Effect of testicular tissue lysate on developmental competence of porcine oocytes matured and fertilized in vitro.

The objective of the present study was to investigate the effect of testicular tissue lysate (TTL) on developmental competence of germinal vesicle (GV) stage porcine oocytes. Two types of TTL were prepared through repeated freeze-thaw in liquid nitrogen, one from whole testicular tissue (wTTL) and other from either of four different sections of testes, namely just beneath the tunica albuginea (TA), from the transitional area between the seminiferous cord/tubules and the mediastinum testis (TR) and from the intermediate area (parenchymal tissue origin) and CE (cauda epididymis origin). The whole or section-wise TTL treatments were given for 44 hr during in vitro maturation (IVM). Oocyte maturation was done in either of the two media, namely defined (high-performance basic medium for porcine oocyte maturation, commercially available) and serum containing (TCM199). After maturation, oocytes were co-incubated with fresh spermatozoa for 6 hr and then transferred to embryo culture media. Treatment of GV stage oocytes with wTTL (1 mg/ml) increased the cleavage and morula percentage rate (69.23 ± 6.23 and 48.15 ± 6.77, respectively) than that of their control (58.33 ± 8.08 and 32.54 ± 5.53, respectively) in defined media, and in serum-containing media, cleavage and morula percentage rate were almost equal in both treatment (54.56 ± 7.79 and 34.70 ± 6.78, respectively) and control (59.52 ± 8.21 and 38.52 ± 6.54, respectively). However, effect of wTTL was not significant. In case of section-wise TTL supplements, TR section significantly (p < .01) improved cleavage and morula rate (58.43 ± 7.98 and 36.14 ± 6.89, respectively) followed by TA. In conclusion, present study indicates that IVM, in vitro fertilization and in vitro culture of embryo are improved in the presence of TTL, particularly its TR section. Further study is expected to reveal the principal components of TTL which may prove useful for IVM.

Effect of holding of semen and washing of seminal plasma on quality and fertility of Hampshire boar semen preserved at liquid state.

The present study was aimed to reveal the effect on keeping quality of boar semen on holding or not holding at an elevated temperature than that used for preservation when combined with washing or not washing of seminal plasma. Twenty ejaculates, four from each of five Hampshire boars were used to hold for 0 and 4h in GEPS extender at 22°C and subsequently washed (1500×g for 10min) of seminal plasma or left unwashed and preserved at 15°C for 72h after extending with the same extender. The seminal parameters in terms of sperm motility, live spermatozoa, and live spermatozoa with intact acrosome (LIA) were evaluated at 0h-(immediately after extension) and thereafter at 24h intervals. The mean percentage of sperm motility was significantly (P<0.01) higher in unwashed than washed semen at both 0h and 4h of holding irrespective of preservation period. It was significantly (P<0.01) higher in semen held for 4h than 0h irrespective of washing and significantly (P<0.01) lower in washed than in unwashed semen irrespective of holding during preservation. Irrespective of preservation period the mean percentage of live spermatozoa was significantly (P<0.01) higher with 4h than 0h of holding in both unwashed and washed semen and was significantly (P<0.01) higher in unwashed than washed semen at both 0h and 4h of holding. It was significantly (P<0.01) higher for 4h held semen irrespective of washing and was significantly (P<0.01) lower in washed than in unwashed semen irrespective of holding during preservation. The mean percentage of LIA was significantly (P<0.01) higher with 4h than with 0h holding in both unwashed and washed semen and was significantly (P<0.01) higher in unwashed than in washed semen at both 0h and 4h of holding irrespective of preservation period. It was significantly (P<0.01) higher for 4h held as compared to unheld semen irrespective of washing and was significantly (P<0.01) lower in washed than unwashed semen irrespective of holding during preservation. The mean percentage of sperm motility, live spermatozoa and LIA decreased significantly (P<0.01) in 0h and 4h holding irrespective of washing and in unwashed and washed semen irrespective of holding with increase in preservation period. Among all the treatments unwashed semen held for 4h yielded superior sperm quality on preservation. A total of 32 female pigs were inseminated using preserved semen obtained with the best processing technique found in the study. The conception rate, farrowing rate and litter size at birth were recorded to be 81.25%, 78.13% and 7.96 respectively as compared to 73.38%, 67.57% and 6.68 respectively in the control group. It could be concluded that unwashed Hampshire boar semen held for 4h, extended with GEPS and preserved at 15°C for 72h was conducive to obtain optimum fertility and fecundity in females when used for artificial insemination.