SPECTROPHOTOMETRIC METHOD OF ESTIMATION OF TANNIN ACYL HYDROLASE ACTIVITY IN RUMEN DIGESTA

摘要

This invention relates to a spectrophotometric method of estimation of Tannin acyl hydrolase activity in rumen digesta, comprising, supplementing animals with two types of total mixed ration containing 0 and 4 percent tannin, wherein Acacia nilotica is used as a source of tannin, estimating the enzyme activity at 0 and 4 hr post feeding after a preliminary feeding of 21 days, collecting rumen digesta (strained Rumen liquor) at 0 and 4 hr post feeding using vacuum suction from rumen fistulated cattle, immediately after filtration through muslin cloth, rumen digesta is collected and preserved in an ice chamber, then sonicated for 5 minutes, centrifuged at 8000 rpm at 4°C for 15 min., dissolving tannic acid in cool citrate buffer on the day of use wherein different substrate concentrations of 0.3, 0.5, 0.7 and 1.0% are used, adding sample rumen digesta to citrate buffer, incubating at 38°C for 40 minutes and stopping reaction by adding 3ml of DNS solution, keeping the tubes in boiling water for 15 minutes, adding 1 ml Na-K tartarate solution to each tube, cooling the tube to room temperature, final volume made up to 20 ml and O D taken at 575 nm.