Astaxanthin supplementation ameliorates simulated heat stress by regulating physio-biochemical responses in Sirohi goats.

The present study was undertaken to assess the ameliorative effect of dietary supplementation of astaxanthin in Sirohi goats under simulated heat stress conditions. Eighteen healthy female Sirohi goats were divided equally into three groups (n = 6): Heat-Stressed Control (HSC), Treatment 1 (T1), and Treatment 2 (T2). During the experiment, goats in the T1 group were supplemented with astaxanthin at the rate of 25 mg/animal/day, while those in the T2 group received supplementation of 50 mg/animal/day. The experiment was conducted for 42 days: 14 days of acclimatization period, next 21 days animals were exposed to 42ºC for 6 h from 09:00 h to 15:00 h and 7 days of recovery period. On a daily basis, we recorded the physiological responses of goats and collected environmental data at the experimental site. Blood samples were collected 0 and 14th days of acclimatization, on 1st, 6th, 11th, 16th and 21st day of heat exposure and on the 7th day of the recovery period. The rectal temperature and respiration rates of the treatment groups were lower than those of the HSC group during the exposure period. Heat stress in the supplemented groups was associated with reduced levels of hepatic enzymes such as AST and ALT. Serum urea, creatinine and albumin levels were significantly (P < 0.05) different between control and treatment groups. It was thus concluded that dietary inclusion of antioxidant astaxanthin can ameliorate induced thermal load as evident from changes in physio-biochemical parameters in the Sirohi goats, that was more prominent at 50 mg/ animal/day than 25 mg/ animal/day.

Thyroid hormone dynamics of Tharparkar and Sahiwal cattle during induced heat stress.

Thyroid hormones and Cortisol level are the essential biomarkers in the assessment of stress condition. This study was done to estimate the metabolic hormonal profile of Tharparkar and Sahiwal during heat stress condition. The experiment was conducted on two groups consisting of Tharparkar and Sahiwal animals (5 in each group) and the experimental period comprised a 7-day acclimatization period, a heat exposure period of 21 days at control (25 °C), moderate (35 °C) and severe (42 °C) heat stress within a 9-10-day recovery period between each exposure. The hormonal concentrations of T3, T4 and cortisol were determined in serum. The serum concentration of Thyroxine (T4) and tri-iodothyronine (T3) decreases whereas cortisol level increases in both the breeds when subjected to heat stress. However, the serum level of T4 was significantly (p < 0.05) more declined in Sahiwal as compared to Tharparkar but there was no significant difference found between the two breeds in serum T3 levels. The cortisol levels were elevated in both breeds during heat stress but significantly (p < 0.05) more elevated in the Sahiwal. Hence, observations of these hormonal profiles suggest a better thermo-adaptability in Tharparkar as compared to Sahiwal.

Expression and functional role of IGFs during early pregnancy in placenta of water buffalo.

Adequate vascularisation is a key factor for successful fetal development. We hypothesized that Insulin-Like Growth Factor (IGF) family members regulate angiogenesis along with promoting fetal development and growth. In this experiment, we determined the expression and functional role of IGF family in placental compartments (caruncle; CAR, cotyledon; COT) during different stages of early pregnancy in the water buffalo (Bubalus bubalis). Samples were collected from early pregnancy 1 (EP1, 28-45 days), early pregnancy 2 (EP2, 45-90 days), and third stage of estrous cycle (11-16 days), which was taken as control. In addition, the role of IGF1 on mRNA expression of vWF, StAR, CYP11A1, 3ßHSD, PCNA, and BAX were elucidated in cultured trophoblast cells (TCC) obtained from EP2. Quantitative real-time PCR (q-PCR), westernblot, and immunohistochemistry were done to investigate the gene expression, protein expression, and localization of examined factors, and RIA was also done to assess progesterone (P4) concentration. Expression of IGFs, its receptors and binding proteins were found to be significantly higher (p < 0.05) in both CAR and COT as compared to control during early pregnancy, except binding proteins IGFBP1, 3 and 4 which were significantly (p < 0.05) downregulated in COT with advancement of pregnancy. mRNA expression was consistent with the findings of immunoblotting and immunolocalization experiments. Trophoblasts cell culture (TCC) study showed a significant time and dose-dependent effect of IGF1 onsteroidogenic transcript, which was found to be maximum at 100 ng/ml that paralleled with P4 accretion in the media (p < 0.05). Further, IGF1 upregulated the transcripts of vWF, PCNA, and downregulated BAX at the same concentration (p < 0.05). Overall, our results demonstrated that the expression of IGFs is a site-specific phenomenon in placentome, which indicates autocrine/paracrine and endocrine function. Our in-vitro finding support that IGF1 plays a critical role in placental development by promoting angiogenesis, steroid synthesis, and cell proliferation during early pregnancy.

Influence of weather patterns and air quality on ecological population dynamics of ectoparasites in goats.

Ectoparasitism has a damaging impact on the economy of goat production in India, but the factors influencing its distribution and dynamics are less explored. The present study was designed to investigate the influence of environmental factors like weather and air quality parameters on the occurrence of different types of ectoparasites in goats of two agro-climatic regions of India, viz. the Upper Gangetic Plain (UGP) and the Western Himalayas (WH). The prevalence survey for ectoparasitism among goats was conducted during the four distinct climatic seasons (winter, summer, monsoon, autumn) in both regions. The season-wise data of weather parameters (maximum and minimum temperature, relative humidity in morning and evening, sunrise and sunset time, mean daily temperature and relative humidity, daily variation in temperature and relative humidity, and day length) and air quality parameters (air quality index (AQI), particulate matter 2.5 µm (PM2.5), particulate matter 10 µm (PM10)) of both regions were analyzed in relation with the ectoparasitic prevalence pattern of corresponding regions. The results depict a noticeable correlation between the studied parameters and seasonal variation in the occurrence of each type of ectoparasites. This outcome on the interaction of studied parameters and ectoparasitism is intriguing and it opens a huge scope for future studies on the biometeorological aspects of host-parasite ecological interplay and evolutionary biology. The better understanding of climatological aspects of ectoparasite occurrences helps goat farmers in formulating appropriate timely intervention strategies for the economic control of ectoparasites, which in turn tackles ectoparasiticidal drug resistance and reduces threat of vector-borne diseases.

Expression and functional role of bone morphogenetic proteins (BMPs) in placenta during different stages of pregnancy in water buffalo (Bubalus bubalis).

The objective of this study was to document the expression and functional role of BMPs in the placental (caruncle; CAR, cotyledon; COT) during different stages of pregnancy in water buffalo. Samples collected from Early pregnancy 1 (EP1); Early pregnancy 2 (EP2), Mid pregnancy (MP), Late pregnancy (LP) while the third stage of oestrus cycle (NP) was taken as control. Also, the synergistic role of BMP4/BMP7 or combination on mRNA expression of vWF, PCNA, StAR, CYP11A1, 3ßHSD, and BAX were studied in trophoblast cells cultured (TCC) during an early stage. The qPCR and immunoblotting studies revealed that BMP2, BMPR1A, BMPR1B, and BMPR2 mRNA level was significantly (p < 0.05) upregulated during early pregnancy in COTs while in CARs it was significantly upregulated (p < 0.05) during all the stages of pregnancy.BMP4 mRNA level was significantly upregulated (p < 0.05) during early pregnancy in COTs as well as in CARs. BMP6 expression was significantly upregulated (p < 0.05) during early and late stages of pregnancy. BMP7 mRNA level was upregulated (p < 0.05) during the late stage of pregnancy in COTs. At 100 ng/ml, the BMP4 maximally stimulated the transcripts of StAR, CYP11A1, and 3ßHSD while BMP7 maximally stimulated the transcripts of 3ßHSD that paralleled with P4 accretion in the media (P < 0.05). BMP4 as well as BMP7 upregulated the transcripts of PCNA, vWF, and downregulated BAX in the TCC (P < 0.05). In conclusion, BMPs are expressed in a regulated manner with stage-specific differences in the placenta and promotes the angiogenesis, proliferation, cell survivability, and steroidogenesis thereby regulating placental function in an autocrine/paracrine manner in water buffalo.

Expression and functional role of fibroblast growth factors (FGF) in placenta during different stages of pregnancy in water buffalo (Bubalus bubalis).

The present study documented the expression and functional role of Fibroblast growth factors (FGFs) family and their receptors (Fibroblast growth factor receptor, FGFRs) in placenta (Cotyledon; COT, Caruncle; CAR) during different stages of pregnancy in water buffalo. Samples were collected from Early pregnancy 1 (EP1); Early pregnancy 2 (EP2); Mid pregnancy (MP) and Late pregnancy (LP) while diestrus stage of oestrus cycle (NP) was taken as control. In addition, modulatory role of FGF2 on mRNA expression of von Willebrand factor (vWF), Proliferating cell nuclear antigen (PCNA), steroidogenic acute regulatory protein (StAR), cytochrome P450 cholesterol side-chain cleavage (CYP11A1), 3ß-hydroxysteroid dehydrogenase (3ßHSD) and BCL2 Associated X (BAX) were studied in cultured trophoblast cells (TCC), obtained from EP2. Real-time PCR (qPCR), Western blot, and immunohistochemistry were applied to investigate mRNA and protein expressions, and the localization of examined factors whereas, P4 secretion was assessed by RIA. The mRNA and protein expression of FGFs and its receptors were maximum (P < 0.05) during EP (EP1 and EP2) in COT. However, FGFR1 and FGFR4 were upregulated (P < 0.05) during EP2 and MP in COT. Similarly, the mRNA and protein expression of FGFs and its receptors were upregulated (P < 0.05) during all stages of pregnancy in CAR. FGF family members were localized in the cytoplasm of trophoblast cells as well as in fetal blood vessels. At 100 ng/ml dosage, FGF2 stimulated the transcript of vWF maximally (P < 0.05). P4 secretion in trophoblast cells treated with FGF2 was maximum with the highest dose at 72 h. These findings corroborate that FGF acts locally in the trophoblast cells to modulate steroid hormone viz. progesterone synthesis, promote angiogenesis and favors cell survivability indicating that this factor may play an essential role in the regulation of placental formation and function in buffalo.

Alpha lipoic acid supplementation ameliorates the wrath of simulated tropical heat and humidity stress in male Murrah buffaloes.

A supplement which ameliorates temperature-humidity menace in food producing livestock is a prerequisite to develop climate smart agricultural packages. A study was conducted to investigate the heat stress ameliorative efficacy of alpha lipoic acid (ALA) in male Murrah water buffaloes (Bubalus bubalis). Eighteen animals (293.61 ± 4.66Kg Bwt) were randomly allocated into three groups (n = 6); NHSC (non-heat-stressed control), HS (heat-stressed) and HSLA (heat-stressed-supplemented with ALA@32 mg/kg Bwt orally) based on the temperature humidity index (THI) and ALA supplementation. HS and HSLA were exposed to simulated heat challenge in a climatically controlled chamber (40 °C) for 21 consecutive days, 6 h daily. Physiological responses viz. Respiration rate (RR), Pulse rate (PR) and Rectal temperature (RT) were recorded daily before and after heat exposure. Blood samples were collected at the end of heat exposure on days 1, 6, 11, 16, and 21 and on day 28 (7th day post exposure which is considered as recovery) for peripheral blood mononuclear cells (PBMCs) separation, followed by RNA and Protein extraction for Real time quantitative PCR and Western blot analysis respectively, of heat shock proteins (HSPs). Two-way repeated measure ANOVA was performed between groups at different experimental periods. RR (post exposure) in HS and HSLA was significantly higher (P < 0.05) than NHSC from day 1 onwards but HSLA varied significantly from the HS 8th day onwards. Post exposure RT and PR in both HS and HSLA varied (P < 0.05) from NHSC throughout the study; but between HS and HSLA, RT significantly varied on initial 2 days and last 6 days (from days 16 to 21). HSP70 mRNA expression significantly up regulated in high THI groups with respect to the low THI group throughout the experimental period. During chronic stress (days 16 and 21) HSP70 significantly (P < 0.05) increased in HS but not in HSLA (P > 0.05) with respect to NHSC. ALA supplementation up-regulates and sustains (P < 0.05) the expression of HSP90 in HSLA in comparison to the HS and NHSC. HSP105 expression was significantly up-regulated (P < 0.05) in HS on days 16 and 21 (during long-term exposure) but only on day 21 (P < 0.05) in HSLA. HSP70, HSP90, and HSP105 protein expression dynamics were akin to the mRNA transcript data between the study groups. In conclusion, supplementing ALA ameliorates the deleterious effect of heat stress as reflected by improved physiological and cellular responses. ALA supplementation improved cellular antioxidant status and sustained otherwise easily decaying heat shock responses which concertedly hasten the baton change from a limited window of thermo tolerance to long run acclimatization.

Milk progesterone profiles in various reproductive states in dairy buffaloes under field conditions.

Fifty-one dairy buffaloes in the last two months of gestation were selected at seven private peri-urban farms in the Peshawar district. Observations were recorded in buffaloes during normal (NBS, August to January) and low breeding seasons (LBS, February to July). After parturition, rectal examination of reproductive organs was carried out. Estrus detection was made through visual observation and the use of intact bull. Postpartum ovulation was confirmed by ovarian palpation per rectum and milk progesterone levels (MPL), determined through radio-immunoassay. MPL was higher (p < 0.01) at various intervals in NBS calves (1.97 +/- 0.30 ng/ml) as compared to LBS calves (0.68 +/- 0.08 ng/ml). During LBS, MPL remained < 0.30 ng/ml up to the third fortnight and started rising later, reaching a peak of 1.27 ng/ml during the sixth fortnight. During NBS, there was a sharp rise in MPL during the second fortnight, reaching 3.64 ng/ml during the sixth fortnight. MPL was significantly different on different experimental farms (p < 0.01). MPL reached the lowest levels on the day of estrus (0.10 ng/ml), reached it's peak on day 7 and started declining on day 17 of estrus. MPL showed two postpartum elevations. In true anestrus buffaloes, MPL remained consistently low. However, in the anestrus period, silent ovulations were also noted, as reflected by increasing MPL without estrus signs. In pregnant buffaloes, MPL remained > 1 ng/ml. Results of the study showed that the low postpartum reproductive performance in dairy buffaloes during LBS was primarily due to inadequate functioning of the corpus luteum in secreting optimum concentrations of progesterone. The higher incidence of silent estrus during LBS indicated improved management for the detection of estrus.

A simple enzymeimmunoassay of milk progesterone for monitoring fertility in dairy buffaloes.

A simple, sensitive, direct (without extraction) enzymeimmunoassay (EIA) was usec to determine progesterone levels in whole milk samples of 400 Nili-Ravi dairy buffaloes. The mean progesterone values 22 d after A.I. were significantly higher in pregnant (16.6 +/- 9.2 ng/ml) than nonpregnant (below 5 ng/ml) animals. The mean progesterone values were below 0.34 +/- 0.12 (the detection limit) both at estrus and in cases of clinically diagnosed inactive ovaries, 3.18 +/- 1.9 at proestrus, 2.25 +/- 1.2 postestrus and 13.22 +/- 6.74 at Day 10 of the estrous cycle. Twenty buffaloes confirmed pregnant for 2 to 3 mo, had a mean value of 20.3 +/- 4.5 ng/ml. The EIA test is very reliable in the selection of nonpregnant buffaloes (100 %) and the confirmation of inactive ovaries and of estrus. Differential diagnosis of inactive or active ovaries can be made by analyzing two milk samples at a 7-d interval.