Interferon-tau stimulated gene expression: A proxy to predict embryonic mortality in dairy cows.

The embryonic mortality in cows is a growing concern for an ever-expanding dairy industry. The current study was an attempt to shorten the open period of dairy cows having suffered embryonic loss by diagnosing them at an earlier stage. The blood samples were collected from the Karan Fries (KF) cows on days 0 (day of AI/estrus), 4, 8, 12, 14, 16, 18, 21, 24, 28, 35 and 42 post insemination. The experimental animals were then categorized into pregnant (P), conception failure/early embryonic mortality (EEM) and late embryonic mortality cows (LEM), based on progesterone assay, ultrasonography and per-rectal palpation. There were 6 animals in each group. The plasma progesterone was higher in pregnant than EEM and LEM cows. Plasma Interferon-tau concentration was significantly (p < 0.05) lower in LEM than pregnant cows where it could be detected from day 14-21 but was non-detectable in EEM cows. The mRNA expression of ISG15, OAS1, MX1 and MX2 in blood neutrophils was significantly (p < 0.05) higher from day 8-42 as against day 0 in pregnant cows. The highest expression was observed around day 18-21 in pregnant cows. The ISG15, OAS1, MX1 and MX2 mRNA expression was significantly (p < 0.05) higher from day 4-42 as compared to day 0 in LEM cows, whereas in EEM cows the expression stayed close to that of day 0 (1.00 ±â€¯0.00). The mRNA expression of ISG15, OAS1, MX1 and MX2 started to decline from day 24 onwards. The degree of expression of Interferon-tau stimulated genes was higher in pregnant and LEM cows than EEM cows. The study reveals that the Interferon tau stimulated gene expression in neutrophils can act as peripheral biomarkers for detecting the embryonic mortality in dairy cows.

Impact of heat stress and hypercapnia on physiological, hematological, and behavioral profile of Tharparkar and Karan Fries heifers.

AIM: The present investigation was undertaken to study the impact of heat stress and hypercapnia on physiological, hematological, and behavioral profile of Tharparkar and Karan Fries (KF) heifers. MATERIALS AND METHODS: The animals of both the breeds of Tharparkar and KF were exposed at different temperatures and CO2 levels. Exposure conditions of 25°C, 400 ppm CO2 level, and 60% relative humidity (RH) were taken as a control condition. The exposure conditions 40°C with two levels of CO2 500 ppm and 600 ppm with RH 55±5% and exposure conditions 42°C with two levels of CO2 500 ppm and 600 ppm with RH 55±5% were taken as treatments. The exposure period in each condition was 4 h daily for 5 consecutive days. RESULTS: Physiological responses (respiration rate [RR], pulse rate [PR], and rectal temperature [RT]) were significantly (p<0.01) higher and different during all exposure conditions compared to control condition in both the breeds of cattle. KF heifers had higher RR, PR, and RT than Tharparkar heifers. Hematological parameters, namely, red blood cell, hemoglobin, and packed cell volume were significantly higher and different during all exposure condition than control in both the breeds, whereas no significant changes were observed in total leukocyte count and differential leukocyte count. Blood pH increased with increase in temperature and CO2 levels and was significantly higher than control conditions. PCO2 and base excess were significantly (p<0.05) lower, and PO2 was higher during different exposure conditions than control in both breeds. Restlessness and excitement signs were observed in all the exposure conditions as compared to control condition in both the breeds. CONCLUSION: Changes in physiological responses, behavioral pattern, and hematological parameters reflect the current functional status of the body system, and it can be used as an index for assessing the adaptation capacity of cattle to predict changes occurring in climate variables due to increasing CO2 levels and environmental temperature.

Expression of HSP70 genes in skin of zebu (Tharparkar) and crossbred (Karan Fries) cattle during different seasons under tropical climatic conditions.

Skin is most important environmental interface providing a protective envelope to animals. It's always under the influence of both internal and external stressors. Heat shock proteins (HSP) are highly conserved stress proteins which play crucial roles in environmental stress tolerance and thermal adaptation. Present study was planned to observe the relative mRNA expression of inducible (HSP70.1 and HSP70.2) and constitutive (HSP70.8) HSP in skin of zebu (Tharparkar) and crossbred (Karan Fries) cattle during different seasons. Skin biopsies were collected from rump region of each animal, aseptically during winter, spring and summer season. Quantitative real time polymerase chain reaction was performed to examine the gene expression of constitutive (HSP70.8) and inducible (HSP70.1 and HSP70.2) HSP in skin of both the breeds during different seasons. Present study observed higher expression of both constitutive and inducible HSP genes in both the breeds during summer and winter than spring season, but magnitude of increase was higher during summer than winter. During summer season, expression pattern of HSPs in skin showed breed differences, where constitutive HSP expression was higher in Tharparkar than Karan Fries and that of inducible HSP was higher in Karan Fries than Tharparkar. Hence, present study suggested that HSP may be conveniently used as biomarkers for assessing protective response of skin against heat stress in zebu and crossbred cattle. Variation in expression between breeds is associated with their heat tolerance and thermal adaptability. In summary, skin of zebu cattle (Tharparkar) is more resistant to summer stress than crossbred (Karan Fries), providing greater protection against heat stress during summer season. Superior skin protective mechanism of zebu (Tharparkar) than crossbred (Karan-Fries) cattle against heat stress may contribute to superior adaptability of zebu cattle to tropical climatic conditions than crossbreed.

Estimation of metabolic heat production and methane emission in Sahiwal and Karan Fries heifers under different feeding regimes.

AIM: The objective of this study was designed to estimate the metabolic heat production and methane emission in Sahiwal and Karan Fries (Holstein-Friesian X Tharparkar) heifers under two different feeding regimes, i.e., feeding regime-1 as per the National Research Council (NRC) (2001) and feeding regime-2 having 15% higher energy (supplementation of molasses) than NRC (2001). MATERIALS AND METHODS: Six (n = 6) healthy heifers of Sahiwal and Karan Fries with 18-24 months of age were selected from Indian Council of Agricultural Research-National Dairy Research Institute, Karnal. An initial 15 days was maintained under feeding regime-1 and feeding regime-2 as adaptation period; actual experiment was conducted from 16(th) day onward for next 15 days. At the end of feeding regimes (on day 15(th) and 16(th)), expired air and volume were collected in Douglas bag for two consecutive days (morning [6:00 am] and evening [4:00 pm]). The fraction of methane and expired air volume were measured by methane analyzer and wet test meter, respectively. The oxygen consumption and carbon dioxide production were measured by iWorx LabScribe2. RESULTS: The heat production (kcal/day) was significantly (p<0.05) higher in feeding regime-2 as compared to feeding regimen-1 in both breeds. The heat production per unit metabolic body weight was numerically higher in feeding regime-1 than feeding regime-2; however, the values were found statistically non-significant (p>0.05). The energy loss as methane (%) from total heat production was significantly (p<0.05) higher in feeding regime-1. The body weight (kg), metabolic body weight (W(0.75)), and basal metabolic rate (kcal/kg(0.75)) were significantly (p<0.05) higher in feeding regime-2 in both breeds. CONCLUSIONS: This study indicates that higher energy diet by supplementing molasses may reduce energy loss as methane and enhance the growth of Sahiwal and Karan Fries heifers.

Influence of induced heat stress on HSP70 in buffalo lymphocytes.

Heat stress in farm animals, such as cattle and buffalo during summer and post-summer seasons is a problem for livestock producers. The effect of heat stress becomes pronounced when heat stress is accompanied with ambient humidity impairing the immune status, growth, production and reproductive performance of animals. Increase in HSP70 levels from cell cultures in presence of different stressors often does not reflect the physiological adaptability of animals governing thermal regulation. In this study we directly compared the effect of different heat stress conditions with the immune status and HSP70 expression patterns from buffalo lymphocytes both in vivo and in vitro. Murrah buffalo calves were exposed to induced heat stress with two experimental treatments: hot-dry (42 °C with existing relative humidity) or hot humid (35 °C with 70% relative humidity) condition in psychometric chamber, 4 h daily for 12 days and compared with control animals maintained in an experimental shed under natural conditions. There was >200-fold increase in serum-HSP70 levels in both heat stress conditions compared with control. Furthermore, the immune status of the calves failed to activate the level of HSP70 expression in serum lymphocytes. Lymphocytes cultured in vitro at higher temperature exert 2.5-fold increase in HSP70 concentration. This study is the first of its kind to demonstrate more complex expression pattern of buffalo serum-HSP70 level as a thermo adaptive response compared with in vitro treated cells. Results from this study indicate that serum-HSP70 levels could be used as a sensitive biomarker for heat stress management in large farm animals.

Peritoneal fluid from rabbits or goats as media for in vitro maturation, fertilization and initial culture of caprine oocytes.

The efficacy of peritoneal fluid from rabbit and goat for in vitro maturation, fertilization and initial culture of embryos from caprine oocytes was evaluated. Peritoneal fluid was collected from adult female goats (n = 9) or rabbits (n = 9). Good quality oocytes were subjected to in vitro maturation and fertilization in three different media viz. Tissue Culture Medium (TCM-199), goat Peritoneal Fluid (gPF) and rabbit Peritoneal fluid (rPF). Maturation rates were 74.7+/-2.07% and 63.6+/-5.28% in TCM-199, gPF 65.8+/-2.54% and 55.6+/-3.79%, and rPF 57.7+/-1.78% and 44.6+/-3.01% when evaluated on the basis of cumulus cell expansion and the achievement of metaphase-II stage, respectively. However, no significant differences were observed in respect of maturation rate between the control and gPF and between gPF and rPF groups. Freshly ejaculated buck semen was treated with heparin (10 microg/ml) and after 45 min incubation with heparin, 8.0% sperm were live and acrosome reacted. The proportions of fertilized oocytes based on male and female pronuclei formation or on cleavage development were 50.5+/-5.03, 42.3+/-3.15 and 34.2+/-1.98%; 31.0+/-2.80, 27.9+/-2.12 and 21.8+/-1.69% for TCM, gPF and rPF, respectively. It was concluded that peritoneal fluids either from goats or rabbits could be used as an alternative medium to TCM-199. However, further research is required to confirm its efficacy for embryo development up to the blastocyst stage.