Effect of Fresh and Frozen Semen on in vitro Fertilization and Subsequent Development of Goat Embryos
محورهای موضوعی : Camelاس. ساها 1 , ام.آ.ام.ی. خاندوکر 2 , ال.ی. اسد 3 , آ.ام.ام.تی. رضا 4 , آ. هوکیو 5
1 - Department of Biotechnology, Bangladesh Agricultural University, Mymensingh, Bangladesh
2 - Department of Animal Breeding and Genetics, Bangladesh Agricultural University, Mymensingh, Bangladesh
3 - Department of Animal Breeding and Genetics, Bangladesh Agricultural University, Mymensingh, Bangladesh
Department of Animal Nutrition, Genetics and Breeding, Sher-e-Bangla Agricultural University, Dhaka, Bangladesh
4 - Department of Animal Breeding and Genetics, Bangladesh Agricultural University, Mymensingh, Bangladesh
5 - Department of Animal Breeding and Genetics, Bangladesh Agricultural University, Mymensingh, Bangladesh
کلید واژه: Embryo, fresh semen, Frozen semen, <i>in vitro</i> fertilization,
چکیده مقاله :
This study was undertaken to compare the efficiency of fresh and frozen-thawed buck semen on in vitro fertilization (IVF) of goat oocytes. Cumulus oocytes complexes (COCs) were collected by aspiration of 2-6 mm diameter follicles, which were obtained from slaughterhouse. Upon grading, only normal quality COCs were maturated in TCM-199 for 48 hours. The percentage of COCs reached to the M-II stage was 61.41 ± 1.97%. The matured COCs were fertilized for 5 hours in Brackett and Oliphant media using fresh and frozen thawed semen separately. After fertilization the oocytes were cultured in TCM-199 for 48 hours to observe the cleavage rate. The maturation, fertilization and culture were performed in an incubator at 38.5 ˚C with 5% CO2 in humidified air. After fertilization cleavage rates were observed to check the fitness of zygotes to be morula and blastocyst. It was observed that the rates of normal fertilization (2 PN formations) for fresh and frozen semen were 36.02 ± 2.79 and 34.73 ± 2.58%, respectively and the cleavage rates were 25.19 ± 2.5 and 21.01 ± 2.8%, respectively. No significant differences (P>0.05) was observed between fresh and frozen semen in the efficiency of in vitro fertilization and subsequent development of goat embryos. It can be concluded that, both fresh and frozen semen can be used for IVF and subsequent development of goat embryos.
این مطالعه برای مقایسه راندمان اسپرم تازه و اسپرم منجمد یخ گشایی شده روی باروری آزمایشگاهی (IVF) تخمک‌های بز انجام شد. مجموعه‌های اووسیتی مورد استفاده توسط مکش 2 الی 6 میلی‌لیتر از محتویات تخمدان از کشتارگاه بدست آمدند. در زمان درجه‌بندی اووسیت‌ها، منحصراً مجموعه اووسیت‌های طبیعی برای 48 ساعت مورد اندازه‌گیری قرار گرفتند. مجموعه‌های اووسیتی رسیده به مرحله M-II، 97/1±41/61 درصد بود. مجموعه‌های اووسیتی بالغ برای 5 ساعت در محیط‌های براکت و اولیفانت با استفاده از اسپرم تازه و اسپرم منجمد یخ گشایی شده به طور مجزا بارور شدند. بعد از باروری، تخمک‌ها به مدت 48 ساعت برای مشاهده سرعت تقسیم پذیری کشت داده شدند. بلوغ، باروری و کشت در دمای 5/38 درجه سانتی‌گراد در یک انکوباتور حاوی 5 درصد دی اکسید کربن در هوای مرطوب انجام شد. بعد از باروری، سرعت تقسیم برای مناسب بودن تخمک‌ها برای تبدیل به مرولا و بلاستوسیت مورد مشاهده قرار گرفتند. برای اسپرم تازه و منجمد میزان باروری طبیعی (اشکال دو هسته‌ای) به ترتیب 58/2±02/36 و 58/2±73/34 درصد و سرعت تقسیم سلولی به ترتیب 5/2±19/25 و 8/2±01/21 درصد بودند. تفاوت معنی‌داری بین اسپرم تازه و منجمد در راندمان باروری آزمایشگاهی و توسعه بعدی جنین‌های بز مشاهده نشد (05/0P>). نتیجه این‌که، هردو نوع اسپرم (هم تازه و هم منجمد) می‌تواند برای IVF و توسعه بعدی جنین‌های بز استفاده شوند.
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