Types of bacteria found in milk:

Milk supports the growth of a variety of bacteria including pathogenic one

The different types of bacteria present in milk are, as follows:
1. Acid-fonning bacteria, such as
o Streptococcus lactis
o Str. faecalis
o Lactobacilli
These ferment lactose, forming lactic acid, and lead to the formation of curd.
2. Alkali-fonning bacteria, such as
o Alkaligenes sp.
o Achromobacter
o Aerobic spore-forming bacilli
These render the milk alkaline.
3. Gas-forming bacteria, such as
o Coliform bacteria
o Cl. peifringens
o Cl. butyricum
These produce acid and gas.
4. Proteolytic bacteria, such as
o Bacillus subtilis
o B. cereus
o Proteus vulgaris
o Staphylococci
o Micrococci
These bacteria are responsible for proteolytic activity.
5. Inert bacteria, such as
o Achromobacter
o Pathogenic bacteria
o Cocci
They do not produce any visible change.

Diseases and infections transmitted through milk are:

I. Infections of animals transmitted to man
o Tuberculosis
o Brucellosis
o Streptococcal and staphylococcal infections
o Salmonellosis
o Q fever
o Anthrax
o Leptospirosis
o Cowpox and milker's nodes
 o Foot and mouth disease
o Tickborne
o viral encephalitis
2. Infection due to ingestion of milk contaminated with
excreta of small mammals
o Streptobacillus moniliformis
o C. jejuni
o Y. enterocolitica

3. Infections primarily of man transmitted through milk

o S. typhi
o Paratyphoid bacilli
o holera vibrio
o Shigella
o E. coli
o Streptococcal and staphylococcal infections
o Tubercle bacilli
o Hepatitis virus
o Diphtheria bacilli

The tests used for examining bacteria present in milk are:

1. Viable count
· It is detected by plate counts with serial dilutions of the milk samples

2. Coliform count
o Dilutions of milk are inoculated into MacConkey's liquid medium
o The production of acid and gas is noted after incubation-positive test indicates
presence of coliform bacilli (in two tubes out of three)
o All coliforms are killed by pasteurization
o The presence of coliforms in pasteurized milk indicates improper pasteurization or
post-pasteurization contamination

3. Methylene blue reduction test

o An indirect method for detection of microorganisms in milk
o An economical substitute for viable count
o Procedure: 1 ml of methylene blue is mixed with 10 ml of milk in a test tube and
o incubated in dark at 37°C
o Milk is considered satisfactory if it fails to decolourize methylene blue III 30 minutes

4. Resazurin test
It is also a dye reduction test similar to methylene blue dye test. Reduction of resazurin by
bacteria passes through a series of color changes-blue to pink to colorless, which depends
upon degree of contamination
5. Phosphatase test
Phosphatase is an enzyme normally present in milk, which gets inactivated by pas
teurization of milk.
Its presence in milk after pasteurization indicates improper/incomplete pasteurization. Its
presence is
detected by adding disodium phenyl phosphate to 1 ml of milk in a test tube and
incubating in a water
bath at 37°C for 2 hours.
Development of yellow color indicates presence of phosphatase

6. Turbidity test
This test is used to check sterilization of milk. When milk is sterilized properly by heating
at 100°C for
5 minutes, all coagulable proteins are precipitated and when ammonium sulphate is
added to this
milk, no turbidity results because of denaturation of proteins. This indicates that milk has
been heated
at 100°C for 5 minutes

Specific tests include:

1. Test for tubercle bacilli

The milk is centrifuged at 3000 rpm for 30 minutes and sediment is inoculated into two
guinea pigs and two L-J media. L-J media are observed for growth and guinea pigs are
observed for a period of 3 months for tuberculosis. Confirmation is done by using
biochemical tests.

2. Tests for Brucella bacilli

Isolation of brucellae is attempted by inoculating cream from the milk sample on serum
glucose agar and then injecting centrifuged deposit of the milk sample intramuscularly in
guinea pigs. Guinea pig can be sacrificed after 6 weeks and the spleen used for culture of
brucella and serum used for demonstration of Abs by milk ring test.
Milk ring test is the highly sensitive test for demonstration of brucella Abs in the milk of
infected cows. The test is used for diagnosis of brucellosis in animals. The positive milk
ring test is confirmed by Whey's agglutination test
In case of suspected food poisoning, the sediment of centrifuged milk should also be
examined for:
o Staph. aureus
o Salmonella sp.
o Campylobacter sp.
o Y. enterocolitica