اثر افزودن 4- هیدروکسی تمپو (تمپول) به رقیق کننده تریس- اسید سیتریک بر قابلیت نگهداری اسپرماتوزوئیدهای گاومیش

محورهای موضوعی : پژوهش های بالینی دام های بزرگ

تاریخ دریافت : 1400/05/31 تاریخ پذیرش : 1400/05/31 تاریخ انتشار : 1393/06/01

کلید واژه: آنتی اکسیدان, گاومیش, منی, 4-هیدروکسی تمپو,

چکیده مقاله :

هدف این تحقیق بررسی اثر افزودن تمپول به رقیق کننده در اسپرماتوزوئیدهای گاومیش قبل و بعد از انجماد- ذوب است. بیست و پنج نمونه منی از 5 گاومیش نر سالم ( 5 نمونه از هر گاومیش در 5 زمان مختلف) مورد استفاده قرار گرفت. هر نمونه در C°37 در رقیق کننده تریس-اسید سیتریک (بدون زرده تخم مرغ) حاوی 0 (شاهد)، 5/0، 1، 4، 8 و 12 میلی مول تمپول رقیق گردید. تحرک روبه جلو و میزان زنده ماندن اسپرماتوزوئیدهای منی رقیق شده در 0 (T0) (بلافاصله بعد از رقیق کردن)، 60 (T1) و 120 (T2) دقیقه بعد از رقیق کردن اندازه گیری شد. در مرحله بعد همین دوزهای تمپول، زرده تخم مرغ و گلیسرول به رقیق کننده فوق افزوده، نمونه های منی در آن رقیق شد و ظرف 2 ساعت تا C°4 سرد گردید، به مدت 4 ساعت در آن دما نگهداشته شد تا به حالت تعادل درآید و سنجه های منی (تحرک روبه جلو، زنده ماندن، یکپارچگی غشا و میزان آسیب دیدن DNA) ارزیابی شد. منی متعادل شده در پایدتهای فرانسوی 5/0 میلی لیتری بسته بندی و در ازت مایع منجمد گردید. بعد، منی را ذوب کرده همان سنجه ها به اضافه توان آنتی اکسیدانی تام آن اندازه گیری شد. نتایج نشان داد افزودن 5/0 و 1 میلی مول تمپول به رقیق کننده منی در قیاس با نمونه های شاهد تحرک روبه جلو اسپرماتوزوئیدها را در نمونه رقیق و متعادل شده بدون تاثیر بر بقیه سنجه ها بهبود می بخشد. در منی منجمد- ذوب شده در رقیق کننده حاوی 5/0 و 1 میلی مول تمپول، تحرک رو به جلو، زنده ماندن، یکپارچگی غشا و توان آنتی اکسیدانی تام اسپرماتوزوئیدها بالاتر ومیزان آسیب دیدن DNA کمتری از شاهد مشاهده شد. نتیجه آن که، افزودن 5/0 و 1 میلی مول تمپول به رقیق کننده، کیفیت منی گاومیش را هنگام انجماد – ذوب حفظ می‌کند.

چکیده انگلیسی:

The aim of the present study is to investigate the effects of in vitro supplementation of Tempol on spermatozoa of buffalo bulls before and after freeze-thawing. A total of 25 samples from five healthy buffalo bulls (5 ejaculates from each bull on 5 different occasions) were used. Each ejaculate was diluted at 37°C in tris-citric acid based extender (without egg yolk) containing 0 (control), 0.5, 1, 4, 8 and 12 mM Tempol. Sperm progressive motility and viability were evaluated in diluted semen at 0 (T0) (immediately after dilution), 60 (T1) and 120 (T2) minutes after semen dilution. In the next step, the same doses of Tempol, egg yolk and glycerol were added to the above extender and semen samples diluted, cooled to 4˚C within 2 h, equilibrated for 4 hours and sperm parameters (progressive motility, viability, membrane integrity and DNA damage) estimated. The equilibrated semen was packed into 0.5 mL French straws and frozen in liquid nitrogen. Later, the semen was thawed and analyzed for the same parameters, as well as total antioxidant capacity (TAC). Results showed that adding 0.5 and 1 mM Tempol to the semen extender preserved the sperm progressive motility in diluted and equilibrated semen compared to the control without affecting other parameters. However, in frozen-thawed semen, extenders containing 0.5 and 1 mM Tempol significantly preserved sperm progressive motility, viability, membrane integrity and semen total antioxidant capacity and also resulted in lower DNA damaged sperm counts. It was concluded that addition of 0.5 or 1mM tempol to the extender would preserve semen quality during freeze-thawing.

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