Investigation of pheromones in cervical and vaginal secrations at different terms of estrous cows

69 7.SUMMARY The aim of this study was to determine the presence and biochemical sturucture of pheromones that are belived to function as a signal of estrus for male animals. Furthermore the physical and chemical properties of cervico-vaginal secretions may influence the pheromones so physical and chemical properties of these secretions were also studied. Cervico-vaginal mucus samples from 104 oestrus cows out of! 97 were collected by rectal massage between early heat, mid heat and late heat of estrus. After these samples were bioassayed by bulls, they were analysed for colour, pH, viscosity, water-dry matter contents, Ca, Na, Mg and K levels. Afterwards, the remaining samples were stored at between -15 to -20° C until analysed. Choloform/methanol, diethyl ether extractions, dialysis, coloumn chromatography (filled with slica gel) of cervico-vaginal mucus were also performed. To determine the chemical structure of the extracts from the positive samples spectrums were determined using Ultraviole-visible, Infrared, TH-Nuclear Magnetic Rezonans and Gas-Mass spectrometers. In 104 cervico-vaginal mucus samples obtained from different period of estrous cows, 78.84 % was transparent colourless, 2.88 % dirty colourless, 7.69 % yellowish, 6.73 % white and 3.84 % reddish. In 104 of samples,obtained different period of estrous cows, the viscosity ranged between 0 and 4 mean 2.37 ± 0.09, pH value was between 6.7 and 9.2 mean 7.46 ± 0.06, amount of dry matter varied between 0.82 % and 5.02 % mean 1.62 % ±0.06, water content ranged between 94.08 % and 99.18 % mean 98.37 % ± 0.07, Ca concentration70 ranged between 9.3 and 12 mg/100 ml mean 10.06 ± 0.06 mg/100 ml, Na concentration ranged between 10.51 and 13.1 mEq/l mean 12.11 ± 0.05 mEq/l, K level ranged between 110 and 160 mEq/l mean 1 18.04 ± mEq/l and Mg amount ranged between 2.6 and 4.3 mg/100 ml mean 3.96 ±0.03 mg/100ml. The differences in physical properties and chemical contents of mucus samples collected during various phases of estrus were found to be statistically insignificant ( P>0.05). When 104 mucus samples obtained from different phases of estrus cows, were tested by 3 bulls sniffing, chin resting, licking, flehmen and penile erection reactions were recorded and the mean values for the reactions were fond to be 91.34, 54.48, 7.04,68.25 and 3.82 %, respectively. The result of bull assay carried out as the extracts obtained from the analysis of colon chromatography, dialysis and chloroform/methanol extractions was negative and also no organic substance was detected by spectroscopic analysis. But, after diethyl ether extraction, the samples still induced bull reaction and following spectroscopy, in these extracts from the functional groups of dialkil substitüed benzene, tri substitued phenol, ester, alkene and steroid, 1 6 organic substance were found. The results of these studies indicate that bovine cervico- vaginal mucus secretions during estrus, contain pheromones. Additionally it was noticed that pheromones could be extracted with solents such as diethyl ether which can evaporise at low temparatures (10-15 °C). For testing the presence of pheromones in biological secretions, bull assay still appears to be the most reliable method.71 8. KAYNAKLAR 1 -Adams, M.G.(1 980). Odour-Producing Organs of Mammals. Symposia of the Zoological Society of London. 45, 57-86. 2- Agrawal, S.C. and Datta I.C. (1977). Physico-Chemical Properties of Bovine Cervical Mucus at Estrus. Indian. J. Exp. Biol. 1 5, 417-419. 3- Alan, M. (1990). İneklerde Östrusun Belirlenmesi ve Fertilite Yönünden Önemi. TİGEM. 5 : 28, 25-29. 4-Albone, E. S., Blazquez, N.B., French, J., Long, S. E. and Perry, GX. (1986). Mammalian Semiochemistry Issues and Futures,with Some Examples From a Study of Chemical Signaling in Cattle. 27-36. Ed. D. Duval, D. Muller-Schwarze and B.M. Silverstein. Chemical Signals in Vertebrates. 4th Ed. Plenum Press. New York. 5- Aron,C. (1979). Mechanism of Control of the Reproductive Function by Olfactory Stimuli in Female Mammals. Physiological Reviews. 59 :2, 229-284. 6- Arya, S. P. and Jain, Y. C. (1985). Genital Secration (Cervico-Vaginal Mucus) of Jersey Cows : Dry Matter, Specific Gravity and Hydrogen ion Concentration as indicators of Heat and Early Pregrancy. Indian Journal of Animal Science. 55 : 11, 948-950. 7- Asotra, S., Asotra, K. and Jain, V.C.(1 985). Changes in Quantity, Dry Matter, Colour, Fibrosity, pH and Contents of Carbonhydrates and Lipids of Cervico-Vaginal Mucus of Cows During Oestrous Cycle and Early Pregnancy. Indian Journal of Animal Sciences. 55:11, 918-931.