BEHAVIOURAL SIGNS OF ESTRUS IN DIFFERENT PARITY OF MURRAH BUFFALOES (BUBALUS BUBALIS): A COMPARATIVE STUDY

Indian J. Anim. Res., 48 (6) : 620-624, 2014 doi:10.5958/0976-0555.2014.00043.0 AGRICULTURAL RESEARCH COMMUNICATION CENTRE www.arccjournals.com / www.ijaronline.in BEHAVIOURAL SIGNS OF ESTRUS IN DIFFERENT PARITY OF MURRAH BUFFALOES (BUBALUS BUBALIS): A COMPARATIVE STUDY K.K. Verma*, Shiv Prasad, T.K. Mohanty, A. Kumaresan, S.S. Layek, T.K. Patbandha 1 and S.C. Kantwa Livestock Research Centre National Dairy Research Institute, Karnal- 132 001, India Received: 11-02-2013 Accepted: 13-09-2013 ABSTRACT In dairy bovines, estrus detection is the key determinant of the herd fertility. However, owing to silent heat in buffaloes, it is difficult to detect estrus at proper time which in turn affects the estrus detection efficiency and reduces conception rate. Further, it is not clear whether the buffaloes during different parities have differential expression of estrus behaviour. Therefore, the present study was designed to study the behavioural signs of estrus expressed by Murrah buffaloes (n= 94) belonging to different parities. Estrus detection was done by parading teaser bulls (4 times bull parading in a day), heat expectancy chart and visual observation of behavioural signs of estrus. The study revealed significantly (P< 0.05) higher bellowing/vocalization behaviour in primiparous (77.27%) and pluriparous buffaloes (72.88%) than heifers (38.46%). Micturition behaviour was significantly (P< 0.05) more pronounced in pluriparous (93.22%) and primiparous buffaloes (90.91%) as compared to heifers (69.23%). In pluriparous and primiparous buffaloes, tail raising behaviour during estrus was also expressed by higher(p< 0.05) proportion of buffaloes (96.61 and 90.91%, respectively) as compared to heifers (76.92%). However, there were no differences in tumefaction of vulval lips, reddening of vulval mucus membrane characteristics, standing to be mounted behaviour, chin resting and Flehmen reaction among pluriparous, primiparous and heifer buffalos. These results indicated that intensity of bellowing/vocalization, micturition and tail raising behaviours during estrus were poorly expressed in heifers and more focussed attention is needed to detect buffalo heifers in estrus. Key words: Bellowing, Bull parading, Micturition, Murrah buffaloes, Tail raising,. INTRODUCTION In dairy industry, inadequate heat detection has been considered as the primary constrain for herd reproductive performance over past decades. In a seasonally calving herd for each missed heat there is loss of 21 days production and also loss of one complete estrus cycle. In organised dairy herd, approximately 50% of the heats go undetected and 20.75% buffaloes are inseminated at improper time (Kumaresan et al., 2001). Further, silent heat is considered as one of the major obstacle in understanding reproductive parameters and assisted reproduction in buffaloes (Mondal et al., 2008). Buffaloes mostly express the behavioural signs of estrus during late night and early morning, thus ignored by most of the dairy farmers. A major difference between buffalo and cattle in regard of estrus expression, in buffaloes behavioural signs of estrus are less overt and homosexual behaviour rarely observed (Vale et al., 1990). The main behavioural signs of estrus in buffaloes are restlessness, bellowing and frequent voiding of small quantities of urine generally not or poorly exhibited in the absence of bull. In the herd, bull shows increasing interest in a buffalo that is approaching oestrus and the buffalo stands to be mounted by the bull during estrus. Buffalo exhibited the signs of estrus in the early morning (3.00 9.00 h) and in the evening (15.00 to 21.00 h) with a peak in early morning (about 37%), while lower percentage was at the noon (El-Wardani and El- Asheeri, 2000). * Corresponding author s -email: drkkvermavet@gmail.com, and address: IVRI Izatnagar, 1 Junagadh Agricultural University, Junagadh-362 001, Gujarat, India

Mostly in dairy bovines estrus is observed through visual appraisal of its cardinal behavioural signs. Among behavioural signs of estrus acceptance of male is considered as the most reliable estrus indicator in buffaloes. Frequent micturition, bellowing, vulval swelling, reddening of mucus membrane and mucus discharge are salient signs during estrus but their expression is poor and varies season to season. In Murrah buffaloes, other signs of estrus observed are restlessness, inappetence, nervousness, reduction in milk yield and crutching the back and lumber region etc. Due to less overt signs of estrus, heat detection is quiet inefficient in buffaloes, so to overcome this problem a keen knowledge of estrus symptoms is of paramount importance Thus for improving heat detection, a combination of estrus detection methods may beusedforcorrectidentificationofheatinmurrah buffaloes (Rameshet al., 2002). Furthermore, among different parity the expression of estrus behaviour is not understood so far, which may contribute to the poor estrus detection in buffaloes. Thus keepi ng these intricaci es the present investigation was designed with an objective to study the behavioural signs of estrus in different parity of Murrah buffaloes. MATERIALS AND METHODS Experimental animals and management: The present study was conducted on Murrah buffaloes (n= 94) maintained at Livestock Research Centre (LRC) of National Dairy Research Institute Karnal, Haryana for a period of 4 months (November 2011 - February 2012). In the present study, we selected both buffalo heifers (n= 13) with body weight (BW) not less than 347 kg (427± 23) and age more than 27 months (39.37± 4.28) and buffaloes (n= 81) 1-6 parity with BW and age 580± 8.8 kg and 80± 2.84 months, respectively. Murrah buffaloes taken in the study were free from anatomical deformity, reproductive dis orders and also from any other diseases. The buffaloes were maintained in loose housing system with group management practice and for easy identification animals were tagged with numbered plastic ear tags. Experimental buffaloes were supplied with ad lib seasonal green fodder along with 1.5-2.0 kg concentrate as maintenance ration. Most of the milking buffaloes were in first and second month of lactation. Milking buffaloes were given Vol. 48, No. 6, 2014 621 additional concentrate at the rate of 1.0 kg for every 2.0 kg milk production, above 5.0 kg milk yield. The concentrate to the milking animals was fed in divided allowances twice during the time of milking. Estrus detection in buffaloes: In the present study, estrus detection was carried using a teaser bull (4- times bull parading in a day), heat expectancy chart along with visual observation of cardinal signs of estrus. The teaser bull of 5 year age with BW 600 kg and bull parading was done four times in a day i.e. at morning (6:00 am), forenoon (10:00 am), afternoon (2:00 pm) and in the evening (6:00 pm). Teaser bull was paraded in each shed of buffaloes for 30 minutes. In the presence of estrus buffalo teaser bull showed the Flehmen reaction, chin resting and mounting behaviours and buffaloes identified in estrus were separated from the herd. Estrus detection was carried out by visual observation of behavioural signs of estrus using bull parading four times a day as stated above. The observers recorded the onset of estrus based on the already proven cardinal signs and the frequency of a particular sign over different time periods. The cardinal signs that were used for assessing the onset and intensity of estrus were bellowing, frequent micturition, tail raising, mucus discharge, swollen vulva, standing to be mounted by teaser bull, chin resting and Flehmen reaction or sniffing/licking of vulva. The heat expectancy chart contains the records of animals about previous estrus. In the present study, heat expectancy chart was used to know the expected date of coming estrus for individual animal in the buffalo herd. Statistical analysis: In the present study Chi-square test was performed to determine difference between heifers, primiparous and pluriparous buffaloes for behavioural signs of estrus, changes in genitalia and mucus discharge during time of estrus. Statistical analysis was performed using SigmaPlot 11 software (Systat software Inc. USA). RESULTS AND DISCUSSION In the present study, behavioural signs of estrus in Murrah buffaloes were observed using teaser bull, heat expectancy chart and visual observation of behavioural signs of estrus viz. standing to be mounted by teaser bull, tail raising, micturition,

622 INDIAN JOURNAL OF ANIMAL RESEARCH tumefaction of vulva, mucus discharge, reddening of vulval mucus membrane and vocalization/ bellowing. The behavioural signs of estrus in response to teaser bull and changes in genitalia and mucus discharge during estrus are presented in Table 1 and Table 2. The present study revealed significant difference (P< 0.05) in tail raising, micturition and bellowing behaviour among different parity of buffaloes. Though the expression of standing to be mounted by teaser bull behaviour did not differ among parity but majority of buffaloes (90.43%) were found to be static during the time of mounting by teaser bull. Almost similar findings were reported by Patel (1979) who observed that 90% buffaloes showed standing to be mounted by teaser bull. In the same way Gunasekaran et al. (2007) also reported slightly higher proportion (95.77%) of buffaloes showed this behaviour in response to teaser bull. Contrary to our findings, Singh et al. (1984) reported that 83.69% buffaloes were found to be static towards teaser bull. Tail raising is very common behaviour during estrus and intensity of this behaviour increases with the approach of teaser bull. The overall tail raising behaviour was observed in 92.55% estrus in presence of teaser bull. Significantly (P< 0.05) higher proportion of pluriparous (96.61%) and primiparous (90.91%) buffaloes had shown the tail raising behaviour in response to teaser bull as compared to heifer (76.92%). The proportion of buffaloes exhibiting tail raising behaviour in the present study was higher than the proportion reported by Patel (1979) in buffaloes (21.25%), Gunasekaran et al. (2007) in Murrah buffaloes (9.86%), Mangal (2009) in Sahiwal cows (47.27%) and Gunasekaran et al. (2008) in crossbred cows (23.53%). The higher expression may be due to time of estrus detection by bull parading, environmental conditions and duration of estrus detection. Frequent micturition is one of the most important behavioural sign in estrus buffaloes. Majority of buffaloes (88.30%) showed this particular behaviour during estrus in response to teaser bull. Like tail raising behaviour, expression of micturition behaviour among different parity of buffaloes differed significantly (P< 0.05) with higher proportion in pluriparous (93.22%) and primiparous (90.91%) compared to heifers (69.23%). The result of the present study is similar to the finding by Patel (1979) in buffaloes (90%) whereas higher compared to the result observed by Singh et al. (1984) in buffaloes and Gunasekaran et al. (2007) in Murrah buffaloes (34.78 and 23.94%, respectively). Variation in our finding and other workers may be TABLE 1: Behavioural signs of estrus in Murrah buffaloes (n= 94) in presence of teaser bull during estrus Parameters Heifer(%) Primiparous Pluriparous Total(%) P- value (n= 13) (%)(n= 22) (%)(n= 59) (n= 94) Standing to be mounted by teaser bull 84.62 90.91 91.53 90.43 P > 0.05 Tail raising during estrus 76.92 90.91 96.61 92.55 P < 0.05 Micturition during estrus 69.23 90.91 93.22 88.30 P < 0.05 Bellowing during estrus 38.46 77.27 72.88 69.15 P < 0.05 Sniffing/licking (Flehmen reaction) by teaser bull 76.92 95.46 94.92 92.55 P > 0.05 Chin resting by teaser bull 76.92 86.36 88.14 86.17 P > 0.05 TABLE 2: Changes in the genitalia and mucus discharge in Murrah buffaloes (n= 94) during estrus Parameters Heifer(%) Primiparous Pluriparous Total(%) P- value (n= 13) (%)(n= 22) (%) (n= 59) (n= 94) Amount of mucus discharge Copious 30.78 40.92 67.79 56.38 P > 0.05 Moderate 53.84 45.45 23.73 32.97 Scanty 15.38 13.63 8.48 10.63 Tumefaction of vulva P > 0.05 Intense 7.69 4.54 11.86 9.57 Moderate 69.23 72.73 64.41 67.02 Weak 23.08 22.73 23.2 23.41 Reddening of vulval mucus membrane P > 0.05 Intense 7.69 36.36 44.07 37.23 Moderate 84.62 50.00 47.46 53.20 Pale 7.69 13.64 8.47 9.57

due to difference in the season of estrus detection because micturition may be some what lesser frequent during hot summer days in animals. Duration and time of estrus detection may be a probable cause for the differences in the results observed by other researchers. Bellowing/vocalization is an important behavioural sign during the time of estrus in Murrah buffaloes. In this study, overall 69.15% of buffaloes had shown vocalization behaviour during the time of estrus. Significantly (P< 0.05) higher proportion of primiparous (77.27%) and pluriparous (72.88%) buffaloes showed vocalization as compared to heifers (38.46%). These findings are supported by Patel (1979) in buffaloes (77.50%) while lower values were reported by Rao (1976), Gunasekaran et al. (2007) and Gill et al. (1973) in buffaloes (56.70, 49.30 and 25.16%, respectively).on the contrary to our findings the higher values (80.43%) were reported by Singh et al. (1984) in buffaloes. The lower proportions of heifers had shown the standing to be mounted behaviour, tail raising, micturition and bellowing may be their inexperience or shyness. Heifer buffaloes learn these behaviours of estrus by looking the experienced (primiparous and pluriparous) buffaloes. The variation in the findings may be due to the observation schedule followed in the study (difference in the time of detection of estrus and duration of observation of cardinal signs of estrus). Flehmen reaction and chin resting are the characteristic sexual behaviour of bull in response to estrus female in dairy animals. In the present study, teaser bull sowed Flehmen response in 92.55% estruses. Though, higher proportions of primiparous (95.46%), pluriparous (94.92%) buffaloes were found with teaser bull showing Flehmen reaction as compared to heifers (76.92%) but statistically no significance difference was observed. The present study revealed a higher value of Flehmen response behaviour by teaser bull in presence of female buffaloes compared to Gunasekaran et al. (2007) who observed Flehmen response in 40.85% buffaloes. Chin resting behaviour was found in majority of buffaloes (86.17%). Similar to Flehmen response, chin resting behaviour did not differ among different parity; however, compared to heifers (76.92%) higher proportion of pluriparous (88.14%) Vol. 48, No. 6, 2014 623 and primiparous (86.36%) buffaloes allowed chin resting by teaser bull. The findings in the present study are lower than reported by Patel (1979) in buffaloes (90%), whereas lower values were reported by Gunasekaran et al. (2007) in Murrah buffaloes (18.31%). The probable cause of lower incidence of Flehmen reaction by teaser bull in heifers may be lower concentration of pheromones in the urine of estrus heifers due to lower concentration of estrogen as compared to pri miparous and pluriparous buffaloes. Other probable causes for lower incidence of chin resting and Flehmen reaction by teaser bull may be due to shyness and fear to the teaser bull and inadequate learning of behaviour in buffalo heifers. The variation in the findings of different authors may be due the different number of animals in sexually active group (SAG) because for the display of cardinal signs of estrus more number of buffaloes should be present in the SAG. The present study revealed that 56.38% estruses buffaloes at the time of artificial insemination had copious mucus discharge. The higher proportion (67.79%) of copious discharge was found in pluriparous buffalo cows as compared to primiparous (40.90%) and heifers (30.76%). The findings of this study observed are lower than reported by Layek et al. (2013) in Sahiwal cows (70.17%). Majority of buffaloes (67.02%) had shown the moderate tumefaction of vulval lips whereas intense vulval swelling was found in very less number of ani mals (9.57%). H i gher proportion of primiparous cows (72.73%) and heifers (69.23%) showed the moderate tumefaction of vulva as compared to pluriparous buffaloes (64.41%). Our findings are more or less similar to those observed by Gunasekaran et al. (2007) in Murrah buffaloes (70.42%) and lower compared to Patel (1979), who reported higher value i.e. 88.75%. In the present study, most of buffaloes (90.43%) showed reddening of vulval mucus membrane. Higher proportions of heifers (84.62%) were observed with moderate reddening of vulval mucus membrane as compared to primiparous (50.00%) and pluriparous (47.46%) buffaloes. Our results are in agreement with the results observed by

624 INDIAN JOURNAL OF ANIMAL RESEARCH Patel, (1979) and Gunasekaranet al. (2007) (> 90% and 90.34%, respectively); whereas lower values (64.78%) were reported by Gill et al. (1973) in buffaloes. Among dairy bovines estrogen is the single most responsible hormone for different estrus behaviours (Lyimo et al., 2000).During the time of estrus estrogen hormone is found to be at peak. Under the influence of increased estrogen there is increase in the blood supply towards the genital organs in estrus female which causes the swelling of vulva, reddening of vulval mucus membrane. The different stage of estrus detection and individual perception may be the contributing factors for variation of result compared to other studies. CONCLUSION From the findings of the present study it may be concluded that i ntensity of bellowing/ vocalization, micturition and tail raising behaviours during estrus vary greatly among different parity of Murrah buffaloes. The difference in the intensities of estrus behaviour in heifers may be due to shyness, inexperience and inadequate learning of these behaviours. ACKNOWLEDGEMENT First author is very thankful to Indian Council of Agricultural Research (ICAR) for providing Junior Research Fellowship (JRF). The authors are also thankful to Director, National Dairy Research Institute, Karnal for providing all research facilities. REFERENCES El-Wardani, M.A. and El-Asheeri, K. (2000). Influence of season and number of heat checks on detecting of ovulatory estrus in Egyptian buffaloes. Egyptian J. Anim. Prod., 37(1): 1-8. fertility in Murrah buffaloes. Indian J. Dairy Sci., 60(5): 348 351. Gill, R.S., Gangwar, P.C. and Kooner, D.S. (1973). Studies on the oestrous behaviour in buffaloes. Indian J. Anim. Sci.,43(6): 472-475. Gunasekaran, M., Singh, C. and Gupta, A. K. (2007). Effect of oestrus behaviour on fertility in Murrah buffaloes. Indian J. Dairy Sci., 60(5): 348 351. Gunasekaran, M., Singh, C. and Gupta, A. K. (2008). Effect of estrus behaviour on fertility in crossbred cattle. Indian Vet. J.,85: 159-163. Kumaresan, A., Ansari, M. R. and Sanwal, P. C. (2001). Assessment of the accuracy of oestrus detection by progesterone assay in cattle and buffaloes. Indian J. Anim. Sci., 71(8): 34-36. Layek, S.S., Mohanty, T.K., Kumaresan, A., Behera, K. and Chand, S. (2013). Cervical mucus characteristics and periestrual hormone concentration in relation to ovulation time in Zebu (Sahiwal) cattle. Livestock Science, http://dx.doi.org/10.1016/ j.livsci.2012.12.023. Lyimo, Z.C., Nielen, M., Ouweltjes, W., Kruip, T A. and van Eerdenburg, F.J.C.M. (2000). Relationship among estradiol, cortisol and intensity of estrousbehavior in dairy cattle. Theriogenology, 53: 1783-1795. Mondal, S., B. S. Prakash and P. Palta. (2008). Peripheral plasma FSH concentrations in relation to expression of oestrus in Murrah buffaloes (Bubalus bubalis). Buff. Bulletin 27(4):258-262. Mangal, V. (2009). Study on Estrous Behaviour, Cervical Mucus Characteristics and Fertility in Sahiwal Cattle. M.V.Sc. Thesis, NDRI, Karnal, Haryana, India. Patel, J.M. (1979). Investigations of heat detection and estrus behaviour in buffaloes. M.Sc. Thesis., Kurukshetra University, Kurukshetra. Ramesh, V., ThangaThaminVanan, A. Varadhrajan (2002). Improvement of reproductive performances of buffaloes. Pashudhan 17(01): 1-4. Rao, M.M. (1976). M.V.Sc. Thesis, Gujarat Agrl. Univ. (Vide Patel, J.M. 1979. M. Sc. Thesis, Kurukshetra). Singh, G., Singh, B.C., Sharma, S.S. and Sharma, R.D. (1984). Studies on oestrous symptoms of buffalo heifers. Theriogenology, 21(6): 849-858. Vale, W.G., Ohashi, O.M., Sousay, J.S. and Ribeiro, H.F.L. (1990). Studies on the reproduction of water buffalo in the Amazon basin. In: Livestock Reproduction in Latin America. International Atomic Energy Agency, Vienna, Austria, pp. 201-210.