Shabir Ahmad Lone

Partial deoxygenation of extender improves sperm quality, reduces lipid peroxidation and reactive oxygen species during cryopreservation of buffalo (Bubalus bubalis) semen

The present study was designed to investigate the effect of partial deoxygenation of extender on sperm quality, lipid peroxidation (LPO) and reactive oxygen species (ROS) in buffalo (Bubalus bubalis) during cryopreservation of semen. Semen extender was prepared freshly and split into three sub-extenders [Extender I: control (non-deoxygenated), Extender II (partially deoxygenated by using LN2 flushing) and Extender III (partially deoxygenated mechanically by vacuum pump)]. Amounts of dissolved oxygen (DO) were determined in all the three extenders and also in post-thaw semen. Ejaculates with mass motility of  ≥3+ and individual progressive motility of 70% or greater were collected from Murrah buffalo bulls and utilized in the study. Each semen sample was divided into Groups I (diluted with Extender I), II (diluted with Extender II) and III (diluted Extender III) with a maximum of 60 × 106 sperm/mL. French mini straws (0.25 mL) were filled with the extended semen samples, sealed with polyvinyl alcohol powder, kept for 3 h at 5 °C for equilibration and then stored in an automatic programmable freezer until the temperature of straws reached -145 °C followed by plunging the straws into liquid nitrogen (-196 °C). Semen samples were evaluated at pre-freeze and post-thaw stages for various variables [sperm motility, live sperm count, acrosomal integrity, hypo-osmotic swelling (HOS) response, LPO and ROS concentrations]. The mean DO was less (P < 0.05) in Extender II as compared to I and III. The DO was less (P < 0.05) in Group II (semen extended with Extender II) as compared with III (semen extended with Extender III) and I (semen extended with Extender I). The percentages for sperm motility, viability and intact acrosomes (PIA) were greater (P < 0.05) in Groups II and III as compared to the control group at the pre-freeze stage, while at the post-thaw stage, percentages of sperm motility, viability, PIA and HOS response were greater (P < 0.05) in Group II as compared with the control group and Group III. Pre-freeze HOS response (%) was greater (P < 0.05) in Group II as compared with the control and Group III. At the pre-freeze stage, sperm LPO and ROS were less (P < 0.05) in Groups II and III as compared with the control and at post-thaw stage, spermatic LPO and ROS concentrations were less (P < 0.05) in Group II than in the control group and Group III. In conclusion, partial deoxygenation of extender improves sperm quality, reduces sperm LPO and ROS concentrations in buffalo during cryopreservation. Partial deoxygenation of the extender with LN2 flushing may be one of the ways for improving quality and fertility of frozen-thawed buffalo sperm.

Effect of incubation on freezability of cholesterol-loaded cyclodextrin treated buffalo (Bubalus bubalis) spermatozoa

Aim: The aim of this study was to investigate the effect of incubation on freezability of cholesterol loaded cyclodextrin (CLC) treated buffalo spermatozoa. Materials and Methods: Semen samples with mass motility of 3+ and greater, collected from Murrah buffalo bulls were utilized. Immediately after collection, four equal groups of semen sample were made. Group I was kept as control and diluted with Tris upto concentration of 60×106 sperm/ml, where as Groups II, III, and IV were treated with CLC at 3 mg/120× 106 spermatozoa, incubated at 37°C for action of CLC for 10, 15 and 20 min, respectively, and diluted with tris upto concentration of 60×106 sperm/ml. All groups were subjected to equilibration and freezing. The evaluation of semen samples from all groups was carried out at fresh, pre-freeze and post-thaw stage for progressive motility, viability and hypo-osmotic swelling response (HOS response). Results: At the pre-freeze stage, significantly (p<0.05) higher percentage of progressive motility and viability was observed in treatment groups as compared to control with no significant difference among treatment groups. HOS response was significantly (p<0.05) higher in treatment groups as compared to control at pre-freeze stage. At post-thaw stage, significantly (p<0.05) higher percentage of progressive motility, viability and HOS response was recorded in Group II as compared to control and other treatment groups (III and IV). Group II retained significant post-thaw motility and viability at various post-thaw incubation periods. Conclusion: Incubation period of 10 min for CLC treated buffalo spermatozoa yielded significantly higher results in terms of freezability as compared to incubation for 15 and 20 min.

Influence of stocking density on growth performance of vencobb broiler.

The objective of the study was to evaluate the influence of stocking density on growth performance of vencobb commercial broiler reared upto 46 days under deep litter system. The birds were fed adlib with commercial broiler feed. Deep litter rearing arrangements were prepared 2 days prior to the arrival of chicks. Vaccination, housing, feeding and watering was done as per standard. A total of 120, day old chicks were divided in three equal groups (I, II and III) with 40 birds in each group. The birds were reared under stocking densities of 16 birds/m 2 (group I), 12 birds/m 2 (group II) and 8 birds/m 2 (group III). Feed consumption was significantly (p<0.01) higher under stocking densities of 12 birds/m 2 and 8 birds/m 2 as compared to stocking density of 16 birds/m 2 . Average live weight and carcass weight were significantly (p<0.01) higher under stocking density of 12 birds/m 2 . There was no significant difference in the feed conversion ratio (FCR) among all three groups. Dressing percentage was better with low stocking density of 12 birds/m 2 (71.05±2.98) and 8 birds/m 2 (72.29±1.04) as compared to high stocking density (16 birds/m 2 ). In group I, feed consumption was positively correlated with live weight (r=0.87), FCR (r=0.67) and dressing percentage (r=0.90) and carcass weight (r=0.91). Live weight was positively correlated with FCR (r=0.79), dressing percentage (r=0.90) and carcass weight (r=0.86). FCR was positively correlated with dressing percentage (r=0.90) and carcass weight (r=0.84). Dressing percentage was positively correlated with carcass weight (r=0.92). In group II, feed consumption was positively correlated (p<0.01) with live weight (r=0.97), FCR (r=0.98) and dressing percentage (r=0.96) and carcass weight (r=0.91). Live weight was positively correlated with FCR (r=0.97), dressing percentage (r=0.90) and carcass weight (r=0.93). FCR was positively correlated with dressing percentage (r=0.90) and carcass weight (r=0.92). Dressing percentage was positively correlated with carcass weight (r=0.98). In group III, feed consumption was positively correlated (p<0.01) with live weight (r=0.82), FCR (r=0.79) and dressing percentage (r=0.92) and carcass weight (r=0.91). Live weight was positively correlated with FCR (r=0.81), dressing percentage (r=0.79) and carcass weight (r=0.81). FCR was positively correlated with dressing percentage (r=0.83) and carcass weight (r=0.81). Dressing percentage was positively correlated with carcass weight (r=0.89).

Possible mechanisms of cholesterol-loaded cyclodextrin action on sperm during cryopreservation

Artificial insemination (AI) with cryopreserved semen has a pivotal role in wider dissemination of germplasm of elite livestock and also for conservation of various endangered species. Cryopreservation allows storage of semen for a prolonged period of time and facilitates greater exchange of genetic material among distant populations. Cryopreservation, however, leads to certain deleterious effects on sperm including premature induction of the acrosome reaction, reduced sperm motility and viability, and impaired sperm DNA integrity and fertility. During cooling procedures, membrane phase transitions take place, which result in micro-domain formation from aggregation of lipids, leading to impaired functions of the sperm membrane, and gap formation between gel and fluid domains. Cyclodextrins are produced by enzymatic degradation of starch and possess a unique feature, that when added alone to sperm cyclodextrins facilitate the removal of cholesterol from the membrane. When preloaded with cholesterol, however, cyclodextrins stimulate the insertion of cholesterol into the sperm membrane due to presence of a hydrophobic core in addition to an outer hydrophilic face. Treating sperm of various species with cholesterol-loaded cyclodextrin improves the quality of sperm during cryopreservation. It is still not clearly known how cholesterol-loaded cyclodextrin functions at sperm cells to enhance the survival during cryopreservation. The present review, therefore, highlights possible mechanisms of cholesterol-loaded cyclodextrin action on sperm during cryopreservation.

Effect of egg yolk powder on freezability of Murrah buffalo (Bubalus bubalis) semen.

Aim: The aim of this study was to investigate the effect of commercial egg yolk powder as an alternative to fresh egg yolk on freezability of Murrah buffalo semen. Materials and Methods: Semen samples (12) from 3 Murrah buffaloes (4 from each bull) with mass motility (≥3+) and total motility (70% and above) were utilized in this study. Immediately after collection, each sample was divided into four groups. Groups I was diluted up to 60×106 sperm/ml with tris extender containing 10% fresh egg yolk and Groups II, III, and IV were diluted up to 60×106 sperm/ml with tris extender containing 2%, 4%, and 6% egg yolk powder, respectively. Semen samples were processed and cryopreserved followed by examination of frozen semen samples after 24 h. Semen samples from each group were evaluated for total motility, viability, acrosomal integrity, abnormality, and hypo-osmotic swelling test (HOST) response after dilution, pre-freeze, and post-thaw stage. Results: Pre-freeze total motility was significantly (p<0.05) higher in Groups III and IV as compared to Groups I and II, and post-thaw total motility was significantly (p<0.01) higher in Group III as compared to other three groups. Viability was significantly (p<0.05) higher in Groups II, III, and IV than Group I at the pre-freeze stage. Significantly (p<0.01) higher viability and acrosomal integrity were recorded in Group III as compared to other three groups at the post-thaw stage. Abnormality was significantly (p<0.05) higher in Group IV than other three groups. HOST response was significantly (p<0.05) higher in Groups II and III than Groups I and IV at the pre-freeze and post-thaw stages. Conclusion: Addition of egg yolk powder at 4% level yielded significantly better results in terms of post-thaw semen quality as compared to the fresh egg yolk and other concentrations of egg yolk powder (2% and 6%).

Effect of Management Systems on Seminal Characteristics of Jakhrana Bucks

A study was carried out with an objective to determine effect of management systems on seminal characteristics of Jakhrana bucks. Twenty-four Jakhrana bucks (one year-old with mean body weight of 30.0±2.0 kg) were randomly allotted into two groups as Gr-I (intensive system, n=12) and Gr-ll (semi-intensive system, n=12). A total of 336 ejaculates (168 from each group) were collected (twice weekly) using artificial vagina. Each ejaculate was evaluated for volume, mass motility, pH, progressive motility, sperm concentration, viability, hypo-osmotic swelling (HOS) response and acrosomal integrity. The mean semen volume was significantly (P<0.05) higher in Gr-I, but the sperm mass motility, initial progressive motility, sperm concentration, viability and acrosomal integrity were significantly (P<0.05) higher in Gr-ll. However, non-significant difference was observed in mean pH and HOS responsive spermatozoa. The study indicated that semi-intensive system of management is better for producing quality semen from Jakhrana bucks.

The Effects of Different Treatments of Pre-Milking Manual Tactile Teat Stimulation on Day-to-Day Variation in Milk Yield, Milk Components, Main Milking Phase, Total Milking Time and Average Milk Flow Rate in Crossbred Cattle

The present study was carried out to investigate the effect of different treatments of udder stimulation on day to day variations in milk parameters in cross bred cows. In group T1, milk yield was positively (P<0.01) correlated with main milking phase, total milking time and average milk flow rate. Milking duration was significantly (P<0.01) correlated with total milking time and average milk flow rate. Total milking time was showed positive (P<0.01) correlation with average milk flow rate. Fat percentage had positive (P<0.05) correlation with SNF (r=0.46). Significantly (P<0.05) positive correlation was observed between total solids with lactose and ash content. SNF was significantly (P<0.05) correlated with acidity. Specific gravity was positively (r=0.59) correlated with total solids. In T2, milk yield was significantly (P<0.01) correlated with main milking phase, total milking time, average milk flow rate (r = 0.99) and negatively (P<0.05) correlated with fat percentage. Main milking phase had positive correlation with total milking time and average milk flow rate and negative correlation with fat percentage. Total milking time was significantly correlated with average milk flow rate. Average milk flow rate had negative correlation with fat percentage (r=-0.71). In T3, milk yield was significantly correlated with main milking phase, total milking time and average milk flow rate and negatively with fat percentage (r=-0.55). Total milking time and average milk flow rate were positively correlated with main milking phase and milking duration was negatively correlated with fat percentage (r=-0.54). Total milking time was positively correlated with average milk flow rate and negatively correlated with fat. Fat percentage was negatively correlated with average milk flow rate was with. Ash content and acidity were positively correlated with total solids.