title: Royal jelly supplementation in semen extender enhances post-thaw quality and fertility of Nili-Ravi buffalo bull sperm.
abstract: Two experiments were conducted to evaluate the effect of royal jelly (RJ) on post-thaw sperm quality, in vitro and in vivo fertility rate of cryopreserved buffalo bull sperm. The semen was collected from three mature regular donor buffalo bulls, ejaculates were pooled and semen evaluated initially. In Experiment 1, the ejaculates were extended in tris-citric acid diluter supplemented with different RJ concentrations (0, 0.05, 0.1, 0.2, 0.3 or 0.4%). The diluted semen was cooled to 4°C, packaged into 0.5 mL straws and frozen using standard procedure. The straws were thawed and assessed for sperm progressive motility, viability, plasma membrane, acrosome, and chromatin integrity. The results indicated that sperm progressive motility was significantly greater (P<0.05) in 0.05, 0.1, 0.2 and 0.3% RJ than 0.4% RJ supplemented and control groups. The sperm viability, plasma membrane and acrosome integrity were significantly improved (P<0.05) in 0.1% RJ supplemented group the compared to other treatment groups. In Experiment 2, cryopreserved sperm with 0.1% RJ supplementation and control (without RJ supplementation) were used to observe the in vitro fertilizing potential and in vivo fertility. In vitro fertilization method was applied to assess the cleavage rate; whereas, AI was performed in buffalo during in vivo fertility trial. The buffaloes were inseminated 12h after standing estrus and pregnancy diagnosis was performed through ultrasonography. The results revealed that the cleavage rate was higher (P<0.05) in 0.1% RJ as compared to control group. However, the pregnancy rate was similar (P>0.05) between 0.1% RJ supplemented and control groups. It is concluded that supplementation of RJ in freezing extender can improve the cryosurvival rate and in vitro fertilizing capacity of buffalo bull sperm.
authors.affiliation: Reproduction Division, Buffalo Research Institute Pattoki, Kasur 55050, Pakistan. Electronic address: raoqaisarshahzad@gmail.com.
authors.forename: Qaisar
authors.initials: Q
authors.lastname: Shahzad
authors.affiliation: Department of Theriogenology, Faculty of Veterinary Sciences, University of Veterinary and Animal Sciences, Lahore 54000, Pakistan.
authors.forename: Muhammad Usman
authors.initials: MU
authors.lastname: Mehmood
authors.affiliation: Department of Animal Health, University of Agriculture, Peshawar 25120, Pakistan.
authors.forename: Hamayun
authors.initials: H
authors.lastname: Khan
authors.affiliation: Department of Zoology, Pir Mehr Ali Shah Arid Agriculture University, Rawalpindi 46000, Pakistan.
authors.forename: Asma
authors.initials: A
authors.lastname: ul Husna
authors.affiliation: Department of Zoology, Pir Mehr Ali Shah Arid Agriculture University, Rawalpindi 46000, Pakistan.
authors.forename: Saima
authors.initials: S
authors.lastname: Qadeer
authors.affiliation: Department of Zoology, Pir Mehr Ali Shah Arid Agriculture University, Rawalpindi 46000, Pakistan.
authors.forename: Asima
authors.initials: A
authors.lastname: Azam
authors.affiliation: Department of Reproduction and Artificial Insemination, Faculty of Veterinary Medicine, Adnan Menderes University, Aydin 09016, Turkey.
authors.forename: Zahid
authors.initials: Z
authors.lastname: Naseer
authors.affiliation: Department of Clinical Sciences, Faculty of Veterinary Sciences, Bahauddin Zakariya University, Multan 54000, Pakistan.
authors.forename: Ejaz
authors.initials: E
authors.lastname: Ahmad
authors.affiliation: Department of Medical Biology and Genetics, Gaziantep University, Gaziantep 27310, Turkey.
authors.forename: Muhammad
authors.initials: M
authors.lastname: Safdar
authors.affiliation: Department of Theriogenology, Faculty of Veterinary Sciences, University of Veterinary and Animal Sciences, Lahore 54000, Pakistan.
authors.forename: Mushtaq
authors.initials: M
authors.lastname: Ahmad
mesh_terms: D000818:Animals; D002020:Buffaloes / Q000502:physiology*; D015925:Cryopreservation / Q000379:methods / Q000662:veterinary*; D005227:Fatty Acids / Q000494:pharmacology*; D005260:Female; D005298:Fertility; D008297:Male; D011247:Pregnancy; D012662:Semen Preservation / Q000379:methods / Q000662:veterinary*; D013081:Sperm Motility; D013094:Spermatozoa / Q000187:drug effects*; D000099219:Royal Jelly
chemical_list: D005227:Fatty Acids; D000099219:Royal Jelly
keywords: Buffalo bull; Cryopreservation; Fertility; In vitro fertilization; Royal jelly