Reduced sperm number of murrah (Bubalus bubalis) bull semen in french mini-straw affects kinetic and functional competence after cryopreservation.

BACKGROUND: Extensive dilution of cattle semen with tris-based extender compromises certain sperm kinetic and functional traits following cryopreservation. OBJECTIVE: To study sperm functions of buffalo bulls under high dilution rates. MATERIALS AND METHODS: Twenty-four ejaculates were harvested twice a week from four buffalo bulls, and diluted to sperm concentrations of 80, 60, 40 and 20 million/mL. Diluted samples were filled in straws, equilibrated at refrigeration temperature for 4 h, and frozen in liquid nitrogen. Frozen sperm samples were thawed for evaluation of kinetic and functional attributes. RESULTS: Compared to 20 million/mL (million/mL) sperm sample, the total motility, progressive motility and rapid motility were reduced (P < 0.05) in 5 million/mL sample. The proportion of live sperm were significantly (P < 0.05) higher in 10, 15 and 20 million/mL samples than in 5 million/mL sample. The percentage of moribund sperm, dead sperm, and sperm with lipid per oxidation increased significantly (P < 0.05) in 5 million/mL sample. CONCLUSION: The reduction of sperm concentrations to < 10 million/mL affects post-thaw Buffalo sperm kinetic and functional attributes.. https://doi.org/10.54680/fr24110110712.

Exploration of urinary metabolite dynamicity for early detection of pregnancy in water buffaloes.

Early and precise pregnancy diagnosis can reduce the calving interval by minimizing postpartum period. The present study explored the differential urinary metabolites between pregnant and non-pregnant Murrah buffaloes (Bubalus bubalis) during early gestation to identify potential pregnancy detection biomarkers. Urine samples were collected on day 0, 10, 18, 35 and 42 of gestation from the pregnant (n = 6) and on day 0, 10 and 18 post-insemination from the non-pregnant (n = 6) animals. 1H-NMR-based untargeted metabolomics followed by multivariate analysis initially identified twenty-four differentially expressed metabolites, among them 3-Hydroxykynurenine, Anthranilate, Tyrosine and 5-Hydroxytryptophan depicted consistent trends and matched the selection criteria of potential biomarkers. Predictive ability of these individual biomarkers through ROC curve analyses yielded AUC values of 0.6-0.8. Subsequently, a logistic regression model was constructed using the most suitable metabolite combination to improve diagnostic accuracy. The combination of Anthranilate, 3-Hydroxykynurenine, and Tyrosine yielded the best AUC value of 0.804. Aromatic amino acid biosynthesis, Tryptophan metabolism, Phenylalanine and Tyrosine metabolism were identified as potential pathway modulations during early gestation. The identified biomarkers were either precursors or products of these metabolic pathways, thus justifying their relevance. The study facilitates precise non-invassive urinary metabolite-based pen-side early pregnancy diagnostics in buffaloes, eminently before 21 days post-insemination.

Demographic pattern of A1/A2 beta casein variants indicates conservation of A2 type haplotype across native cattle breeds (Bos indicus) of India.

Genetic variations of the beta casein gene hold importance because of their probable association with human health. Comparative sequence analysis of ß-casein gene across Indian native, crossbred and exotic breeds in India revealed 15 SNPs and 4 INDELs corresponding to 14 haplotypes. The frequency of A2 type haplotype was maximum (0.941) across all Indian native breeds. Among the 15 variants reported for taurine breeds, only three (A1, A2 and B) were observed in analysed populations. Allelic profiling of A1/A2 ß-casein variants in ~ 4000 animals belonging to three cattle types and breeding bulls also revealed the predominance of A2 allele (0.95) in Indian cattle. The high proportion of A2 allele/haplotype indicates that Indian native cattle are the best suited to meet the demands for A2 milk globally. However, a higher percentage of heterozygous genotype (A1A2) in breeding bulls warrants the need to screen sire lines so as to drift the herd towards A2. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-022-03232-0.

Distribution, annual committed effective dose, and health safety assessment of 210Po in marine biota from Kalpakkam coast, Bay of Bengal.

Seafood, intertidal biota, beach sediment, and seawater from Kalpakkam coast, Bay of Bengal were analyzed for 210Po to evaluate the internal exposure and other radiological safety aspects. Kalpakkam houses various nuclear power generation facilities on its coast. The activity concentration of 210Po was more pronounced in the intertidal organisms. Pelagic planktivorous fishes have the highest activity of the non-technogenic radionuclide, followed by the detrital feeders, benthic planktivores, benthic carnivores, and pelagic carnivore fishes. The affinity of 210Po to organic detrital matter and planktonic organisms has led to a higher accumulation of radionuclide in planktivorous fishes. Activity concentration of 210Po in seafood ranged between 1.13 ± 0.3 and 96.71 ± 1.6 Bq kg-1 (Becquerel/kilogram). In seaweeds and gastropods, it ranged from 2.09 ± 0.2 to 8.21 ± 0.6 and from 9.31 ± 0.7 to 21.58 ± 1.2 Bq kg-1, respectively. The committed effective dose (CED) of 210Po in seafood varied from 31.18 to 456.68 µSv yr-1 (microSievert/year). Radiological hazard parameters, such as activity intake, CED in consumption, of the seafood from this coast are within the acceptable levels prescribed by the International Commission on Radiological Protection and US Environmental Protection Agency.

Spatial and seasonal variations in coastal water characteristics at Kalpakkam, western Bay of Bengal, Southeast India: a multivariate statistical approach.

A study was carried out in the coastal waters of Kalpakkam with the objectives to evaluate the seasonality in hydrobiological parameters in surface and bottom waters, and assess the anthropogenic stress and monsoonal flux on a spatiotemporal scale. The study covered an area of approximately 100 km2 in the coastal environment. Relatively high values for pH, temperature, and TP were observed during the post-monsoon (POM) season. The monsoon (MON) season was linked with TN, ammonia, and DO concentrations as all these parameters have shown increased values during this season due to freshwater input. The summer (SUM) season was characterized by salinity, turbidity, nitrate, phosphate, and silicate, indicating a true marine environmental condition for plankton production. Principal component analysis (PCA) and cluster analysis (CA) indicated the presence of distinct coastal water masses with respect to seasons and sampling regions. The spatial pattern indicated the distinctness of the coastal nearshore water (CNW) and coastal offshore water (COW) with respect to water quality. The CNW was more dynamic due to direct external influence as compared to the relatively stable COW environment. Similarly, the study region in the northern part, which is continuously exposed to the backwater inputs and tourism activities, was statistically different from the southern part.

Intense bloom of the diatom Hemidiscus hardmanianus (Greville) in relation to water quality and plankton communities in Tuticorin coast, Gulf of Mannar, India.

The present study reports a dense bloom of the marine-diatom Hemidiscus hardmanianus observed off the Tuticorin coast in the Gulf of Mannar (GoM), India. The surface water discoloration (pale green) was observed during a coastal survey conducted in the initial period of the northeast monsoon (October 2018). The bloom extended over an area of approximately 5 km2 around the Tuticorin harbor. Distribution and relative abundance of the phytoplankton and zooplankton species together with the water quality and Chlorophyll-a were studied in the area of bloom. H. hardmanianus density was maximum (10.57 × 104 cells L-1) in the bloom site, which was almost 97% of the total phytoplankton population. The present report is the first record of H. hardmanianus bloom in the Gulf of Mannar. The chain-forming diatom Biddulphia biddulphiana was also observed in strong numbers (802 and 432 cells L-1), which has been rarely reported from the Indian coastal waters.

Distribution and ecological- and health-risk assessment of heavy metals in the seawater of the southeast coast of India.

The objective of the present study was to conduct an ecological and health risk assessment of heavy metals in the seawater of the southeast coast of India. The distribution profile of heavy metals in the surface seawater was Fe (79.60 ± 21.57 µg/L) > Zn (9.31 ± 1.33 µg/L) > Cu (5.19 ± 2.00 µg/L) > Ni (2.45 ± 0.76 µg/L) > Mn (1.20 ± 1.00 µg/L) > U (0.44 ± 0.23 µg/L) > Pb (0.36 ± 0.06 µg/L) > Cr (0.31 ± 0.57 µg/L) > Cd (0.11 ± 0.05 µg/L) > Co (0.07 ± 0.20 µg/L). Cu level for most of the samples exceeded the USEPA criteria for acute CMC (criterion maximum concentration) and chronic CCC (criterion continuous concentration). Other studied metals, Cd, Cr, Pb, and Ni, remained below the acute CMC and chronic CCC guidelines. The seawater pollution index (Iwp) of Cr, Ni, Zn, Cd, and Pb complied with the category-I seawater (<1, unpolluted). The ERI values (0.46-3.99) of the seawater of the studied coast mostly fell under the ecologically low risk category with respect to heavy metals. Dermal Hazard index values were orders of magnitude lower than one, indicating no potential health concern due to dermal exposure.

Comparative proteomic analysis of Salmonella Typhimurium wild type and its isogenic fnr null mutant during anaerobiosis reveals new insight into bacterial metabolism and virulence.

AIM: The aim of this study was to understand the role of anaerobic regulator FNR (Fumarate Nitrate Reduction) in Salmonella Typhimurium through proteomic approach. METHODS AND RESULTS: We did label free quantitative proteomic analysis of Salmonella Typhimurium PM45 wild type and the fnr null mutant cultured under anaerobic conditions. The data revealed 153 significantly differentially expressed proteins (DEPs) in the mutant out of 1798 total proteins identified. Out of 153 DEPs, 94 proteins were up-regulated (repressed by FNR) and 59 proteins were down-regulated (activated by FNR) in the mutant. The network analysis indicated up-regulation of TCA cycle, electron transport chain and ethanolamine metabolism and down regulation of pyruvate metabolism and glycerol and glycerophospholipid metabolism. CONCLUSIONS: Our study showed that FNR represses ethanolamine utilization. The different metabolic pathways such as pyruvate metabolism, glycerol metabolism and glycerophospholipid metabolism were activated by FNR. Further, FNR positively regulated the DNA binding protein Fis, one of the global regulators of virulence in Salmonella Typhimurium. Thus, our finding highlights the pivotal role of FNR in regulating bacterial metabolism and virulence during anaerobiosis for systemic infection of the host.

Ameliorative effect of fermentable fibres on adiposity and insulin resistance in C57BL/6 mice fed a high-fat and sucrose diet.

The consumption of diets rich in fat and refined sugars is recognized to be one of the causes of lifestyle disorders, and dietary fibres are being advocated to ameliorate the complications associated with these disorders. In the present study, the effects of two soluble fermentable fibres, viz., gum acacia and inulin on the progression of adiposity, insulin resistance, and the expression of genes related to metabolism were examined in C57BL/6 mice fed a high-fat and sucrose diet for 18 weeks. The feeding of either type of fibre resulted in decrease in body weight, epididymal fat mass, adipocyte size, hyperlipidaemia, hyperglycaemia and hyperinsulinemia. In the fibre-fed groups, the expressions of adiponectin and glucose transporter 4 in the epididymal fat increased significantly, while the expressions of leptin, interleukin 6 and tumor necrosis factor alpha decreased significantly. Moreover, the expressions of genes related to beta-oxidation, viz., carnitine palmitoyltransferase 1, peroxisome proliferator-activated receptor gamma co-activator 1ß, and peroxisome proliferator-activated receptor alpha in the liver tissue of the fibre-fed groups enhanced significantly. Furthermore, due to the feeding of either type of fibre, the expressions of zonula occludens 1 and fasting-induced adipose factor in the distal ileum and proglucagon in the colon increased significantly. From the results of the present study, it can be concluded that the beneficial effects of the fibres are mediated due to enhanced energy expenditure, improved intestinal integrity, and reduced inflammation.

Monitoring chlorine residual and trihalomethanes in the chlorinated seawater effluent of a nuclear power plant.

Periodic sampling of the discharged seawater effluent from Madras Atomic Power Station (Kalpakkam, Tamil Nadu, India) was carried out during 2013-2017 to assess the residual chlorine and trihalomethanes content in the outfall discharge water. The variations in dissolved oxygen, temperature, and pH were correlated with the residual chlorine and trihalomethanes content in the discharged effluent. The difference in temperature (ΔT) between influent and effluent seawater samples ranged from 1.95 to 11.0 °C (6.47 ± 1.87). More than 95% of the ΔT values were within the guideline value of 7 °C. The discharge water was associated with a marginal reduction in dissolved oxygen and a marginal increase in conductivity values. The total residual chlorine content in the discharged seawater at outfall ranged from 0.06 to 0.42 (0.16 ± 0.08) mg/L, which was within the stipulated values of 0.5 mg/L. Trihalomethanes values ranged from 0.04 to 65.03 (13.06 ± 14.38) µg/L. In addition to bromoform as the major constituent, occurrence of significant amount chloroform of was occasionally observed in the discharge water.

Targeted transcript analysis revealed association of suboptimal expression of certain endometrial immunity-related genes with disparate uterine diseases in zebu cows.

The onset of uterine infection during postpartum period compromises uterine health, fertility, and productivity of dairy cattle. Endometrial innate immunity plays a key role in eliminating uterine infection and keeping the uterus healthy. Hence, the present study has been designed with the hypothesis that altered endometrial immune response around calving may compromise uterine health during postpartum period. Expression of interleukins (IL-1ß, IL-6, IL-8, IL-10, and TNF-α), prostaglandin synthase (PGFS, PGES), and antimicrobial peptides (beta-defensins (BDEF-4, BDEF-5), lingual antimicrobial peptide (LAP), and calcium-binding proteins (S100A8, S100A9, and S100A12) in endometrial tissues on the day of calving was studied using qRT-PCR, and circulating concentrations of prostaglandin E and F metabolites (PGEM and PGFM) during peripartum period (on days - 7, - 4, - 1 (before calving), 0 (on the day of calving), + 1, + 4, and + 7 (post calving)) of normal (healthy) cows (n = 11) that did not develop postpartum uterine infection and cows that developed puerperal metritis (n = 7) and clinical endometritis (n = 6) were studied. Endometrial expression of IL-1ß, TNF-α, BDEF-4, BDEF-5, S100A8, S100A12, and PGFS was higher (P < 0.05), and expression of IL-6, IL-8, IL-10, and PGES was lower (P < 0.05) in normal (healthy) cows than puerperal metritic and clinical endometritic cows. The PGFM concentration in serum was high (P < 0.05) on days 0, + 1, and + 4 of calving in puerperal metritic cows followed by normal and clinical endometritic cows. However, PGEM concentration in serum was high (P < 0.05) during peripartum period in uterine-infected (puerperal metritic and clinical endometritic) cows compared with normal cows. From the above findings, it is concluded that higher constitutive expression of IL-1ß, TNF-α, PGFS, BDEF-4, BDEF-5, S100A8, and S100A12 genes in the endometrium and lower concentration of PGEM during the period immediate to calving might be beneficial for uterine health of cows.

Evaluation of some in vitro probiotic properties of Lactobacillus fermentum Strains.

This study aimed to check the in vitro probiotic properties of eleven Lactobacillus fermentum strains previously isolated from fermented dairy products and infant faeces. These cultures were tested for their tolerance to different pH such as 2.0, 2.5, 3.0, 3.5 and 6.5, bile salt hydrolysis and cell surface hydrophobicity. All the strains were persistent at pH 3.5 for 3 h whereas only faecal origin isolates such as L. fermentum BIF-19, BIF-20, BIF-18 and MTCC 8711 had shown considerable growth at pH 2.5. The strains NCDC-400, MTCC 8711, BIF-18, BIF-19 and BIF-20 showed slight to intense precipitation zone of bile salt hydrolase activity by agar plate assay. The strain L. fermentum BIF-19 exhibited best preliminary probiotic properties was selected for the adhesion to Caco-2 cell lines, which shows similar adhesion to that observed for standard probiotic Lactobacillus rhamnosus GG.

Effect of tropical thermal stress on peri-implantation immune responses in cows.

Pregnancy losses during the peri-implantation period cause a negative impact on the reproductive and economic performance of dairy herds. In this study, we investigated the possible immunological factors which may contribute to pregnancy loss during the peri-implantation period under different seasons of tropical conditions. Eighteen Karan Fries cows, six cows in each season (W: winter; HH: hot-humid; HD: hot-dry) were selected. These cows exhibited heat and were brought for artificial insemination (AI; day 0). Blood was collected on days 0, 10, 14, 16, 18, 21 and 28 post-AI. Pregnancy was confirmed by non-return to heat, progesterone assay and ultrasonography. Blood neutrophils were isolated and tested for their number, myeloperoxidase (MPO) concentrations and expression of cell adhesion molecules (CD11b, CD14, CD25, CD47), interferon tau stimulated genes (ISG15, MX1, OAS1) and chemokine receptors (CXCR1, CCL2). Plasma cortisol, progesterone, IL-2 and IL-10 were also estimated. Neutrophil number, MPO levels, the relative expression of various neutrophil receptors and plasma IL-2 were low between days 14-21 post-AI in all seasons. However, plasma cortisol and IL-10 were higher during the same period. The inflammatory activity of neutrophils, plasma IL-2 and cortisol were highest in HH, intermediate in HD and lowest in W season. However, plasma progesterone and IL-10 were highest in W season and lowest in HH season. Our results show that blood neutrophils sense the implanting embryo and downregulate their activity to ensure successful implantation; however, under harsh environmental conditions, it is a great challenge for the immune system to maintain such balance and thus it may negatively affect the outcome of pregnancy.

Global proteomic analysis of water buffalo (Bubalus bubalis) saliva at different stages of estrous cycle using high throughput mass spectrometry.

Accurate and efficient detection of estrus is one of the major constraints for exploitation of the production potential of buffalo owing to its poor manifestation of estrus signs, seasonal differences in expression and higher incidences of silent estrus (29%). The current study focused on identification of estrus specific candidate proteins in saliva of buffaloes. Estrus was detected based on behavioral signs in response to the teaser and changes in reproductive organs and confirmed by per-rectal examination, trans-rectal USG of reproductive organs, cervico-vaginal mucus characteristics and blood serum progesterone estimation. Day of onset of estrus was considered as day 0 and day -3, +3, +10 were considered as proestrus, metestrus and diestrus stage of the estrous cycle respectively. A total of 19 animals and their 38 estrous cycles (two from each) were included in this study. Saliva was collected from these animals during different stages of estrous cycle. Out of these, 08 animals were selected for global proteome analysis of saliva using in-solution digestion and nano-LC-MS/MS. A total of 275, 371, 304 and 565 proteins were identified with ≥2 peptides during proestrus, estrus, metestrus and diestrus stages of estrous cycle. Among the identified proteins 31, 62, 32 and 104 proteins were found specific to proestrus, estrus, metestrus and diestrus stage of the estrous cycle. Few salivary proteins such as Cullin-associated NEDD8-dissociated protein 1, Heat shock 70 kDa protein 1A, 17-beta-hydroxysteroid dehydrogenase type 1, Inhibin beta A chain, testin were identified as estrus specific and are important for estrus physiology. Taken together, these estrus specific proteins could be considered as the candidate biomarker for detection and confirmation of estrus in buffalo after thorough validation.

Impingement of marine organisms in a tropical atomic power plant cooling water system.

A one-year impingement monitoring was conducted at Madras Atomic Power Station (MAPS), Kalpakkam, southeastern coast of India and identified a total of 67 species of marine organisms in the cooling water system. Estimates of total annual impingement contributed about 1.47×106 individuals and 142.5t of biomass. Jellyfish contributed about 6.8×105 individuals and 135.6t of biomass. Crabs, shrimps and fish were the most vulnerable organisms contributing about 4.29×105 individuals, 1.39×105 individuals and 2.16×105 individuals respectively. Commercially important species namely Trichiurus lepturus, Sardinella longiceps and Portunus pelagicus were found to be impinged 1.88% and 0.29% by number and weight of the total biomass respectively. Out of ~327 fish species recorded at Kalpakkam, only about 9.4% of species were impinged at MAPS. Multispecies impingement at MAPS poses the problem of finding the best mitigation options for tropical conditions.

Changes in sub-soil river water quality upon its open storage-a case study.

A study was carried out to investigate the changes in the physicochemical and biological properties of sub-soil river water upon its storage in a man-made reservoir. Palar sub-soil and reservoir water samples were collated fortnightly for a period of 5 years (2010-2014). The open reservoir is used as a reliable raw water source for condenser cooling systems and for the demineralizing (DM) plant input of Fast Breeder Test Reactor (FBTR), Madras Atomic Power Station (MAPS), and other laboratories at Kalpakkam, southeast coast of India. Relatively high nutrient concentration was observed in the Palar sub-soil water, and a significant reduction in average concentration (µmol l-1) of phosphate (Palar 1.92; open reservoir 1.54) and nitrate (Palar 9.78; open reservoir 5.67) was observed from Palar to open reservoir. Substantial increase in pH (Palar 8.05; open reservoir 8.45), dissolved oxygen (mg l-1) (Palar 6.07; open reservoir 8.47), and chlorophyll-a (mg m-3) (Palar 1.66; open reservoir 8.43) values were noticed from the Palar sub-soil water to open reservoir water. It is concluded that sub-soil water with higher nutrient concentrations when stored openly, exposing to the sun, resulted in growth of plants, planktonic, and macrophytes, which led to substantial deterioration in water quality from its utility point of view as a condenser cooling medium and raw water input for DM plant.

Elemental distribution and trace metal contamination in the surface sediment of south east coast of India.

Spatial distribution and potential ecological risk of trace metals in the surface sediment of south east coast of India covering eight different ecosystems was studied. The concentration of major elements viz. Ca, Mg, K, Ti and trace metals viz. Cr, Mn, Co, Al, Fe, Ni, Cu, Zn, Cd and Pb were analysed by energy dispersive X-ray fluorescence technique. Contamination factor, geo-accumulation index, probable effect level, enrichment factor and pollution load index were calculated to evaluate the pollution status. Except cadmium, CF values for all the metals ranged between 1≤CF≤3 indicating moderate metal contaminations along the coast. Mean PEL quotient (Qm-PEL) indicated toxicity probability to be below 21%. Fe, Cu, Zn and Co showed significant positive correlation (p<0.01) with clay. Chromium was the only metal that demonstrated strong negative correlation with clay (p<0.01) and positive correlation (p<0.01) with sand content.

Transcriptional abundance of antioxidant enzymes in endometrium and their circulating levels in Zebu cows with and without uterine infection.

Oxidative stress during peripartum period may compromise the uterine immunity. In the present study, we assessed the oxidative stress and antioxidant status during peripartum period and studied their relationship with postpartum uterine infection in dairy cows. Peripheral blood concentrations of total antioxidant capacity (TAC), malondialdehyde (MDA) and nitric oxide (NO) were determined (day -21, -7, on the day of calving and day +7, +21, +35) in normal (n=11), puerperal metritic (n=7) and clinical endometritic (n=6) cows. Endometrial biopsy was performed on the day of calving and expression of CAT, GPx4 and SOD2 genes was studied using qRT-PCR. Puerperal metritic cows had significantly (P<0.05) lower TAC (on day -7, day 0, day +7, +21 & +35), higher MDA (on day -21, -7 & on the day of calving) and NO (on day 0, +7 & day +35) concentrations compared to normal cows. Similarly, clinical endometritic cows had significantly (P<0.05) lower TAC (on day -7, 0, +7 & +21), higher MDA (on day -21, -7, +7 and +35) and NO (on day +7, +21 & +35) concentrations compared to normal cows. The expression of CAT and GPx4 genes was lower (P<0.05) and SOD2 gene was higher (P<0.05) in endometrial tissue of cows that developed uterine infection compared to normal cows. The relationship of peripheral levels of MDA and NO with antioxidant enzymes expression in endometrial tissue was found significant. Receiver operator characteristic analysis revealed that the concentrations of TAC on day -7 to day +35, MDA on day -21 to day +7 and NO on the day of calving to day +35 were highly correlated to the development of postpartum uterine infection in cows. It may be inferred that the low serum TAC level and high level of lipid peroxidation and NO during peripartum period influenced the endometrial expression of anitioxidative genes that compromised the uterine health during postpartum period.

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.

Serum MX2 Protein as Candidate Biomarker for Early Pregnancy Diagnosis in Buffalo.

Interferon-tau (IFN-τ)-induced molecular markers such as ubiquitin-like modifier (ISG15), 2',5'-oligoadenylate synthetase 1 (OAS1) and myxovirus resistance genes (MX1 and MX2) have generated immense attention towards developing diagnostic tools for early diagnosis of pregnancy in bovine. These molecules are expressed at transcriptional level in peripheral nucleated cells. However, their presence in the serum is still a question mark. This study reports sequential changes in expression of MX2 transcript in whole blood and serum MX2 protein level on days 0, 7, 14, 21, 28 and 35 in pregnant (n = 9) buffalo heifers, and on days 0, 7 and 14 in non-inseminated (n = 8) and inseminated non-pregnant (n = 10) control animals. In non-inseminated and inseminated non-pregnant heifers, the differential expression of MX2 transcript and MX2 protein level remained similar between day 7 and 14 post-oestrus. However, in pregnant heifers, on 14th and 28th day post-insemination MX2 transcript was 16.38 ± 1.57 and 28.16 ± 1.91 times upregulated as compared to day 0. Similarly, serum MX2 protein concentration followed analogous trend as MX2 transcript and increased gradually with the progression of pregnancy. Correlation analysis between expression of MX2 transcript and its serum protein level showed a significant positive correlation in pregnant animals, while it was random in other two groups. Therefore, MX2 surge at transcriptional and serum protein level after day 14-28 of pregnancy in buffalo holds potential for its use in early pregnancy detection.