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光照周期和卵泡波出现方式对尼里-拉菲水牛通过采卵获得的卵泡更新、卵母细胞回收与质量以及胚胎早期体外发育能力的影响:初步证据

Effect of photoperiodicity and methods of follicular wave emergence on follicle turn-over, recovery and quality of oocytes, and early in-vitro developmental competence of embryos using ovum pick-up in Nili-Ravi buffaloes: Preliminary evidence.

作者信息

Sagheer Masroor, Ullah Farid, Arshad Usman, Saleem Muhammad, Nawaz Mudassar, Sarwar Zaeem, Saad Muhammad, Riaz Amjad, Naveed Ul Haque Muhammad, Basheer Atia, Ahmad Nasim

机构信息

Department of Theriogenology, Faculty of Veterinary Science, University of Veterinary and Animal Sciences, Lahore, 54000, Pakistan.

Department of Animal Sciences, University of Florida, Gainesville, 32611, USA.

出版信息

Theriogenology. 2020 Nov;157:508-516. doi: 10.1016/j.theriogenology.2020.07.021. Epub 2020 Jul 28.

Abstract

The objectives of the present study were to evaluate the effects of photoperiodicity, gauge (G) of ovum pick-up (OPU) needle, and two methods of follicular wave emergence on follicular turn-over, oocyte recovery (OR), quality of the oocytes (OQ), and early in-vitro developmental competence of embryos in Nili-Ravi buffaloes (n = 20). In 1 experiment, buffaloes (n = 12; 4 buffaloes/season) were randomly assigned to optimize the OPU's (n) either with 17 G or 18 G needle in one of the following seasons: 1) peak breeding season (PBS; Sep-Nov; n = 31), 2) transition breeding season (TBS; Dec-Feb; n = 32), and 3) low breeding season (LBS; Apr-June; n = 32). During 2 experiment, buffaloes (n = 8) were enrolled randomly in a 2 × 2 cross-over design to compare the two methods of wave emergence either using follicular ablation (FA; n = 4), or synchronization protocol (CIDR-EB; n = 4) during PBS. In FA method, the ovarian follicles were aspirated (week -1), and subsequently repeated OPU's (n = 55) were performed for 7 weeks. However, in CIDR-EB synchronized buffaloes, a progesterone device (CIDR) was inserted in the anterior vagina and a single dose of estradiol benzoate (2 mg) and prostaglandin (150 μg) were administered i.m. on d 1. The CIDR was removed on d 7 and repeated OPU's (n = 56) were performed. In both experiments, a 7-d resting period was provided between each OPU session. Data on the follicular turn-over, OR, OQ, and early stages of embryonic development were analyzed with mixed models using the PROC MIXED and GLIMMIX procedures of SAS. Results revealed that the number of medium sized follicles (LSM ± SEM) and rate of OR (%) were greater (P < 0.05) during PBS (2.76 ± 0.40; 61%) and TBS (1.73 ± 0.30; 54%) as compared to LBS (0.68 ± 0.30; 31%) in buffaloes, respectively. The OR rate was also greater (P < 0.05) using 17 G as opposed to 18 G needle (62% vs. 35%), however, there was no effect (P > 0.05) of season and G of needle on OQ in buffaloes. In experiment 2, the number of small (3.50 ± 0.63 vs.2.69 ± 0.60) sized follicles was higher (P < 0.05) in FA, whereas, medium (1.13 ± 0.45 vs. 1.59 ± 0.45) sized follicles were greater (P < 0.05) in CIDR-EB synchronized buffaloes. The OR rate (67% vs. 53%), and OQ (Grade I_+_II; 15% vs. 16% and Grade III_+_IV; 85% vs. 84%), remained similar (P > 0.05) in FA and CIDR-EB treated buffaloes, respectively. Similarly, rates of in-vitro maturation (66% vs. 59%), cleavage (42% vs. 53%), 4-cell (27% vs. 32%), 8-cell staged embryos (23% vs. 25%), and morula (13% vs. 8%) did not differ (P > 0.05) between FA and CIDR-EB methods of follicular wave emergence, respectively. Taken together, it is concluded that peak or transition breeding seasons are suitable to perform OPU using 17 G needle for maximum follicular turn-over, and OR in buffaloes. However, the two fundamental methods of synchronization of wave emergence resulted in similar efficiency for OPU derived in-vitro embryo production in Nili-Ravi buffaloes.

摘要

本研究的目的是评估光照周期、采卵(OPU)针的规格(G)以及两种卵泡波出现方法对尼里-拉菲水牛(n = 20)卵泡更新、卵母细胞回收(OR)、卵母细胞质量(OQ)和胚胎早期体外发育能力的影响。在1个实验中,将水牛(n = 12;每个季节4头水牛)随机分配,在以下季节之一使用17G或18G针来优化OPU的次数:1)繁殖旺季(PBS;9月至11月;n = 31),2)繁殖过渡季(TBS;12月至2月;n = 32),以及3)繁殖淡季(LBS;4月至6月;n = 32)。在2个实验中,将水牛(n = 8)随机纳入2×2交叉设计,以比较在PBS期间使用卵泡消融(FA;n = 4)或同步方案(CIDR-EB;n = 4)这两种卵泡波出现方法。在FA方法中,抽吸卵巢卵泡(第-1周),随后在7周内进行重复OPU(n = 55)。然而,在CIDR-EB同步处理的水牛中,在前庭阴道插入孕酮装置(CIDR),并在第1天肌肉注射单剂量苯甲酸雌二醇(2mg)和前列腺素(150μg)。在第7天取出CIDR,并进行重复OPU(n = 56)。在两个实验中,每次OPU之间都有7天的休息期。使用SAS的PROC MIXED和GLIMMIX程序,通过混合模型分析卵泡更新、OR、OQ和胚胎发育早期阶段的数据。结果显示,与LBS(0.68±0.30;31%)相比,水牛在PBS(2.76±0.40;61%)和TBS(1.73±0.30;54%)期间,中等大小卵泡的数量(LSM±SEM)和OR率(%)更高(P < 0.05)。使用17G针时的OR率也高于18G针(62%对35%,P < 0.05),然而,季节和针的规格对水牛的OQ没有影响(P > 0.05)。在实验2中,FA组中小(3.50±0.63对2.69±0.60)卵泡的数量更高(P < 0.05),而CIDR-EB同步处理的水牛中,中(1.13±0.45对1.59±0.45)卵泡更多(P < 0.05)。FA组和CIDR-EB组处理的水牛中,OR率(67%对53%)和OQ(I+II级;15%对16%以及III+IV级;85%对84%)分别相似(P > 0.05)。同样,FA组和CIDR-EB组卵泡波出现方法在体外成熟率(66%对59%)、卵裂率(42%对53%)、4细胞(27%对32%)、8细胞期胚胎(23%对25%)和桑葚胚率(13%对8%)方面没有差异(P > 0.05)。综上所述,得出结论:繁殖旺季或过渡季适合使用17G针进行OPU,以实现水牛最大的卵泡更新和OR。然而这两种卵泡波同步的基本方法在尼里-拉菲水牛的OPU体外胚胎生产中效率相似。

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