Seven phenolic compounds (phenol, 3- and 4-cresols, 3- and 4-ethylphenols, and 3- and 4-n-propylphenols) previously shown to be components of a fraction of an extract of buffalo urine which was active as a tsetse attractant, were evaluated in field experiments, individually at different concentrations, and in blends. The results indicate that 4-cresol and 3-n-propylphenol are the most important components for the of the attractancy phenolic mixture.

Behavioural-antennal responses of tsetse to buffalo urine, which has been identified as a potent olfactory attractant, were investigated. Tsetse Glossina morsitans morsitans rapidly raised their antennae upon stimulation with the odour of buffalo urine. Flies showed a clear preference for the undiluted urine, though antennal responses even to 100,000 × diluted urine were significantly different from control, indicating the high efficacy of the attracting compounds present in it. Fresh buffalo urine was the most attracting as compared to 4- and 8-day-old urine. The dichloromethane extract of fresh urine was as active as the natural product, indicating that the attracting compounds were extractable into dichloromethane. Under laboratory conditions, responses of males to the urine were significantly more than females.

Urine from the buffalo, Syncerus caffer, a natural host of the tsetse, has been shown previously to be a potent olfactory attractant for tsetse in the field. The attractant compounds have been found to be extractable into dichloromethane. Fractionation of the extracts by liquid flash chromatography yielded four fractions one of which gave a seven-fold increase in trap catches in the field compared to the control traps. Gas Chromatographie and gas chromatography–mass spectrometric analysis of this fraction revealed the presence of seven simple phenols including phenol itself.

Field studies were done to elucidate the effectiveness of buffalo urine as a tsetse attractant under a variety of test conditions. When traps were baited with the test sample of urine placed in large-necked dispensers allowing evaporation of 2 to 2.9 ml/hr, they performed better than traps baited with similar samples placed in narrow-necked dispensers with evaporation rate of 0.2 to 0.4 ml/hr. Buffalo urine was compared with samples of cow urine: the catch size increased 1.8 times in traps with cow urine when compared to that of the control trap, whereas buffalo urine was 9.6 times more attractive than control. Acetone added to the test sample, even in very small quantity, boosted the effectiveness, increasing the catch size 2.8 times more. Urine samples that had stayed on the laboratory shelves for some weeks or months were more effective than freshly obtained ones.