Asit Jain

Molecular characterization and expression profile of uterine serpin (SERPINA14) during different reproductive phases in water buffalo (Bubalus bubalis).

Uterine serpins (SERPINA14) play important roles during pregnancy in the farm animals. In this study, we have cloned and characterized cDNA sequence encoding the bubaline SERPINA14. We also studied its spatio-temporal expression in the uterine endometrium. The bubaline SERPINA14 has an open reading frame of 1299bp. Itshares ∼90% identity with the SERPINA14 of other ruminant livestock species. Phylogenetically, bubaline SERPINA14 has been placed in the same clade that contained other mammalian homologues with a maximum identity to bovine SERPINA14. Using an anti-ovine monoclonal antibody, Western blot analysis of the uterine fluid of buffalo during the early stage of pregnancy confirmed the presence of SERPINA14 of about 48,000Da. The results of quantitative real time PCR (RT-qPCR) as well as in situ hybridization demonstrated a stage and tissue specific expression of bubaline SERPINA14. The level of SERPINA14 mRNA was low during stage I (Days 3-5), which increased (P<0.05) during stage II (Days 6-15) and then subsequently declined during stage III (Days 16-21) of the estrus cycle. During early pregnancy (Days ∼30 of gestation) the level of SERPINA14 mRNA was as high as that during stage II of the estrus cycle. The SERPINA14 mRNA was localized in the glandular epithelium. The differential spatio-temporal expression of SERPINA14 in the uterine endometrium of buffalo suggests its plausible important roles in reproduction.

Interferon stimulated gene 15 (ISG15): Molecular characterization and expression profile in endometrium of buffalo (Bubalus bubalis)

Interferon stimulated gene 15 (ISG15), one of the several proteins induced by conceptus derived Type I and/or a Type II interferon (IFN), is implicated as an important factor in determining the uterine receptivity and conceptus development. However, presence as well as specific role of the ISG15 in buffalo (Bubalus bubalis) reproduction is yet to be elucidated. In the present study, both genomic and cDNA sequences of bubaline (bu) ISG15 were cloned and investigated for its expression in different tissues of female reproductive tract of buffalo. Sequence analysis revealed 100% identity among the genomic sequences (1014 bp) of buISG15 from three different breeds of buffalo (viz., Murrah: Acc. No. DQ118137, Mehsana: Acc. No. DQ118138, and Nagpuri: Acc. No. DQ118136) and cDNAs (Acc. Nos. HM543268-HM543270). As in cattle, the buISG15 was comprised of two exons of 57 bp and 520 bp encoding a peptide of 154 amino acids. Moreover, the buISG15 cDNA sequence exhibited 98.3% and 98.5% identity with that of taurine and indicine cattle, respectively. Subsequent reverse transcription PCR analysis revealed expression of the buISG15 in the uterine endometrium, corpus luteum (CL), corpus hemorrhagicum and oviduct. Quantitative Real Time PCR (RTqPCR) analysis also confirmed the constitutive expression of the buISG15 in the uterine endometrium during different stages (i.e. estrus, diestrus and proestrus) of estrous cycle and also during early (∼d 30-40) pregnancy. Western blot analysis of the endometrial extract from both estrous cyclic as well as pregnant buffalo demonstrated the presence of only conjugated ISG15 which was >40 kDa. ISG15 mRNA and immune-reactive proteins were localized in the stromal as well as glandular epithelial cells of the uterine endometrium of estrous cyclic as well as pregnant buffalo. However, there was no significant difference in amount of ISG15 mRNA across the different reproductive phases. To conclude, this study will be helpful for the further understanding of the roles of the ISG15 in pregnancy of buffalo cows.

Molecular characterization and expression profile of ghrelin gene during different reproductive phases in buffalo (Bubalus bubalis)

Ghrelin, a novel motilin-related endogenous ligand for growth hormone secretagouge receptor, is implicated in various biological functions, including regulation of female reproduction. But the presence of ghrelin and its role in reproductive functions in buffalo, a species with poor reproductive efficiency, is not known. In the present study full-length ghrelin cDNA was isolated from bubaline abomasum, which encodes the entire prepropeptide of 116 amino acids. The deduced amino acid sequence of ghrelin of buffalo showed >95% and 31% identity with that of ruminants (cattle, sheep, and goat) and humans, respectively. Analysis of synonymous and nonsynonymous nucleotide substitutions in the coding region of ghrelin indicated that these sequences of different species have been under purifying selection. The 3995-bp amplicon of ghrelin gene consisting of 4 exons and 3 introns was cloned with genomic DNA from buffalo. Further, ghrelin expression was determined by quantitative real-time PCR, in situ hybridization, and immunohistochemistry in bubaline endometrial tissues at different stages of the estrous cycle and early pregnancy. Our results indicated the persistent expression of ghrelin mRNA and protein in the endometrium during stage I (day 3-5), stage II (day 6-15), and stage III (day 16-21) of the estrous cycle and also during early (~day 30-40) pregnancy. Immunohistochemistry and quantitative real-time PCR experiments indicated the relatively higher expression of ghrelin in the endometrium during stage II (day 6-15) of the estrous cycle and early pregnancy than during stage I (day 3-5) and stage III (day 16-21) of the estrous cycle, but no statistically significant difference in ghrelin expression was observed among stages. To conclude, the results of the present study indicate the persistent expression of ghrelin in the uterine endometrium throughout the estrous cycle and in early pregnancy which might be helpful in determining its role in buffalo reproduction.

Effect of Age and Sex on Haematological Profile of Kadaknath Fowl Reared Under Intensive System

The study was designed to investigate the haematological parameters at different age groups of male and female Kadaknath fowl, maintained in the poultry unit of College of Veterinary Science & Animal Husbandry, Anjora, Durg Chhattisgarh, reared under intensive farming system using standard feeding and management practices. For this study the blood samples were taken from 15 male and 15 female birds at 8, 12, 24, 40 and 48 weeks of age. The values for packed cell volume (PCV), erythrocyte sedimentation rate (ESR), total erythrocyte count (TEC), total leucocytes count (TLC), Haemoglobin (Hb), mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH), mean corpuscular haemoglobin concentration (MCHC) and differential leukocyte count (DLC) were assessed. Effect of age in PCV, TEC, TLC, Lymphocyte, Heterophil, Monocyte, Eosinophil, Hb, MCV, MCH and MCHC were significant (p0.05) age difference was observed in basophil. In general in all the haematological parameters there is an increasing trend with advancement of age. Effect of sex was found significant for eosinophil, lymphocyte, Hb, MCHC, PCV and heterophil. In general, these values are higher in males than in females.

Socio-Economic Status and Perceptions of Kosali Cattle Keepers and Constraints to Cattle Production in Central Plain Region of Chhattisgarh state -

The present study was conducted to find out the socio-economic status and constraints to cattle production in Central Plain Region of Chhattisgarh state. Total 800 respondents from three districts of the region were selected. Sex, age, family size, education, land and livestock holding, annual income, occupation and herd size factors were considered to study the socio-economic status. It was observed that majority (95.42%) of the respondents had male headed ownership, more than 50 per cent in above 30 years of age category. The average family size observed was 6.21 persons. It was found that only 66.56 per cent of respondents were literate. The percent of landless, sub-marginal and marginal farmers is more than 50% and have less than 0.5 hectare land. Most of the respondents (47%) found in the category of annual income less than rupees 20000. The main profession of livestock owners was agriculture (68.44%) followed by agriculture and some other small work. Most of the respondents had small herd size. Shrinkage of grazing land, scarcity of green fodder and concentrate and poor management were the major constraints to cattle or livestock productivity.