Indicator Microorganisms and

Microbiological Criteria
~ Why we need to know microbiological criteria?
To distinguish between an acceptable and
unacceptable products, between acceptable and
unacceptable food processing practices.

~ Indicator microorganisms may be used to assess

either the microbiological quality or safety.

~ Microbiological criteria: the safety of food,

adherence to good manufacturing practice, the
keeping quality (shelf life of perishable food), the
suitability of a food or ingredient for a particular
purpose.
The Importance of Microbiological Criteria
1. Evidence of a health hazard based on epidemiological data
or hazard analysis.

2. The nature of the food’s normal microbial make up and the

ability of the food to support microbial growth.

3. The effect of processing on the microflora of the food.

4. The potential for microbial contamination and/or growth

during processing, handling, storage and distribution.

5. Spoilage potential, utility and GMPs.

 Decision on Establishing Microbiological
Criteria
International
~ Joint Food and Agricultural Organization (FAO),
World Health Organization (WHO), Codex
Alimentarius International Food Standards
Program.

Malaysia
~ Veterinary Department, FAMA, MARDI etc.
SAMPLES OF MILK PRODUCTS
LABELLING
 SAMPLING PROCEDURE

~ SAMPLING PLAN must include the sampling

procedure and decision criteria.
eg: representative sample from a lot. Let say 10
samples were taken and decision criteria was to
reject the lot if more than 2 showed positive
results containing bacteria.
Then the whole lot must be rejected if more than
2 give positive result.
HOW TO CHECK BATCH NUMBER
~ESTABLISHING LIMITS depends on knowledge
on raw materials and the effects of processing,
product handling, storage, and the end use of the
products.

~ Perishable products shelf life depend on number

of microorganisms initially present. General rule; a
food containing a large population of spoilage
organisms will have shorter shelf life.

~ Foods produced and stored under GMPs may be

expected to have better microbiological profile than
those produced and stored under inferior places.
~ Use of quantitative risk assessment techniques
to scientifically determine the probability of
occurrence of known human exposure to food
borne hazards.

~ The process consists of hazard identification,

hazard characterization, exposure assessment
and risk characterization.
~ INDICATORS OF MICROBIOLOGICAL QUALITY can
provide simple, rapid information, reliable results, post
processing contamination, contamination from the
environment and the general hygiene under which the
food was processed and stored.

~ Ideal indicators of product quality or shelf life should

meet these criteria:
1. Should be present and detectable in all foods whose
quality to be assessed.
2. Their growth and numbers should have a direct
negative correlation with product quality.
3. They should be easily detected, enumerated and
clearly distinguishable from other microorganisms
4. They should be enumerable in short period of time,
ideally within 24 hours.
~ Indicator microorganisms show that their present
suggest a microbial hazard. e.g: presence of Escherichia
coli in a sample indicates possible fecal contamination,
Bacillus sp. In bread dough, lactic acid bacteria in beers
and wines, Lactobacillus lactis in raw milk (refrigerated),
yeasts in fruit juice concentrates.

~ Aerobic plate counts (APC) use to determine the total

number of microorganisms in food products. APC is a
component of assessing food quality when monitoring
foods for compliance with standard guidelines set by
regulatory agencies, monitoring foods for compliance
with purchase specifications and monitoring adherence
to GMPs.
~ However APC measures only live cells. APC
may be lower when spoilage microorganisms
died. APC are of little value in assessing sensory
quality.

~ Alternatively, use direct microscopic count

(DMC), use to estimate both viable and non
viable cells in sample containing large number
of microorganisms (ie > 105 CFU/mL), Howard
mold count, yeast and mold count, thermophilic
mold count.
~ Metabolic products can be used to estimate
bacterial levels in food products. e.g. cadaverine
and putrescine in vacuum packaged beef,
histamine in canned tuna, lactic acid in canned
vegetables, total volatile bases in seafoods and
volatile fatty acids in butter and cream.
Indicators of Food borne Pathogens and
Toxins
~ Microbiological criteria should be developed in order to
reduce or eliminate a food borne hazard AND to ensure
product safety.

~ Must be developed for each food type and evaluated

through a risk assessment to determine the potential
hazards and their significance to consumers.

~ Microbiological criteria apply to milk, shellfish and

vegetables. Contamination of food with pathogens that
cannot grow to a harmful level does not need
microbiological criteria.
~ Criteria may be appropriate for enteric pathogens on
produce since many outbreaks occur as foodborne
illness on fresh produce.

~ Depend on the pathogen, low level of microorganism in

the food product may or may not be of concern.

~ However, some microorganisms have a low infectious

dose that their presence create public health risk.
e.g: we assumed that certain acidity in fermented food
product to be sufficient for pathogen control. However,
while the growth and toxin production of S. aureus might
be prevented, enteric pathogens such as E. coli 0157:H7
could survive and produce a product that is unsafe for
consumption.
~ More rigid microbiological requirement may be
needed if the food to be consumed by infants,
the elderly or immuno compromised people.

~ The stringency of sampling plans is based on

either the hazard to consumer from the
pathogens and their toxins or toxic metabolites
or the potential for quality deterioration.

~ Foodborne pathogens are grouped into three

categories.
 Category 1 (Severe hazards)

Clostridium botulinum type A, B,E and F

Shigella dysenteriae

Salmonella enteritica serovars Paratyphi A and B

Vibrio cholerae O1

Hepatitis A and E viruses

Taenia solium

Enterohemorrhagic E. coli
Category II (moderate hazards, potentially extensive spread)

Listeria monocytogenes

Salmonella sp.

Shigella sp.

Other enterovirulent E. coli

Rotavirus

Entamoeba histolytica

Ascaris lumbricoides
 Category III (moderate hazards, limited spread)

Bacillus cereus

Camphylobacter jejuni

Clostridium perfrigens

Staphylococcus aureus

Vibrio cholerae, non O1

Vibrio paraheamolyticus

Yersenia enterolytica

Giardia lamblia

Taenia saginata
~ Microbial indicators should have characteristics such as:

1. Easily and rapidly detectable.

2. Readily distinguishable from microbes commonly associated with the food.

3. A history of constant association with the pathogens whose presence it is to

indicate.

4. Present when the pathogen of concern is present.

5. Number that ideally correlate with those of the pathogen of concern.

6. Growth requirements and growth rate must be equal to those of pathogen.

7. A die off rate that at least parallels that of pathogen and has a slightly longer
persistence than the pathogen of concern.

8. Absence from foods that are free of the pathogen except at certain minimum
numbers.
Case example:
~ Fecal coliforms and E. coli are easily destroyed by heat and
may die during freezing and storage of frozen food.

~ Microbial criteria involving E.coli are useful in cases where it

is desirable to determine if fecal contamination may have
occurred.

~ Contamination of a food with E.coli implies a risk that other

enteric pathogens may also present.

~ Use as microbial standards to monitor the wholesomeness

of shellfish and the quality of shellfish growing waters and to
reduce the chances of harvesting shellfish from water
polluted with fecal material.
~ Fecal coliforms include Klebsiella, Enterobacter and
Citrobacter species but these are known as false
positive indicators of fecal contamination since they can
grow in non fecal niches such as water, food and waste.

~As a result, only E.coli appropriate to be used as

indicator for fecal contamination.

~ In the Ready to eat food (RTE) with heat processed

procedure, the presence of E.coli means either process
failure or post processing contamination. In this case,
coliforms are a better indicator of process integrity (poor
sanitation, poor temperature control) since the level of
coliforms do not increase over time when the product is
properly stored.