Reliability criteria of a bioassay using rats trained to detect estrus-specific odor in cow urine

An odor discrimination apparatus was used to quantify the reaction of rats against odor differences of estrous and diestrous urine of cows. Rats were trained to release impulses from a microswitch when they came into contact with estrous but not diestrous urine. After training, the discrimination ability was highly significant (P< or = 0.001) between both urine types. The reaction was specific for cow urine, and no significant discrimination occurred for estrous and diestrous urine from pigs or goats, or that of women. The discrimination of urine diluted with water was still highly significant up to 1:25. Another group of rats was trained to detect camphor. Decreasing concentrations were detected down to 0.1/microg/ml when diluted in water, and down to 1/microg/ml when diluted in estrous urine. Once trained to discriminate between estrous and diestrous urine, rats maintained this ability for 12 mo. Thus, a bioassay was set up to accompany fractionation steps in the laboratory which could lead to the isolation of estrous pheromones in cow urine.