Riverine Buffalo Research Articles

Identification of transcription factor binding sites of <i>HSP60</i> and <i>HSP90B1</i> Genes’ promoter in Bubalus bubalis

The gene expression determines cellular homeostasis while spatio-temporal nature of gene expression is regulated by several cis regulatory elements, such as promoter regions which bind to transcription factors. The ever-changing global climatic conditions have raised a necessity to scan heat tolerant livestock especially the riverine buffalo in tropics. The study focuses on characterization of the promoter region of HSP60 and HSP90B1 and looking for the binding sites of transcription factors in Bubalus bubalis and comparison with the nearest relative Bos taurus to gauge the interspecies variations. The results indicated no transcription binding factors in HSP90B1 promoter (550 nucleotides) within the region analyzed except for the Elk-1 and COMP1 in the cattle while promoter region of HSP60 gene (971 nucleotides) varied highly. Nine and seven transcription factor binding sites were specifically unique in Bubalus bubalis and in Bos taurus respectively. Occurrence of 1-Oct, BR-C Z4, Hairy, and HSF was higher in buffalo. Overall, this study provided the framework to explore cis regulatory elements and contributes to decipher the transcriptional regulatory elements of HSP60 and HSP90B1 genes in Bubalus bubalis.

Detection of Polymorphisms in FASN, DGAT1, and PPARGC1A Genes and Their Association with Milk Yield and Composition Traits in River Buffalo of Bangladesh.

This study aimed to identify SNPs in the intron, exon, and UTR regions of the FASN, DGAT1, and PPARGC1A genes and to investigate their possible association with milk yield and composition traits in the riverine buffalo of Bangladesh. A total of 150 DNA samples from riverine buffalo were used for PCR amplification with five pairs of primers, followed by association studies using a generalized linear model in R. SNP genotyping was performed by direct sequencing of the respective amplicon. Traits analyzed included DMY, fat%, protein%, and SNF%. This study identified 8 SNPs in FASN (g.7163G>A and g.7271C>T), DGAT1 (g.7809C>T and g.8525C>T) and PPARGC1A (g.387642C>T, g.387758A>G, g.409354A>G, and g.409452G>A). Genotypic and allelic frequencies differed significantly for each SNP genotype and did not follow the Hardy-Weinberg principle (p < 0.01 or p < 0.001) in most cases. The g.7163G>A and g.7271C>T SNP genotypes of the FASN gene were significantly associated with milk fat%, with the latter also significantly associated with SNF%. The g.8525C>T polymorphism of the DGAT1 gene significantly affected protein% (p < 0.01). Additionally, PPARGC1A gene polymorphisms showed significant associations: g.387642C>T with fat% (p < 0.05); g.387758A>G and g.409354A>G with protein% (p < 0.001) and SNF% (p < 0.01); and g.409452G>A with DMY (p < 0.001), fat% (p < 0.05), and protein% (p < 0.01). Reconstructed haplotypes of the PPARGC1A gene were significantly associated (p < 0.01) with all traits except SNF%. These findings suggest that polymorphisms in these three candidate genes have the potential as molecular markers for improving milk yield and composition traits in the riverine buffalo of Bangladesh.

Maternal genetic diversity and phylogenetic analysis of Indian riverine and swamp buffaloes: insights from complete mitochondrial genomes.

This study explored the genetic diversity and evolutionary history of riverine and swamp buffaloes in India, utilizing complete mitochondrial genome sequences. Through comprehensive sampling across varied agro-climatic zones, including 91 riverine buffaloes from 12 breeds and 6 non-descript populations, along with 16 swamp buffaloes of the Luit breed, this study employed next-generation sequencing techniques to map the mitogenomic landscape of these subspecies. Sequence alignments were performed with the buffalo mitochondrial reference genome to identify mitochondrial DNA (mtDNA) variations and distinct maternal haplogroups among Indian buffaloes. The results uncovered the existence of 212 variable sites in riverine buffaloes, yielding 67 haplotypes with high haplotype diversity (0.991), and in swamp buffaloes, 194 variable sites resulting in 12 haplotypes, displaying haplotype diversity of 0.950. Phylogenetic analyses elucidated the genetic relationships between Indian buffaloes and the recognized global haplogroups, categorizing Indian swamp buffaloes predominantly into the SA haplogroup. Intriguingly, the haplogroup SB2b was observed for the first time in swamp buffaloes. Conversely, riverine buffaloes conformed to established sub-haplogroups RB1, RB2, and RB3, underscoring the notion of Northwestern India as a pivotal domestication site for riverine buffaloes. The study supports the hypothesis of independent domestication events for riverine and swamp buffaloes, highlighting the critical role of genetic analysis in unraveling the complex evolutionary pathways of domestic animals. This investigation contributes to the global understanding of buffalo mitogenome diversity, offering insights into this important livestock species' domestication and dispersal patterns.

Comparative transcriptome profile of embryos at different developmental stages derived from somatic cell nuclear transfer (SCNT) and in-vitro fertilization (IVF) in riverine buffalo (Bubalus bubalis).

Somatic cell nuclear transfer (SCNT) is a very important reproductive technology with many diverse applications, such as fast multiplication of elite animals, the production of transgenic animals and embryonic stem (ES) cells. However, low cloning efficiency, a low live birth rate and the abnormally high incidence of abnormalities in the offspring born are attributed to incomplete or aberrant nuclear reprogramming. In SCNT embryos, the aberrant expression pattern of the genes throughout embryonic development is responsible for the incomplete nuclear reprogramming. The present study was carried out to identify the differential gene expression (DEGs) profile and molecular pathways of the SCNT and IVF embryos at different developmental stages (2 cell, 8 cell and blastocyst stages). In the present study, 1164 (2 cell), 1004 (8 cell) and 530 (blastocyst stage) DEGs were identified in the SCNT embryos as compared to IVF embryos. In addition, several genes such as ZEB1, GDF1, HSF5, PDE3B, VIM, TNNC, HSD3B1, TAGLN, ITGA4 and AGMAT were affecting the development of SCNT embryos as compared to IVF embryos. Further, Gene Ontology (GO) and molecular pathways analysis suggested, SCNT embryos exhibit variations compared to their IVF counterparts and affected the development of embryos throughout the different developmental stages. Apart from this, q-PCR analysis of the GDF1, TMEM114, and IGSF22 genes were utilized to validate the RNA-seq data. These findings contribute valuable insights about the different genes and molecular pathways underlying SCNT embryo development and offer crucial information for improving SCNT efficiency.

Influence of Ovarian Follicle Sizes and Estrous Signs on Pregnancy Following Progesterone-Based Fixed Time Artificial Insemination in Water Buffaloes

The objectives of the present study were to elucidate the importance of follicle sizes and estrous signs during Controlled Internal Drug Release-Synch-human Chorionic Gonadotropin (CIDR-Synch-hCG) protocol for Fixed Time Artificial Insemination (FTAI) and to evaluate their association with pregnancy in water buffaloes. Data from riverine buffaloes (n = 207) under the CIDR-Synch-hCG protocol were analyzed. Buffaloes were administered with Gonadotropin-Releasing Hormone (GnRH) with insertion of CIDR on Day 0. Prostaglandin (PGF2α) was given on Day 7 with the removal of CIDR. hCG was given on Day 9, and AI was performed on Day 10. Follicle measurements by ultrasonography were done on Days 0, 7, and 10, and follicle sizes on those days were categorized into I, II, and III. Estrus signs were taken on the day of AI. The pregnancy diagnosis was done on Day 30-35 post-AI. The average size of follicles in Category III is significantly higher than those of Categories I and II, regardless of the Days of the protocol. Pregnancy is significantly (P&lt;0.001) associated with Pre-Ovulatory Follicle (POF) size and uterine tonicity on the Day of AI but not with follicle sizes on Days 0 and 7, nor with mucus discharge discharge (P&gt;0.05). The overall pregnancy rate is 44.44% while performing AI with POF size ≥12.0 mm increased the probability of pregnancy rate to 56.25%. In conclusion, the present study demonstrated a follicle size-based CIDR-Synch-hCG protocol providing new fertility indicators to improve FTAI efficiency in buffaloes with huge application in other livestock species.

Unlocking physiological transformations in riverine buffaloes following uterine torsion correction

This study aimed to examine the physiological alterations in riverine buffaloes with incompletely dilated cervix after the uterine torsion correction and the main objective was to understand the hormonal changes that lead to complete cervical dilatation following detorsion, facilitating per-vaginal birth. Torsion characteristics in buffaloes (n=24) were examined, and detorsion was performed using Sharma’s Modified Schaffer’s approach. Blood samples were collected pre-detorsion, post-detorsion, and at parturition. The average cervical dilation rate was 1.22±0.16 cm/h, and detorsion-to-parturition duration averaged 17.38±1.44 h. There were no significant differences between haematological and biochemical parameters at three-time intervals except serum calcium. Progesterone levelsdecreased progressively (1.43±0.01 ng/mL, 1.37±0.01 ng/mL, 1.34±0.01 ng/mL), while estradiol levels increased significantly (65.86±2.05 pg/mL, 77.08±2.05 pg/mL, 91.8±2.55 pg/mL) from pre-detorsion to post-detorsion and parturition. The levels of cortisol were significantly higher pre-detorsion (24.3±2.07 ng/mL) and post-detorsion (27.1±2.5 ng/mL) than at parturition (18.31±1.76 ng/mL). In conclusion, elevated estradiol levels, coupled with reduced progesterone, post-detorsion, trigger complete cervical dilatation, facilitating per-vaginal birth. This study sheds light on the hormonal dynamics driving the birthing process in buffaloes after uterine torsion correction.

Guardians of the Herd: Molecular Surveillance of Tick Vectors Uncovers Theileriosis Perils in Large Ruminants.

Theileria sp. (Piroplasmida: Theileriidae) is one of the most widely known infections transmitted by hard ticks (Acari: Ixodidae) and has been linked to significant economic losses across the globe. The study's main emphasis was theileriosis, a disease that is common in Pakistan and has an incidence ranging from 0.6% to 33%. Through DNA screening of the vector ticks and host blood, this study sought to determine the risk of tick-borne theileriosis in populations of buffalos (Bubalus bubalis) and cattle (Bos indicus) in Toba Tek Singh district of Punjab, Pakistan. Identified tick species include Hyalomma anatolicum (35.4%), Rhipicephalus (Boophilus) microplus (30.2%), and R. sanguineus (25%). Tick specimens were collected from animals and their respective microenvironments. PCR assays targeting Theileria annulata were used to investigate the infection in the DNA extracted from the collected blood samples from large ruminants and salivary glands (SGs) of the Hyalomma ticks. The 18S rRNA of T. annulata was amplified using specific primers. Positive T. annulata amplicons were sequenced and verified using BLAST analysis. Overall, 50% of SGs contained T. annulate DNA. Female ticks, and those collected from cattle and from riverine environments had significantly higher (p < 0.05) rates of Theileria infection in their acini. Overall prevalence of Theileria infection was 35.9% in blood collected from large ruminants. Cattle had a substantially greater frequency of bovine theileriosis (43.2%) than buffalos (28.7%). Age and sex of large ruminants were significantly positively associated (p < 0.05) with Theileria infection. Furthermore, compared to non-riverine cattle (35%) and buffalo (19.5%), riverine cattle (52.2%) and buffalo (36.2%) showed a considerably higher prevalence. The results of this study, which is the first in Pakistan to examine the blood of large ruminants and vectorial function of Ixodid ticks in the transmission of T. annulata along with associated risk factors, offer an important insight for risk assessment of Theileria infection in livestock using vectorial infectivity.

Direct – Maternal Genetic Parameters Estimated for Real-Time Ultrasound Scan Measures of Eye Muscle Area, Rib Fat Depth, and their Correlation with Body Weight in Male Philippine Riverine Buffaloes

Background: Ultrasound scanning is a non-invasive technique for evaluating animals based on carcass yield and meat quality traits. It has been incorporated into the beef cattle breeding program. However, little has been done in buffaloes. Thus, this study was conducted to estimate the genetic parameters for growth traits- body weights (BW) and a real-time ultrasound scan of eye muscle area (EMA) and rib fat (RF) to determine their suitability as a tool for selection.&#x0D; Methods: Four hundred thirty-eight (438) male Philippine riverine buffaloes with 1,535 BW records and 417 real-time ultrasound scans of EMA and RF scans at the 12th rib were available for the study. BW was taken and adjusted at birth, 6, 12, 18, 24, 30, and 36 months while RF and EMA scans were taken at 12 &amp; 18 mos. Genetic parameters were estimated with a pedigree-based multi-trait animal model using ASREML software.&#x0D; Results: The average BW of buffaloes adjusted to 6, 12, 18, 24, 30, and 36 months of age were 135kg, 230.8kg, 323.0kg, 382.7, 453.6, and 502.4kg, respectively. The mean EMA and RF at 12 months were 29.7 cm2 and 3.2mm, respectively. Direct genetic heritabilities were 0.26±0.16, 0.29±0.12, 0.35±0.19 and 0.67±0.32 for birth, BW12mos, EMA12mos and RF12mos, respectively. Maternal genetic heritability for birth weight. was 0.12±0.09. Genetic correlations among traits were positive except between birth and RF.&#x0D; Conclusions: Results indicate that ultrasound scanning can be used for selection together with BW at 12mos. and suggest that early performance is an excellent predictor of performance at a later age.

Molecular Data and Karyotype Revealed Two Distinct Species of Domesticated Water Buffaloes in the Philippines

The six species of the genus Bubalus include the lowland and mountain anoa, Asian wild buffalo, tamaraw, swamp buffalo, and riverine buffalo. Few studies were conducted on the matrilineal phylogeny and genetic diversity of Bubalus species. In addition, there is controversy regarding the identity of the two domesticated water buffaloes in the Philippines – the introduced riverine and the native swamp buffalo. Thus, this study aimed to determine the genetic distance and phylogenetic relationship of Bubalus species using the mitochondrial cytochrome b (cytb) variation and to re-evaluate the taxonomic species designation of Bubalus bubalis. A total of499 Philippine cytb sequences from this study (n = 179), as well as other Asian countries (n = 319)and Syncerus caffer (n = 1), from NCBI were included in the analysis. The highest interspecies genetic distance (d) (0.040) was between the tamaraw and lowland anoa, whereas the lowest genetic distance (0.020) was between mountain and lowland anoa. Moreover, the phylogenetic tree indicated 52 haplotypes that delineated three groups, including lowland anoa, mountain anoa, and tamaraw with two domesticated buffaloes. The latter group further separated into three species – tamaraw, riverine, and swamp buffaloes. The analyses of genetic distance,phylogeny, and median-joining network structure based on cytb confirmed that the swamp buffaloes from Occidental Mindoro and South Cotabato, Philippines belong to maternal lineage B. Karyotype analysis consistently confirms that riverine buffaloes possess 50 chromosomes, whereas swamp buffaloes have 48 chromosomes, aligning with previous findings. Moreover, the swamp and riverine buffaloes appear to be distinct species, with the swamp buffalo having a closer affinity to the tamaraw than to the riverine buffalo. We, therefore, propose to revive the species assignment of the Bubalus kerabau Fitzinger, 1860 as a distinct species to the swamp buffalo. Furthermore, a common maternal ancestry was confirmed between the tamaraw and two domesticated buffaloes – the swamp and riverine. Research findings provided valuable information on understanding and monitoring the matrilineal genetic diversity of the genus Bubalus toward conservation and management.

A comparative study on effect of heat stress on physiological and cellular responses of crossbred cattle and riverine buffalo in subtropical region of India.

The study was carried out to compare the in vitro and in vivo heat shock responses of cattle and buffaloes. The expression of heat responsive genes (HSP70 and HSF family) were studied in vitro in peripheral blood mononuclear cells (PBMCs) of cattle and buffalo. In vivo observations on animals were carried out to investigate the physiological responses of cattle and buffalo at different THI over a period of 14months. The study indicated that onset and severity of heat stress at different THI varied significantly between cattle and buffalo. Rectal temperature (RT) showed a significant (p < 0.05) increase at THI 67 in buffaloes and at THI 68 in cattle. Significant (p < 0.01) differences in RT between the species were observed at THI 71, 72, and 73. Respiration rate (RR) significantly (p < 0.05) increased at THI 70 in both the species and significant (p < 0.05) differences in RR were observed between the species at THI 65, 68, 69, and 74. THI had significant (p < 0.05) effect on blood glucose and blood electrolytes of the species with increased levels at higher THI. Serum AST and ALT levels showed less pronounced changes over increasing THI. Heat stress-associated expressions of HSP 70 genes followed temporal changes with incremental THI. The expression of HSPA8 was consistent at lower THI whereas upregulation of HSPA1A and HSPA1L was evident at higher THI. The study concludes that changes in physiological parameters such as RT and RR occur in a phasic pattern in both species and onset of heat stress was early in buffalo as compared to cattle.

Expression of heat shock and apoptosis genes in riverine buffalo (Bubalus bubalis) cumulus–oocyte complexes during in-vitro maturation under thermal stress conditions

Expression of heat shock and apoptosis genes in riverine buffalo (Bubalus bubalis) cumulus–oocyte complexes during in-vitro maturation under thermal stress conditions

Detection of genome-wide copy number variation in Murrah buffaloes

Riverine Buffaloes, especially the Murrah breed because of their adaptability to harsh climatic conditions, is farmed in many countries to convert low-quality feed into valuable dairy products and meat. Here, we investigated the copy number variations (CNVs) in 296 Murrah buffalo using the Axiom® Buffalo Genotyping Array 90K (Affymetrix, Santa Clara, CA, USA). The CNVs were detected on the autosomes, using the Copy Number Analysis Module (CNAM) using the univariate analysis. 7937 CNVs were detected in 279 Buffaloes, the average length of the CNVs was 119,048.87 bp that ranged between 7800 and 4,561,030 bp. These CNVs were accounting for 10.33% of the buffalo genome, which was comparable to cattle, sheep, and goat CNV analyses. Further, CNVs were merged and 1541 CNVRs were detected using the Bedtools-mergeBed command. 485 genes were annotated within 196 CNVRs that were identified in at least 10 animals of Murrah population. Out of these, 40 CNVRs contained 59 different genes that were associated with 69 different traits. Overall, the study identified a significant number of CNVs and CNVRs in the Murrah breed of buffalo, with a wide range of lengths and frequencies across the autosomes. The identified CNVRs contained genes associated with important traits related to production and reproduction, making them potentially important targets for future breeding and genetic improvement efforts.

Impact of Cysteine as a Semen Additive on Mitigating the Decline in Sperm Quality due to Summer Stress in Riverine Buffaloes (Bubalus bubalis)

The seasonal variations such as temperature, rain and humid environment greatly influences reproduction, and semen quality particularly in buffalo bulls, either positively or negatively. The heat stress coupled with freezing and thawing processes generates reactive oxygen species (ROS), which further reduce post-thaw spermatozoa quality. Application of cysteine as a semen additive have been reported to alleviate such damaging effect in canine, buck, bull, human, boar, and rooster. Therefore, objective of this study was to investigate ameliorating effect of cysteine on summer stress on buffalo spermatozoa. For the experiment, 24 ejaculates with individual progressive motility &gt;70, volume &gt;1mL and sperm concentration 1000 million/mL from four Murrah buffalo bulls were collected and divided into four groups as Control (Group-I), and Group-II, III, and IV (4, 8, and 12 mmol cysteine, respectively) containing 80 million spermatozoa per mL. Semen was filled (French mini straw), sealed and equilibrated at 40⸰C and frozen into LN2. The group containing L-cysteine at 4 mmol concentration had higher significant (p&lt;0.05) difference in post thaw motility, viability, plasma membrane integrity and acrosome integrity, TAC, DNA integrity and lower significant (p&lt;0.05) difference in spermatozoa abnormality, LPO and intracellular ROS. Moreover, cysteine concentration at 8 mmol has no beneficial effect compared to the Control, whereas Group-IV had greater adverse effect. This may be due to its toxicity at this concentration. It is evident from the study that the cysteine molecules at 4 mmol concentration produced more pronounced improvement in the freezability potential of buffalo bull spermatozoa.

Genetic Diversity Relationship in Azakheli Buffalo Inferred from mtDNA and MC1R Sequences Comparison.

Azakheli is relatively smaller riverine breed with a very peculiar characteristics kept under unique traditional husbandry practices in comparison with rest of the Pakistani buffalo breeds; however, milk production is comparable. The present study was conducted to evaluate the genetic diversity of Azakheli breed. A total of sixty-six blood samples were collected for the amplification of mtDNA D-loop region and MC1R gene sequencing analysis. Median-joining network analysis of 191 mtDNA D-loop sequences of Azakheli and eight Indian riverine buffalo breeds clustered into three clades. Ancient Azakheli Region 1 clade was the oldest with the highest mutation steps and was present close to the root of UPGMA phylogenetic tree. There was 5 mutated lines distance between Pakistan buffalo and Indian riverine buffaloes. The populations of neighboring countries did not share any haplotypes with Azakheli buffalo of Pakistan. Possibly, residing for so long in the cold atmosphere and high elevation regions caused the mutation in mtDNA D-loop, though these conditions did not affect the overall performance of Azakheli as milch buffalo breed of Pakistan. MC1R analyses showed high mutations in Azakheli of Albino phenotype and all the black phenotype individuals of Azakheli buffalo share haplotypes with dominant Chinese and Indian black phenotypes buffaloes in MC1R median-joining network, indicating the reason of black coat color is due to MC1R gene. The haplotype diversity and nucleotide diversity was (H. 0.923, Pi: 0.00895) in Azakheli. Current results illustrated Asian ancestry for Azakheli buffalo, and mtDNA and MC1R analyses provided further evidence. Additional genetic analyses and archeological studies may provide further insight into the domestication period and history of Azakheli buffalo breed. The further studies are required on different coat colors with different genes on Azakheli buffalo to understand the phenotype variation.

Signatures of selection in riverine buffalo populations revealed by genome-wide SNP data

The detection of selection signatures assists in understanding domestication, evolution, and the identification of genomic regions related to adaptation and production traits in buffaloes. The emergence of high-throughput technologies like Next Generation Sequencing and SNP genotyping had expanded our ability to detect these signatures of selection. In this study, we sought to identify signatures of selection in five buffalo populations (Brazilian Murrah, Bulgarian Murrah, Indian Murrah, Nili-Ravi, and Kundi) using Axiom Buffalo 90 K Genotyping Array data. Using seven different methodologies (Tajima’s D, CLR, ROH, iHS, FST, FLK and hapFLK), we identified selection signatures in 374 genomic regions, spanning a total of 381 genes and 350 quantitative trait loci (QTLs). Among these, several candidate genes were associated with QTLs for milk production, reproduction, growth and carcass traits. The genes and QTLs reported in this study provide insight into selection signals shaping the genome of buffalo breeds. Our findings can aid in further genomic association studies, genomic prediction, and the implementation of breeding programmes in Indian buffaloes.

Performance of Illumina® BovineHD BeadChip in genotyping Indian riverine buffalo breeds

The current study tested the suitability of Illumina® BovineHD BeadChip in genotyping three riverine breeds of buffalo viz. Jaffarabadi, Murrah and Mehsana. Of the 777962 SNPs, 670955 SNPs were called in all animals and only 13150 SNPs were found to be polymorphic at a call rate of 90%, and had Minor Allele Frequency (MAF) greater than 0.05. The average Minor Allele Frequency (MAF) of polymorphic SNPs was 0.23 and average expected heterozygosity (HE) of 0.321. Principal Component Analysis (PCA) revealed 2 different clusters where Murrah and Mehsana breeds clustered together, while Jaffarabadi breed formed a distinct cluster. Result obtained in this study indicated that the SNPs available in Illumina BovineHD BeadChip may not be good enough for studying population structure and genetic analyses in Indian buffalo breeds. Hence it might be necessary to develop a custom SNP marker panel for Indian riverine buffaloes.

Identification of novel polymorphism in mammary-derived growth inhibitor gene of water buffalo and its expression analysis in the mammary gland

Mammary-derived growth inhibitor (MDGI), a member of the lipophilic family of fatty acid-binding proteins, plays an important role in the development, regulation, and differentiation of the mammary gland. The aim of the study was to identify polymorphism in the MDGI gene and its expression analysis in the mammary gland at various stages of lactation, in Indian buffalo. Nucleotide sequence analysis of MDGI gene in different breeds of riverine and swamp buffaloes revealed a total of 16 polymorphic sites and one Indel. Different transcription factor binding sites were predicted for buffalo MDGI gene promoter sequence, using online tools and in-silico analysis indicating that the SNPs in this region can impact the gene expression regulation. Phylogenetic analysis exhibited the MDGI of buffalo being closer to other ruminants like cattle, yak, sheep, and goats. Further, the expression analysis revealed that buffalo MDGI being highly expressed in well-developed mammary glands of lactating buffalo as compared to involution/non-lactating and before functional development to start the milk production stage in heifers. Stage-specific variation in expression levels signifies the important functional role of the MDGI gene in mammary gland development and milk production in buffalo, an important dairy species in Southeast Asia.

Antibiogram and virulence profiling reveals multidrug resistant Staphylococcus aureus as the predominant aetiology of subclinical mastitis in riverine buffaloes.

Staphylococcus spp. are the major causal agents of mastitis in dairy animals worldwide leading to profound economic losses and public health threats. Recently, Staphylococcus aureus has emerged as a multidrug resistant and zoonotic pathogen. This study aimed to characterize S. aureus in subclinical mastitis (SCM) milk samples of riverine buffaloes in Bangladesh through antibiogram and virulence gene(s) profiling, and 16S rRNA gene sequencing. We characterized S. aureus in SCM milk samples (N = 500) of riverine buffaloes through antibiogram and virulence gene(s) profiling, and 16S rRNA gene sequencing. Out of 500 milk samples tested, 188 (37.6%) were found positive for SCM. From 188 SCM samples, 291 isolates were obtained with a prevalence of S. aureus in 37.4% (109/291) isolates. Phylogenetic analysis revealed the evolutionary divergence of S. aureus isolates in bubaline SCM milk samples. The antibiogram profiling showed that about 96.0% S. aureus isolates were multidrug resistant (MDR). Notably, 29 and 16 isolates harboured methicillin-resistant (mecA) and panton-valentine leucocidin (pvl) genes, respectively, and 46 plasmid-bearing isolates were MDR. Nine Staphylococcal enterotoxins (SEs/SEls) including sea (11.9%), sec (7.4%), sed (4.6%), seg (3.7%), and seh (3.7%) were detected with 72.48% toxinotypes comprising a single gene. This study therefore suggests S. aureus as the single-most aetiology (∼37.0%) of SCM in riverine buffaloes, and emergence of MDR, enterotoxin producing, and virulent S. aureus strains could impose potential threats to animal welfare and public health.

Genomic analysis of arginine vasopressin gene in riverine buffalo reveals its potential association with silent estrus behavior.

Poor estrus expression behavior causes suboptimal reproductive efficiency through poor conception rate. Various signaling pathways are involved in estrus expression but arginine vasopressin (AVP) gene with oxytocin predominantly regulates estrus behavior. This study aimed to perform genomic characterization and evolutionary dynamics of AVP gene through association testing of the novel polymorphic loci and comparative genomic analysis to explore the potential effect of AVP gene on estrus behavior of Nili-Ravi buffaloes. 198 Nili-Ravi buffaloes were screened for the quest of novel polymorphism in the AVP gene. In exon-1, five polymorphic sites were detected including deletion of two (c.47delA and c.57delA) nucleotides that caused drastic variation in subsequent amino acid sequence due to frame shift including functional short peptide of nine residues. The 3-D structure revealed a loss of transmembrane loop between 16 and 31 residues in Nili-Ravi buffalo AVP protein sequence, suggesting that missing loop apparently reduced the gene functionality in Nili-Ravi buffalo by inhibiting cellular reactions and muting the animal estrus cyclicity. Three polymorphisms detected in AVP gene were significantly associated with silent estrus (P <0.05). The comparative genomic analysis revealed that AVP gene is present on chromosome 14 having one conserved motif (Neurohypophysial) in buffalo. This study suggested the potential use of polymorphic sites as promising genetic markers for selection of buffaloes with pronounced estrus expression.

Colostrum feeding induced alkaline indigestion syndrome in riverine buffaloes- clinical assessment and therapeutic management

The present study was undertaken to elucidate the alkaline indigestion syndrome in riverine buffaloes caused by inadvertent colostrum feeding, clinical pattern and its therapeutic management. Fifteen recently parturited buffaloes with history of feeding average 2.96 kg (1 to 5 kg) of colostrum immediately after parturition followed by sudden onset of anorexia, decrease in milk yield and nervous signs were investigated. Clinical examination showed normal body temperature (100.26±0.42 vs 100.62±0.18), respiration (23.73±3.53 vs 24.13±0.66) and heart rate (57.26±3.30 vs 53.20±1.79) compared to healthy buffaloes. Examination of rumen revealed impaction with highly significant (P&lt;0.01) reduction in rumen motility (0.80±0.24 vs 3.80±0.17 per five minutes). Similarly, highly significant (P&lt;0.01) increase in rumen pH (8.10±0.16 vs 6.25±0.07) and decrease in protozoal density (5.00±0.88 vs 32.93±1.72) with sluggish (+) to no (-) protozoal motility was observed in ailing buffaloes. Haematological analysis showed normal blood profile with significant (P&lt;0.05) increase only in neutrophil count (51.60±5.1 vs 42.06±3.4). Prominent clinical signs of anorexia, congested conjunctival mucosae, decreased milk yield and varied nervous signs like restlessness, head pressing, staggering gait, incoordination, circling, convulsions, dummy syndrome, apparent blindness and coma were observed in affected buffaloes. Postural abnormalities like sternal / lateral recumbency and lateral deviation of neck were also observed in two buffaloes. All the ailing buffaloes were treated with Dextrose Normal Saline, single dose of preparation containing calcium, phosphorus and magnesium, vitamin B complex, antihistaminic, sedatives, laxatives, pre, probiotics, and oral antibiotics. All the treated buffaloes showed satisfactory improvement from 1st to 3rd day with complete clinical recovery by 4.93 (3 to 9) days of treatment. Buffalo owners should be made aware of unscientific practice of colostrum feeding in adult animals and accidentally intoxicated buffaloes could be successfully treated with the standardized treatment protocol.