Functional transcriptome analysis revealed upregulation of MAPK-SMAD signalling pathways in chronic heat stress in crossbred cattle.

Animal geneticists and breeders have the impending challenge of enhancing the resilience of Indian livestock to heat stress through better selection strategies. Climate change's impact on livestock is more intense in tropical countries like India where dairy cattle crossbreeds are more sensitive to heat stress. The main reason for this study was to find the missing relative changes in transcript levels in thermo-neutral and heat stress conditions in crossbred cattle through whole-transcriptome analysis of RNA-Seq data. Differentially expressed genes (DEGs) identified based on the minimum log twofold change value and false discovery rate 0.05 revealed 468 up-regulated genes and 2273 down-regulated significant genes. Functional annotation and pathway analysis of these significant DEGs were compared based on Gene Ontology (Biological process), Kyoto Encyclopedia of Genes and Genome (KEGG), and Reactome pathways using g: Profiler, ShinyGO v0.76, and iDEP.951 web tools. On finding network visualization, the most over-represented and correlated pathways were neuronal and sensory organ development, calcium signalling pathway, Mitogen-activated protein kinase (MAPK) and Smad signalling pathway, Ras-proximate-1, or Ras-related protein 1 (Rap 1) signalling pathway, apoptosis, and oxidative stress. Similarly, down-regulated genes were most expressed in mRNA processing, immune system, B-cell receptor signalling pathway, Nucleotide oligomerization domain (NOD)-like receptors (NLRs) signalling pathway and nonsense-mediated decay (NMD) pathway. The heat stress-responsive genes identified in this study will facilitate our understanding of the molecular basis for climate resilience and heat tolerance in Indian dairy crossbreeds.

Weighted single step GWAS reveals genomic regions associated with economic traits in Murrah buffaloes.

The objective of the present study was to identify genomic regions influencing economic traits in Murrah buffaloes using weighted single step Genome Wide Association Analysis (WssGWAS). Data on 2000 animals, out of which 120 were genotyped using a double digest Restriction site Associated DNA (ddRAD) sequencing approach. The phenotypic data were collected from NDRI, India, on growth traits, viz., body weight at 6M (month), 12M, 18M and 24M, production traits like 305D (day) milk yield, lactation length (LL) and dry period (DP) and reproduction traits like age at first calving (AFC), calving interval (CI) and first service period (FSP). The biallelic genotypic data consisted of 49353 markers post-quality check. The heritability estimates were moderate to high, low to moderate, low for growth, production, reproduction traits, respectively. Important genomic regions explaining more than 0.5% of the total additive genetic variance explained by 30 adjacent SNPs were selected for further analysis of candidate genes. In this study, 105 genomic regions were associated with growth, 35 genomic regions with production and 42 window regions with reproduction traits. Different candidate genes were identified in these genomic regions, of which important are OSBPL8, NAP1L1 for growth, CNTNAP2 for production and ILDR2, TADA1 and POGK for reproduction traits.

Discriminant analysis reveals differences in breed structure in river buffaloes of Maharashtra (India).

Buffaloes are a major milch species of India, and owning a true-to-breed buffalo herd is a reverie for livestock keepers, as it substantiates their incomes. Multivariate discriminate analysis is an efficient methodology to classify different farm animal populations based on canonical discriminant function. The present study was conducted on 392 adult buffaloes using 18 body biometric traits. Except for body height and body length, all the body characters revealed significant differences amongst the four buffalo populations. Unlike cattle, distance between horns and rump width were more discriminating factors, compared to body height, leg length, chest girth, and body length. We report the most suitable discriminate function which can discriminate with maximum between-group variation in four studied populations. Mahalanobis distances were calculated between populations and they ranged from 0.786 to 1.227. Purnathadi has revealed a distinct phenotypic structure, and these buffaloes have a distinct native breeding region comprising Akola, Amravati, and Buldhana districts of Maharashtra state. On the other hand, Elachpuri animals failed to depict physical and biometric discriminations from Nagpuri, Marathwadi, and Purnathadi buffalo populations, and Elachpuri can be considered as a strain of Nagpuri buffalo.

Modelling the lactation curve in Alpine × Beetal crossbred dairy goats using random regression models fitted with Legendre polynomial and B-spline functions.

The current study sought to genetically assess the lactation curve of Alpine × Beetal crossbred goats through the application of random regression models (RRM). The objective was to estimate genetic parameters of the first lactation test-day milk yield (TDMY) for devising a practical breeding strategy within the nucleus breeding programme. In order to model variations in lactation curves, 25,998 TDMY records were used in this study. For the purpose of estimating genetic parameters, orthogonal Legendre polynomials (LEG) and B-splines (BS) were examined in order to generate suitable and parsimonious models. A single-trait RRM technique was used for the analysis. The average first lactation TDMY was 1.22 ± 0.03 kg and peak yield (1.35 ± 0.02 kg) was achieved around the 7th test day (TD). The present investigation has demonstrated the superiority of the B-spline model for the genetic evaluation of Alpine × Beetal dairy goats. The optimal random regression model was identified as a quadratic B-spline function, characterized by six knots to represent the central trend. This model effectively captured the patterns of additive genetic influences, animal-specific permanent environmental effects (c2 ) and 22 distinct classes of (heterogeneous) residual variance. Additive variances and heritability (h2 ) estimates were lower in the early lactation, however, moderate across most parts of the lactation studied, ranging from 0.09 ± 0.04 to 0.33 ± 0.06. The moderate heritability estimates indicate the potential for selection using favourable combinations of test days throughout the lactation period. It was also observed that a high proportion of total variance was attributed to the animal's permanent environment. Positive genetic correlations were observed for adjacent TDMY values, while the correlations became less pronounced for more distant TDMY values. Considering better fitting of the lactation curve, the use of B-spline functions for genetic evaluation of Alpine × Beetal goats using RRM is recommended.

Diversity assessment of a lesser known buffalo population from Central India and its comparative evaluation reveals presence of sufficient genetic variation and absence of selection.

Planned breeding and conservation strategies for a lesser-known population require an assessment of complete genetic diversity and population structure analysis in addition to its morphometric characteristics. In the present study, a comparative analysis of the genetic structure of a rare buffalo population, namely Chhattisgarhi, was extensively studied using a panel of FAO-recommended microsatellite markers along with well-established breeds namely Murrah, Nili-Ravi, Gojri, Kalahandi, and Nagpuri. Mode shift analysis indicated the absence of genetic bottleneck in the recent past. Assessment of genetic diversity indices across all loci indicated the presence of sufficient genetic variation within and between populations. Analysis of molecular variance between the six different buffalo populations attributed 19.05% of the variations to between-population differentiation. Cluster analyses using DAPC and Bayesian approach along with the phylogenetic tree based on UPGMA grouped six populations into three groups. The Chhattisgarhi population was revealed to be genetically closer to Nagpuri and Kalahandi populations. The study reveals the presence of sufficient genetic diversity within the Chhattisgarhi population and indicates the absence of a systematic selection program. We suggest improvement and conservation programs should be planned for this breed in the near future through short-term selection.

Exome-wide comparative analyses revealed differentiating genomic regions for performance traits in Indian native buffaloes.

In India, 20 breeds of buffalo have been identified and registered, yet limited studies have been conducted to explore the performance potential of these breeds, especially in the Indian native breeds. This study is a maiden attempt to delineate the important variants and unique genes through exome sequencing for milk yield, milk composition, fertility, and adaptation traits in Indian local breeds of buffalo. In the present study, whole exome sequencing was performed on Chhattisgarhi (n = 3), Chilika (n = 4), Gojri (n = 3), and Murrah (n = 4) buffalo breeds and after stringent quality control, 4333, 6829, 4130, and 4854 InDels were revealed, respectively. Exome-wide FST along 100-kb sliding windows detected 27, 98, 38, and 35 outlier windows in Chhattisgarhi, Chilika, Gojri, and Murrah, respectively. The comparative exome analysis of InDels and subsequent gene ontology revealed unique breed specific genes for milk yield (CAMSAP3), milk composition (CLCN1, NUDT3), fertility (PTGER3) and adaptation (KCNA3, TH) traits. Study provides insight into mechanism of how these breeds have evolved under natural selection, the impact of these events on their respective genomes, and their importance in maintaining purity of these breeds for the traits under study. Additionally, this result will underwrite to the genetic acquaintance of these breeds for breeding application, and in understanding of evolution of these Indian local breeds.

Optimizing selection strategy for enhancing reproduction efficiency in Indian crossbred goats using milk productivity as a selection criterion.

The present study was aimed at optimizing the selection strategy for enhancing reproductive efficiency and milk productivity of Alpine × Beetal crossbred goats. The data set included 2949 milk trait records across parities and 1389 milk records from first parity and corresponding reproductive traits. The traits included for analysis were 150-day milk yield (150DMY), days in milk (DIM), peak yield (PY) and total milk yield (TMY). The litter size (LS) and litter weight (LW) were used for specifically formulating selection plan using indirect selection. The least squares mean for lactation traits during the first parity were 150DMY: 195.32 ± 2.09 kg, DIM: 236.42 ± 3.04 days, PY: 1.82 ± 0.02 kg, TMY: 269.62 ± 4.52 kg. Notably, Alpine × Beetal goats demonstrated genetic superiority pan India for milk productivity as compared to other native goat breeds. The least squares mean for 150DMY across all parities was 236 ± 3.13 kg. An animal model employing average information restricted maximum likelihood was used for (co)variance component estimation to get the genetic parameters. The analysis revealed total heritability estimates for 150DMY, DIM, PY and TMY as 0.18 ± 0.06, 0.04 ± 0.04, 0.12 ± 0.06 and 0.08 ± 0.05, respectively. Repeatability estimates for 150DMY, DIM, and TMY were 0.28 ± 0.04, 0.21 ± 0.03 and 0.37 ± 0.03, respectively. Bivariate analysis of 150DMY with reproductive traits revealed heritability for LS and LW as 0.05 ± 0.01 and 0.10 ± 0.01, respectively using Gibbs sampling. Strong and positive genetic correlations of 150DMY with other production and reproduction traits was observed, such as DIM (0.72), PY (0.98), TMY (0.88), LS (0.57) and LW (0.33). Moderate heritability and repeatability estimate of 150DMY, along with its positive correlation with production and reproductive traits suggested it as a suitable selection criterion for early selection and overall genetic progress of lactation traits. The genetic trend analysis showed an overall improvement in all these traits, with observed gain of 98.4 g per year for 150DMY, 0.04 days per year for DIM, 0.5 g per year for PY and 220.5 g per year for TMY. We observed that selecting based on 150DMY would lead to a favourable indirect improvement for LW as 79 g and LS 0.04 units per generation. We, therefore, recommend employing 150DMY as the single trait selection criteria to enhance both milk productivity and reproductive potential of Alpine × Beetal goats.

TWAS revealed significant causal loci for milk production and its composition in Murrah buffaloes.

Milk yield is the most complex trait in dairy animals, and mapping all causal variants even with smallest effect sizes has been difficult with the genome-wide association study (GWAS) sample sizes available in geographical regions with small livestock holdings such as Indian sub-continent. However, Transcriptome-wide association studies (TWAS) could serve as an alternate for fine mapping of expression quantitative trait loci (eQTLs). This is a maiden attempt to identify milk production and its composition related genes using TWAS in Murrah buffaloes (Bubalus bubalis). TWAS was conducted on a test (N = 136) set of Murrah buffaloes genotyped through ddRAD sequencing. Their gene expression level was predicted using reference (N = 8) animals having both genotype and mammary epithelial cell (MEC) transcriptome information. Gene expression prediction was performed using Elastic-Net and Dirichlet Process Regression (DPR) model with fivefold cross-validation and without any cross-validation. DPR model without cross-validation predicted 80.92% of the total genes in the test group of Murrah buffaloes which was highest compared to other methods. TWAS in test individuals based on predicted gene expression, identified a significant association of one unique gene for Fat%, and two for SNF% at Bonferroni corrected threshold. The false discovery rates (FDR) corrected P-values of the top ten SNPs identified through GWAS were comparatively higher than TWAS. Gene ontology of TWAS-identified genes was performed to understand the function of these genes, it was revealed that milk production and composition genes were mainly involved in Relaxin, AMPK, and JAK-STAT signaling pathway, along with CCRI, and several key metabolic processes. The present study indicates that TWAS offers a lower false discovery rate and higher significant hits than GWAS for milk production and its composition traits. Hence, it is concluded that TWAS can be effectively used to identify genes and cis-SNPs in a population, which can be used for fabricating a low-density genomic chip for predicting milk production in Murrah buffaloes.

Explicating the genetic diversity and population structure of Saanen × Beetal goats using pedigree analysis.

Pedigree analysis is required to assess the genetic diversity and population structure of a close breeding population in order to effectively manage the breeding program and keep inbreeding under acceptable limits. Saanen × Beetal is a crossbred population of goats, reared at the National Dairy Research Institute (NDRI), Karnal, for the last five decades. This germplasm has been acclimated to a tropical climate and has a higher milk potential and prolificacy. The objective of this study was to elucidate the genetic diversity, population structure, and inbreeding in the flock of the Saanen × Beetal goats. The data were collected from the Animal Genetics and Breeding Division of ICAR-NDRI, Karnal, for 2603 animals from the year 1971 to 2021. Animals born between 2014 and 2017 were considered as a reference cohort. Results revealed that the average generation interval was 3.44 years for the complete pedigree. The average inbreeding coefficient and the average relatedness were 4.20% and 6.87%, respectively, for the complete pedigree and 10.78% and 10.80% for the reference population. Higher inbreeding coefficient and average relatedness in the reference cohort demonstrated the impact of the enclosed gene pool and demands immediate intervention for managing diversity in the closed nucleus under study. Ancestors contributing 50% of the gene pool were 8 and 3 for the complete pedigree and reference cohort, respectively, which illustrates the fact that very few ancestors were responsible for genetic diversity in the flock, which results in the decline of effective population size. Effective numbers of founders (fe), ancestors (fa), and founder genome equivalents (fg) were 15, 7, and 3.11, respectively. The (fe/fa) ratio in the reference population was 2.14, indicating the occurrence of the bottleneck effect in the flock. We observed that inbreeding was non-significant for all reproductive traits except for age at first service and age at first kidding. To lessen inbreeding and augment genetic diversity in the flock, the stratified breeding plan needs to be followed, where mate selection would be based on relatedness. Furthermore, the introduction of unrelated Saanen and Beetal crosses will help alleviate the inbreeding accumulation.

Estimation of genetic parameters and genetic change of first parity reproductive traits in Alpine × Beetal goats.

The study of reproductive traits is crucial for improving genetic potential of goats because of their significant utility in meat production. Hence, genetic analysis was conducted for reproductive traits on Alpine × Beetal goats using animal model for first parity data. Information on 1462 reproductive records were collected over five decades from ICAR-National Dairy Research Institute, Karnal, Haryana (1971-2021). Single-trait and multi-trait animal models were used for genetic analysis. Estimates of (co)variance components and genetic parameters were obtained using Gibbs Sampler for Animal Model due to non-normal distribution of data. Six single-trait animal models (including or excluding maternal and environmental effects) were fitted and best models were determined based on Deviance Convergence Criterion values. The prolificacy for the A × B goats for first parity data was 32%, having 68% single births, 31% twins and 1% triplets/quadruplets. The least squares mean for age at first service (AFS), age at first kidding (AFK), service period (SP), dry period (DP), gestation length (GL), kidding interval (KI), litter weight (LW), number of kids born (NKB) and number of females kids born (NFKB) in first parity were 546.15 ± 4.10 days, 679.05 ± 4.07 days, 226.51 ± 4.02 days, 67.96 ± 2.76 days, 150.74 ± 0.13 days, 362.53 ± 3.35 days, 3.99 ± 0.04 kg, 1.32 ± 0.02 and 0.64 ± 0.02, respectively. The heritability estimates obtained from best model for AFS, AFK, GL, KI, SP, and DP were 0.12 ± 0.00, 0.10 ± 0.00, 0.09 ± 0.01, 0.03 ± 0.00, 0.04 ± 0.00, and 0.05 ± 0.00, respectively. For NKB, NFKB and LW, heritability estimates were 0.16 ± 0.01, 0.03 ± 0.03, and 0.04 ± 0.00, respectively. These results imply lower heritability estimates for reproductive traits, and hence meagre scope for selection for further improvement. Maternal effects contributed significantly for traits such as GL, NKB and NFKB. Genetic correlation for number of female kids born was negative with SP and DP which is favourable. Furthermore, genetic correlation was negative for dry period and litter weight which is favourable as number of kids born and litter weight are traits of direct economic importance. Results reveal high genetic potential of this breed for meat industry owing to high prolificacy, provided consistent efforts are made for the genetic improvement of this germplasm.

Genetic improvement of economic traits in Murrah buffalo using significant SNPs from genome-wide association study.

GWAS helps to identify QTL and candidate genes of specific traits. Buffalo breeding has primarily focused on milk production, but its negative correlation with reproduction traits resulted in unfavorable decline of reproductive performance among buffaloes. A genome wide scan was performed on a total of 120 Murrah buffaloes genotyped by ddRAD sequencing for 13 traits related to female fertility, production, and growth. The identified 25 significant single nucleotide polymorphisms (SNPs) (P <1×106) are associated with age at first calving (AFC), age at first service (AFS), period from calving to 1st Artifical Insemination (AI), service period (SP) and 6 month body weight (6M). Fifteen genetic variants overlapped with different QTL regions of reported studies. Among the associated loci, outstanding candidate genes for fertility, including AQP1, TRNAE-CUC, NRIP1, CPNE4, and VOPP1, have effect in different fertility traits. AQP1 gene is expressed in ovulatory phase and various stages of pregnancy. TRNAE-CUC gene is associated with AFC and number . of calvings after 4 years of age. Glycogen content-associated gene CPNE4 regulates muscle glycogen and is upregulated during early pregnancy. NRIP1 generegulates ovulation, corpus luteum at pregnancy, and mammary gland development. The objective is to identify potential genomic regions and genetic variants associated with economic traits and to select the most significant SNP which have positive effect on all the traits.

ASIP gene polymorphism associated with black coat and skin color in Murrah buffalo.

The melanogenesis pathway regulates pigmentation through the synergic action of various genes. We are interested in analyzing the genetic variations in the ASIP which determine eumelanin production in the dermis layer. In the present study, the ASIP gene was characterized in buffalo and 268 genetically unrelated buffaloes belonging to 10 different populations were genotyped for the non-synonymous SNP (c.292C>T) identified in the exon 3 region of the gene using Tetra-ARMS-PCR. The TT genotype occurred at a higher rate in Murrah, followed by Nili Ravi, Tripura, and Paralakhemundi (42.63%, 19.30%, 3.45%, and 3.33%). These results convey the association of the black coat color of Murrah with the ASIP gene TT genotype and the lighter shades of black coat (brown and grayish-black) color phenotype in other breeds with the CC genotype.

Comparative analysis of various methods for estimating expected genetic gain of production and reproduction traits in Murrah buffaloes.

The present study was conducted to assess the expected genetic gain for first lactation production and reproduction traits in Murrah buffaloes, in addition to optimization of progenies/sire. Data for period 1971-2020 were used from National Dairy Research Institute. Performance traits considered were 305 days milk yield (305DMY), average daily milk yield (ADMY), peak yield (PY), lactation length (LL), calving to first insemination interval (CFI), days open (DO), and calving interval (CI). Expected ΔG was estimated and compared by three different methods; method I involved heritability and selection differential; method II involved selection intensity, phenotypic standard deviation, and heritability; method III involved estimation of ΔG through four paths of inheritance. Initially, eleven progenies/sire were utilized for assessing expected ΔG by method III, and expected ΔG was found as 34.33, 0.12, 0.12 kg, 2.63, 1.51, 2.74, and 2.80 days/year for 305DMY, ADMY, PY, LL, CFI, DO, and CI, respectively. Additionally, there was a significant increase in expected ΔG on increasing progenies/sire from 6 to 11 while subsequent increase upto 16 had little effect on expected ΔG. These findings will be helpful in formulating breeding strategies worldwide in small buffalo herds to obtain sustainable ΔG in production and reproduction traits.

Genomic clues of association between clinical mastitis and SNPs identified by ddRAD sequencing in Murrah buffaloes.

The total milk production of India is 209.96 MT out of which 45% is contributed by the indigenous buffalo and due to their high producing virtue, the prevalence of mastitis is 5-20%. Despite the increasing level of technological advancement, mastitis is still an issue of concern for dairy industry in India as well as across the world. Therefore, the present study aimed to identify the SNPs and associate them with the incidence of clinical mastitis in Murrah buffalo using the ddRAD sequencing approach taking mastitis incidence data of 96 Murrah buffaloes. A total of 246 million quality controlled reads were obtained with an average alignment rate of 99.01% and at a read depth of 10, quality controlled SNPs obtained were 18,056. The logistic regression model was used and a total of seven SNPs were found significantly associated (p < 0.001) with mastitis incidence and seven genes were identified viz., NCBP1, FOXN3, TPK1, XYLT2, CPXM2, HERC1, and OPCML. The majority of them were having tumor suppressing action, related to immunogenetics or glycolytic and energy production. Conclusively, the SNPs identified in this study may be useful for future studies on mastitis incidence in Murrah buffalo and the SNP associations can be further validated.

Genetic analysis of milk and milk composition traits in Murrah buffaloes using Bayesian inference.

Accurate and unbiased assessment of genetic parameters of milk and milk composition traits play an important role in formulating breeding program for genetic improvement of Murrah buffaloes. In this study, data spread over 28 years were utilized to estimate genetic parameters of traits viz., 305 d milk yield (305MY), 305 d fat yield (305FY), 305 d solid not fat yield (305SNFY), milk fat percentage (fat%) and solid not fat percentage (SNF) percentage (SNF%) in Murrah buffaloes kept at ICAR-National Dairy Research Institute, Karnal. Bayesian multiple-trait analysis was done using animal model and Gibbs sampling to estimate (co)variance components. Posterior means of heritability and posterior standard deviation for 305MY, 305FY, 305SNFY, fat% and SNF% were 0.18 ± 0.05, 0.17 ± 0.05, 0.18 ± 0.05, 0.07 ± 0.03 and 0.15 ± 0.06 and posterior means of repeatability estimates along with posterior standard deviation for corresponding traits were 0.33 ± 0.04, 0.32 ± 0.04, 0.33 ± 0.04, 0.14 ± 0.02 and 0.30 ± 0.04, respectively. Estimates of genetic correlation varied from -0.080 (305MY and fat %) to 0.999 (305MY and 305SNFY). Permanent environmental correlations varied from -0.060 (305MY and SNF%) to 0.999 (305FY and 305SNFY). This study indicated that all considered traits except fat% have ample genetic variability which can be exploited for selection and genetic improvement of Murrah buffaloes.

Sustainable multi-trait selection index based on production, reproduction, and health traits for genetic improvement of Murrah buffaloes.

Selection of Murrah buffaloes based on production, reproduction, and health traits play an important role in sustainable genetic improvement. With the aim to construct an index comprising of production, reproduction and health traits of Murrah buffaloes, the production and reproduction data during a 50-year span (1971-2020) and for health traits data of 27 years (1994-2020) were analyzed. Various three, four, five, and six trait combinations of production, reproduction, and health traits were explored for construction of performance indices. The income and expenditure method was used to calculate relative economic values (REV) for traits under consideration. A total of 37 performance indices were constructed using a multi-trait combination of production, reproduction, and health traits. Out of 37 indices, the (ADMY-PY-DO-AFC-FF) based index was identified as the best, with 78%, 2%, and 20% weightage given to production, reproduction, and female fertility, respectively. Furthermore, when REVs of traits were increased by 25% and 50%, there was no change observed in the relative importance of traits in the best index, indicating that the index was robust to substantial increases in economic values. Therefore, it is recommended that the above constructed composite selection index can be employed in selection of female buffaloes to achieve sustainable genetic gain.

Identification of differential methylome signatures of white pigmented skin patches in Nili Ravi buffalo of India.

The DNA methylation events mark a major epigenetic change in the genome, reflecting non-genetic disease developments and varied phenotypes. The water buffalo is a dairy production animal with wide agro-climatic distribution in India. Breed-wise the coat color of water buffalo varies from ash-gray to jet black. A typical pigmentation pattern is found in one of the breeds of North India, Nili Ravi, with variedly distributed white patches. The DNA methylation pattern could potentially reveal the epigenetic factors responsible for the pigmentation patterns. To address this question, the DNA isolated from the skin tissues of Nili Ravi with varied white pigmentation and black Murrah buffaloes was subjected to reduced representation bisulfite sequencing. DNA methylation analysis revealed, 68.44%, 63.39%, and 47.94% of the promoter regions were hypermethylated in Nili Ravi over-white versus Murrah, Nili Ravi under-white versus Murrah, and Nili Ravi under-white versus Nili Ravi over-white, respectively. Major genes identified to be differentially methylated among over-white and under-white skin tissues in Nili Ravi included TBX2, SNAI2, HERC2, and CITED1. Overall the results have indicated differential methylation patterns to be potentially involved in hyper or hypopigmentation in Nili Ravi and Murrah buffaloes.

Impact and utility of shallow pedigree using single-step genomic BLUP for prediction of unbiased genomic breeding values.

In unstructured dairy programs, pedigree is usually shallow, which leads to biased prediction of breeding values using best linear unbiased prediction (BLUP). The objective of this study was to come out with a genomic prediction strategy that can utilize shallow pedigree information and predict unbiased and more accurate GEBV for sex-limited traits in a small population using single-step GBLUP (ssGBLUP). The data and models for a population under selection were simulated. Out of current 10 generations, 10th generation with 1000 candidates served as validation population. For the complete pedigree scenario, pedigree (P)BLUP estimated breeding values (EBV) were unbiased with accuracy (r) of 0.35 ± 0.02 and 0.26 ± 0.01 for 0.3 and 0.1 h2 scenario, respectively. For the shallow pedigree, biased prediction of breeding values and low accuracies were obtained with linear decline in the accuracy of EBV for removal of information on more distant pedigree. Accuracy and bias (ρ) for scenario with removing 4 distant generations from pedigree were 0.30 ± 0.02 and 0.55 ± 0.03, respectively, in moderate h2 scenario. Use of Genomic (G)BLUP, especially with "extreme phenotypic contrast selective genotyping," (TB) resulted in higher accuracy for a small reference of females; however, GEBV were highly biased. We observed that ssGBLUPF, where the numerator relationship matrix is corrected for inbreeding, resulted in more accurate and unbiased estimates of GEBV across shallow pedigree scenario, with TB all female reference (missing 4 distant generations: r = 0.50 ± 0.02; ρ = 0.96 ± 0.02). We recommend use of ssGBLUPF with two tailed selectively genotyped all female reference in shallow pedigree scenarios, to obtain unbiased and accurate GEBV for sex-limited traits, when resources are limited.

Genetic admixture and population structure analysis of Indian water buffaloes (Bubalus bubalis) using STR markers.

BACKGROUND: India has a vast riverine and swamp buffalo diversity adapted to various agro-ecological conditions. In the present study, genetic diversity data for 10 different buffalo populations of India, using 20 highly polymorphic microsatellite markers has been generated for the genetic diversity analysis. The buffalo populations of Eastern Odisha state, were the primary focus. METHODS AND RESULTS: The minimal spanning network based on Bruvo's distance, PCA (Principal Component Analysis) based on the Fst (Fixation Index) values, and genetic admixture analysis using both the STRUCTURE and 'snapclust' were performed. The analysis could identify the Manda population as distinct from other Odisha buffalo breeds as well as adjoining Chhattisgarhi buffalo breeds. The total observed number of alleles ranged between 143 (Manda) and 301 (Paralakhemundi) with an average of 204 alleles per breed. The Sambhalpuri buffalo population also clustered into two separate subpopulations, half of the unique sub-population located geographically south-wards, displayed no admixture with any of the adjacent buffalo populations. The Manda buffalo population has shown sufficient allelic richness and heterozygosity under random mating being practiced in the field conditions. CONCLUSIONS: The study has led to the identification of the Manda as a distinct buffalo population, and the germplasm has been registered as a new Indian buffalo breed. Whereas, the Sambhalpuri population requires elaborate analysis to confirm the existence of two distinct sub-populations.

Current status and unique attributes of Indian Chilika buffalo for adaptation to brackish water ecology.

Chilika buffalo is native to the Eastern coast of India and well adapted to the largest coastal brackish water lagoon of Asia, Chilika Lake. We present here a report on the Chilika buffalo breed emphasizing the conservational urgency based on unique biochemical and molecular evidence related to liver and kidney functions while comparing it with tropically adapted other water buffalo breeds (Bubalus bubalis) of India. It is found that the Chilika buffalo breed has a better ability to withstand a long dehydration period as evident from its better glomerular filtration and higher expression of the ion transport channel. Mitochondrial D-loop sequencing results have shown these buffaloes being closer to swamp-type buffaloes of Bangladesh and northeast India and represent a unique "hybrid zone" on the eastern coast of India. Conservation of such uniquely adapted germplasm is crucial owing to the current global trend, where the introduction of exotic breeds has negatively impact "sui-generis" germplasm and they require higher managerial resource consumption for maintaining higher productivity. Further, the introduction of unconventional fisheries activities has proved detrimental to the lagoon ecosystem, potentially causing more threat to the buffalo's population.