Population-wide genetic analysis of Ovar-DQA1 and DQA2 loci across sheep breeds in India revealed their evolutionary importance and fitness of sheep in a tropical climate.

Genetic variability at the major histocompatibility complex (MHC) is important in any species due to significant role played by MHC for antigen presentation. DQA locus has not been studied for its genetic variability across sheep population in India. In the present study, MHC of sheep at DQA1 and DQA2 loci were evaluated across 17 Indian sheep breeds. Results revealed high degree of heterozygosity (10.34% to 100% for DQA1 and 37.39 to 100% for DQA2). 18 DQA1 alleles and 22 DQA2 alleles were isolated in different breeds. Nucleotide content for DQA region revealed richness of AT content (54.85% for DQA1 and 53.89% for DQA2). DQA1 and DQA2 sequences clustered independently. We could see evidence of divergence of DQA as DQA1 and DQA2 across sheep breeds. Wu-Kabat variability index revealed vast genetic variation across DQA1 and DQA2, specifically at peptide binding sites (PBS) that consisted 21 residues for DQA1 and 17 residues for DQA2. Evolutionary analysis revealed the presence of positive and balancing selection for DQA1 locus, however DQA2 was under purifying selection across sheep breeds. Higher heterozygosity and large diversity at both loci especially at PBS indicated the fitness of the sheep population for evading pathogens and adapt to the harsh tropical climate.

Cross-population genetic analysis revealed genetic variation and selection in the Ovar-DRB1 gene of Indian sheep breeds.

In sheep, MHC variability is studied widely to explore disease association. The aim of the current study was to explore the genetic diversity of Ovar-DRB diversity across sheep breeds of India. Here, Ovar-DRB1 locus was studied across 20 sheep breeds. DRB1 was amplified (301 bp) and sequenced using a PCR-sequence-based typing approach. Results revealed a high degree of heterozygosity across breeds (mean: 73.99%). Overall mean distance for DRB1 was highest in Sangamneri (0.18) and lowest in Madgyal sheep (0.10). There was a higher rate of transition, across breeds. Further, 39 alleles were isolated in different breeds, out of which 10 were new. To allow easy access and use of the immune-polymorphic database, an online database management system was launched (http://www.mhcdbms.in/). Nucleotide content across breeds for the DRB1 region revealed the richness of GC content (59.26%). Wu-Kabat index revealed vast genetic variation across peptide binding sites (PBS) of DRB1. Residues 6, 66, 69, 52, and 81, were polymorphic showing utility for antigen presentation. All breeds were under positive selection for DRB1 locus (dN > dS). Study revealed the importance of DRB locus diversity for beta chain specifically at PBS across sheep breeds of the Indian subcontinent and presented evidence of positive selection for DRB owing to its evolutionary significance.

Farmer's participatory approach for breed improvement in Malpura sheep.

The objective of the study was to propose a model for a community-based breeding programme (CBBP) in Malpura sheep. The study was carried out for nine years (2012-2018) in 31 villages covering 77 flocks. The average flock size was 54.39 ± 0.65. The study revealed that the Malpura sheep is a long-day breeder with 72.1% of lambing during August to January. The higher concentration of lambing in this period is to avoid the stress of harsh summer and scarcity of feed. The disposal pattern of male lambs was stringent due to selection. Only 3.1% of male lambs reach to adult stage. For females, the replacement rate was 34.11%. The live weights (kg) from 8,263 lambs were 3.34 ± 0.01, 14.61 ± 0.04, 20.48 ± 0.07 and 30.01 ± 0.16, for birth, three-, six- and 12-month age, respectively. Non-genetic factors such as sex, season and year of birth and flock significantly influenced growth. The heritability was 0.20 ± 0.03, 0.20 ± 0.008, 0.23 ± 0.004 and 0.17 ± 0.009 for birth, three-, six- and 12-month weight, respectively. This indicated scope for genetic improvement, given a systematic breeding programme, is in place. The present model followed by sheep breeders is highly dependent upon the Government aided Central Sheep and Wool Research Institute (CSWRI) for supply of breeding rams, data collection and health care. Alternate self-sustainable model of CBBP is proposed wherein a breeder's organization leads the way to establish a ram-rearing centre (RRC), teams of experts for ram selection, data collection and market linkage. CSWRI will continue to assist for selection decisions and real-time data entry using a cloud-based "Avimegh" platform. The early selection of 5% male lambs at weaning and rearing them at RRC until final selection at 12-month will provide unbiased selection of rams under CBBP. The surplus rams will be sold by RRC for profit. Several key measures such as micro-finance, co-ordination between stakeholders, market linkage and technical input for data collection and selection have been suggested to make the CBBP self-sustainable.

Selecting sheep for Haemonchus contortus resistance and susceptibility: Flock dynamics and genetic architecture.

The study was carried out in the selected strains of Malpura and Avikalin sheep for nematode resistance versus susceptibility (Malpura resistant: MR, Malpura susceptible: MS, Avikalin resistant: AR and Avikalin susceptible: AS). Strains were selected since 2004 and inter-se breeding followed since year 2007. The objective was to decipher the genetic structure of strains and to estimate genetic parameters for fecal egg count (FEC). The population size was small (107-279); however, all the four flocks had sizable effective population size (fe) for founders. The ratio for fe to effective number of ancestors (fa) was nearly 1 indicating genetic stability. Inbreeding was under control. The log transformed FEC (LFEC) was significantly (P < .05) affected by sex, year and month of recording for all 4 strains. Generation had desired genetic trend. There was low heritability for LFEC in MR(0.02), MS(0.06), AR(0.07) and AS(0.02). The estimate of the permanent environment effect (c2) was low. Repeatability (ρ) for LFEC were 0.05, 0.11, 0.07 and 0.06 for MR, MS, AR and AS, respectively. Pruning the data for repeated records in peak infection period (1: "Aug-Sep" and 2: only "Sep") resulted in considerable increase in ρ for MR (Aug-Sep: 0.07 ± 0.03 and Sep: 0.12 ± 0.07) and AR (Aug-Sep: 0.05 ± 0.04 and Sep: 0.09 ± 0.10). Results revealed genotype x environment interaction for repeated measures across months. Low flock size and low genetic variability are hurdles for improvement in R strains. It is recommended to incorporate new candidates post selection from the other breeding flocks and admit them in the R strains for selective breeding.

The breeding structure for the small ruminant resources in India.

Intense selection for a few desired traits has resulted in reduction of the effective population size (Ne) in most of the plant and livestock populations across the world. The objective of the research was to assess the impact of Ne on the genetic architecture of the population in a simulated data with variable Ne for general population under selection. Along with this, the estimate of Ne and its ratio to adult breeding population (NB) in the census data of small ruminants of India were also investigated. Results indicated that the average inbreeding ([Formula: see text]) decreases with increase in Ne; similarly, increase in [Formula: see text] per generation was highest in population with lowest Ne. Correlation of estimated breeding value (EBV) with true breeding value (TBV) was not much affected with effective population size. An effective number of chromosome segments (Me) in the populations under selection were significantly affected by magnitude of Ne, with linear positive relation between Ne and Me. Results on livestock census data revealed that all the sheep and goat breeds have sufficiently large Ne based on derived and actual census data. The median for ratio of effective population size to adult census size in sheep breeds was 0.120 and for goat breeds was 0.131. Karnah and Poonchi sheep shares the status of endangered breeds due to a smaller number of breeding female population and hence need attention for conservation. The Ne was large in sheep and goat due to less selection pressure as a result of low coverage of breed improvement programs, availability of large number of breeding males, and absence of artificial insemination (AI) in the field flocks. The estimates of Ne and its ratio to the adult census size (NB) excluded several factors such as fluctuating population size and overlapping generations. Study revealed introspection from most of the industrial breeding programs on the issue of Ne for populations under selection. Similarly, in small ruminants, large Ne indicates huge genetic diversity and scope of improvement in the productivity in near future.

Survival analysis of mortality in pre-weaning kids of Sirohi goat.

Pre-weaning animals exit a flock through death induced by various reasons, causing significant economic losses to the goat producers. In this study, we investigated the survival from birth to weaning of Sirohi goat kids within framework of the survival analysis. Kid records were accessed from 1997 to 2017, with the information on 4417 pre-weaning animals of farmed Sirohi goat native to the Rajasthan State of India. A multivariable Cox regression was fitted to the data after checking the assumptions of regression. The explanatory variables were sex, type of birth, season of birth, birthweight, doe weight at kidding and year of birth. Model selection eliminated doe weight from the model, and sex, type of birth, season of birth, birthweight and year of birth were retained in the model. With model calibration also, these five covariates were retained in the model. The mortality on the first day after birth was 0.3%, constituting 3.5% of all pre-weaning mortality. The mortality until the end of weaning period was 7.8%. Regression analysis revealed that the higher birthweight at kidding was associated with reduced hazard of death among the kids. Male kids had higher hazards of death compared with female kids. The single-born kids had lower risks of death compared with twin-born kids after accounting for heterogeneity. The winter season had a very high adverse effect on the survival of the kids. With each passing year, risks of death decreased. The results of this study indicate that better survival of kids can be achieved by controlling both environmental and animal-related factors.

Genetic parameter estimates for fecal egg counts and their relationship with growth in Avikalin and Malpura sheep.

Breeding for resistance against nematodes has become the need of the hour due to emergence of anthelmintic resistant strains of major pathogenic nematodes of economic importance and rising demand for chemical residue free food by consumers. ICAR-Central Sheep and Wool Research Institute Avikanagar (Rajasthan) has developed Haemonchus contortus resistant lines of sheep in Avikalin and Malpura breeds by harvesting benefits of over-dispersion in fecal egg counts (FEC) through executing a breeding program since year 2004. Aim of the present study was to assess the genetic parameters for nematode resistance in these lines and also to develop suitable criteria for selection targeting resistance as well as growth improvement in these two lines. The data on 1240 Avikalin and 2172 Malpura sheep generated over 13 years (2004-16) for FEC along with deep pedigree and growth records for live weight at 6 (6WT) and 12 month were used for study. Data were analyzed using Average Information Restricted Maximum Likelihood (AIREML) approach. Results revealed moderate heritability (h2) for pre-drench log transformed fecal egg count (LFEC) in Avikalin (0.21±0.06) and Malpura (0.18±0.04) sheep. The post-drench h2 for LFEC was low in Avikalin (0.04±0.03) and Malpura (0.11±0.03) sheep. Effective selection program can be carried out for further improving the resistance against H. contortus in both the breeds using pre-drench LFEC estimates. The genetic correlation between the pre-drench LFEC and growth traits was not in the desired direction. Existence of substantial genotype × environment (G×E) interaction was seen in Malpura sheep, where major shift in ranks of sheep based on pre-drench LFEC as that of post-drench LFEC was observed owing to genetic correlation of 0.65±0.15. The G×E was absent in Avikalin sheep. Unreliable genetic correlation between growth and LFEC does not warrant a multi trait selection index development and its utilization in breeding program. The independent selection for LFEC followed by corrected 6WT can precisely help in achieving the goal of improving growth in nematode resistant sheep.

Genetic diversity of Cahi DRB and DQB genes of caprine MHC class II in Sirohi goat.

The objective of this study was to assess the genetic diversity of the Sirohi goat for DQB and DQB1 loci, and to study their association with antibody response induced by the Peste des petits ruminants (PPR) vaccine. A total of 360 Sirohi kids were studied using single stranded confirmation polymorphism (SSCP) followed by polymerase chain reaction sequence-based typing (PCR-SBT) for DQB and DQB1 diversities. The competitive enzyme-linked immuno-sorbent assay (C-ELISA) was used to evaluate immune response post-PPR vaccination. Study revealed rich diversity of major histocompatibility complex (MHC) region in goat. A total of 18 DQB and 15 DQB1 alleles were obtained which were new. Alleles DRB*0104 and DQB1*0101 were the most common. The approach of SSCP combined with PCR-SBT reflects cost-effective and most powerful approach to decipher the genetic diversity in complex MHC region. Study revealed variation in DQB and DQB1 genes in Sirohi flock along with high Wu-Kabat index. A total of 16 of the 89 amino acid residue sites in DQB and 19 of 86 residue sites in DQB1 had more than three amino acid substitutions. Positive evolutionary selection was evident in Sirohi for MHC region. Nonsignificant association of DQB and DQB1 genotypes with PPR virus (PPRV) vaccine response revealed complexity of the phenotype and importance of other factors for vaccine response. Rich diversity of DQB and DQB1 genes reflects the fitness of the population and importance of this locus for future selection programmes.

Polymorphism study of growth differentiation factor 9B (GDF9B) gene and its association with reproductive traits in sheep.

GDF9B protein plays a critical role in growth and differentiation of early ovarian follicles. In Inverdale and Hanna sheep, mutations in exon-2 of GDF9B gene have been recorded to show increased ovulation rate in heterozygous condition whereas homozygotes are infertile. Present screen study was carried out to explore the presence of these reported mutations in Corriedale and Local Kashmir Valley sheep with high rate of twinning. Exon-2 of GDF9B gene was amplified and the polymorphism was explored by SSCP technique. In the process three different bandings were observed. Later on these patterns corresponded with three different allelic forms on nucleotide sequencing. Phylogenetic analysis revealed that the nucleotide sequences of alleles observed in the present study and that of a published sequence of sheep were having the same point of origin. The results were also compared with goats, large ruminants and humans. The allelic frequencies of allele A and B were 0.64 and 0.36, respectively in Corriedale sheep whereas the allelic frequencies of all the three alleles in Kashmir Valley sheep were 0.60, 0.34 and 0.06. SNP "C" of the designated genotype AC was observed to pronounce a significant effect on litter size with average litter size going up by 0.63 as compared with the nearest genotype AB wherein the litter size was 1.29±0.05. The average litter size between AA and AB genotypes did not vary significantly.

Variability of milk fat globule membrane protein gene between cattle and riverine buffalo.

A study on butyrophilin (BTN) gene was conducted to detect variability at nucleotide level between cattle and buffalo. Hae III PCR-RFLP was carried out in crossbred cattle and it revealed polymorphism at this locus. Three genotypes namely, AA, BB and AB and two alleles were observed with frequencies 0.78, 0.17, 0.04 and 0.87, 0.13, respectively. The sequences of different cattle, buffalo and sheep breeds have been reported in the EMBL gene bank with accession numbers: AY491468 to AY491475. The nucleotides, which have been substituted from allele A to B, were found to be C to G (71st nucleotide), C to T (86th nucleotide), A to T (217th nucleotide), G to A (258th nucleotide), A to C (371st nucleotide) and C to T (377th nucleotide). The nucleotide substitution at 71st, 86th and 377th position of the fragment were expected to be a silent mutation where as nucleotide changes at 217th, 258th and 371st positions were expected to be substituted by lysine with arginine, valine with isoleucine and leucine with proline in allele B. The differences of nucleotides and amino acids between cattle, buffalo and sheep breeds have been revealed and on the basis of nucleotide as well as protein variability the phylogenetic diagram have been developed indicating closeness between cattle and buffalo.