اثر روش‌های مختلف یخ گشایی بر فراسنجه های اسپرم منجمد قوچ قزل

نوع مقاله: مقاله پژوهشی

نویسندگان

1 گروه علوم دامی دانشکده کشاورزی دانشگاه تبریز

2 گروه علوم دامی دانشگاه تبریز

3 استاد دانشگاه تبریز

چکیده

سابقه و هدف: امروزه عمل تلقیح مصنوعی حایگاه ویژه ای در بهبود راندمان تولید و اصلاح و بهبود پیشرفت ژنتیکی بازی می کند. به منظور تحقق بخشیدن به بسیاری از مزایای بالقوه تلقیح مصنوعی، ذخیره سازی منی برای بلند مدت یک امر ضروری محسوب می شود. این امر توسط فرایند انجماد محقق می شود. مراحل انجماد از جمله فرایند یخ گشایی منجر به تغییرات مورفولوژیکی، آسیب به عملکردهای طبیعی اسپرم و در نهایت کاهش باروری می شود. بنابراین، بهبود روش یخ گشایی منی قوچ با شاخص‌های کیفی اسپرم ارتباط داشته و از اهمیت قابل‌توجهی برخورداراست. هدف از این مطالعه مقایسه اثرات دو نرخ یخ گشایی بر پارامترهای میکروسکوپی و فعالیت های آنزیمی منی قوچ پس از فرایند انجماد - یخ گشایی بود.

مواد و روش ها :در این مطالعه، نمونه‌های منی از پنج رأس قوچ قزل، با استفاده از مهبل مصنوعی، دو بار در هفته جمع آوری شد. برای از بین بردن اثرات فردی دام‌های نر، نمونه های منی در هر بار اسپرم گیری با هم مخلوط شدندو سپس مورداستفاده قرار گرفت. نمونه ها با رقیق کننده تریس حاوی 5/1% لستین سویا و 7% گلیسرول، رقیق شدند. نمونه‌های رقیق شده در معرض بخار ازت منجمد گردیده و نهایتا تا زمان ارزیابی در تانک حاوی ازت مایع ذخیره شدند. پایوت های حاوی منی با حجم ml 25/0 در حمام آب گرم توسط دو پروتکل (i)ºc 37 بمدت s30 و (ii)ºc 60 بمدت s6 یخ گشایی شدند. بدنبال فرایند انجماد-یخ گشایی، اثرات زمان و دمای یخ‌گشایی بر ویژگی‌های جنبایی (CASA)، زنده‌مانی، یکپارچگی غشاء پلاسمایی (HOS test)، ناهنجاری‌های مورفولوژیکی، غلظت مالون دی آلدهید (MDA)، فعالیت‌های آنتی اکسیدانی (GPx ،SOD) و ظرفیت تام آنتی اکسیدان منی (TAC) ارزیابی شدند.آزمایش به صورت طرح کاملا تصادفی انجام شد و مدل آماری شامل اثر دما و زمان یخ گشایی است. تجزیه و تحلیل داده ها داده‌ها به کمک رویه Proc GLM انجام شد و میانگین‌ها به روش توکی مورد مقایسه قرار گرفتند. اختلاف معنی‌داری در سطح پنج درصد(P<0/05) گزارش شد.

یافتهها: نتایج نشان داد که فراسنجه های حرکتی اسپرم همچون درصد تحرک پیش‌رونده (PM)، سرعت در مسیر مستقیم (VSL)، سرعت در مسیر منحنی (VCL) در دمای ºc37 بهبود یافت، اما این افزایش از نظر آماری معنادار نبود. درصد زنده‌مانی و یکپارچگی غشاء پلاسمایی در ºc37 بمدت s30 نسبت ºc60 بمدت s6 بهبود یافت اما این افزایش نیز غیرمعنی‌دار بود. همچنین میزان اسپرم‌های ناهنجار در نرخ یخ گشایی بالا نسبت به نرخ پایین بهبود نیافت. علاوه براین نتایج حاصل از مقایسه پراکسیداسیون لیپید (MDA)، فعالیت آنزیم گلوتاتیون پراکسیداز GPx))، سوپر اکسید دیسموتاز (SOD) و ظرفیت آنتی‌اکسیدانی کل ((TAC در دو نرخ یخ گشایی معنی دار نبود.

نتیجه گیری :با توجه به نتایج بدست آمده از روش‌های یخ گشایی، نرخ یخ‌گشایی ºc60 بمدت s6 نمی تواند جایگزین مناسبی برای نرخ یخ گشایی ºc37 بمدت 30 ثانیه باشد.

کلیدواژه‌ها

موضوعات


عنوان مقاله [English]

Effect of different thawing procedures on frozen semen quality of Ghezel rams

نویسندگان [English]

  • Hossein Daghigh Kia 1
  • S Vatankhah 1
  • M Ebrahimi 2
  • Gholamali Moghaddam 3
1 ِDepartment of Animal Science, Faculty of Agriculture, University of Tabriz
2 Department of Animal Science, University of Tabriz
3 University of Tabriz
چکیده [English]

Background and objectives: Today, artificial insemination plays an important role in improving production efficiency and improving genetic progress. In order to realize many of the potential benefits of artificial insemination, semen storage for a long-term is considered a necessary step by freezing process. The freezing process, including the freezing process, leads to morphological changes, damage to the natural functions of the sperm and ultimately reduced fertility. Therefore, improving the thawing method of ram semen is related to the sperm quality indicators and is of considerable importance. The purpose of this study was to compare the effects of two thawing rates on microscopic parameters and enzyme activity of ram semen after freeze-thawing process.

Materials and methods: In this study, semen samples were collected from five Ghezel ram using an artificial vagina twice a week. To eliminate the individual effects of animals, semen samples were pooled after each sampling. Then diluted with Tris diluent. The diluted semen samples were frozen in nitrogen vapor and eventually stored in a tank containing liquid nitrogen until evaluation. The straws containing 0.25 ml semen were thawed at water bath temperatures at (i) 37ºC for 30s and (ii) 60ºC for 6 s. Following the freeze-thawing process, the effects of time exposure and thawing temperature on the kinematic features (CASA), viability, plasma membrane integrity (HOS test), morphological abnormalities, malondialdehyde (MDA) concentration, antioxidant activity (GPx, SOD) and total antioxidant capacity (TAC) were evaluated. The experiment was conducted as a completely randomized and the statistical model includes the effect of temperature and thawing time. The data were analyzed by the general linear model (GLM) procedure and statistical differences between the various group means were determined by Tukey's test. Significant differences were reported at the level of (P<0/05).

Results: The results showed that sperm motility parameters such as PM, VSL, VCL were improved at 37°C, but this increase was not statistically significant. The viability and plasma membrane integrity was improved at 37°C for 30 s compared 60°C for 6 s, but it was no significant. Also, total abnormality did not improve at high thawing rates than low rates. In addition, the results of the comparison of lipid peroxidation (MDA), glutathione peroxidase (GPX), superoxide dismutase (SOD) and total antioxidant capacity (TAC) were no significant at two thawing rates.

Conclusion: According to the results obtained from thawing methods, the thawing at 60°C for 6s cannot be a suitable alternative for semen thawing at 37°C for 30 seconds.

کلیدواژه‌ها [English]

  • Thawing rates
  • Motility parameters
  • Frozen semen
  • Ram
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