Integrity of sperm plasmalemma plays a pivotal role in the process of fertilization. The present study was undertaken to investigate the plasma membrane integrity by Hypo-osmotic swelling test (HOST) of Deccani ram sperm during liquid storage of semen (5ºC) and its inter-relation with other seminal characteristics (Acrosomal integrity, livability, motility, sperm abnormalities and methyl blue reduction test (MBRT). Semen was collected from 8 Deccani rams (18 ejaculates/ram) and diluted (1:4; 200millions/ml) with egg yolk citrate extender (EYC) after assessment of semen volume (ml), concentration (millions/ml), colour, consistency and mass activity (0-5 scale). Seminal characteristics were evaluated immediately after dilution, after 24 and 48 h of storage at 5ºC. The Pearson's (r) correlation test was used to calculate correlations between Hos-test and other sperm parameters (Motility, Livability, Acrosome integrity, abnormalities and MBRT). The findings of the present investigation showed highly significant positive correlation between HOS coiling and motility (P<0.01), sperm livability (P<0.01) and acrosome integrity (P<0.01) which indicate its efficiency as feasible screening method for evaluation of male fertility prior to Artificial insemination.
To explore the effect of kisspeptin on in vitro maturation of buffalo oocytes under different hormonal combinations. For the present study, Kp@10 µg/ml was selected as optimum by assessing the proportion of Cumulus Cell Expansion (CCE) and extrusion of 1st polar body (PB) from the earlier study conducted. Media containing TCM 199 (supplemented with Gentamicin) and oocytes were used as control. Overall, oocytes were allowed to mature in 12 different maturation media viz., Control (T1), Kp at 10 µg/ml (T2), FSH alone (T3), FSH +Kp (T4), LH (T5), LH +Kp (T6), E2 (T7), E2+Kp (T8), FSH+LH+E2 (T9), FSH+LH+E2+Kp (T10), FSH+LH+E2+FCS+BSA (T11) and FSH+LH+E2+FCS+BSA+Kp (T12), incubated at 38.5°C and 5% CO2 under humidified atmosphere for 22 hours. The results showed significantly (P £ 0.5) higher percentages of CCE and PB in groups supplemented with Kp (T4, T6, T8, T10 and T12,) compared to control (T1) and other treatment groups supplemented with individual hormones either alone (T3, T5 and T7) or combination of hormones (T9) and IVM alone (T11). Therefore, it has been clearly demonstrated that addition of Kp (10 µg/ml) to FSH, LH, E2 supplemented media, in various combinations enhances buffalo oocyte maturation rates in vitro.
The present investigation was aimed to study the sequential changes in the sperm cell deterioration dur-ing liquid storage of Deccani sheep breed semen from dilution to 48 h of storage along with its seminal characteris-tics and sperm morphometric measurement. Thus the two Deccani adult rams (aged 2 years), were selected (six ejaculates/each ram) and the collected semen was diluted with Egg yolk citrate extender (EYC) (final concentration - 400 million spermatozoa/0.2 ml semen).Seminal characteristics were assessed along with sperm morphologi-cal changes by Electron microscopy immediately after dilution, at 24 and 48 h of storage, respectively. Sperm morphometry was analysed by Image analysis.The percentage of Individual motility, Live spermatozoa, Acrosomal integrity and Hos-test reactive sperm decreased significantly (P<0.05) from 80.41 to 49.16%, 82.75 to 51.25%, 94.16 to 83% and 76 to 48.58%, respectively during liquid storage of semen from initial dilution to 48 h of storage. The sperm head length (μm), Head width, sperm head area (μm2), sperm head perimeter (μm), mid piece length (μm), proximal mid piece width (μm), distal midpiece width (μm), volume of mid piece (μm3) and acrosomal cap length (μm) were 7.80, 4.33, 26.84, 20.63, 14.03, 0.74, 0.51, 4.54 and 5.24, respectively. Electron microscopic qualitative evaluation revealed that the main site of injury is the apical ridge of ram spermatozoa when stored at 5ºC. The electron density of the mitochondria reduced indicating concomittant depletion of ATP and loss of motility resulting in reduction of fertility.
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