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Milk
Examination
In veterinary
By Dr. Shashank
(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.
(2) Storage
✔Milk must be refrigerated if it is
to be transported to a distant
place for laboratory tests.
(3)Examination
a)Physical examination
✓ Color
✓ Odour
✓ Consistency
✓ pH
✓ Specific gravity
✓ 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.
Blood stained milk
✓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
✓Consisitency -
Normal: thin of cow milk thick of
buffalo
Abnormally:
#Watery milk (decrease
consisitency): Streptococcal mastitis
#Increased consisitence:
Corynebacterial mastitis
✓ 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.
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.
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.
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:
✓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.
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.
1 2
3 4
✓ Formation of gel indicates +ve reaction
✓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
Thank you
!

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Veterinary laboratory diagnosis

  • 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.
  • 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.
  • 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
  • 18.