Milk

MILK
 Milk is the normal mammary secretion of milking
animals obtained from one or more milkings
without either addition to it or extraction from it,
intended for consumption as liquid milk or for
further processing.
 As an agricultural product, milk is extracted from
mammals (Cattle,Goats,Camels & Sheeps) and
used as food for humans.
ARE YOU SURE ABOUT
YOUR MILK??
It is forbidden to deliver or receive
milk
 from diseased cattle (goats, sheep, buffalos) or during
its treatment.
 after treatment but before the end of used medicines
withdrawal time
 not later than 3 weeks before calving and not earlier
than 6 days after calving
 in case of reception ban established by official
veterinarian
Milk should derived from animals
 healthy
 free of brucellosis and tuberculosis
 free of visible mastitis symptoms
MILK RING TEST
 a special form of agglutination test
done on the pooled milk of many cows,
usually entire herds, for the detection
of herds containing individuals
infected with bovine brucellosis.
Brucellosis in cattle
Brucella abortus
 Caused by bacteria that in gram negative
,facultative, non motile, on spore forming, rod
shaped.
 Mainly among cattle.
 This is a Zoonotic disease.
 Human cause disease – Undulant fever
 Can be transmitted to humans by
* By consuming Unsterilized milk or meat.
* Splashing of infected droplets.
Process
Antigen production
 MRT antigen is prepared from concentrated, killed B.
abortus strain 99 or 1119-3 cell suspension
 It is centrifuged for 10 minutes at 4°C, followed by
resuspension in haematoxylin-staining solution.
 After standing for 30 minutes at room temperature
the deep purple solution is added to 940 ml of 10%
(w/v) ammonium aluminium sulphate in distilled
water.
 Maintain the pH at 3.1
 the solution must be aged by storage at room
temperature in the dark for 45- 90 days.
 The antigen must be stored at 4°C and not frozen.
 Before use, the staining solution is shaken and filtered
through cotton wool
METHOD OF PERFORMING THE
TEST
1. Mix the milk thoroughly and transfer on ml to a
narrow test (11 x 100 mm.)
2. Add one drop (0.03 ml) of the antigen and mix.
3. Incubate at 37o C for one hour and read using a
uniform light source.
4. If the intensity of blue colour in the cream layer is
equal to or deeper than in the skim portion, the test
is considered Positive
5. If the intensity of the colour in the cream layer is less
than in the skim potion, the test is considered
negative
A B
Negative

Ring of cream less colored

than the underlying milk
 Positive

Ring of cream
equal or more
colored than the
underlying milk
Mechanism
 This method used to detect brucella antibodies in
milk samples
 After adding the colored bacterial antigen of
brucella to the milk sample, it form antigen-
antibody complexes based on the principle of
agglutination.
 Those made complexes are progressively carried by
the fat towards the surface of the milk and formed
a blue violet ring.
AFTER THE M.R.T
 After obtaining the positive Results from M.R.T.
It is useful to carry on confirmatory tests such as
ELISA, CFT ,SAT
PRECAUTIONS
 Milk samples should be mixed thoroughly
before applying the test.
 Shake the antigen very well before use.
 Known positive and negative samples
should be used in each test.
Advantages of Milk Ring
Test
 One of the Simple, Cheap Methods to
determine the possible presence of
Brucellosis in a herd of cows
 Not Time consuming
 Easy to perform
 Need less equipments
Limitations of Milk Ring Test
M.R.T Cannot be considered as the best test
to determine Brucellosis in Milk. Because
the tendency to give false positive results is
higher in this method.

 Milk samples from newly vaccinated cows

 Colostrum milk
 From abnormal milk like Mastitis
 Not effective with pasteurized or treated
milk
 Milk of sheep and goats also gives false
observations
 May occur at the periods where a high
proportion of the animals in the herd are
drying off.
References
 Ajogi, I. (1998). Settling the nomads in Wase and Wawa-
Zange grazing reserves in the Sudan Savannah zone of
Nigeria; Strategies for the control of bovine brucellosis.
Nigerian Veterinary Journal 19
 Hamilton A.V. and Hardy, A.V. (1950). The Brucella ring
test; its potential value in the control of brucellosis.
American Journal of Public Health 40, 321
 Corbel, M.J. (1997). Brucellosis: An Overview, Emerging
Infectious Diseases 3.213-221
 JOINT FOOD AND AGRICULTURE ORGANIZATION OF THE
UNITED NATIONS/WORLD HEALTH ORGANIZATION EXPERT
COMMITTEE ON BRUCELLOSIS (1986). Technical Report Series 740,
Sixth Report. WHO, Geneva, Switzerland
THANK YOU