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 4C, 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 4C and not frozen.

Before use, the staining solution is shaken and filtered

through cotton wool

METHOD OF PERFORMING THE TEST

1.
2. 3. 4.

5.

Mix the milk thoroughly and transfer on ml to a narrow test (11 x 100 mm.) Add one drop (0.03 ml) of the antigen and mix. Incubate at 37o C for one hour and read using a uniform light source. 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 If the intensity of the colour in the cream layer is less than in the skim potion, the test is considered negative

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 antigenantibody 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

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