California Mastitis Test Reagent (MASTEST)

Indication :   For performing California Mastitis Test for the detection of subclinical mastitis

Procedure :  California Mastitis Test is usually performed using a white plastic paddle of a special design. About 2ml of milk is taken directly to the receptacles of the plastic paddle from all the four quarters and the test solution is squirted into the pool of milk by squeezing it from a plastic wash bottle. By estimation, the test solution is added in equal quantity to that of milk.The reagent and the milk are mixed by gentle circular rotation of the plastic paddle in a horizontal plane. Since the reagent thickens immediately it is important to have sufficient quantity of the test solution. Hence it is better to have a little excess than too little of the reagent. Insufficient reagent precludes the full development of a positive reaction, while excess do not interfere rather than a waste of reagent.  The paddle is prepared for use on another cow by emptying its contents and rinsing it in clean water.

Kit :  Contains the antigen, positive serum and a negative serum

Testing : Collect the blood and separate the serum. One drop of serum is put on a glass slide. One drop of antigen is then added to the serum. The serum and antigen are then mixed quickly using a tooth pick or a match stick (use separate tooth pick for each serum sample). The slide is held over a white surface and rocked gently from side to side for 4 to 5 minutes. A strongly positive reaction will be visible during first 30 seconds as distinct clumps.

Storage : The antigen should be stored at 4oc.expiry date is 9 months from the date of manufacture

Grading and Interpretation  : Sym bol Suggested Meaning Description Interpretation - Negative Mixture remains liquid with no evidence of formation of a precipitate < 2,00000 cells/ml T Trace A slight precipitate forms and is seen to best advantage by tipping the paddle back and forth as it flows over the bottom of the cup. Trace reaction tend to disappear with continued movement of fluid 150000 to 500000  cells per ml 1 Weak positive A distinct precipitate occurs with no tendency towards gel formation, and with continued movement of the paddle the precipitate may disappear. 400000 – 1500000 cells per ml 2 Distinct positive The mixture thickens immediately with some suggestion of gel formation. As the mixture is caused to swirl it tends to move towards the centre leaving the bottom of the outer edge of cup exposed. As the motion is stopped the mixture levels out again and covers bottom of cup 800000 – 5000000 cells per ml. 3 Strong positive A gel is formed which causes the surface of the mixture to turn convex. There will be a central peak projecting over the main mass after the motion of paddle is stopped. Viscosity is increased and there is a tendency of mass to adhere to bottom of the cup. > 5000000 cells per ml. + Alkaline milk The reaction is distinctly alkaline indicated by a deeper purple colour of mixture pH 7.0 or above        Y Acid milk Brom cresol purple is distinctly yellow at an acidic pH. It indicated fermentation of lactose by bacterial action within the gland        pH 5.0 or below Presentation :  100 ml and 500 ml Technical Collaboration : Department of Preventive Medicine, College of Veterinary & Animal Sciences, Kerala Agricultural University, Thrissur.