Answers to MCB 403 exam questions for 2014/2015

QUESTION 1: Give a detailed description of the various techniques employed in detecting the
presence of microorganism in foods.

ANSWERS:

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 stains 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. 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.
8. Specific staining procedures of food samples to make observations of the
microorganism present easy.
9. 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: -

(a) For direct count, cooked meat media are used.

(b) Liver broth to detect the presence of anaerobes in canned foods.
(c) MacConkey Agar for detection of Salmonella, Shigella in shell fish.
(d) 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.

10. 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.
11. 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.

QUESTION 2: What roles do microorganisms play in the production and preservation of the
following foods?

a) Garri b) Sauerkraut c) breads

Answers

So sorry, answer to the above questions not gotten yet.

QUESTION 3: With four specific examples each, enumerate microbial spoilage associated with
the following foods i) fish and other sea foods ii) Eggs iii) vegetables and fruits

Answers

i) 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 shinning 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 protolysisof 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.

ii) EGGS

(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

Aerobactercloacae gives the odour of hay and Achromobacterperolens gives a musty
odour.

iii) VEGETABLE AND FRUIT

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. Softrot 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 candegrade 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.
iv. iv .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.

QUESTION 4: Describe any 10(ten) different types of microbial food poisoning in a tabular
form, under the following headings :diseases , etiology agent, incubation period and
characteristics

Answers:

Incubation
Disease Etiologic Agent Symptons
Period

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

Nausea, vomiting,
1 to 6 hours; abdominal cramps, diarrhea,
2. Staphylococcal food Staphyloccal
average 3 and acute prostration.
poisoning enterotoxin
hours Temperature subnormal
during acute attack, may be
 elevated later. Rapid
recovery-usually within 1
day.

Abdominal pains, diarrhea,

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

Abdominal cramps, fever,

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

Escherichia coli Usually 10 to Headache, malaise, fever,

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

Abdominal cramps and

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

7. Bacillus cereus food Usually about Similar to Clostridium

Bacillus cereus 12 hours; perfringens poisoning
poisoning
range about 8
 to 16 hours

Abdominal pain, server

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

QUESTION 5a: Briefly discuss food borne illnesses of non microbial origin.

Answers:

Non-microbial food borne illnesses

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.

QUESTION 5b: Describe any five (5) factors that can affect microbial growth in foods

Answers:

The following are the factors that affect microbial growth

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 they 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.

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 Aureobasidiumspp. grow commonly on foods
under refrigerator. Thammedium spp grow readily on cold store, cured, wheat.
Streptococcusthermophiluscan 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. The food processor can feel safe in
the knowledge that foods held above or below the limits in Figure 1 and rotated properly
will remain safe. 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 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.
Pediococcusthalophilusgrow 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.

QUESTION 6: Discuss the various methods of controlling food spoilage.

Answers: 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 (aw) 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 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) Incipentthermophilic
(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 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.