Biostimulation and reproductive performance of artificially inseminated rabbit does (Oryctolagus cuniculus)

F. E. EL-Azzazi, I. M. Hegab, A. M. Hanafy


Biostimulation is a non-hormonal and practical technique that has not yet been widely utilised when applied immediately before insemination to improve reproductive efficiency in livestock species. This study was conducted to determine the influence of short-term male biostimulation on behavioural and reproductive performance of inseminated rabbit does. A total of 142 female New Zealand White rabbits were randomly assigned to 3 groups. Females were either exposed to male odour (Odour group) or an adult aproned male (Male group), while the remaining does that were neither exposed to the male odour nor the adult male are considered the control group. All females were inseminated after the 2 h exposure session. Conception rates were determined by abdominal palpation 12 d after insemination. The results showed that conception rate of the male odour group (79.59%) was greater than that of male presence group (76.09%) and that of the control group (68.09%). Moreover, biostimulated does showed significant behavioural activities during the 2 h exposure session compared to the control group. Although no significant differences were recognised, litter size at birth and at weaning was slightly increased in biostimulated compared to control females. Nor were there any significant difference in serum oestradiol concentrations between treated groups. Conclusively, short-term 2 h biostimulation of rabbit does resulted in the appearance of various behavioural responses followed by differences in conception rates between groups after routine artificial insemination.


rabbit; reproduction; biostimulation; artificial insemination; behavioural; hormonal

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Asaba A., Hattori T., Mogi K., Kikusui T. 2014. Sexual attractiveness of male chemicals and vocalizations in mice. Front. Neurosci., 8: 231.

Baum M.J., Cherry J.A. 2015. Processing by the main olfactory system of chemosignals that facilitate mammalian reproduction. Horm. Behav., 68: 53-64.

Beyer C., Hoffman K.L., González-Flores O. 2007. Neuroendocrine regulation of estrous behavior in the rabbit: Similarities and differences with the rat. Horm. Behav., 52: 2-11.

Bonanno A., Mazza F., Alabiso M., Di Grigoli A., Alicata M.L. 2003. Effects of bio-stimulation induced by the presence of buck on reproductive performance of rabbit does. Ital. J. Anim. Sci., 2:133-135.

Boucaud I.C.A., Valère P.A., Aguirre Smith M.L.N., Doligez B., Cauchard L., Rybak F., Vignal C. 2016. Interactive vocal communication at the nest by parent Great Tits Parus major. Ibis, 158: 630-644.

Brennan P.A. 2010. Pheromones and Mammalian Behavior. In A. Menini (Ed.), The Neurobiology of Olfaction. Boca Raton (FL). Available at: Accessed November 2017.

Caro S.P., Balthazart J., Bonadonna F. 2015. The perfume of reproduction in birds: Chemosignaling in avian social life. Horm. Behav., 68: 25-42.

Chen Y., Li J., Simkin M.E., Yang X. Foote R.H. 1989. Fertility of Fresh and Frozen Rabbit Semen Inseminated at Different Times Is Indicative of Male Differences in Capacitation Time. Biol. Reprod., 41: 848-853.

deCatanzaro D. 2015. Sex steroids as pheromones in mammals: the exceptional role of estradiol. Horm. Behav., 68: 103-116.

Dielenberg R.A., McGregor I.S. 2001. Defensive behavior in rats towards predatory odors: a review. Neurosci. Biobehav. Rev., 25: 597-609.

Eiben C., Tobias G., Kustos K., Godor-Surmann K., Kotany S., Gulyas B., Szira G. 2007. The change of nursing for oestrus induction (biostimulation): Effect of contact between rabbit doe and its young. Livest. Sci., 111: 193-203.

Finn C. A. 1964. Influence of Male on Litter Size in Mice. J. Reprod. Fertil., 7: 107-111.

Fleischman S., Terkel J., Barnea A. 2016. Visual recognition of individual conspecific males by female zebra finches, Taeniopygia guttata. Anim. Behav., 120: 21-30.

Fujiwara M., Nitta A., Chiba A. 2016. Regulation of sexual odor preference by sex steroids in the posterodorsal medial amygdala in female rats. Horm. Behav., 82: 46-55.

Gelez H., Fabre-Nys C. 2004. The “male effect” in sheep and goats: a review of the respective roles of the two olfactory systems. Horm. Behav., 46: 257-271.

González-Mariscal G., García Dalmán C., Jiménez A. 2015. Biostimulation and nursing modify mating-induced c-FOS immunoreactivity in the female rabbit forebrain. Brain Res., 1608: 66-74.

Hegab I.M., Jin Y., Ye M., Wang A., Yin B., Yang S., Wei W. 2014. Defensive responses of Brandt’s voles (Lasiopodomys brandtii ) to stored cat feces. Physiol. Behav., 123: 193-199.

Hegab I.M., Kong S.S., Yang S.M., Mohamaden W.I., Wei, W.H. 2015. The ethological relevance of predator odors to induce changes in prey species. Acta. Ethol., 18: 1-9.

https://doi. org/10.1007/s10211-014-0187-3

Khanh N.P., Rosnina Y., Omar M.A., Dhaliwal G.K., Wahid H., Khumran A.M., Yap K.C., Fahmi M., Azmil A. 2012. Effects of Biostimulation on Oestrus Behaviour, Ovulation Time and Conception Rate in Primiparous and Multiparous Beef Cows. J. Anim. Vet. Adv., 11: 404-411.

Kiyokawa Y., Kikusui T., Takeuchi Y., Mori Y. 2004. Alarm pheromones with different functions are released from different regions of the body surface of male rats. Chem. Senses, 29: 35-40.

Korgan A.C., O’Leary E., Bauer J., Fortier A., Weaver I.C., Perrot T.S. 2016. Effects of Paternal Predation Risk and Rearing Environment on Maternal Investment and Development of Defensive Responses in the Offspring. eNeuro, 3.

Kustos K., Eiben C.S., Szendro Z.S., Theau-Clément M., Gódor S-Né., Jovánczai Z.S. 2000. Effect on reproductive traits of male presence among rabbit does before artificial insemination. In Proc.: 7th World Rabbit Congress, 4-7 July 2000, Valencia, Spain., Vol. A: 161-166.

Lecchini D. 2011. Visual and chemical cues in habitat selection of sepioid larvae. C. R. Biol., 334: 911-915.

López-Sebastian A., González-Bulnes A., Carrizosa, J.A., Urrutia B., Díaz-Delfa C., Santiago-Moreno J., Gómez-Brunet A. 2008. New estrus synchronization and artificial insemination protocol for goats based on male exposure, progesterone and cloprostenol during the non-breeding season. Theriogenology, 69: 651-651.

Marongiu M.L., Dimauro C. 2013. Preliminary study on factors influencing rabbit doe reproductive efficiency: Effect of parity, day of mating and suckling on ovarian status and estrogen levels at day 6 of pregnancy. Can. J. Vet. Res., 77: 126-130. Avaliable at: Accessed November 2017.

Masini C.V., Sauer S., Campeau S. 2005. Ferret odor as a processive stress model in rats: neurochemical, behavioral, and endocrine evidence. Behav. Neurosci., 119: 280-292.

McDonald P.G., Vidal N., Beyer C. 1970. Sexual behavior in the ovariectomized rabbit after treatment with different amounts of gonadal hormones. Horm. Behav., 1: 161-172.

Morrell J.M. 1995. Artificial-Insemination in Rabbits. Br. Vet. J., 151: 477-488.

NRC. 1977. Nutient requirements of domestic animals, No. 9. Nutrient requirements of rabbits. Second Revised Ed. National academy of sciences-National Research Council, Washington, DC.

Ola S.I., Oyegbade M.O. 2012. Buck Effect on Rabbit Oestrous: Vulva Colour, Vaginal Lumen Cells and Ovarian Follicle Populations. World Rabbit Sci., 20: 71-79.

Ola S.I., Olatunbosun O.S. 2013. Effect of buck presence on sexual receptivity and kindling performance of rabbit doe. Livest. Res. Rural Dev,. 25: 135. Available at: Accessed November 2017.

Pallares P., Gonzalez-Bulnes A. 2009. A new method for induction and synchronization of oestrus and fertile ovulations in mice by using exogenous hormones. Lab. Anim., 43:295-299.

Paul M., Patrick B. 2007. Measuring behavior, an introductory guide, 3rd edn. Cambridge University, London.

Prebble J.L., Langford F.M., Shaw D.J., Meredith A.L. 2015. The effect of four different feeding regimes on rabbit behaviour. Appl. Anim. Behav. Sci., 169: 86-92.

Quintela L.A., Peña A.I., Vega M.D., Gullón J., Prieto M.C., Barrio M., Becerra J.J., Maseda F., Herradón P.G. 2004. Ovulation induction in rabbit does submitted to artificial insemination by adding buserelin to the seminal dose. Reprod. Nutr. Dev., 44: 79-88.

Rebollar P., Milanés A., Pereda N., Millán P., Cano P., Esquifino A., Villarroel M., Silván G., Lorenzo P.L. 2006. Oestrus synchronisation of rabbit does at early post-partum by doelitter separation or ECG injection: Reproductive parameters and endocrine profiles. Anim. Reprod. Sci., 93: 218-230.

Richardson H.N., Nelson A.L.A., Ahmed E.I., Parfitt D.B., Romeo R.D., Sisk C.L. 2004. Female pheromones stimulate release of luteinizing hormone and testosterone without altering GnRH mRNA in adult male Syrian hamsters (Mesocricetus auratus). Gen. Comp. Endocrinol., 138: 211-217.

Roberts S.A., Simpson D.M., Armstrong S.D., Davidson A.J., Robertson D.H., McLean L., Beynon R.J., Hurst J.L. 2010. Darcin: a male pheromone that stimulates female memory and sexual attraction to an individual male’s odour. BMC Biol., 8: 75.

Rodríguez-de Lara R., López-Fallas M., Rangel-Santos R., Mariscal-Aguayo V. 2003. Influence of short-term relocation and male exposure on sexual receptivity and reproduction in artificially inseminated lactating doe rabbits. Anim. Reprod. Sci., 78:111-121.

Rodríguez-de Lara R., Noguez-Estrada J., Rangel-Santos R., García-Muñiz J.G., Martínez-Hernández P.A., Fallas-López M., Maldonado-Siman E. 2010. Controlled doe exposure as biostimulation of buck rabbits. Anim. Reprod. Sci., 122: 270-275.

Schmidova J., Milerski M., Svitakova A., Vostry L. 2016. Effects of service ram on litter size in Romanov sheep. Small Rumin. Res., 141: 56-62.

Taylor G.T., Haller J., Regan D. 1982. Female rats prefer an area vacated by a high testosterone male. Physiol. Behav., 28: 953-958.

Theau-Clément M. 2008. Factors of success of rabbit doe insemination and methods for oestrus induction. Prod. Anim., 21: 221-230.

Theau-Clément M., Lebas F., Boiti C., Brecchia G., Mercier P. 2008. Influence of different eCG doses on sexual receptivity and productivity of rabbit does. World Rabbit Sci., 16: 65-72.

Theau-Clément M., Sécula A., Saleil G., Monniaux D., Brecchia G., Boiti C., Bodin L., Brun J.M. 2015. Genetic and Non-Genetic Factors Affecting Rabbit Doe Sexual Receptivity as Estimated from One Generation of Divergent Selection. World Rabbit Sci., 23: 171-179.

Thibert J., Farine J.P., Cortot J., Ferveur J.F. 2016. Drosophila Food-Associated Pheromones: Effect of Experience, Genotype and Antibiotics on Larval Behavior. PLoS One, 11: e0151451.

Ubilla E., Rebollar, P.G. 1995. Influence of the Postpartum Day on Plasma Estradiol-17-Beta Levels, Sexual-Behavior, and Conception Rate, in Artificially Inseminated Lactating Rabbits. Anim. Reprod. Sci., 38: 337-344.

Ubilla E., Rebollar P.G., Pazo D., Esquifino A. I., Alvarino J.M.R. 2000. Pituitary and ovarian response to transient doe-litter separation in nursing rabbits. J. Reprod. Fertil., 118: 361-366.

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