This document summarizes procedures for examining milk samples from cattle. It describes how to collect milk from each mammary quarter, noting the importance of cleaning and labeling samples. Milk should be refrigerated if transported for testing. Physical examination of milk considers attributes like color, odor, consistency, pH and specific gravity. Chemical tests described are the Milk Ring Test to detect Brucella antibodies and the California Mastitis Test to detect somatic cells indicating infection. Somatic cell counts normally below 200,000 cells/ml indicate lack of mastitis.

1. Milk
Examination
In veterinary
By Dr. Shashank

2. (1)Collection of milk sample
a)First wash the udder and the teats with
clean water thoroughly and wipe it with
clean cloth.
b)Swab the teats with 70 per cent alcohol or
tincture iodine.
c)Label sterile tubes/vials as RH, RF, LF & LH.
d)Discard first 2-3 streaks of milk.
e)Then collect 5 ml (approx) of milk from
each quarter in respective tubes/vials.

3. (2) Storage
✔Milk must be refrigerated if it is
to be transported to a distant
place for laboratory tests.

4. (3)Examination
a)Physical examination
✓ Color
✓ Odour
✓ Consistency
✓ pH
✓ Specific gravity

5. ✓ Color of Milk –
Normal: white in buffalo and whitish
yellow in cow
Abnormal:
#Blood stained: Highly produced animal’s
pathological discoularation
#Red: Dicumarol toxicity, Leptospirosis,
Stapyylococcosis, Bacterium
erythrogene
#Green: Corynebacterium
#Yellow: most of bacterial Mastitis.

6. Blood stained milk

7. ✓Odor –
Normal milk: milky odor
Abnormal:
#Fetid (putrefied) odor: Gangrenous mastitis,
Dry cow
#Fecal odor: Coliform mastitis
#Rancid odor: Milking in bad ventilated place
or leaving milk for 2 days in
environment
#Antiseptic odor: Excessive dipping of teat
#Acetone odor: Ketosis

8. ✓Consisitency -
Normal: thin of cow milk thick of
buffalo
Abnormally:
#Watery milk (decrease
consisitency): Streptococcal mastitis
#Increased consisitence:
Corynebacterial mastitis

9. ✓ pH: The pH of normal milk is 6.4 to 6.6.
But in mastitis it becomes alkaline upto 7.4
due to presence of sodium bicarbonate in
the milk. It can be determined by using pH
strips.
✓ Specific gravity: The specific gravity of
normal milk is 1.015 to 1.037 which may
get altered due to mastitis.

10. b)Chemical examination
✓Milk ring test
#The milk ring test is a type of agglutination test
that detects brucella antibodies in milk samples and
is recommended as a screening test for bovine
brucellosis by OIE.
#It consists of Brucella abortus cells stained with
hematoxylin, which are added to a sample of 1 to 2
ml of milk and incubated at 37°C for one hour. If
antibodies for Brucella abortus are present in the
milk the antigen agglutinate with the cells, and float
to the surface where it forms a sharp blue or purple
colored cream layer. The milk column underneath
will remain white.

11. Procedure MRT
✓ Collect milk from an animal, sample is
identified by date of collection and refrigerated
✓Gentle mix the milk in sample tube to ensure
distribution of cream
✓ Place 1 ml of milk in a plastic agglutination
tube to give a column of milk about 2 cm high
✓ Add 1 drop of milk ring test antigen or B.
abortus suspension by pipette into test tube.
✓Gentle mix thoroughly the contents within 1
minute of adding antigens and avoiding
frothing and incubate the mixture at 37 oC for
39-40 minutes and then examine.

12. Positive test result :
cream layer is deeply
coloured and beneath
cream layer is white
Negative test result :
cream layer is white, milk
beaneth is white coloured
Interpretation:

13. ✓Calfornia mastitis test
-It operates by disrupting the cell membrane of any
cells present in the milk sample, allowing the DNA
in those cells to react with the test reagent, forming
a gel.
CMT Reagent:
Solution A: Sodium lauryl sulphate 30 gram
dissolved in 1000 ml distilled water, heated to 50°C
to make a clear solution.
Solution B: Bromocresol purple 0.5g dissolved in
100ml distilled water.Mix 100ml solution A and 2 ml
solution B.

14. Procedure CMT
✓Collection of milk samples from each quarter of udders
in the CMT paddle. The paddle has four shallow cups
marked in different to identify the individual quarter
from which milk sample was obtain.
✓Take about 2 ml or 1 teaspoon of milk from each
quarter.
✓Add an equal amount of CMT solution to each cup in
the paddle.
✓Rotate the CMT Paddle in a circular motion to mix the
milk contents.
✓The result is observed after about 20 seconds.
Note: Cow in early and late stages of lactation may give false +ve reactions.

15. 1 2
3 4

16. ✓ Formation of gel indicates +ve reaction

17. ✓Somatic cell count
✓Somatic cells which found in milk are composed
of white blood cell and epithelial cells. Cell white
blood cell found in normal milk from uninfected
glands include neutrophils, macrophages,
lymphocytes and epithelial cells. SCCs are a
measure of the number of cells present in the milk.
✓The largest factor that infl uences the SCC of milk
is mastitis. The SCC of a cow that is not infected
with mastitis is usually less than 200,000 cells/ ml
and many cows maintain SCC values< 100,000
cells/ml

19. Thank you
!