water and electrolytes, dehydration, shock,
collapse and death can occur without treatment. The diarrhea involved is caused
by the action of an exo-enterotoxin called choleragen produced by the organism.
The disease is transmitted by injection of food or water contaminated by faeces of
infected person or coming into direct contact with an infected person faeces.
Houseflies may also spread the disease. The organism can adhere to intestinal
linings of humans.
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12. Staphylococci food poison: Caused by Staphylococcus aureus. Foods that are
common sources of infection include unrefrigerated leftover meat, fish and dairy
products left at temperature at or above room temperature for several hours.
Enterotoxins are produced which are not inactivated by heat. Symptoms include
abdominal cramps, chills, diarrhea, headache, nausea and vomiting.
13. Mushroom poisoning: This is commonly caused by injecting species or strains of
mushrooms that produce toxins e.g. Amanita phalloides which is called the death
cap in Central Europe. This mushroom bears some resembles to the edible type,
Agaricus and thus are exchanged for each other with death as the result.
Phallotoxins and amatoxins are the general names of toxins produced and their
effect is on the CNS causing hallucinations. Only a few species of mushrooms are
edible. Vomiting and diarrhea are the first noticeable effect of mushroom
poisoning.
14. Ergotism: This is caused by Claviceps purpurea, the ergot fungus. The organism
produces ergot on rye and other grains. Ergot contains alkaloids that are very toxic
but upon purification, the components are of great use in medicine. One of the
components, Ergometrine is useful in inducing early pregnancy or later to hasten
child birth. It is also used to treat migraine. Other examples are ergonine and
ergotamine.
15. Aflatoxins: These are produced by certain strains of Aspergillus flavus and A.
parasiticus. The fungus is common on moldy groundnut. Aflatoxins are of various
types, aflatoxin A, B1, B2, G1, G2, and M1, M2. Aflatoxins are implicated in
human carcinogens. Studies have shown that extensive liver cancers occur in
Asiatic country where mold fermented foods are commonly consumed.
16. Patulin: this is a toxic and carcinogenic compound produced by Penicillium
griseofulvum, P. expansum and Aspergillus terreus, A. gigantus. This toxin has
been found associated with food substances such as moldy bread, grains, sausage
and fruit.

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FOOD QUALITY CONTROL

In food processing factories, adequate measures are taken to ensure that food being
produced and released to the market are of the right quality. There are set regulations in
different countries on the microbiological quality of foods meant for human consumption.
Microbiological quality control therefore involves the examination of food at every stage
of processing for the presence of microbes or their metabolites that

1. Interfere with the processing


2. Cause spoilage
3. Cause diseases

Finished products are also rigorously examined for microbes and their metabolites to
determine the level of contamination by pathogenic or spoilage organisms and compared
with the acceptable number of colony forming unit (CFU). Routine checking are carried
out on foods by visual observations for mechanical defects such as leaks, dents, rusts in
canned foods and bruises, wounds, bleeding in fresh plants or animal foods. If the visual
observation result is greater than 3% the following should be carried out.

1. Direct microscopic examination to identify the microorganisms associated.


2. Perform different isolation techniques to determine the actual types and number of
microbes present.
3. Perform biochemical and physiological characterization to determine the effect of
microbes present on food quality and consumers.
Pasteurized foods are not microbiologically sterile but the number of spoilage
organisms have been reduced while all pathogenic organisms would have been
destroyed by the heat treatment e.g. milk is pasteurized at 71 o C for 15 sec or at 63o
C for 30mins.

Sterilized foods contain no microorganisms at all. Heat and radiations are employed in
food sterilization to destroy all microbes. Higher temperature is made use of in
sterilization than in pasteurization.
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