Journal of Buffalo Science, 2012, 1, 1-4 1 ISSN: 1927-5196 / E-ISSN: 1927-520X/12 © 2012 Lifescience Global Effect of Royal Jelly on the Fertilizing Ability of Buffalo Spermatozoa In Vitro Saber Mohamed Abd-Allah* Faculty of Veterinary Medicine, Beni-Suef University, 62511 Beni-Suef, Egypt Abstract: The aim of the present study was to assess the effect of addition of Royal jelly in presence of heparin on buffalo (Bubalus Bubalis) sperm motility, acrosome reaction and in vitro fertilization (IVF) of buffalo oocytes. Frozen buffalo spermatozoa from five bulls were thawed and motile fraction was obtained by swim up technique. The spermatozoa were washed, treated with100 µg/ml heparin, and then exposed to 0.4% Royal Jelly (RJ) for 3 h. Sperm motility, acrosomal integrity and fertilization rate of matured oocytes were assessed at 1, 2 and 3 h. The percentages of sperm motility, intact acrosome and fertilization rate of matured oocytes were higher (P<0.05) in 0.4% RJ compared to that in the control. After 2 h of incubation the percentage of motility, intact acrosome of spermatozoa and fertilization rate of matured oocytes, respectively, were 93.6 %, 77.6% and 72.6% in 0.4% RJ. These results suggest that treating buffalo sperm with 0.4% RJ in combination with heparin is effective not only to induce sperm acrosome reaction but also is effective for in vitro fertilizing capacity of the cryopreserved buffalo spermatozoa. Keywords: Buffalo, Capacitation, Spermatozoa, IVF, Royal Jelly. INTRODUCTION The enhancement of artificial insemination is a valuable tool in genetic improvement programs [1]. However, the biggest problem to exploiting the cryopreserved buffalo semen is damage of sperm membrane structure during freezing and thawing which lead to a fewer viable and motile cell post-thawing [2] Therefore, An important factor in the efficiency of extender is supplementation with royal jelly to enhance the viability and longevity of post-thawed spermatozoa [3]. In vitro fertilization rate of in vitro matured oocytes using frozen thawed spermatozoa was low due to low fertilizing ability of buffalo spermatozoa [4]. Therefore, there is an urgent need to study the possibilities of improving fertilizing ability of buffalo spermatozoa. Royal jelly is secreted by the hypopharyngeal glands of worker bees to feed young larvae and the adult queen bee. On dry matter basis, royal jelly contains considerable amounts of proteins, lipid, sugars and amino acids. Royal jelly also contains vitamin A, B (pantothenic acid, has antioxidant effect), C, D and E, mineral salts are in descending order: K, Ca, Na, Zn, Fe, Cu, and Mn, enzymes antibiotic components. It also has an abundance of nucleic acid- DNA and RNA. Also it contains sterols, phosphorous compounds and acetylcholine, which is needed to transmit nerve messages from cell to cell [5, 6]. Mammalian sperm must undergo a series of controlled molecular processes called capacitation *Address corresponding to this author at the Faculty of Veterinary Medicine, Beni-Suef University, 62511 Beni-Suef, Egypt; Tel: +201114822589; Fax: +20822327982; E-mail: abdallahsaber49@gmail.com before they are capable of penetrating and fertilizing the oocyte. Although capacitation naturally occurs in the female reproductive tract [7], it can be also induced in vitro using specific media and physical conditions [8]. Capacitation is a complex molecular process that results in changes of calcium concentration, protein phosphorylation, acrosomal matrix and membrane rearrangement. Parrish et al. [9] reported that increased fertilization rates in vitro were obtained when frozen-thawed spermatozoa were treated with heparin which is known as the most potent glycosaminoglycan for inducing capacitation and acrosome reactions Vijayaraghavan et al. [10] reported that the various adenosine analogues contains motility stimulants such as the adenosine monophosphate ((AMP) N (1)-oxide), are already known to enhance the motility of sperm by an inhibiting phosphodiesterase activity, thus enhancing cAMP at the level of the sperm tail. The same author found that AMP N (1)-oxide stimulated the phosphorylation of not only mitogen-activated protein kinase (MAPK) but also that of cAMP/calcium-response element-binding protein (CREB) [10]. In mouse spermatozoa, adenosine modulates the adenylyl cyclase/cAMP pathway [11] and induces sperm response via adenosine A 2 receptors in uncapacitated cells and through adenosine A 1 receptors in capacitated cells [12]. Fertility is equally related to the quality of the spermatozoa and oocytes apart from appropriate conditions for fertilization and embryo development. In recent years, only one study has been focused on