Heat stress has adverse effects on fertility of dairy animals. Decline in fertility is linearly associated with an increase in combination of both temperature and humidity. The purpose of this study was to investigate the relationship between temperature humidity index (THI) and the pregnancy rate of Murrah buffaloes in a subtropical climate. The effects of genetic and non-genetic factors viz., sire, parity, period of calving and age group at first calving were found non-significant on pregnancy rate. The effect of THI was found significant (p<0.001) on pregnancy rate of Murrah buffaloes calved for first time and overall pregnancy rate. The threshold THI affecting the pregnancy rate was identified as THI 75. The months from October to March showed THI<75 and considered as non heat stress zone (NHSZ), while months from April to September were determined as heat stress zone (HSZ) with THI≥75. The lowest overall pregnancy rate (0.25) was obtained in July with THI 80.9, while the highest overall pregnancy rate (0.59) was found in November with THI 66.1. May and June were identified as critical heat stress zone (CHSZ) within the HSZ with maximum decline (−7%) in pregnancy rate with per unit increase in THI. The highest overall pregnancy rate was estimated as 0.45 in NHSZ with THI value 56.7 to 73.2. The pregnancy rate was found to have declined to 0.28 in HSZ with THI 73.5 to 83.7. However, the lowest pregnancy rate was estimated as 0.27 in CHSZ with THI value 80.3 to 81.6.

Aim: This study aimed to study the electrophoretic properties of seminal plasma and sperm proteins of Black Bengal buck semen and their correlation with in vitro sperm characters and freezability. Materials and Methods: Semen ejaculates from nine Black Bengal bucks were collected by artificial vagina (n=20/buck). Ejaculates were evaluated for in vitro sperm characters and electrophoretic profile of seminal protein. In vitro sperm characters were evaluated immediately after collection, after completion of equilibration period, and after freeze-thawing. For seminal protein studies, seminal plasma proteins were precipitated by ice-cold ethanol method, and sperm proteins were extracted by Triton X detergent extraction method. Discontinuous sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was performed to assess the molecular weight of seminal proteins. Correlation between in vitro sperm characters and protein bands was determined by Pearson's correlation coefficient, and two-way ANOVA was applied to find the individual buck differences. Results: Significant difference (p<0.01) among the bucks was noticed in the in vitro sperm characters evaluated at all the three stages of semen evaluation such as immediately after collection, after completion of equilibration period, and post-freeze thawing. Progressive loss of sperm motility, membrane integrity, and other in vitro sperm characters were noticed during cryopreservation. A total of ten protein bands in the molecular weight ranging from 17 to 180 kDa were found in the SDS-PAGE of seminal plasma proteins, while nine bands of 17-134 kDa were observed in sperm proteins. Seminal plasma proteins of molecular weight 75, 62-49, 20, and 17 kDa and sperm proteins of 75, 20, and 17 kDa were present in all the nine bucks (100%) screened, and variation among the bucks was noticed for the presence of other proteins. Seminal plasma protein of 180-134 kDa showed a negative correlation with individual motility (−0.716) and functional membrane integrity of sperm cells (−0.724) in post-freeze-thaw analysis and 48 kDa protein had a positive correlation with individual motility (0.649) and functional membrane integrity of sperm cells (0.664) in post-thaw analysis. Sperm proteins of 63 kDa had a negative correlation (−0.616) with sperm concentration in neat semen. Conclusion: Variation among the bucks was noticed in the in vitro sperm characters and semen freezability. Correlation between seminal proteins and in vitro sperm characters and semen freezability had been found which might be useful as a tool to select breeding bucks.

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The present study was conducted to identify the most suitable temperature humidity index (THI) model among seven reported THI models for analyzing the impact of thermal stress on monthly test day fat % (MTDF%), monthly test day SNF% (MTSNF%), monthly test day fat yield (MTDFY) and monthly test day SNF yield (MTDSNFY) of Murrah buffaloes at subtropical climatic conditions of Karnal, India. A total of 8868 MTDF% and 8606 MTDSNF% records from 1107 lactational records of Murrah buffaloes under five parities were included in the present study and weather information on dry bulb temperature (T db ), wet bulb temperature (T wb ) and relative humidity (RH in %) for the corresponding period of 20 years (March 1994-December 2013 were collected from ICAR-NDRI and ICAR-CSSRI, Karnal, respectively. The overall leastsquares means for MTDF% ranged from 7.71 ± 0.067 in TD1 to 8.10 ± 0.08 in TD 9 and MTDSNF% ranged from 9.61 ± 0.01 in TD5 and TD 6 to 9.65 ± 0.01 in TD 8. The overall least squares means of MTDFY (g) ranged from 411.23 ± 14.74 to 745.98 ± 13.57 while for MTDSNFY (g) the value ranged from 491.90 ± 17.21 to 922.16 ± 15.17. Monthly average THI was computed for each of the seven models. The lowest monthly average THI value was found in January, while either May, June or July showed the highest average THI value for all seven THI models. Regression analysis was performed for identifying the best THI to assess the impact of heat stress on milk constituent traits under study anda negative association was found between the milk constituent traits and monthly average THI values.The THI model[THI = (0.55 × T db + 0.2 × T dp ) × 1.8 + 32 + 17.5]developed by NRC(1971)was identified as the most suitable THI model to assess the impact of heat stress on milk composition traits of Murrah indicating maximum decline in MTDF% (-0.005), MTDFY (-0.68 g),MTDSNF% (b=-0.0008) and MTDSNFY (-2.25 g) per unit rise in THI.

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