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In the present study, the effect of genetic polymorphism of WD-repeat containing protein 41 (WDR41) and Ankyrin repeat domain containing protein 31 (ANKRD31) gene on 17 traits related to udder and teat type and mastitis in 123 Karan Fries cows was studied. Restriction Fragment Length Polymorphism (RFLP) was used to identify the SNP (410 bp and 475 bp) in PCR amplified product of intron 4 and exon 10 in WDR41 gene. Both of them were polymorphic with Guanine to Adenine transition, and three genotypes namely AA, AG and GG were observed. In ANKRD31 gene, RFLP was used to identify the SNP in 513 bp PCR amplified product of intron 24 and two SNPs were found. We report for the first time that intron 4 and exon 10 of WDR41 gene is significantly associated with udder depth (UD), udder balance (UB), central ligament (CL), teat circumference (TC), SFF and tear diameter (TD), rear udder width (RUW), and mastitis. SNP rs110012582 in intron 24 of ANKRD31 gene is associated significantly with UD, distance between left and right teat (DLR), SFF, and SNP rs473512406 with udder length (UL), udder width (UW), udder circumference (UC), fore teat length (FTL), rear teat length (RTL), distance between fore and rear teat (DFR), DLR and mastitis. This information can augment future studies to determine the role of WDR41 and ANKRD31 genes as a candidate gene marker with desired udder and teat conformation and mastitis. Thus, it is essential to work for better udder health and prevent the incidence of mastitis in the herd.

Mastitis in cattle is a prevalent mammary gland disease that contributes significantly to the increase in veterinary expenditures in the dairy sector. KCNB1 (Potassium voltage-gated channel, subfamily B member 1) gene is involved in regulating apoptosis, cell proliferation and differentiation, udder epithelial tissue maintenance and repair, mammary gland development and recommended as a candidate gene for production related traits in cattle. The purpose of this research was to detect the genetic variants of KCNB1 gene in Sahiwal cattle and to analyze the association between polymorphisms with milk production traits, udder traits, and teat traits in Sahiwal cattle. A total of 87 cattle were genotyped by polymerase chain reaction-restriction fragment length polymorphism technique. Two single nucleotide polymorphisms within the non-coding sequence of KCNB1 gene were identified (g.78216220G>A and g.78216335A>G). Analysis of productivity traits within the genotyped animals revealed that the SNP1-Msp1 locus (g.78216220G>A) located at intron 1 was associated with milk production traits, but the SNP2-BspHI locus (g.78216335A>G) had no association with milk production. Significant associations were also observed between SNP1-Msp1 and SNP2-BspHI loci with both udder and teat traits. Our results demonstrate that polymorphisms in the cattle KCNB1 gene were associated with milk production, udder and teat traits and might be utilized as a genetic marker for marker-assisted selection in cattle breeding programs.

Heat shock proteins (HSPs) are known to modulate cellular response during summer stress in dairy cattle. Among different classes of HSPs, heat shock protein 20 (HSPB6) is a member of the small HSP family protein, the role of which has not been fully characterized in the context of heat stress in cattle. This study identified single nucleotide polymorphisms (SNPs) in the HSPB6 gene in Sahiwal cattle and their associations with heat tolerance traits (RR, RT and HTC). Three SNPs (SNP 1-3) were reported, which included two transitions, viz. SNP1-g.436G>A (Intron 1) and SNP2-g.2152A>G (3′-UTR) and one transversion, viz. SNP3-g.2417A>T (3′-UTR). The association analysis revealed that SNPs loci, viz. SNP1-g.436G>A and SNP2-g.2152A>G were significantly associated with heat tolerance traits. The GG genotype of SNP2-g.2152A>G was significantly associated with heat tolerance traits in Sahiwal cattle. The association analysis of four available haplotypes, viz. Hap1 (GGA), Hap2 (AAA), Hap3 (GAA), and Hap4 (AAT) of HSPB6 gene with heat tolerance traits did not differ significantly with any haplotype in Sahiwal cattle. This study provides the first association analyses between the SNPs of HSPB6 gene and heat tolerance traits in Sahiwal cattle, which could be used as effective SNP markers in genetic selection for heat tolerance in cattle breeding programs.

An understanding of genetic principles and environment factors affecting the growth traits is needed to implement optimal breeding and selection programs. In this study, total of 18,989 records of body weight were used to estimate genetic parameters of body weight at birth (BW), 3-month (3BW), 6-month (6BW), 9-month (9BW),12-month (12BW), 18-month (18BW), 24-month (24 BW), 30-month (3BW) and 36-month (36BW) of Murrah buffalo at ICAR-NDRI Karnal, Haryana, for the period 1974–2019. The genetic parameters were estimated using average information restricted maximum likelihood (AIREML) procedure by excluding or including maternal effects. Six analytical models were fitted in order to optimize the model for each trait. Most appropriate univariate model was selected based on Log likelihood ratio test (LRT). Influencing factors like calf sex, period of birth, season of birth and dam’s parity were investigated. The results showed that the maternal genetic effects, in addition to direct additive effect are also important determinants of the genetic parameter estimates for growth traits in Murrah buffaloes. Total heritability estimates h2T1 for BW, 3BW, 6BW, 9BW, 12BW, 18BW, 24BW, 30BW and 36BW were 0.25, 0.04, 0.14, 0.16, 0.10, 0.15, 0.21, 0.24, and 0.23 respectively. Maternal effect was important for birth weight and accounted for 13% variation through maternal genetic and 5% variability through maternal permanent environmental effect. Maternal genetic effect was also important and interfered with the estimates of variance ratios in live weight traits owing to large and negative covariance between direct and maternal genetic effects. Direct genetic correlation between body weight traits were positive and high ranging from 0.10 to 0.94. Results revealed that the Murrah herd has a sizable genetic variability for growth traits and hence there is sufficient scope for selection for achieving better growth rate if selection in this direction is applied. Owing to higher positive genetic correlation of 6BW with later ages, the scope of indirect selection for optimum growth in later ages can be aimed at by selecting animals for their higher 6-month live weight.

Heat shock proteins (HSPs) act as a chaperone activity ensuring the folding, unfolding, and refolding of denatured proteins, which help in a protective role during thermal stress in dairy cattle. T...

The aim of this study was to determine the associations of persistency with genetic variants of bovine growth hormone (bGH) gene in Indian Sahiwal cows. The data pertaining to the monthly test day milk yield (MTDMY) and first lactation length (FLL) of 305 lactating Sahiwal cows over a period of 23 years (1993–2016) were subjected to analysis. Persistency of milk yield was estimated by using test day based ratio method and Wilmink b function. Mean LL was 220.06±8.15 days in all studied Sahiwal cows, however, 276.67±10.25 days in cows with ≥100 days of LL and having 500 kg MY. The peak of milk yields was achieved at third TD of the first lactation. Macrophage stimulating protein 1 (MspI) and Arthrobacter luteus1 (AluI) restriction endonucleases were used to digest the amplified regions in intron 3 and exon 5, respectively of bGHgene. The bGHMspI restriction fragment length polymorphism (RFLP) yielded TT, CT and CC genotypes, whereas bGH-AluIRFLP yielded CC, CG and GG genotypes. Significant association was found between first lactation persistency estimated using test day based ratio method and bGH-MspI polymorphism. No significant association was found between first lactation persistency and bGH-AluI polymorphism. In summary, results of present study revealed that the TT genotype of bGH–MspI gene in Sahiwal cows has better lactation persistency than other genotypes.

Lactoferrin is one of the important candidate genes for mastitis resistance in dairy animals. The gene is located on Bos taurus autosome (BTA) 22 and consists of 17 exons spanning over 34.5 kb of genomic DNA. The present study was undertaken to identify allelic variant in exons 6 and 11 of lactoferrin gene and to analyze association with incidence of clinical mastitis in Murrah buffalo. The amplification of exons 6 and 11 of lactoferrin gene yielded 301 and 131 bp amplicon size. Comparison of nucleotide sequence of exonic region of lactoferrin gene with Bos taurus (NCBI accession number AC_000179.1) revealed 6 mutations; among them 3 were in coding DNA sequence and remaining 3 were in flanking intronic region. All these mutations were found in exon 6 and synonymous in nature without affecting the sequence of amino acid. PCR-restriction fragment length polymorphism (RFLP) analysis of 301 bp amplicon using FokI restriction enzyme exhibited polymorphic pattern with two genotypes (AA and AB) with respective frequency of 0.625 and 0.375. The frequencies of two alleles, A and B were estimated as 0.81 and 0.19 respectively. RE Hpy188I and HinfI used for digestion of exon 11 had exhibited monomorphic pattern. The chi-square (χ2) analysis revealed significant association between incidence of clinical mastitis and genetic variant of exon 6 and animals with AA genotype were found to be less susceptible to mastitis. The findings indicate potential scope for incorporation of lactoferrin gene in selection and breeding of Murrah buffaloes for improved genetic resistance to mastitis.

Background: The profitability in the dairy enterprises is strongly affected by udder health and, to a minor extent, by morphometry. The present study was conducted to differentiate mastitis susceptible and resistant animals based on udder and teat type traits infrequently exist. Methods: The study was undertaken in ICAR-NDRI, Karnal of Haryana. A discriminate model was applied to differentiate mastitis susceptible and resistant Karan Fries cows for the study. For meeting the objective; 123 Karan Fries cows were subjected to developing a reliable mathematical function/equation (discriminant functions) for the provision of maximum separation among the mastitis resistant and susceptible animals. The hypothesis was designed to know if these udder and type traits appeal to mastitis-resistant and susceptible animals. The hypothesis for testing the equality of covariance matrices was depicted as: H0: ∑1 = ∑2 Vrs H1: At least one pair of sigma’s ∑ is different. Result: The model included contributory factors such as fore udder length, central ligament, udder balance, shortest distance of floor from fore teat (SDFT), shortest distance of floor from rear teat (SDRT), fore udder attachment (FUA), teat direction, Rear legs rearview, Hock development, udder depth, and udder width constructed and was found to demonstrate 69.1 per cent accuracy with p less than 0.001 and the Karan Fries functions at group centroids are -0.169 and 0.443 respectively for mastitis resistant and mastitis infected animals. The model will facilitate 92.1 per cent of the cases to correctly classify for mastitis resistant animals and the remaining 7.9 per cent being misclassified into the mastitis group.

The main objective of this study was to determine the association of production traits with genetic variants in exon 5 and intron 3 of bovine growth hormone (bGH) gene in Sahiwal cattle. The analyses were based on the detection of single nucleotide polymorphisms (SNPs) in GH-AluI (exon 5) and GH-MspI (intron 3) using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay. The frequency of AluI (L/V) and MspI (T/C) alleles were 86.20/13.70 and 89.80/9.50, respectively. The distribution of the frequency of GH genotypes for LL, LV, and VV were 73.4, 25.5, and 1.0 and for TT, TC, and CC genotypes were 80.9, 17.7, and 13.0, respectively. Season of calving had none but period of calving had significant effect on the studied production traits. The phenotypic data was adjusted for non-genetic factors and regression analysis was done. There was detectable significant effect of the GH-AluI and GH-MspI on the analyzed production traits. The cows with LL genotype had significantly higher milk yield traits than the LV genotype. Similarly, the productive performance of the studied cows with TT genotype was also significantly higher than TC and CC genotypes. Results revealed that the Sahiwal cows with LL and TT genotypes of the bGH locus can be considered to be a favourable genotype for production traits, although these findings need to be confirmed by further research before SNPs can be used in a marker assisted selection program by the animal breeder.

Background: The present study was designed to find out the magnitude of environmental and other non-genetic factors affecting the linear udder and teat type traits in Sahiwal and Karan Fries cows. Methods: Total eight udder morphometric traits and seven teat morphometric traits were measured in 87 Sahiwal and 166 Karan Fries cows. The data regarding milk yield, stage of lactation, parity and season of calving for all Sahiwal and Karan Fries cows were collected. Result: In Karan Fries cows, udder and teat type traits were significantly affected by parity and stage of lactation, while in Sahiwal cow udder and teat type traits were significantly affected by parity only. Teat circumference was significantly affected by season and stage of lactation. The results indicated that parity and stages of lactation were important sources of variation for most of linear udder and teat type traits.

GH has important role in milk production and their polymorphisms may alter quality and quantity of gene's products and affects the phenotype. This study was designed to find the polymorphisms on GH-Msp1 loci and to find its association with milk production traits in Sahiwal cattle. Msp1 polymorphism of the bGH gene was identified in a population of Sahiwal cows. PCR-RFLP genotyping of 329 bp fragment of GH in exon 3 revealed three different allelic variants. The T allele was the major allele in Sahiwal cattle. The TT genotype Sahiwal cows yielded more FL305DMY, FLTMY and also had higher FLL than that of CT and CC genotypes cows. Results of the present findings reveal that TT allele of GH locus should be the favoured genotype in Sahiwal cattle breed and can be used in marker-assisted selection programmes.

Estimates of variance components are needed for implementing genetic selection. This study was conducted to genetic parameters for production and reproductive traits on Indian Karan-Fries cattle using multi-trait repeatability animal model. Data collected from ICAR-National Dairy Research Institute, Karnal, India (from 1988 to 2019) were used. Single-trait and multi-trait repeatability animal models were used for parameter estimation. The posterior mean of Heritability estimates for 305-day milk yield (305-DMY), lactation milk yield (LMY), lactation length (LL) were 0.20 ± 0.03, 0.19 ± 0.03 and 0.06 ± 0.02, respectively. For age at first calving (AFC), calving interval (CI), and days open (DO), the posterior mean of heritability estimates were 0.24 ± 0.08, 0.06 ± 0.01, and 0.07 ± 0.02, respectively. The repeatability estimates for 305-DMY, LMY, LL, CI, and DO were 0.37 ± 0.02, 0.34 ± 0.02, 0.15 ± 0.02, 0.09 ± 0.02, and 0.12 ± 0.02, respectively. Genetic correlation between milk production traits (305-DMY, LMY, and LL) was positive and strong (> 0.80). However, the genetic correlation between milk production trait and AFC ranges from - 0.31 to 0.12. Unfavorable strong genetic correlations were observed between production and reproductive traits (CI and DO) with values ranged from 0.5 to 0.7. Phenotypic correlations among 305-DMY, LMY, and LL were generally positive and high. The moderate heritability estimates and potential genetic variation for 305-DMY, TMY, and AFC suggested that genetic gain can be obtained for these traits through genetic selection. Low heritability estimates found for LL, CI and DO, indicating that the possibility of changing these traits through genetic selection is small. High genetic correlation observed between productive and fertility traits were unfavorable. The existed strong genetic and phenotypic correlation estimates between CI and DO indicates that recording only one of them would be sufficient in the herd. As the multi-trait model showed slight improvements in the h2 as well as r estimates for both productive and reproductive traits over univariate analysis, future selection with a multi-trait animal model applying Bayesian approach would be recommended.

The SPAG11 gene is one of pivotal molecules in reproduction as it takes part in spermatozoa maturation, acquiring motility, capacitation, and egg-sperm interaction as well. The current study was aimed to characterize SPAG11 gene in Indian Murrah bulls through direct DNA sequencing approach. Genomic DNA from Murrah animals were isolated from 130 Murrah bulls and amplified using three sets of forward and reverse primers which were based on reference sequence (Genbank accession no. AC_000164.1) of Bos taurus covering entire coding region of SPAG11B gene. The PCR products of 563, 340 and 373, bp covering exons 1 to 3 were subjected to sequencing and subsequently ClustalW analysis revealed the substitution at 34 positions and a single stretch of 22 bp deletion in comparison to the Bos taurus reference sequence. Total seven novel SNPs were observed as two in the coding region and five in 5ˈUTR. However, only one of SNPs resulted in amino acid substitution viz. p.1279 arginine to tryptophan in translated protein in Murrah buffaloes. Sequence alignment and homology across species for the targeted nucleotide sequence of SPAG11 gene in Murrah bulls was done by nucleotide BLAST (NCBI) that showed maximum identity of 97% with mRNA of Bos taurus and Capra hircus followed by 96% homology with Bos indicus and Bison bison and 95% homology with Ovis aries and Bos mutus.

The present study was undertaken to explore the association of occurrence of clinical mastitis with udder and teat morphometry in cattle. The relationship of nine udder and eight teat morphometric traits with the occurrence of clinical mastitis was analysed using a binomial logistic regression model. A significant association of the udder and teat morphometric traits with the occurrence of mastitis was observed both in Karan Fries and Sahiwal cattle. The rear udder height, rear udder width, udder width and fore udder attachment were found to be the important udder morphometric traits associated with the occurrence of mastitis. In both groups, mastitic animals had broader rear udder width, lower rear udder height and loose fore udder attachment with lengthy and thicker teats. The inclusion of udder and teat morphometric traits in the selection and breeding program of Karan Fries and Sahiwal cattle may help in selection of animals for mastitis resistance.

The study was conducted to understand the effect of season, parity and stage of lactation on productive performance of Sahiwal cattle. The data of productive traits of 133 Sahiwal cattle from 2016 to 2018 maintained at the Livestock Research Center, Karnal, Haryana, were used for this study. The production data were analyzed using least-square procedures based on season, parity and stage of lactation. Results indicated that the parity had significant (p less than 0.05) influence on monthly milk yield (MMY) whereas, the stage of lactation had highly significant (P less than 0.01) influence on monthly milk yield (MMY) and test day milk yield (TDMY). However, there was no significant effect of season on different productive performance traits in Sahiwal cattle. The phenotypic parameter estimates indicated that the parity and stage of lactation of cattle can be used as a selection tool for improvement of productive traits considering its high positive phenotypic correlations with succeeding productive traits in Sahiwal cows.

The present study was conducted to identify polymorphisms in CACNA2D1 gene and their association with clinical mastitis and production traits. Exon 18 and its flanking regions were screened for the presence of SNPs. Statistical analysis was performed to identify association of period of birth, breed, and genotype with mastitis incidence on randomly selected 103 Sahiwal and 102 Karan Fries cattle. PCR-RFLP analysis revealed that g.38819398G A mutation in exon 18 (269 bp amplicon) of CACNA2D1 gene resolved into AA, AG, and GG genotypes in Sahiwal and Karan Fries cattle. Wald chi-square analysis revealed that the period of birth, breed, and genotype were significantly associated with mastitis incidence. GG genotyped cattle were found to be less susceptible to mastitis. Least square analysis revealed that GG and AG genotype animals of G38819398A SNP of CACNA2D1 gene in Sahiwal as well as in Karan Fries cattle were associated with higher average milk yields during 1st, 2nd, and 3rd lactations (P 0.01). These observations and their differential association with the incidence of mastitis and production traits can be utilized as an aid to selection for simultaneous improvement of both antagonistic traits; however, validation of results on large number of animals is warranted.

Heat shock proteins (HSPs) act as molecular chaperones which are preferentially transcribed in response to severe perturbations of the cellular homeostasis such as heat stress. The present study was undertaken for molecular characterization and detection of genetic polymorphisms of HSPB6 gene in 100 Karan Fries Cattle. HSPB6 gene was mapped on Bos taurus autosome 18 (BTA 18), comprising three exons and two introns. Four sets of primers for HSPB6 gene were designed using Primer3 software (version 0.4.0). For detecting single nucleotide polymorphisms (SNPs), sequence data was analyzed using BioEdit software (version 7.2). Comparative sequence analysis of HSPB6 gene showed five nucleotide polymorphisms, which included three transitions viz. g.161A > G, g.436G > A and g.2152A > G and two transversions viz. g.1743C > G and g.2417A > T compared to B. taurus (NCBI GenBank: AC_000175.1). HSPB6 gene of Karan Fries cattle exhibited a high percentage of nucleotide identity (47.0–100.0%) with the corresponding mammalian homologue. The present study indicated a high degree of genetic variability in the HSPB6 gene in the Karan Fries cattle populations.

Leptin an adipokine plays a significant role in several physiological processes and has been indicated as a candidate gene for marker-assisted selection for high-yielding cows. Insulin-like growth factor-1 (IGF-1) hormone plays an important physiological role in growth, development, metabolism, and lactation in bovines. It is believed to be one of the main mediators of energy balance effects on the reproductive performance of dairy cows after calving. The present investigation was carried out to identify the novel polymorphisms in exon 3 region of leptin and exon 3 partial intron 3 of IGF-1 genes and their association with the milk production performance in indicine and taurine crossbred (Karan Fries) cows. Blood samples were collected from 160 apparently healthy Karan Fries (KF) cows. Four SNPs (single-nucleotide polymorphisms) at positions rs29004508 (C > T), rs29004509 (C > T), rs29004510 (T > C), and rs29004511 (T > C) in leptin gene and two SNPs at positions rs133251968 (C > A) and rs137289661 (C > T) in IGF-1 gene were found in KF cows; however, rs29004509 (C > T) had a positive correlation (r = 0.376; P

Objective The present study was aimed at (i) reduction in dimensionality using principal component analysis of 17 linear udder type traits to define those components which best represent udder and teat conformation of Indian Sahiwal cattle and (ii) to identify those components having strongest relationship with milk production traits in Sahiwal cattle. Methods The Principal Component Analysis (PCA) included 17 linear udder type traits using the correlation matrix between the traits to ensure that all traits are standardized in the analysis. Bartlett's sphericity test was used to determine the degree of interrelations between variables and adequacy for use in factor analysis. The relationship of these principal components with milk production traits was analysed using the general linear model. Results The means of linear udder type trait measurements varied between -2.21 for udder balance and 136.35 for udder circumference. Out of 17 principal components, six were extracted using Kaiser Rule criterion. The identified six components could explain 75.777% of the cumulative variance between the linear udder type traits. The communality ranged from 0.476 (rear udder width) to 0.967 (fore teat length) for all these 17 different udder type traits. A relation between first principal component and 305-day milk production was significant with a positive and non-significant effect of 305-day milk-fat yield (305 DF) and 305-day milk-protein yield (305 DP). Results of PCA suggest that the use of orthogonal synthetic variables principal component one (PC1), two (PC2) and three (PC3) provided a means of reduction in the number of linear udder type traits to be recorded in Sahiwal cattle which could be used in explaining the whole udder and teat conformation. The PC1 can be used in selection to explain the mammary system of better milk producing animals to be used in breeding programmes.

Heat shock proteins (HSPs) act as molecular chaperones those are preferentially transcribed in respose to heat stress and the polymorphism in HSP genes associated with heat tolerance traits in cows. HSP90AA1 gene has been mapped on Bos taurus autosome 21 (BTA-21) and spans nearly 5368 bp comprising of 11 exons out of which the first exon does not translate. The present study was done on Karan Fries (5/8 HF × 3/8 Tharparkar) cows reared in tropical climate with the objectives of identifying single-nucleotide polymorphisms (SNPs) in targeted regions (exon 3) of HSP90AA1 gene and analyzing their association with heat tolerance traits in Karan Fries cows. Respiration rate (RR) and rectal temperature (RT) were recorded once daily for four consecutive days during probable extreme hours in different seasons or temperature humidity index (THI), viz., winter, spring, and summer. For detecting single-nucleotide polymorphisms, sequence data were analyzed using BioEdit software (version 7.2). Comparative sequence analysis of HSP90AA1 gene showed point mutation, viz., g.1209AG (exon 3) as compared to Bos taurus (NCBI Ref Seq: AC_000178.1). Association analysis indicated that THI was influenced (P0.01) by RR, RT, and HTC. Similarly, SNPs at locus g.1209AG were categorized into three genotypes, i.e., AA, AG, and GG, and the least squares means (LSMEANS) of RR, RT, and HTC for GG (homozygous) genotype were significantly lower (P0.01) than AA (homozygous) and AG (heterozygous) genotypes. These findings may partly suggest that cows with GG genotypes were favored for heat tolerance trait, which can be used as an aid to selection for thermo-tolerance Karan Fries cows for better adaptation in subtropical and tropical hot climate.

Mastitis is an economically important disease causing great loss to dairy sector owing to its negative effect on production. Its multi factorial etiology makes it difficult to use prophylactic methods for control. Identification of genetic resistance mechanisms and selection for it is a potential strategy for mastitis control. The present study was done in 101 Murrah buffaloes to find the common causative agents of mastitis in buffaloes, to identify variants in exon 2 and 3' UTR of TLR 2 gene and to study their association with intramammary infection. The microbiological examination revealed Staphylococcus aureus to be most common causative organism followed by Streptococcus agalactiae. PCR-RFLP using Sau3AI and HpyCH4V for exon 2 and 3' UTR respectively resulted in polymorphic pattern for exon2, while 3' UTR resulted in monomorphic pattern. PCR RFLP Sau3AI genotypes of TLR2 exon 2 were found to be highly associated with mastitis resistance. AA genotype was found to be least susceptible, while AB genotype was found most frequent among affected buffaloes.

Heat shock protein functions as a molecular chaperone and plays an important role in thermotolerance. The study was undertaken to identify single nucleotide polymorphisms (SNPs) of Hsp90ab1 gene by comparative sequencing and to analyze their association with thermo-physiological parameters, viz. respiration rate (RR), rectal temperature (RT) and heat tolerance coefficient (HTC) in different seasons, viz. winter, spring and summer, at probable extreme hours of the day in Sahiwal and Karan Fries cows. The RR, RT and HTC in summers were significantly (P£0.01) higher than the other two seasons. Polymorphism of Hsp90ab1 gene, evaluated by comparative sequencing revealed six SNPs, viz. T17871421C, C17871485del, C17872061T, T17872112C, T17872148G and A17872199C, two each located in exon 8, intron 10 and exon 11. SNP loci T17871421C and C17871485del were found to be monomorphic for allele C and Deletion (-) respectively in experimental population. Individuals with AA genotype showed significantly (P £0.01) lower RR than AC genotype. While, individuals with CT genotype recorded significantly (P £ 0.01) lower RT than CC genotype. Therefore, it is inferred that propagation of AA and CT genotypes may be an aid to selection and breeding programmes to enhance thermo-tolerance in dairy cattle.

Present investigation was carried out to study the prevalence of bacterial pathogens in clinical and subclinical mastitis in buffaloes during winter season. A total of 118 Murrah buffaloes were screened using California mastitis Test (CMT). Milk samples were collected and analyzed for fat, protein, lactose, SNF, pH, Electrical conductivity (EC) and Somatic Cell Counts (SCC). Out of 118 milk samples, 60 samples (50.84%) were negative for CMT and 58 samples (49.15%) were detected positive for mastitis incidence. Milk pH and EC was significantly higher (p

Calcium channel, voltage-dependent, alpha-2/ delta subunit 1 (CACNA2D1) gene plays an important role in excitation-contraction coupling in muscle-cells, glial cells and neurons. The CACNA2D1 gene may be one of the candidate genes related with some phenotypic traits due to its location in QTLs associated with Somatic cell score (SCS ) and mastitis. A total of 120 Sahiwal cattle were selected to characterize exon 19 of CACNA2D1 gene to identify polymorphism and its association with mastitis susceptibility/resistance. A 249 bp PCR fragment of CACNA2D1 gene encompassing the exon 19 and partial intronic region was amplified and digested with Hae III to screen the SNP which was significantly associated with SCS. Genotype analysis using PCR-RFLP revealed a monomorphic banding pattern. Sequencing was also carried out to explore other SNPs which are deposited in dbSNP in the nucleotide sequence of a particular region. This study provides preliminary information that the targeted region of CACNA2D1 gene in indigenous cattle has no significant association with mastitis resistance which may be a breed specific characteristic.

Present study was done in thirty Karan Fries bulls to characterize the Exon 3 of PROP1 gene and to screen for the H173R polymorphism as well as other variations including reported and novel SNPs in the targeted region. The exon 3 was characterized by sequencing the amplicons obtained after PCR amplification using custom designed primers. The BLAST analysis of the obtained sequence yielded 100% and 99% homology with sequences of Bos taurus and bison respectively. The multiple alignment of the target region sequence with Bos taurus reference sequence revealed that the bulls under the study were free of H173R mutations. No variations were observed thus giving the targeted region a highly conserved one.

Expression of stress genes under thermal stress partially explain the relatively higher thermal adaptability of indigenous breeds compared to exotic breeds of cattle. ATP1A1 gene is one of such gene that encodes for a1 isomer of Na+, K+- ATPase enzyme for Na+- coupled transport of metabolites, nutrients, ions and represents a plausible candidate for heat tolerance traits. Present study was designed to compare SNP variations in ATP1A1 gene and to evaluate their association with respiration rate, rectal temperature and heat tolerance coefficient in Sahiwal (51) and Karan Fries (50) cows maintained at Livestock Research Centre of National Dairy Research Institute, Karnal. Two SNPs (T27008243C and A27008223G) were identified in both the dairy cattle breeds. Association of each SNP genotype was analyzed using Generalized Linear Model procedure in Statistical Analysis System (SAS). Sahiwal cows with TT (15.91±1.89b) at T27008243C locus had lowest RR compared to TC genotypes (18.25±1.77ab) and CC genotype (19.24±1.52a), while in Karan Fries cows RR for AA genotype at A27008223G locus was lower (28.85±1.96b) compared to GG genotype (32.37±2.51a). Thus, the study indicated that the TT genotype at T27008243C locus in Sahiwal and AA genotype at 27008223 locus in Karan Fries cows are desirable genotypes for genetic adaptability under heat stress.

A 37-year old man presented with 5 months history of left scrotal mass and had underwent left orchidectomy following a presumptive diagnosis of testicular tumour. Histopathological diagnosis of testicular tuberculosis was subsequently made, which revealed a case of isolated TB orchitis. It signifies that the careful evaluation of patients with testicular mass is extremely significant for diagnostic accuracy, optimal treatment and avoiding unnecessary surgery in case of testicular tuberculosis. Simultaneously it will also help in early detection of testicular malignancies and vital for the clinicians to avoid negligence charges.

Heat Shock Proteins (HSPs) are group of proteins which are synthesized during heat stress. HSP genes have been reported to be associated with heat tolerance and production performance in cattle. HSP90AA1 gene has been mapped on Bos taurus autosome 21 (BTA 21) and spans nearly 5368 bp comprising 11 exons out of which first exon does not translate. The present study was carried out in Sahiwal cows (n=100) with the objectives to identify SNPs in targeted regions (exon 3, 7, 8 & 11) of HSP90AA1 gene and to analyze their association with heat tolerance traits in Sahiwal cows. Respiration rate (RR) and rectal temperature (RT) was recorded once during probable extreme hours in winter, spring and summer season. Further, heat tolerance coefficient (HTC) also calculated to see the adaptability of the animals during the period of heat stress. On the basis of comparative sequence analysis, total five SNPs were revealed at position of A1209G, A3292C, T4935C, T5218C and A5224C in the targeted region of HSP90AA1 gene. Out of these, only two SNPs at A1209G and A3292C loci were found significantly associated with heat tolerance traits in Sahiwal cows. Whereas, THI has a highly significant associated with RR, RT and HTC in all the seasons. At A1209G locus in Sahiwal cows for traits RR, genotype AA (18.40±0.46a), AG (19.60±0.85b) and GG (21.18± 0.64ab) and trait HTC AA (1.78±0.04a), AG (1.85±0.03b) and GG (1.91± 0.02c) differ significantly (p

The objective of this study was to characterize and validate the candidate point mutation in Calcium channel, voltage-dependent, alpha-2/ delta subunit 1 (CACNA2D1) gene in Karan Fries (Bos taurus x Bos indicus) cattle. The CACNA2D1 gene reported as one of the potential candidate gene influencing Somatic cell Score and Mastitis. A PCR product of 249 bp amplifying the exon 19 and partial 18 and 19 intronic region of CACNA2D1 gene was digested with Hae III restriction enzyme to screen the reported point mutation. A monomorphic banding pattern with genotype AA was found in Karan Fries cattle. Sequencing was also carried out to characterize and explore insilico screened mutation in the nucleotide sequence of a particular region. The result indicates highly conserved sequence in Karan Fries cattle. The Phylogenetic tree revealed that Karan Fries cattle were closer to Bos taurus cattle, Bos mutus (Yak), and Bison bison (American buffalo) compared to other species.

Heat Shock Proteins (HSPs) are expressed in response to heat stress and the polymorphism in HSP genes at single nucleotide level is reported to be associated with heat tolerance and production performance traits in cattle. ATP1B2 gene was mapped to Bos taurus autosome 19 (BTA 19) spanning nearly 4310bp, comprising 7 Exons and 6 Introns. The present study was conducted in Sahiwal cows (n=108) raised under sub-tropical climate with the objectives to identify SNPs in targeted regions (Intron 2,4 and Exon 3,5) of ATP1B2 gene and to analyze their association with heat tolerance traits in Sahiwal cows. Respiration rate (RR) and rectal temperature (RT) were recorded during probable extreme hours in winter, spring and summer seasons. Heat tolerance coefficient (HTC) was also calculated to check the adaptability of the animals during the period of heat stress. The comparative sequence analysis revealed a total of two SNPs in Intron 2 of ATP1B2 gene i.e. g.2243G>A and g.2366T>C. Out of these two identified SNPs, only one SNP i.e. g.2243G>A was found to be significantly (PC, RR, RT and HTC were having non-significant association with the different genotypes i.e. TT and TC. These findings may partly suggest that AA and GA genotype of SNP g.2243G>A of ATP1B2 gene can be utilized as potential marker for propagating thermo-tolerant cattle.

The present study was undertaken with the objectives of molecular characterization and detection of genetic polymorphisms in the HSPB6 gene of Sahiwal cattle. A total of 100 Sahiwal cattle maintained at LRC, NDRI, Karnal (Haryana) were included in the study. HSPB6 gene has been mapped on Bos taurus autosome 18 (BTA 18) and spans nearly 2620 bp comprising 3 exons and 2 introns. Four sets of forward and reverse gene-specific oligonucleotide primers of HSPB6 gene were designed using Primer3 software and amplicon size of 529 bp, 460 bp, 670 bp and 502 bp were ascertained. On the basis of comparative sequence analysis, total three polymorphic loci (SNPs) were detected at locus G436A, A2152G and A2417T in HSPB6 gene as compare to Bos taurus (NCBI GenBank AC_000175.1). Out of these three SNPs, two (A2152G & A2417T) were found in 3′-UTR region and third one (G436A) was detected in intronic region 1. However, monomorphic pattern was found in exon 2 of HSPB6 gene in Sahiwal cattle. Further research need be directed to find out the association of allelic variants in major heat shock protein genes to establish markers for the future genetic improvement of livestock through marker assisted selection.

Heat shock proteins (HSPs) are expressed in response to heat stress, and the polymorphism in HSP genes at single-nucleotide level has been reported to be associated with heat tolerance and production performance traits in cattle. HSPB8 gene has been mapped on Bos taurus autosome 17 (BTA-17) spanning nearly 13,252 bp and comprising three exons and two introns. The present study was conducted in Sahiwal cows (n = 108) reared in subtropical climate with the objectives to identify SNPs in all three exons and part of intron 1 of HSPB8 gene and to analyze their association with heat tolerance traits in Sahiwal cows. Respiration rate (RR) and rectal temperature (RT) were recorded once during probable extreme hours in different seasons or Temperature-Humidity Index (THI), i.e., winter, spring, and summer. Heat tolerance coefficient (HTC) was also calculated to check the adaptability of the animals during the period of heat stress. The comparative sequence analysis revealed a total two single-nucleotide polymorphisms (SNPs), i.e., g.507GA in exon 1 and g.881TC in intron 1 of HSPB8 gene. Out of these two identified SNPs, only one SNP, i.e., g.507GA, was found to be significantly associated with heat tolerance indicator traits (RR, RT, and HTC) in Sahiwal cows. The perusal of results across different seasons showed the significant (P 0.01) difference in RR, RT, and HTC between winter, spring, and summer seasons. RR, RT, and HTC were found to be significantly lower (P 0.01) in GA as compared to GG genotype of g.507GA SNP of HSPB8 gene. However, in case of another SNP, i.e., g.881TC, located on intron 1, the RR, RT, and HTC were having non-significant association with the different genotypes, i.e., TT and TC. These findings may partly suggest that GA genotype of SNP g.507GA of HSPB8 gene has a probable role in heat tolerance in Sahiwal cattle and can therefore be utilized as a marker in propagation of thermo-tolerance cattle in hot tropical and subtropical climate. Nevertheless, the involvement of other regulatory mechanisms cannot be overruled.

Lactoferrin gene is one of the important candidate genes for mastitis resistance. The gene is located on chromosome BTA 22 and consists of 17 exons spanning over 34.5 kb of genomic DNA. The present study was undertaken with the objectives to identify allelic variants in exons 7 and 12 of lactoferrin gene and to analyze association between its genetic variants and incidence of clinical mastitis in Murrah buffalo. The amplification of exons 7 and 12 of lactoferrin gene yielded amplicons of 232- and 461-bp sizes. PCR-restriction fragment length polymorphism (RFLP) analysis of 232-bp amplicon using BccI restriction enzyme revealed three genotypes (AA, AB, and BB) with frequencies of 0.62, 0.22, and 0.16, respectively. The frequencies of two alleles, A and B, were estimated as 0.73 and 0.27. Hpy188I-RFLP for 461-bp amplicon revealed polymorphism with three genotypes, CC, CD, and DD, with respective frequencies of 0.06, 0.39, and 0.56, whereas frequencies for C and D alleles were 0.25 and 0.75. The chi-square (χ(2)) analysis revealed a significant association between incidence of clinical mastitis and genetic variants of exon 7, and animals of AA genotype of exon 7 were found to be least susceptible to mastitis. The findings indicate potential scope for incorporation of lactoferrin gene in selection and breeding of Murrah buffaloes for improved genetic resistance to mastitis.

Prevalence of several diseases especially mastitis in dairy animals causes reduction in milk production leading to heavy economic loss to the dairy farmers. Lactoferrin (Lf) is a minor whey non-heme iron binding protein produced by the mammary gland may serve a dual role, protecting both the mammary gland and the neonatal intestine from infection. The characterization of lactoferrin gene promoter region has been described in Bos taurus cattle, but there are no reports available in Bos indicus cattle which are widely distributed in Indian sub-continent. About 1kb sequence of lactoferrin gene was generated and the BLAST analysis revealed 99% sequence homology with the exotic cattle sequence. The phylogenetic analysis showed clustering of indicus and taurine cattle together in a single node indicating 5’ regulatory region of lactoferrin gene is highly conserved in these two sub-species. A total of 13 putative transcription factor binding sites were indentified. Although it seems unlikely that all the putative transcription factors identified in the promoter region exert their individual effects, some may actually influence the regulation of bovine lactoferrin gene expression in various tissues especially in mammary gland which need to be further assessed for their functional significance.

Cluster of Differentiation (CD14) protein was identified as a differentiation marker on the surface of monocytes and macrophages and was characterized as a receptor for bacterial endotoxin (LPS). It is regarded as the first described pattern-recognition receptor (PRR) and one of the excellent candidates for mastitis resistance in cattle. Present study was carried out with the objective to characterize and identify genetic polymorphism of promoter region of CD14 gene and its association with clinical mastitis in lactating Karan Fries cattle maintained at NDRI Karnal. Twelve SNPs were found in the complete sequence of 553 base pairs of promoter region of CD14 gene of Karan Fries cows. Cows were also screened using PCR-RFLP with Hpy188I restriction enzyme which revealed three genotypes AA, AB and BB. All three genotypes differed significantly regarding mastitis incidence.

The data on body weights of 780 Sahiwal animals from birth to first calving for a period of 47 years (1961- 2007) were adjusted for significant non-genetic factors (season and period) using least squares analysis and the adjusted records were used for subsequent analysis. The season of birth had no significant effect on birth weight, body weights at 24 months age, 30 months age and weight at first calving. Body weight at 12 months of age was significantly influenced (P £ 0.05) by season of birth and the effect of season was highly significant (P £ 0.01) on 6 and 18 months body weights. The period of birth had highly significant (P £ 0.01) influence on all body weights from birth to first calving. The least squares analysis of variance showed the highly significant (P £ 0.01) regression of body weights at 6, 12, 18, 24 and 30 months of age and weight at first calving on birth weight in Sahiwal cattle. The heritability estimates were: 0.13 ± 0.06, 0.15 ± 0.09, 0.13 ± 0.08, 0.11 ± 0.08, 0.27 ± 0.11, 0.31 ± 0.12 and 0.19 ± 0.10 for body weights at birth, 6, 12, 18, 24, 30 and at first calving. The heritability estimate of birth weight was moderate and significant for Sahiwal calves. The estimates of heritability of body weights at six, twelve and eighteen months were not significantly different from zero as standard errors were higher. However, the heritability estimates of twenty four and thirty months body weights were higher and significant in Sahiwal heifers. The estimates of genetic correlations of 6 months body weight with 12 months body weight, 24 months body weight with 30 months body weight and body weight at first calving and 30 months body weight with body weight at first calving were higher in magnitude. Most of the body weight traits showed highly significant (P £ 0.01) phenotypic correlation amongst themselves. However, non-significant phenotypic correlations were observed in case of birth weight with six months body weight and weight at first calving.

Cluster of diffferention-14 (CD14) gene is involved in innate immunity and has been identified as one of the candidate gene for disease resistance. CD14 molecule works as cell surface receptor present on surface of inflammatory cells. Bovine CD14 gene spans over 2630 bp having 2 exons. Present study is based on identification of DNA polymorphism and to analyze the association between CD14 gene variants with incidence of mastitis in Sahiwal cattle. A total of 100 lactating Sahiwal cattle maintained at NDRI, Karnal (Haryana) were included in the present study. PCR-RFLP analysis of contig 4 using HinfI restriction enzyme showed three different genotypic patterns (CC, CD and DD). Remaining contig-enzyme combinations showed monomorphic pattern. Genotypic frequency of CC genotype was found highest (54%). Chi-square (χ2) analysis revealed that there is significant difference between these genotypes regarding mastitis incidence. CC genotyped animals were found to be less susceptible for mastitis.