The Effect of Omega ۳-۶-۹ on CASA Parameters of Buffalo Epididymal Sperm in HTF Cell Culture Medium

Background: Mammalian sperm contains very high proportionof PUFA (polyunsaturated fatty acids), especially Docosahexaenoicacid (DHA) and docosapentaenoic acid. DHA isan important fatty acid for sperm motility and viability. Thereare positive correlations between DHA and the proportion ofmotile spermatozoa in humans, boars, goats, rams and bulls. Ithas been known sperm cells can be damaged by the process ofcooling and freezing in cryopreservation and this is related tothe disruption of membrane lipids resulting in mitochondrialdamage and loss of integrity of acrosomal and plasma membranes.These events cause a loss and decline in motility andviability. Different rations of n-۳/n-۶ PUFAs improved spermquality and increasing n-۳/n-۶ PUFA ratio, sperm density andmotility were increased. In fact, very few studies have beenconducted to examine the effects of the ratio of n-۳/n-۶ PUFAson male sperm cells.Materials and Methods: In this research, sperms were collectedfrom ۲۰ pairs of bmale buffalo testicles. Collection wascarried out in slaughtered animals. Sample processing was performedin a walk-in fridge (۵°C) immediately after its arrival tolaboratory. Then tail of epididymis was fixed between two fingers.Sperm cells were collected performing several incisionsin the caudal epididymis with a surgical blade. Four levels ofomega-۳, ۶, ۹ (۰.۲۵-۰.۵-۱-۳- mM) were added into human tubalfluid containing sperms (۴۰×۱۰۶ sperm/ml), with ۱۰%, bovineserum albumin and were kept for ۳۶ hours at ۵ºC. Sperm motilitywas examined at ۱, ۱۲, ۲۴, ۳۶, hours after kept in refrigeratorat ۵ºC with computer assisted semen analysis (CASA).Results: Data were analyzed using SPSS ۲۵ statistical packageand presented as mean ± SEM. Multiple comparisons by posthoctest such as tukey and tamhane were performed to determinestatistical differences among groups. Results were consideredsignificant at P<۰.۰۵ for all tests. Analysis of data revealedthat major CASA parameters such as rapid progressive motility( Class A %), progressive motility (Class B %), progressivemotility (Class A+B+C %), velocity curvilinear (VCL μm/s),velocity average path (VAP μm/s), straight line velocity (VSLμm/s) and linearity (LIN %) were not significantly changes inomega-۳,۶,۹ fatty acids treated groups as compared to control.Conclusion: These findings demonstrated that omega -۳, ۶, ۹at these levels had no desirable effect on motility patterns ofbuffalo epididymal sperm measured by CASA during storageat ۵ºC .

کلیدواژه ها:

CASA ، Buffalo ، Omega-۳ ، ۶ ، ۹ ، Epididymal Sperm