Temporal Changes in Endogenous Estrogens and Expressionof Behaviors Associated with Estrus during the Periovulatory Period in Doublesynch Treated Murrah Buffaloes (Bubalus bubalis)

Document Type: Research Article


1 Department of Animal Science, Faculty of Agriculture, University of Jiroft, Jiroft, Iran

2 Reproductive Physiology Laboratory, Division of Dairy Cattle Physiology, National Dairy Research Institute, Karnal 132001, Haryana, India


The aims of this study were (a) to establish the occurrence, duration and intensity of behavioral estrus symptoms, and (b) to determine the relationship between behavioral estrus symptoms with the total estrogen profiles in cycling Murrah buffaloes (n=10) subjected to Doublesynch protocol (0 PGF, 2 GnRH, 9 PGF and 11 GnRH). After both GnRH injections, estrus and its behavioral signs were detected at hourly intervals using visual observations, transrectal examination of genitalia, and bull parading twice a day for 30 min each. Frequent urination (100%) and excitement (100%) were the best indicators of estrus followed by swollen vulva (90%) and bellowing (85%). The longest duration of estrus signs viz. swollen vulva, was observed up to 8.9 ± 0.5 h and 11.1 ± 0.3 post first and second GnRH administrations, respectively. The mean intervals between the onset of estrus symptoms appearance and ovulation were 13.8 ± 0.9 h and 13.4 ± 1.0 h post first and second GnRH inactions, respectively. The mean total estrogen concentrations at the time of both GnRH injections were above the basal level (8.4±1.9 pg/mL; range 2.9-18.4 and 7.9±1.9 pg/mL; range 2.9-17.6 at the time of the first and second GnRH injections, respectively), temporarily increased post GnRH treatments and then gradually declined to its basal level during ovulation time window. Frequent urination, excitement and swollen vulva appeared to be the most notable estrus symptoms after GnRH injections. Our findings also suggest that the AI can be done at a fixed time (once 20 to 24 h posts second GnRH) in buffaloes treated with Doublesynch protocol.


Danell B., Gopakumar N., Nair M.C.S. and Rajgopalan K. (1984). Heat detection symptoms in Surti buffalo breeds. Indian J. Anim. Reprod. 5, 1-7.
Gunasekaran M. (1998). Estrous behaviour and fertility in crossbred cattle and Murrah buffaloes. MS Thesis. Deemed Univ., Karnal, India.
Hafez E.S.E. and Hafez B. (2000). Reproduction in Farm Animals. Wiley.
Janakiraman K. (1978). Control of optimizing of reproductive cycle in buffalo. Pp. 220-225 in Proc. Semina. buffalo Reprod. Artif. Insemin, NDRI, Karnal, India.
Kamboj M. and Prakash B.S. (1993). Relationship of progesterone in plasma and whole milk of buffaloes during cyclicity and early pregnancy. Trop. Anim. Health Prod. 25, 185-192.
Kobayashi Y. (1995). Gonadotropin-releasing hormone at estrous in the bovine: changes in characteristics of the preovulatory follicle and corpus luteum and in vitro production of progesterone and oxytocin. MS Thesis. Kansas State Univ., Manhattan, New York.
Madan M.L. and Prakash B.S. (2007). Reproductive endocrinology and biotechnology applications among buffaloes. Soc. Reprod. Fertil. 64, 261-281.
Mirmahmoudi R. and Prakash B.S. (2012). The endocrine changes, timing of ovulation and efficacy of the Doublesynch protocol in the Murrah buffaloes (Bubalus bubalis). Gen. Comp. Endocrinol. 177, 153-159.
Mohan K., Kumar V., Sarkar M. and Prakash B.S. (2010). Temporal changes in endogenous estrogens and expression of behaviors associated with estrus during the periovulaory period in Murrah buffaloes (Bubalus bubalis). Trop. Anim. Health Prod. 42, 21-26.
Mohan K. and Prakash B.S. (2010). Changes in endogenous estrogens and expression of behaviors associated with estrus during the periovulatory period in Heatsynch treated Murrah buffaloes (Bubalus bubalis). Trop. Anim. Health Prod. 42, 947-952.
Mohan K., Sarkar M. and Prakash B.S. (2009). Efficiency of Heatsynch protocol in estrus synchronization, ovulation and conception of dairy buffaloes (Bubalus bubalis). Asian-Australas J. Anim. 22, 774-780.
Moioli B.M., Napolitano F., Puppo S., Braile V.L., Terzano G.M. and Malfatti A. (1998). Patterns of oestrus, time of LH release and ovulation and effects of time of artificial insemination in Mediterranean buffalo cows. Anim. Sci. 66, 87-91.
Mondal M., Rajkhowa C. and Prakash B.S. (2006). Relationship of plasma estradiol-17-β, total estrogen and progesterone to estrus behavior in mithun (Bos frontalis) cows. Horm. Behav. 49, 626-633.
Paul V. (2003). Studies on estrus synchronization methods, timing of ovulation and endocrine profile in buffaloes. Ph D. Thesis. National Dairy Research Institute, Karnal, Haryana, India.
Prakash B.S., Sarkar M., Paul V., Mishra D.P., Mishra A. and Meyer H.H.D. (2005). Postpartum endocrinology and prospects for fertility improvement in the lactating riverine buffalo (Bubalus bubalis) and yak (Poephagus grunniens). Livest. Prod. Sci. 98, 13-23.
SAS Institute. (1996). SAS®/STAT Software, Release 6.11. SAS Institute, Inc., Cary, NC.
Schneider F., Tomek W. and Gründker C. (2006). Gonadotropin-releasing hormone (GnRH) and its natural analogues: a review. Theriogenology. 66, 691-709.
Singh G., Singh G.B., Sharma S.S. and Sharma R.D. (1984). Studies on oestrus symptoms of buffalo heifers. Theriogenology. 21, 849-858.