Separation of deterrents to ingestive behavior of cattle from cattle feces

Feeding-deterrent chemicals were extracted from cattle feces and then separated with three chromatographic methods. Behavioral two-choice test bioassays with cattle were used to examine the deterrent properties of the fractions. Cattle feces were extracted with diethyl ether, and the extracts were separated into neutral, acidic, and basic fractions. Of the three fractions, only the neutral fraction was a deterrent. Separation of the ether-soluble neutral chemicals was conducted with an open column of silica gel using four carrier solutions consisting of pentane and ether. Fraction B (eluted with the carrier solution; pentane:ether = 90:10) was the most effective deterrent among the four fractions. This fraction was divided into 10 fractions by liquid chromatography. Fractions 6, 7, and 8 seemed to deter cattle from feeding. The combined Fractions 6, 7, and 8 were separated into 15 fractions with HPLC. Deterrent activities were detected in Fractions 2, 3, 9, 10, 11, 12, 13, and 14, suggesting that deterrents were separated into two groups using HPLC. These results suggested that several specific chemicals in feces are involved in inhibiting cattle from ingesting grass near cattle feces.