The aim of this study was to investigate the effects of different extenders viz. Tris citric acid fructose egg yolk (TCFEY), Tris citric acid glucose egg yolk (TCGEY), Egg yolk citrate fructose (EYCF) and Egg yolk citrate glucose (EYCG) on the quality of ram spermatozoa during preservation at 4°C. Semen samples showing more than 3+ mass motility and 70% progressive motility were pooled and subsequently divided into four aliquots. Each aliquot was extended separately in four different extenders viz. TCFEY, TCGEY, EYCF and EYCG and stored at 4°C up to 72h. The quality of spermatozoa on the basis of percentage of sperm motility, live sperm, morphological abnormalities, intact acrosome and hypoosmotic swelling test (HOST) reacted spermatozoa was evaluated immediately after extension in particular extenders (0 h), 24 h, 48 h and 72 h after preservation at 4°C. The percent sperm motility was significantly (P<0.01) higher for TCFEY and TCGEY than EYCF and EYCG at 72 h of preservation at 4°C. The percent HOST reacted spermatozoa and intact acrosomes were significantly (P<0.01) higher and morphological abnormalities were significantly (P<0.01) lower for Tris based fructose extender than other three extenders at 72 h at 4°C. In conclusion, Tris citric acid fructose egg yolk (TCFEY) was found the best in maintaining the quality of ejaculated ram spermatozoa during preservation for 72 h at 4°C.
In this study, we tested the hypothesis that use of misoprostol and isosorbide mononitrate would improve cervical penetrability and fertility after artificial insemination (AI) with chilled semen. Twenty four crossbred ewes were randomly assigned to three groups; CON (control), MIS (misoprostol) and IMN (isosorbide mononitrate). Estrus was induced by placing progesterone sponges in vagina for 11 days along with an injection of PGF 2α 24 hours before sponge removal. Cotton sponges loaded with glycerol, misoprostol and isosorbide mononitrate were placed intravaginally in respective groups at 36 hours and then fixed time AI was done twice at 48 and 60 hours after progesterone (P 4) sponge removal using 12-24 hour old chilled semen. Blood samples were collected on the day of sponge insertion, day 0, 15 and 35 for P 4 measurement. The pregnancy and lambing rates tended to be higher (p>0.05) in the IMN group (62.5%) compared to the MIS and CON groups (25% each).Interestingly, low P 4 concentration (p<0.05) was observed in pregnant ewes on days 15 and 35 than day of sponge insertion possibly indicating lower steroidogenic capacity of early pregnancy corpus luteum compared to cyclic corpus luteum. In conclusion, use of NO donor (IMN) intravaginally 12 hours before AI improved fertility.
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