A slight, transient increase in heat shock induces stage-specific detrimental effect on reproductive performance of riverine buffaloes (Bubalus bubalis). The vulnerability of oocytes and early embryos, as well as their ability to recover are both restricted and stage-dependent, consequently form major basis for reproduction failure during heat stress. The current study aimed to identify the most sensitive and susceptible stage of oocytes and early embryos against short thermal stress which hampers the reproduction significantly, and intent to unravel the underlying molecular signals responsible for the poor cellular growth and stunted embryonic development. Six different in vitro experimental groups of oocytes and early embryos were exposed to 40ºC for 4 h during different stages of in vitro maturation (IVM) and in-vitro fertilization (IVF), as well as in vitro culture (IVC) of early embryos to assess the stage dependent effect of heat stress on buffalo oocytes and early embryos. The most detrimental effect of heat stress was observed in group 2 with sharp reduction in morula to blastocysts transition rate (P < 0.05). The underlying molecular signals of HSP8, MnSOD and SIRT3 genes were significantly increased from mid-IVM oocytes to 4-cell embryo. Disruption of molecular signal network of genes viz; eIF1A, U2AF, BAX, BCL2, CASPASE9, TEAD4, CDX2, ZO-1, ETS2 and NKB1 was implicated in cellular damage, inconsistent and embryonic development, consequently leading to formation of poor quality of blastocysts with reduced inner cell mass (ICM) and trophectoderm (TE) cells in the affected groups. The current study provides insight into the molecular and cellular effects of short thermal stress on buffaloes oocytes and early embryos, demonstrating development potential and identifying buffalo oocytes and pre-morulae embryos as the most vulnerable stages.

The current study aimed to investigate whether cathepsin B (cath B) inhibition in cumulus cells of morphologically graded buffalo cumulus–oocyte complexes (COCs) during in vitro maturation (IVM) could modulate apoptotic markers and enhance developmental potential. Cumulus cells from grade A and B buffalo COCs were collected at 0, 12, and 24 h of IVM, with or without supplementation of the optimized cysteine protease inhibitor E-64. Cathepsin B and selected apoptotic regulators were quantified by qRT-PCR. Cathepsin B protein levels and apoptotic signals were assessed by immunostaining and TUNEL assay, respectively. Following 24 h of IVM, in vitro fertilization (IVF) and in vitro culture (IVC) were performed, and developmental competence was evaluated based on oocyte maturation, cleavage rate, blastocyst formation rate, and blastocyst quality. Expression of cath B and pro-apoptotic genes (BID, BCL2, BAX, caspase-3) was found higher along with lower expression of anti-apoptotic genes (BCL2 and XIAP) in the cumulus cells of B grade COCs. The addition of 10 µM E-64 significantly (P < 0.05) reduced cath B and pro-apoptotic gene expression, lowered the BAX/BCL2 ratio, and decreased apoptosis signals in cumulus cells of both COC grades. E-64 supplementation improved blastocyst yield from grade A COCs and enhanced blastocyst quality in both grades, without affecting maturation and cleavage rates. E-64 supplementation during IVM modulates apoptotic markers in cumulus cells of different grades of buffalo COCs by inhibiting cath B, thereby enhancing the developmental competence and offering valuable insights for improving in vitro embryo production programs in this species.

Goats provide significant contribution to the human health, nutrition and income generation. The oocyte-derived, growth differentiation factor 9 (GDF-9) is important for fertility and fecundity in mammalian species. Species-specific different expression pattern of GDF-9 indicates its involvement in regulating follicular development and female reproduction in various mammalian species. The present study was undertaken to amplify the full length coding DNA sequence (CDS) of GDF-9 from corpus luteum of goat. Mid-luteal stage was collected for RNA isolation. Quality and quantity of RNA was determined by agarose gel electrophoresis and nanodrop spectrophotometer, respectively. cDNA was prepared from total RNA using commercially available kit and used subsequently. PCR amplification from cDNA using gene specific primers revealed full length product of size 1362 bp. Two exons and one intron comprise the GDF-9 gene structure. Several synonymous and non-synonymous mutations with other selected species were revealed in the GDF-9 CDS. In conclusion, GDF-9 full length cDNA sequence showed high homology with those of other species indicating conserved nature of GDF-9 in mammals.

Interferon-stimulated gene 15 (ISG15) is induced by conceptus-derived interferon tau (IFNT) in the endometrium during early pregnancy in ruminants, including goats. The expression of ISG15 in extra-uterine tissues, such as peripheral blood leukocytes (PBLs), suggests its potential as a novel pregnancy biomarker. The onset and length of the breeding season in goats are influenced by various factors such as breed, latitude, climate, photoperiod, etc. The Osmanabadi, an Indian goat breed, known for its early maturity, prolificacy, and good dressing percentage, was the focus of the current study. The present investigation was designed to study the messenger ribonucleic acid (mRNA) profile of Caprine ISG15 (cpISG15) in PBLs using real-time reverse transcription PCR (qPCR) on days 0, 13, 17, 21, 25 and 30 post-service in this breed. Pregnancies were confirmed by measuring plasma progesterone (P4) concentration and conducting trans-abdominal ultrasound scanning. The study revealed that the expression of cpISG15 mRNA was 5 to 8-fold higher (P < 0.05) during early pregnancy on days 17, 21, and 25 compared to day 0. There was no significant difference in the expression of cpISG15 mRNA between days 0 and 30 post-service. Progesterone concentration was higher on days 17, 21, and 25 in pregnant does compared to day 0. The presence of fetal parts was observed by ultrasound between 45 and 90 d of pregnancy. It is concluded that the detection of elevated ISG15 expression in PBLs during the early stages of pregnancy may be used as a marker for pregnancy detection in does.

Background: Tick infestation in cattle is commonly controlled using chemical acaricides producing acaricidal resistance, environmental pollution and hazards to human health. Biological control of ticks using entomopathogenic fungi can be an alternative to chemical acaricides and can be used in conjunction with chemical control in integrated pest management. Methods: The isolation of entomopathogenic fungi was carried out from naturally infected ticks in Durg district of Chhattisgarh. The species confirmation of fungal isolates was conducted by Polymerase chain reaction (PCR) using ITS gene as a molecular marker. In vitro efficacy of entomopathogenic fungi at various concentrations against engorged (adult) female ticks and unfed (Nymphs), egg laying capacity and egg hatchability of treated Rhipicephalus microplus was assessed in laboratory. Result: The entomopathogenic fungi isolated from naturally infected ticks was confirmed as Fusarium beomiforme by morpho- molecular analysis. The efficacy of F. beomiforme against adult female and nymphs of R. microplus at the concentration of n x 108 spores ml-1 showed 100% mortality. The 100% reduction of egg laying capacity of treated female ticks and no hatchability of eggs treated with F. beomiforme at the concentration of n x 107 spores ml-1 and n x 108 spores ml-1 was observed.

In the present study, genetic screening of Sahiwal and Holstein Friesian (HF) cattle and its crosses was conducted using the PCR-RFLP technique to detect the presence of CVM and DUMPS disorders. Genomic DNA was extracted from 50 Sahiwal and 100 HF animals and its crosses using commercially available kits. Gene-specific primers were utilized for in vitro amplification of SLC35A3 and UMPS genes by PCR, followed by RFLP analysis using PstI and AvaI restriction endonucleases for CVM and DUMPS, respectively. Subsequently, the PCR products were purified and sequenced to confirm any mutations in the DNA target sequences. Notably, no affected or carrier animals for CVM and DUMPS disorders were identified among the screened animals. Thus, the gene and genotype frequency of CVM and DUMPS carriers was zero, indicating a 100% frequency of normal animals. The study underscores the importance of continued screening for these genetic disorders, involving a larger sample size across the state, and advocates for mandatory screening before utilizing animals for breeding purposes, in adherence to minimum standard protocols.

Growth differentiation factor 9 (GDF9) is an oocyte-specific paracrine factor involved in bidirectional communication, which plays an important role in oocyte developmental competence. In spite of its vital role in reproduction, there is insufficient information about exact transcriptional control mechanism of GDF9. Hence, present study was undertaken with the aim to study the expression of basic helix-loop-helix (bHLH) transcription factors (TFs) such as the factor in the germline alpha (FIGLA), twist-related protein 1 (TWIST1) and upstream stimulating factor 1 and 2 (USF1 and USF2), and nuclear receptor (NR) superfamily TFs like germ cell nuclear factor (GCNF) and oestrogen receptor 2 (ESR2) under three different in vitro maturation (IVM) groups [follicle-stimulating hormone (FSH), insulin-like growth factor-1 (IGF1) and oestradiol)] along with all supplementation group as positive control, to understand their role in regulation of GDF9 expression. Buffalo cumulus-oocyte complexes were aspirated from abattoir-derived ovaries and matured in different IVM groups. Following maturation, TFs expression was studied at 8 h of maturation in all four different IVM groups and correlated with GDF9 expression. USF1 displayed positive whereas GCNF, TWIST1 and ESR2 revealed negative correlation with GDF9 expression. TWIST1 & ESR2 revealing negative correlation with GDF9 expression were found to be positively correlated amongst themselves also. GCNF & USF1 revealing highly significant correlation with GDF9 expression in an opposite manner were found to be negatively correlated. The present study concludes that the expression of GDF9 in buffalo oocytes remains under control through the involvement of NR and bHLH TFs.

To evaluate the effect of diet supplementation with Moringa oleifera leaf meal (MOLM) on egg quality traits of Japanese quails, a study was conducted for 16 weeks. For this purpose, a total of 270 adult female quails and 90 adult male quails of seven weeks of age were divided into 5 groups (M0, M0.5, M1, M1.5, and M2) of 3 replicates of 24 birds (18 females and 6 males) each. The groups corresponded to 0, 0.5, 1, 1.5, and 2% inclusion levels of MOLM. The egg quality traits were studied fortnightly in four randomly collected eggs per replicate. An improvement (P≤0.05) in the egg quality traits, particularly egg shape index, albumen index, yolk colour score, and Haugh unit was recorded in the quails fed MOLM-based diets which denote better egg quality and higher chances of acceptability of the eggs by the consumer. It is concluded that MOLM can be safely included at 2% in the diets of Japanese quails.

Mineral composition in milk can affect its nutritional as well as physio-chemical properties of milk and is considered an important trait in the evaluation of milk quality. The composition and concentration of milk minerals could be altered with natural source of variation including nutrition and genetics. The effect of diet on milk minerals is well studied. However, genetic effects on the milk minerals have recently gained the attention. This review provides an overview of the genetic variation of milk minerals, and the genomic regions associated with mineral concentration in the milk are also discussed. The difference of milk minerals between breeds and the genetic parameters including heritability estimates and correlation among minerals indicates that milk minerals are under strong genetic control. Recently, the genome-wide association study (GWAS) has explored several regions associated with milk minerals and thus provides a new genetic source for improving the milk quality through genomics-assisted breeding. Hence, a combination of the qualitative and molecular approaches can be exploited to improving the nutritional quality of cattle milk in terms of its mineral composition.

Milk fat composition is an important trait for the dairy industry as it directly influences the nutritional and technological properties of milk and other dairy products. The synthesis of milk fat is a complex process regulated by a network of genes. Thus, understanding the genetic variation and molecular mechanisms regulating milk fat synthesis will help to improve the nutritional quality of dairy products. In this review, we provide an overview of milk fat synthesis in bovines along with the candidate genes involved in the pathway. We also discuss de novo synthesis of fatty acids (ACSS, ACACA, FASN), uptake of FAs (FATP, FAT, LPL), intracellular activation and channelling of FAs (ACSL, FABP), elongation (EVOLV6), desaturation (SCD, FADS), formation of triglycerides (GPAM, AGPAT, LIPIN, DGAT), and milk lipid secretion (BTN1A1, XDH, PLIN2). The genetic variability of individual fatty acids will help to develop selection strategies for obtaining a healthier milk fat profile in bovines. Thus, this review will offer a potential understanding of the molecular mechanisms that regulate milk fat synthesis in bovines.

An interferon-inducible gene, 2'-5'-oligoadenylate synthetase-1 (OAS1), plays an essential role in uterine receptivity and conceptus development by controlling cell growth and differentiation in addition to anti-viral activities. As OAS1 gene has not yet been studied in caprine (cp), so present study was designed with the aim to amplify, sequence, characterize and in-silico analyze the coding sequence of the cpOAS1. Further, expression profile of cpOAS1 was performed by quantitative real-time PCR and western blot in the endometrium of pregnant and cyclic does. An 890 bp fragment of the cpOAS1 was amplified and sequenced. Nucleotide and deduced amino acid sequences revealed 99.6-72.3% identities with that of ruminants and non-ruminants. A constructed phylogenetic tree revealed that Ovis aries and Capra hircus differ from large ungulates. Various post-translational modifications (PTMs), 21 phosphorylation, two sumoylation, eight cysteines and 14 immunogenic sites were found in the cpOAS1. The domain, OAS1_C, is found in the cpOAS1 which carries anti-viral enzymatic activity, cell growth, and differentiation. Among the interacted proteins with cpOAS1, Mx1 and ISG17 well-known proteins are found that have anti-viral activity and play an important role during early pregnancy in rumi-nants. CpOAS1 protein (42/46 kDa and/or 69/71 kDa) was detected in the endometrium of pregnant and cyclic does. Both cpOAS1 mRNA and protein were expressed maximally (P<0.05) in the endometrium during preg-nancy as compared to cyclic does. In conclusion, the cpOAS1 sequence is almost similar in structure and probably in function also to other species along with its higher expression during early pregnancy.

An interferon-inducible gene, 2’-5’-oligoadenylate synthetase-1 (OAS1), plays an essential role in uterine receptivity and conceptus development by controlling cell growth and differentiation in addition to its anti-viral activities. However, the specific part of OAS1 is not yet documented in the goat. Therefore, in the present study, the coding sequence (CDS) of the OAS1 gene was amplified, sequenced and characterized through various Bioinformatics tools. Its temporal expression profile was also examined in the endometrium of caprine (cp) for the first time. A fragment of the cpOAS1 gene, 890 bp in length, was amplified from complementary DNA (cDNA). Nucleotide and deduced amino acid sequences revealed 99.6 to 80.4 % and 99.3 to 72.3 % identities with that of ruminants and non-ruminants, respectively. A constructed phylogenetic tree revealed that sheep and goats belong to the same clade but differ from large ruminants. Besides the Physico-chemical characteristics, its localization, various posttranslational modifications, immunogenic sites, possible domains, various motifs, and interactions with other probable proteins were predicted. In addition to the secondary structure, the three-dimensional structure was predicted using homology modelling servers and a refined model, generated by the Swiss model was the best one as based on stereochemistry validated with different programs. Further, relative expression of cpOAS1 mRNA and protein was determined by quantitative real-time PCR (qPCR) and western blot, respectively, in the endometrium of pregnant and cyclic does. Caprine OAS1 protein (42/46kDa) was detected in the endometrium of pregnant and cyclic does. Both cpOAS1 mRNA and protein were expressed maximally (P<0.05) in the endometrium during pregnancy as compared to cyclic does. It is concluded that the cpOAS1 sequence is conserved and integrative Bioinformatics analysis provides a sound foundation for further structural and functional characterization of cpOAS1 protein in the establishment of pregnancy in goats.

The corpus luteum (CL) is a dynamic and transient endocrine gland. It secretes variety of protein hormonesand have an important role in survivability of the embryo during early pregnancy. Among the interferon stimulatedgenes (ISGs), interferon-stimulated gene 15 (ISG15) is one of the gene having important role in maintaining corpusluteum in different ruminant species, but in goat (Capra hircus) its role is not yet elucidated. In the present study,temporal expression profile of cpISG15 mRNA and protein was examined by quantitative real-time PCR (qPCR)and western blot techniques, respectively, in the CL of cyclic and pregnant does. Both cpISG15 mRNA and proteinwere expressed maximally in the CL during early stage of pregnancy (16 to 24 days) as compared to cyclic does.But significant difference was not found in the expression of cpISG15 mRNA and protein in the CL between thelater stage of pregnancy (25 to 40 days) and cyclic does. Finding of significant upregulation of ISG15 in CL duringearly pregnancy at mRNA as well as protein level suggest its probable role in maintaining CL and its function at thisstage in caprine.

The emergence and spread of antimicrobial resistance have become a major global public health concern. A component of this problem is the spread of antibiotic-resistant bacteria. Flies move freely between habitats of food-producing animals and human beings and thus have great potential for dissemination of antimicrobial-resistant bacteria from a contaminated environment to milk and meat markets, posing potential hazards for consumers. During the present study, a total of 150 houseflies were captured from milk and meat shops located in Durg and Raipur city of Chhattisgarh, India. The Escherichia coli were isolated from houseflies and characterized on the basis of cultural and molecular tests. Further, the isolates were subjected to antimicrobial susceptibility testing against frequently used antibiotics using the disk diffusion method. The antibiotic resistance genes and int1 gene were detected using polymerase chain reaction (PCR). A total of 45 E. coli isolates were obtained from the fly samples with an overall prevalence rate of 30·0%. Antibiogram results confirmed that E. coli isolates were resistant to multiple antibiotics. Out of the (45) isolates of E. coli, 17 (37·8%) isolates were extended-spectrum beta-lactamase (ESBL) producer and multi-drug-resistant (MDR). Out of the ESBL and MDR E. coli isolates, blaCTX-M (24·4%), blaTEM (11·1%), tetA (28·8%), tetB (26·7%), gyrA (26·7%), parC (31. 1%) and int1 genes (15·5%) were detected but none of the isolates were found positive for blaSHV gene. Findings of the present study confirm that MDR E. coli are widely distributed in houseflies and play an important role in the transmission of antibiotic-resistant bacteria from contaminated environments to milk and meat shop environment.

The study was aimed to investigate the response of different levels of Moringa oleifera leaf meal on the quality attributes of Japanese quail meat and eggs. The study comprised of 300 day-old Japanese quail chicks reared up to 6 weeks for study on the quality attributes of broiler Japanese quail meat, and 360 adult Japanese quails reared from 7-22 weeks for study on the quality attributes of layer Japanese quail meat and eggs. The birds were allocated arbitrarily and uniformly in five different treatments: T0 (Control), T0.5, T1, T1.5, and T2 based on 0, 0.5, 1, 1.5, and 2% inclusion levels of M. oleifera leaf meal, respectively. The study revealed improvements in the quality attributes of meat and eggs, claiming meat to be of good quality, and eggs to be slightly desirable to desirable in M. oleifera leaf meal based treatments. The best level of M. oleifera leaf meal inclusion in the diet was 1% in the study. A 1% level of inclusion of M. oleifera leaf meal can be recommended in the diets of broiler and layer Japanese quails for enhancement in quality and consumer acceptance of meat and eggs.