Indian Zebu Research Articles

Genetic variations in HsP90AA1 chaperone gene across diverse Indian native cattle (<i>Bos indicus</i>) breeds adapted to varied agroclimatic regions

Indian zebu cattle during their separate evolution from Bos taurus, have acquired certain changes in a set of genes that regulate the body temperature in response to heat stress. Heat shock proteins (HSPs), the abundant and ubiquitous molecular chaperones play essential role in many cellular biological processes, and are preferentially transcribed in response to heat stress. The objective of this study was to comparatively screen variants/ single nucleotide polymorphisms (SNPs) in the HSP90AA1 gene in cattle breeds, viz., Gir, Kankrej, Ladakhi, Nagori, Nimari, Ongole, Rathi and Sahiwal, adapted to different agroclimatic regions of India including the exotic breed Holstein Friesian and yak (Bos grunniens). Sanger sequencing was carried out in all the translatable exons of the gene. The analysis revealed 6 SNPs – 3 were located in exon 3, 1 SNP each in exon 4 and 6, and another SNP in exon 9. We also identified a novel SNP 2937 (G>T) nucleotide position of exon 9. Interestingly, 2 variants detected at position 1924 (A>G) of exon 3 and 2343 (T>C) of exon 6 with a frequency of 0.9 and 0.73, respectively, were found to be almost fixed in nearly all of the Indian native cattle breeds. These SNPs could potentially be utilized to undertake genotype-phenotype association studies for superior heat stress tolerance trait in Indian native cattle.

"In vitro antimicrobial and synergistic effect of fermented Indian zebu (Sahiwal) cow colostrum whey derived peptides with Lactobacillus rhamnosus against pathogenic bacteria".

The online version contains supplementary material available at 10.1007/s13197-023-05776-2.

Understanding of molecular basis of histological graded horn cancer by transcriptome profiling

Horn cancer is most devastating and prominent cancer in Indian zebu cattle that affects socio-economic condition of small-scale farmers who depends on their cattle for farm work. Development in the field for genomics through next generation sequencing and bioinformatics advancement have helped to identify genes which have a role in horn cancer development. Histopathological examination of cancerous tissues of horn revealed myxomatous changes, well, moderate and poorly differentiated squamous cell carcinoma. Differential gene expression analysis showed 40, 11, 66 and 29 upregulated genes and 10, 14, 08 and 07 down-regulated genes in myxomatous, well, moderate and poorly differentiated squamous cell carcinoma as compared to normal. Significant differentially expressed genes are related to cell development, cell proliferation, cell–cell communication, cell signaling and angiogenesis which are linked to Akt pathway, mTOR pathway and Wnt pathway. Activity of these genes and related pathways have already been established about their role in development of cancer. Among the candidate genes; keratin family, keratin family related gene, chemokine signaling and cytokines signaling associated genes could be a prominent target for the development of stage specific prognosis marker after further detailed study at large sample population level. CSTA, PTN, SPP1 genes have upregulation in all stages of cancer and they have enrolled as biomarkers for horn cancer.

Assessment of Mutation Drift Equilibrium and the Occurrence of a Recent Genetic Bottleneck in South Indian Zebu Cattle.

Simple SummaryThis study was conducted on eight Indian zebu breeds with an objective to reveal any cryptic genetic bottlenecks in the population. The alarming reduction in the effective population size of these breeds was the major reason for conducting this study. Genotyping of 27 FAO/ISAG-recommended microsatellite markers and further statistical analysis using allele frequency data under three different models of mutation revealed the absence of recent genetic bottlenecks in any of the populations under study. The results from the qualitative test of mode shift distortion were in accordance with the above findings. Even though the results of the present study indicated little or no critical loss of rare alleles in any of the breeds understudy, the chances of this happening cannot be completely ignored. One of the plausible explanations for this scenario could be the potential gene flow from commercial taurine cattle through indiscriminate crossbreeding in the native tract of these breeds. Thus, we here emphasize the need of further initiatives for improving selective breeding practices in order to conserve and effectively utilize the existing South Indian zebu cattle genetic diversity.During the last few decades, the effective population size of indigenous zebu cattle breeds has declined drastically, resulting in the classification of some of them into the vulnerable, endangered, or critically endangered category. Drastic reductions in the effective size of a population may result in genetic bottlenecks and can affect within-breed genetic variability and its viability. The present study was undertaken with the objective of evaluating South Indian zebu cattle populations for mutation drift equilibrium and to detect the occurrence of recent genetic bottleneck events. A total of 293 cattle from eight indigenous breeds were genotyped at 27 FAO/ISAG-recommended microsatellite marker loci. Three different statistical tests, viz., the sign test, standardized differences test, and Wilcoxon sign rank test were performed using allele frequency data to detect loci with heterozygosity excess under the infinite alleles, stepwise, and two-phase mutation models. Under the infinite alleles model, the observed number of loci with heterozygosity excess (He > Heq) ranged between 10 and 19 among the investigated cattle breeds. However, the observed heterozygosity excess was not statistically significant (p > 0.05) in any of the studied breeds. Similarly, the standardized differences test and Wilcoxon sign rank test revealed no concrete evidence for the occurrence of a recent genetic bottleneck in South Indian zebu cattle breeds. The qualitative test for mode-shift distortion revealed a normal L-shaped distribution of allele frequencies, suggesting a lack of evidence for the loss of low-frequency alleles in all the investigated South Indian zebu cattle breeds.

Whole-Genome Analysis Deciphers Population Structure and Genetic Introgression Among Bovine Species.

It is known that throughout history and presently, taurine (Bos taurus) and indicine/zebu (Bos indicus) cattle were crossed with other bovine species (e.g., gayal, gaur, banteng, yak, wisent, and bison). Information on the role of interspecific hybridization to facilitate faster adaptation of the newly arrived domestic species to new environments is poorly known. Herein, we collected 266 samples of bovine species of the taurine, zebu, yak, and gaur from West Europe, Southwest China, Indian subcontinent, and Southeast Asia to conduct the principal component analysis (PCA), admixture, gene flow, and selection signature analyses by using SNPs distributed across the bovine autosomes. The results showed that the genetic relationships between the zebu, yak, and gaur mirrored their geographical origins. Three ancestral components of the European taurine, East Asian taurine, and Indian zebu were found in domestic cattle, and the bidirectional genetic introgression between the Diqing cattle and Zhongdian yak was also detected. Simultaneously, the introgressed genes from the Zhongdian yak to the Diqing cattle were mainly enriched with immune-related pathways, and the ENPEP, FLT1, and PIK3CA genes related to the adaptation of high-altitude hypoxia were detected. Additionally, we found the genetic components of the Zhongdian yak had introgressed into Tibetan cattle. The 30 selected genes were detected in Tibetan cattle, which were significantly enriched in the chemokine signaling pathway. Interestingly, some genes (CDC42, SLC39A2, and EPAS1) associated with hypoxia response were discovered, in which CDC42 and SLC39A2 played important roles in angiogenesis and erythropoiesis, and heart function, respectively. This result showed that genetic introgression was one of the important ways for the environmental adaptation of domestic cattle.

The genetic history of Mayotte and Madagascar cattle breeds mirrors the complex pattern of human exchanges in Western Indian Ocean.

Despite their central economic and cultural role, the origin of cattle populations living in Indian Ocean islands still remains poorly documented. Here, we unravel the demographic and adaptive histories of the extant Zebus from the Mayotte and Madagascar islands using high-density SNP genotyping data. We found that these populations are very closely related and both display a predominant indicine ancestry. They diverged in the 16th century at the arrival of European people who transformed the trade network in the area. Their common ancestral cattle population originates from an admixture between an admixed African zebu population and an Indian zebu that occurred around the 12th century at the time of the earliest contacts between human African populations of the Swahili corridor and Austronesian people from Southeast Asia in Comoros and Madagascar. A steep increase in the estimated population sizes from the beginning of the 16th to the 17th century coincides with the expansion of the cattle trade. By carrying out genome scans for recent selection in the two cattle populations from Mayotte and Madagascar, we identified sets of candidate genes involved in biological functions (cancer, skin structure, and UV-protection, nervous system and behavior, organ development, metabolism, and immune response) broadly representative of the physiological adaptation to tropical conditions. Overall, the origin of the cattle populations from Western Indian Ocean islands mirrors the complex history of human migrations and trade in this area.

Assessing Genomic Diversity and Productivity Signatures in Dianzhong Cattle by Whole-Genome Scanning.

Dianzhong cattle is a classic Chinese indigenous cattle breed with historical records dating back to 200 BC. But with its genomic differences having not been clearly elucidated, the quest for genomic characterization will be an essential step towards understanding the genomic basis of productivity and adaptation to survival under Chinese farming systems. Here we compared 10 Dianzhong cattle (four newly sequenced and six downloaded) with 29 published genomes of three underlying ancestral populations (Chinese zebu, Indian zebu, and Yanbian cattle) to characterize the genomic variations of Dianzhong cattle. Dianzhong cattle has a high nucleotide diversity (0.0034), second only to Chinese zebu. Together with analyses of linkage disequilibrium decay and runs of homozygosity, Dianzhong cattle displayed higher genomic diversity and weaker artificial selection compared with Yanbian cattle. From a selective sweep analysis by four methods (Fst, π-ratio, XP-CLR, and XP-EHH), the positive selective signals were mainly manifested in candidate genes and pathways related to heat resistance, growth and development, fat deposition, and male reproduction. Missense mutations were detected in candidate genes, SDS (c.944C > A and p.Ala315Glu), PDGFD (c.473A > G and p.Lys158Arg), and DDX4 (rs460251486, rs722912933, and rs517668236), which related to heat resistance, fat deposition, and spermatogenesis, respectively. Our findings unravel, at the genome-wide level, the unique diversity of Dianzhong cattle while emphasizing the opportunities for improvement of livestock productivity in further breeding programs.

South Indian zebu dwarf cattle show higher natural resistance to tick infestation compared to crossbreds: preliminary observations.

The present study aimed to evaluate the influence of various factors on the host resistance of cattle against tick infestation in Kerala State, South India. The number of naturally infested semi-engorged female ticks present on the cattle was counted on a sample cattle population across the four regions of Kerala. Then, the animals were grouped based on the tick count. The chi-square test and Fisher exact test were used to identify the effects of genetic and non-genetic factors on tick burden in cattle. It was found that the Holstein Friesian crossbred and Jersey crossbred cattle had a significantly higher level of tick infestation than the indigenous Vechur breed (Bos indicus). No tick infestation was observed in indigenous Kasaragod Dwarf cattle (B. indicus). Tick infestation was more in heifers and dry animals than lactating animals. However, the sex of the animal and the stage of lactation did not influence the tick load. Tick infestation was also influenced by parity, grazing, and region.

Legacy of draught cattle breeds of South India: Insights into population structure, genetic admixture and maternal origin.

The present study is the first comprehensive report on diversity, population structure, genetic admixture and mitochondrial DNA variation in South Indian draught type zebu cattle. The diversity of South Indian cattle was moderately high. A significantly strong negative correlation coefficient of -0.674 (P<0.05) was observed between the effective population size of different breeds and their estimated FIS. The genetic structure analysis revealed the distinctness of Kangayam, Vechur and Punganur cattle from the rest of the zebu breeds. The results showed the influence of Hallikar breed in the development of most Mysore type cattle breeds of South India with the exception of Kangayam. Bayesian clustering analysis was performed to assess the taurine admixture in South Indian zebu cattle using purebred Jersey and Holstein-Friesian as reference genotypes. Relatively high levels of taurine admixture (>6.25%) was observed in Punganur, Vechur, Umblachery and Pulikulam cattle breeds. Two major maternal haplogroups, I1 and I2, typical of zebu cattle were observed, with the former being predominant than the later. The pairwise differences among the I2 haplotypes of South Indian cattle were relatively higher than West Indian (Indus valley site) zebu cattle. The results indicated the need for additional sampling and comprehensive analysis of mtDNA control region variations to unravel the probable location of origin and domestication of I2 zebu lineage. The present study also revealed major concerns on South Indian zebu cattle (i) risk of endangerment due to small effective population size and high rate of inbreeding (ii) lack of sufficient purebred zebu bulls for breeding and (iii) increasing level of taurine admixture in zebu cattle. Availability of purebred semen for artificial insemination, incorporation of genomic/molecular information to identify purebred animals and increased awareness among farmers will help to maintain breed purity, conserve and improve these important draught cattle germplasms of South India.

A Study on β-Casein Gene Polymorphism in Crossbred Cattle and Murrah / Graded Murrah Buffalo in Tamil Nadu

Background: Majority of the people believed that, only the native breeds have A2 milk than exotic or crossbred animals. There is no enough research on milk protein variants have been carried out in Indian zebu cattle. Present study was conducted to screen more number of crossbred cattles and buffaloes in Tamil Nadu to identify the frequency of A1 and A2 alleles in the population.Methods: The study was conducted on 68 cattle and 172 murrah / graded murrah buffaloes to explore the polymorphic variants of β-casein gene. Genomic DNA was extracted by phenol-chloroform method. Polymerase Chain Reaction (PCR) was performed with allele specific primers to amplify a 244 bp long fragment of beta-casein gene and visualized in 2% agarose gel electrophoresis. The population genetic indices were calculated based on the formulas.Result: In the present study revealed higher level of A2A2 genotype frequency (1.00) and fixation of A2 allele in kangayam cattle and murrah / graded murrah buffalo. The observed frequency of A1A1, A1A2 and A2A2 genotypes were 0.38,0.62 and 0.00 for jersey crossbred and 0.29, 0.71 and 0.00 for HF crossbred cattle. The range of Expected homozygosity (0.50 to 1.00), polymorphism information content (0.30 to 0.38), effective number of alleles (1.00 to 2.00) and level of possible variability realization value (44.54% to 100%) reflected existence of medium genetic variability in studied population.

Detection of A1 and A2 Alleles at Beta-casein Locus in Bargur and Umblachery (Indian Zebu) Cattle Breeds by Allele-specific PCR

Background: Many genetic variants of beta-casein in different breeds of cattle have been reported. The A1 and A2 are the most common variants. The breeds of Zebu cattle have high frequency of A2 allele or monomorphic for A2 allele. The current study aimed to screen Indian Zebu cattle breeds, Bargur and Umblachery, for A1 and A2 alleles at beta-casein locus.Methods: A total of 48 Bargur and 42 Umblachery cattle were genotyped for β-casein (CSN2) gene using allele-specific PCR. The gene and genotype frequencies were estimated. The theoretical heterozygosity (Heexp), experimental heterozygosity (Heobs), polymorphism information content (PIC), expected homozygosity (E), effective number of alleles (ENA) and level of possible variability realization (V%) were calculated.Result: The investigation revealed the presence of both A1 and A2 alleles at beta-casein locus in both Bargur and Umblachery cattle breeds. The A1A1 genotype was not observed in both the breeds. The frequencies of A1A2 and A2A2 genotypes were 0.125 and 0.875 respectively in Bargur and 0.050 and 0.950 respectively in Umblachery breed. The study indicated the predominance of A2 variant in both the breeds. The frequencies of A1 and A2 alleles were 0.063 and 0.937 respectively in Bargur and 0.02 and 0.98 respectively in Umblachery breed. The values of experimental heterozygosity (Heobs), theoretical heterozygosity (Heexp), polymorphism information content (PIC), expected homozygosity (E), effective number of alleles (ENA), level of possible variability realization (V%) were 0.125, 0.1163, 0.1095,0.8837, 1.131 and 11.88 respectively in Bargur breed. These values were 0.048, 0.0468, 0.0458, 0.9532, 1.049 and 4.79 respectively in Umblachery population. The observed heterozygosity and PIC values revealed the existence of very low genetic variability in the tested populations. The present work will be a contribution to the study on beta-casein locus in Indian zebu cattle.

Sequence Characterization of Forebrain Embryonic Zinc Finger-like (FEZL) Gene in Indian Zebu (Bos indicus) Cattle and their Crossbreds

Background: Forebrain embryonic zinc finger-like (FEZL) gene is an important candidate associated with mastitis resistance in dairy cattle. FEZL is involved in transcriptional regulation of neuronal development and there exists a crosstalk between neuronal development and immunity via downstream cytokine expression. A single glycine insertion into glycine stretch of FEZL gene has large effect on downstream cytokine pathway making the cows susceptible to mastitis. The present study was aimed to sequence characterize FEZL gene in Sahiwal (Bos indicus) and Karan Fries (Bos inidcus X Bos taurus) cattle.Methods: Sequence characterization of bovine FEZL gene was carried out by primer walking method. Ten sets of oligonucleotide primers were designed to synthesize overlapping fragments and generate the complete sequence of about 3.7 kb covering all exons and 5’ upstream regulatory and flanking regions.Result: A total of eight nucleotide variations including three INDELS and five substitution mutations were observed among FEZL gene sequences of Bos taurus, Bos indicus (Sahiwal) and Bos taurus X Bos indicus (Karan Fries) cattle. The conceptualized amino acid sequence of bovine FEZL gene in Sahiwal and Karan Fries cattle was found to have 13 tandem Glycine residues and a serine to proline change within exon 1 region. The percent identity of FEZL gene of Sahiwal and Karan Fries cattle was 99% with that of Bos taurus, 95% with dog, horse and pig, 94% with human, 93% with rabbit, 92% with marmoset, 89% with rat and 79% with chicken. Sequence characterization of ~0.7 kb 5’ flanking region showed that it is highly conserved among bovines and resulted in prediction of six putative sites for binding of transcription factors (including Elk-1, Oct-1, HNF4, Lmo2 complex, GATA-3 and Nkx2-5). Elucidation of Bos indicus FEZL gene will further form the basis to identify candidate gene markers for association with mastitis resistance/susceptibility in cattle.

High-Resolution Mass Spectrometer-Based Ultra-Deep Profile of Milk Whey Proteome in Indian Zebu (Sahiwal) Cattle.

Milk serves as a mode of protection to neonate through transferring the host defense proteins from mother to offspring. It also guards the mammary gland against various types of infections. Along with the presence of six vital proteins, bovine milk (whey) contains a massive class of minor proteins, not all of which have been comprehensively reported. In this study, we performed an LC-MS/MS-based ultra-deep identification of the milk whey proteome of Indian zebu (Sahiwal) cattle. Three independent search engines that are Comet, Tandem, and Mascot-based analysis resulted in the discovery of over 6,210 non-redundant proteins commonly identified. Genome-wise mapping revealed that chromosome 1 showed a minimum expression of 14 proteins, whereas chromosome 19 expressed 250 maximum proteins in milk whey. These results demonstrate that milk proteome in Sahiwal cattle is quite complicated, and minor milk fractions play a significant role in host defense.

Haemato-biochemical profile, mineral and electrolyte concentration, and antioxidant status of Zobawng cattle of Mizoram

Blood biochemical profile plays a pivotal role in assessing health status of animals. The present study was conducted to evaluate haematological and biochemical parameters, trace element and electrolyte concentration, oxidant and antioxidant status in local cattle (Zobawng) of Mizoram. The values recorded for Hb (12.61±0.40 g/dL), RBC (9.10±0.50 106/μL), WBC (11.52+1.13 103/μL), PCV (43.36±0.99%), total protein (8.30±0.17 g/dL), globulin, BUN (10.68±0.68 mg/dL) were on higher side than the exotic cattle breeds. Creatinine (1.96±0.10 mg/dL) and GGT (19.22+1.29 U/L) were also towards upper limit. Calcium (9.78+0.13 mmol/ L), magnesium (2.43+0.07 mmol/L) and inorganic phosphorus (6.76+0.27 mmol/L) values were higher than the exotic breeds and also higher than the Indian Zebu cattle. Total antioxidant was higher and the oxidative stress index was lower as compared to the crossbred or exotic cattle. This study provides first insight into the blood biochemistry of these indigenous cattle indicating its higher disease resistance and will be useful for its better management.

Four Novel SNPs of MYO1A Gene Associated with Heat-Tolerance in Chinese Cattle.

Simple SummaryChinese indigenous cattle were clustered into two groups of southern and northern breeds. The populations distributed in the hot south are mainly dominated by Bos indicus, and the populations distributed in the cold north are mainly dominated by Bos taurus. Myosin-1a (MYO1A) gene is a candidate gene related to pigmentation, which may be closely related to the heat tolerance traits of cattle. Therefore, our study aimed to explore the relationship between MYO1A gene and heat tolerance in Chinese indigenous cattle. Based on the previous GWAS research related to bovine heat tolerance trait, this study aimed to explore the effect of myosin-1a (MYO1A) gene on bovine heat tolerance trait, and find the molecular markers related to the heat tolerance of Chinese cattle. In our study, four novel candidate SNPs highly conserved in B. indicus breeds but barely existed in B. taurus were identified in MYO1A gene according to Bovine Genome Variation Database and Selective Signatures (BGVD). PCR and DNA sequencing were used to genotype 1072 individuals including 34 Chinese indigenous cattle breeds as well as Angus and Indian zebu. Two synonymous mutations (rs208210464 and rs110123931), one missense mutation (rs209999142; Phe172Ser), and one intron mutation (rs135771836) were detected. The frequencies of mutant alleles of the four SNPs gradually increased from northern groups to southern groups of Chinese cattle, which was consistent with the distribution of various climatic conditions of China. Additionally, four SNPs were significantly associated with four climatic conditions including annual mean temperature (T), relative humidity (H), temperature-humidity index (THI), and average annual sunshine hours (100-cloudiness) (SR). Among these, rs209999142 and Hap 1/1 had better performance than others. Our results suggested that rs209999142 was associated with heat-tolerance trait and rs208210464, rs110123931, and rs135771836 showed high phenotypic effect on heat-tolerance trait because of the strong linkage with rs209999142. These SNPs could be used as candidates for marker-assisted selection (MAS) in cattle breeding.

Transcriptomic analysis to affirm the regulatory role of long non-coding RNA in horn cancer of Indian zebu cattle breed Kankrej (Bos indicus).

Long non-coding RNA (lncRNA) was previously considered as a non-functional transcript, which now established as part of regulatory elements of biological events such as chromosome structure, remodeling, and regulation of gene expression. The study presented here showed the role of lncRNA through differential expression analysis on cancer-related coding genes in horn squamous cell carcinoma of Indian zebu cattle. A total of 10,360 candidate lncRNAs were identified and further analyzed for its coding potential ability using three tools (CPC, CPAT, and PLEK) that provide 8862 common lncRNAs. Pfam analysis of these common lncRNAs gave 8612 potential candidates for lncRNA differential expression analysis. Differential expression analysis showed a total of 59 significantly differentially expressed genes and 19 lncRNAs. Pearson's correlation analysis was used to identify co-expressed mRNA-lncRNAs to established relation of the regulatory role of lncRNAs in horn cancer. We established a positive relation of seven upregulated (XLOC_000016, XLOC_002198, XLOC_002851, XLOC_ 007383, XLOC_010701, XLOC_010272, and XLOC_011517) and one downregulated (XLOC_011302) lncRNAs with eleven genes that are related to keratin family protein, keratin-associated protein family, cornifelin, corneodesmosin, serpin family protein, and metallothionein that have well-established role in squamous cell carcinoma through cellular communication, cell growth, cell invasion, and cell migration. These biological events were found to be related to the MAPK pathway of cell cycle regulation indicating the role of lncRNAs in manipulating cell cycle regulation during horn squamous cell carcinomas that will be useful in identifying molecular portraits related to the development of horn cancer.

The Indian zebu in Brazilian lands

In this article we will examine the history of Brazilian cattle breeding in the early twentieth century, focusing on the transitional period of intensification of production, when the racial standardization of cattle began to be considered key to industrializing breeding practices. To this end, we consider the initiatives of a group of farmers from the far east of Minas Gerais (Mineiro Triangle), who travelled to Northern India between 1913 and 1917 to acquire bulls and cows of the zebu breed. Retracing the routes of their voyages to India thanks to the diaries and letters between these farmers in Brazil and India, we attempt to understand the role that Indian bovine cattle played in Brazil. The first farmers to make this trip moved in the sphere of the influential elites of the oligarchic Republic that was established in Brazil in 1889. This article argues that these farmers were driven by diverse factors, which could not be reduced to the single issue of an increase in economic productivity. In truth, there were political interests mainly related to the strengthening of personal prestige and the construction of a class character as “innovative cattle farmers” in a context of intra-oligarchic disputes over exerting influence in the Republican state.

Comparative blood haemato-biochemical variations in Indian Zebu cattle breeds during early summer

Haematological and blood chemistry profile plays an important role in breed-specific performance of the animals and it varies significantly between different cattle breeds. The present study was undertaken to compare haematological and blood chemistry profile of apparently healthy cattle breeds of India namely Gir, Kangayam, Ongole, Tharparkar, Rathi, Deoni and Sahiwal during early summer. There was significant variation in all the haematological indices except total leukocyte count (TLC) and basophils. Among the blood chemistry parameters, total protein, globulin, blood urea nitrogen, creatinine and calcium varied significantly between breeds whereas variation in albumin and phosphorus concentrations was non-significant. Globulin content was very high in Rathi breed and it correlated positively with TLC. Creatinine concentration was high in milch breeds (Gir and Sahiwal) with the exception of Deoni, followed by draught breeds (Kangayam) and dual-purpose breeds (Ongole, Rathi and Tharparkar).

Distribution of single nucleotide polymorphisms and protein domain architecture of toll-like receptor-2 in Pahari cattle (Indian non-descript indigenous breed)

Distribution of polymorphism in TLR2 gene was studied in Pahari (Indian non-descript indigenous breed) cattle of Himachal Pradesh. The complete sequence of TLR2 gene consisted of 136bp 5'-UTR, 2355bp CDS and 1316bp 3'UTR. The phylogenetic analysis revealed clustering of Indian Zebu with Bos taurus as the nearest neighbour. The nucleotide sequences of TLR2 revealed the occurrence of 19 SNPs including 47% non-synonymous and 53% synonymous SNPs within TLR domain among all bovines. The dS/dN ratio (ω)<1 at polymorphic sites revealed purifying selection and thus, indicating the presence of highly conserved domains. Two SNPs in the extracellular domain of TLR2 were predicted to have damaging effect whereas the rest have benign effect. The amino acid sequence of the coding region corresponds to a protein of 784 amino acids long with predicted molecular weight 104kDa and 6.97 pI value. The amino acid sequence analysis showed extracellular domain composed of leucine rich repeats (LRR), trans- membrane domain and Toll-IL receptor domain in the protein. The 3D structure of TLR2 is a solenoid-like having active sites buried in the concave side and forming a pocket for ligand binding. The substitution of G with T in nucleotide sequence leads to the replacement of amino acid W↔L at position 119 which has not been reported so far. These findings indicate that polymorphism may be associated with PAMPs mediated differential TLR signaling of bovine immunity and could determine the outcome of infection.

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