Intrauterine proteolytic enzymes therapy hastens expulsion of fetal membranes in dystocia affected buffaloes.

Background: Difficult calving (dystocia) in buffalo cows is a major obstetrical problem which further leads to metritis complex, encompassing the retention of fetal membranes (RFM), puerperal metritis, endometritis and pyometra with impaired future fertility. Aims: The current study aimed to evaluate the effect of the administration of intrauterine proteolytic enzymes on the expulsion of fetal membranes and postpartum fertility in dystociac buffaloes. Methods: Proteolytic enzymes consisting of Trypsin (16 mg), Chymotrypsin (16 mg), and Papain (8 mg) were dissolved in 500 ml normal saline were administered after 1 h of assisted delivery in dystociac buffaloes along with the conventional therapy. Results: The treated animals (n=15) expelled fetal membranes within a shorter period of time (P=0.043) compared to the control group (n=15) with none in the treatment group retaining it for more than 24 hours. Fewer (26.67 vs 73.33%; P=0.027) postpartum uterine infections developed in the treated animals compared to the control group. The interval between first postpartum estrus (P=0.067), service period (P=0.554), and open days (P=0.557) was shorter in the treatment group compared to the control group where postpartum anestrus developed less frequently (26.67 vs 66.67%; P=0.066) in the animals treated with enzymatic therapy. Systemic illness (neutrophillia) was reduced in the treatment group compared to the control on day 20 (64.55 ± 1.14% vs 70.23 ± 0.99%; P=0.001) and 45 (55.05 ± 1.63% vs 64.92 ± 1.45%; P<0.001) postpartum. Conclusion: It is concluded that proteolytic enzymes therapy after assisted delivery in dystociac buffalo cows could help in the early expulsion of fetal membranes and reduce uterine infections with decreased neutrophils count.

Effect of Cryopreservation on Semen Quality Parameters in Relation to Lipid Peroxidation and Antioxidant Profile in Indian Buffalo.

BACKGROUND: Lipid peroxidation (LPO) due to oxidative stress leads to structural and functional changes in spermatozoa. OBJECTIVE: To evaluate any association of various seminal characteristics at the pre- and post-cryopreservation stages with LPO and total antioxidant capacity (TAC) in Murrah buffalo semen samples. MATERIALS AND METHODS: Sixty-five ejaculates from seven bulls were processed for cryopreservation in liquid nitrogen. RESULTS: Only 31 (47.7%) samples were found satisfactory for inclusion in the further artificial insemination. A strong negative correlation was observed between LPO and individual progressive motility, TAC, viability, plasma membrane integrity as well as acrosome integrity of fresh spermatozoa. At the post-thaw stage, post-thaw motility, viability, plasma membrane integrity and acrosome integrity had strong positive correlation with TAC. CONCLUSION: The effort to minimize LPO and enhance TAC shall play a pivotal role in improving buffalo semen quality upon cryopreservation.

Effect of Mito-TEMPO Incorporated Semen Extender on Physico-morphological Attributes and Functional Membrane Integrity of Frozen Thawed Buffalo Spermatozoa.

BACKGROUND: Sperm mitochondria are the major site of reactive oxygen species (ROS) production and excess production during freezing-thawing process inflicts oxidative damages to spermatozoa. Buffalo spermatozoa are more prone to oxidative damage due to inherently more polyunsaturated fatty acids and low cholesterol to phospholipids ratio in the plasma membrane. A mitochondrial targeted antioxidant, Mito-TEMPO was used in this study. OBJECTIVE: To study the effect of Mito-TEMPO incorporated semen extender on the post-thaw semen quality in buffalo. MATERIALS AND METHODS: A total of 18 ejaculates from three murrah buffalo bulls with ≥70% individual progressive motility were utilized for the study. Each semen sample was equally divided and extended with five groups: Group I (Control, without Mito-TEMPO addition); Group II (10 µM Mito-TEMPO); Group III (50 µM Mito-TEMPO); Group IV (100 µM Mito-TEMPO); Group V (500 µM Mito-TEMPO) to have 80×106 progressive motile sperm/mL of extender, filled and sealed in French mini straws (0.25 mL) and frozen following equilibration. The effect of Mito-TEMPO was assessed at fresh/post-dilution and post-thaw stages by evaluating physico-morphological attributes and functional membrane integrity such as hypo-osmotic swelling test (HOST). RESULTS: Initial progressive motility, viability, acrosomal integrity and HOS response was significantly (p<0.05) improved and sperm abnormality was significantly (p<0.05) reduced in extended semen with Mito-TEMPO (50 µM) compared to control at post-thaw stage, although improvement was also observed at 10 and 100 µM in post-thaw samples. CONCLUSION: Mito-TEMPO incorporated semen extender at 50 µM concentration, could be part of a rationale for improving post-thaw semen quality in buffalo.