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ThesisItem Open Access A COMPARATIVE STUDY ON CYTOGENETIC PROFILE OF CROSSBRED AND INDIGENOUS PIGS(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2017-02) HARSHINI, V; SAKUNTHALA DEVI, K(MAJOR); PUNYA KUMARI, B; SURESH, JABSTRACT: The present analysis was carried out on 30 blood samples of LWY crossbred pigs (15 females and 15 males) maintained at All India Coordinated Research Project on pig (AICRP), College of Veterinary Science, Tirupati, Chittor District of Andhra Pradesh and on 30 blood samples of indigenous pigs (15 females and 15 males) maintained by the farmers in and around Tirupati to study the cytogenetic profile of both the breeds and their morphometric measurements using short term leukocyte culture technique. The diploid chromosome number (2n) in both the breeds was 38 with XX and XY complement in females and males respectively and a fundamental number of 64 as in exotic pigs. Among 18 pairs of autosomes, first 5 pairs were sub-metacentric followed by 6th and 7th pair as sub-telocentric, next 5 pairs (8th to 12th pairs) were metacentric and remaining six pairs (13th to 18th) were telocentric. Among these one pair of allosomes i.e. X-chromosome was large metacentric, while Y-chromosome was metacentric and smallest chromosome of the karyotype. The least squares mean relative length of autosomes in LWY crossbred pigs ranged from 2.79±0.02 to 10.83±0.05 in females and 2.81±0.05 to 10.87±0.07 in males. The least squares mean relative length values for non-descript pigs ranged from 2.84±0.03 to 11.01±0.07 in females and 2.86±0.03 to 11.04±0.05 in males. The relative contribution of X-chromosome in LWY crossbred female and male pig was 4.67±0.01 and 4.65±0.04, whereas the values were 4.82±0.03 and 4.80±0.04 in non-descript pigs respectively. The mean relative contribution of Y-chromosome were 2.05±0.10 and 2.19±0.13 in LWY crossbred and non-descript pigs respectively. Genetic group had significant effect on the relative length of all autosomes and allosomes except 17th and 18th chromosomes, whereas sex had no significant effect on relative length of all chromosomes in both the breeds. The overall mean arm ratio of 12 pairs of autosomes ranged from 1.01±0.03 to 2.45±0.03 and 1.02±0.01 to 2.43±0.02 in LWY crossbred females and males respectively, whereas these values varied from 1.03±0.02 to 2.51±0.06 in females and 1.04±0.06 to 2.52±0.03 in males of non-descript pigs. The mean arm ratio values of X-chromosome was 1.15±0.04 in females and 1.13±0.03 in males of LWY crossbred pigs, while the respective mean values in non-descript pigs were 1.14±0.03 and 1.12±0.03 respectively. The arm ratio values of Y-chromosome were 1.17±0.02 and 1.18±0.02 in LWY crossbred and non-descript pigs. The mean centromeric indices of autosomes (1 to 12 pairs) differed from 0.50±0.04 to 0.71±0.02 and 0.52±0.03 to 0.72±0.03 in LWY crossbred female and male pigs, whereas it ranged from 0.51±0.01 to 0.82±0.01 and 0.53±0.05 to 0.83±0.02 respectively in non-descript pigs. The X-chromosome had mean centromeric index values of 0.53±0.03 and 0.52±0.03 in LWY crossbred female and male pigs, while the value for Y-chromosome was 0.56±0.02. The mean centromeric index values of X-chromosome in non-descript pigs were 0.52±0.02 and 0.54±0.02 in females and males, whereas for Y-chromosome, it was recorded as 0.55±0.01 respectively. The average morphological index value of autosomes ranged from 2.43±0.06 to 7.29±0.11 in females and 2.41±0.05 to 7.31±0.12 in males of LWY crossbred pigs, while X-chromosome had 5.85±0.05 and 5.84±0.05 respectively. The mean values for autosomes ranged in non-descript pigs were 2.42±0.02 to 7.96±0.14 and 2.44±0.04 to 7.98±0.13 in females and males, whereas X-chromosome had an index value 6.01±0.02 and 5.99±0.03 respectively. The values for Y-chromosome were 2.01±0.02 and 2.09±0.01 in LWY crossbred and non-descript pigs. In the present investigation with 128 metaphase spreads, a total of 311 NORs were detected in LWY crossbred and non-descript pigs. The mean number of NORs per metaphase was 2.67 and 2.20 in both breeds respectively. The highest frequency of metaphases (47.61% and 81.54%) had 2 number of NORs, whereas lowest frequency of metaphases (7.94% and 3.07%) had 4 number of NORs. The percentage share of silver deposits on 10th pair of chromosome accounts for 96.67 and 68.33 in LWY crossbred and non-descript pigs, whereas on 8th chromosome pair the percentage share of deposits were 44.16 and 51.67 respectively. The outcome of present study have revealed that the morphology, number and various morphometric measurements of the chromosomes in both the breeds were analogous to those reported in literature for various pig breeds like Canadian Lacombe, Landrace, Duroc, Thai wild boar, Nigerian indigenous pig, Niang Megha pig, Korean native pig, Polish Landrace, Hampshire, Large White Yorkshire, Turkish sus scrofa. Therefore, the efforts should be undertaken to recognize the non-descript pigs as a distinct breed of Andhra Pradesh.ThesisItem Open Access A STUDY ON MORPHOLOGICAL AND GENETIC VARIATION IN NELLORE PALLA SHEEP(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2017-01) SAI HARINI, K; Punya kumara, B(MAJOR); Vinoo, R; Suresh Babu, DABSTRACT: The southern peninsular region of India is home to fourteen descriptive mutton breeds, of which Nellore is the country's tallest sheep, predominantly distributed in areas of Nellore and Prakasam districts of Andhra Pradesh. Based on coat colour three varieties were seen: Jodipi, Brown (Dora) and Palla. Nellore Jodipi is predominant in its population followed by Brown and Palla. Nellore palla sheep are confined to only certain areas of Nellore district and very scanty information is available on the performance of the breed. The population of Nellore Palla is under threat (<1500). Hence, genetic and phenotypic evaluation of the breed is important not only from conservation point of view, but also to assess the level of inbreeding in the population. The present study was carried out to figure out productive and reproductive performance in Nellore Palla sheep under field conditions and also to assess the genetic structure of Nellore Palla sheep at molecular level using twenty four microsatellite markers recommended by ISAG/FAO for genetic diversity studies in sheep. A total of 347 sheep of different age groups were used to study the morphometric measurements of Nellore palla sheep and 50 blood samples from unrelated sheep flocks were used for molecular studies. The overall least squares means of body weights (kg), height at withers, chest girth, paunch girth and body length were 35.18 ± 0.27, 78.47 ± 1.55, 80.82 ± 0.66, 80.93 ± 1.37 and 69.90 ± 0.83 at 2- teeth age, 39.65 ± 0.33, 82.45 ± 0.98, 84.57 ± 0.72, 84.11 ± 1.34 and 72.23 ± 1.02 at 4- teeth age, 41.18 ± 0.56, 83.63 ± 0.64, 86.32 ± 0.74, 85.88 ± 1.23 and 74.55 ± 0.81 at 6- teeth age, 45.00 ± 0.46, 85.22 ± 1.97, 87.17 ± 0.58, 86.14 ± 0.52 and 76.17 ± 0.33 at 8- teeth of age respectively. Villages and sex showed a significant (p<0.01) effect at all ages and the parameters studied. The overall least squares means of age at first mating in males, age at first mating in females, age at first lambing, tupping and lambing percentages were 598.03 ± 3.02 days, 361.54 ± 1.09 days, 509.32 ± 1.00 days, 81.06 ± 1.61 and 73.37 ± 1.53 percent, respectively. The coefficient of correlations between body weights with the height at withers, chest girth, paunch girth and body length varied from 0.58(PG) to 1.00(CG) in males and 0.16(PG) to 0.69(HW) in females. A perfect phonotypic correlation (1.00) was observed for body weights and height at withers at in 8- teeth age males and for body weights and chest girth for males at 6- teeth age. All the correlations were positive and highly significant (p<0.01) in majority of the age groups studied. The standard Phenol-Chloroform method was used for isolation of DNA from 50 blood samples. The quality and quantity of DNA was checked by agarose gel electrophoresis and Nanodrop spectrophotometer, respectively. The allele frequencies, observed number of alleles (Na), effective number of alleles (Ne), observed (Ho) and expected (He) heterozygosity, within population inbreeding estimates (FIS) were computed using POPGENE32 software. A total of 189 alleles were found across the 24 microsatellite loci studied. The number of alleles at each locus ranged from five (BM6506, CSSM31 and HUJ616) to a maximum of thirteen (BM827 and OARHH64) with a mean of 7.8 alleles across all loci. The allele sizes ranged from a minimum of 81 bp (OarJMP8) to a maximum of 258 bp in MAF214, while, the allele frequencies ranged from a minimum of 0.010 (BM827, INRA63, OarFCB48 and OarHH64) to a maximum of 0.40 (OarFCB128). The mean effective number of alleles was 6.6452 ± 0.3933 which was less than the observed number of alleles and ranged from 3.4916 (CSSM31) to 11.574 (BM827). The observed heterozygosity (Ho) and expected heterozygosity (He) ranged from 0.00 (BM1314, BM6506, BM8125, HUJ616, OARCP20, OARFCB128, OARJMP8) to 0.16 (OARHH64) and from 0.7208 (CSSM31) to 0.9228 (BM827) with an overall value of 0.0626 ± 0.010 and 0.8361 ± 0.009, respectively. All the twenty four microsatellite loci were found to be highly polymorphic and the PIC values ranged from 0.6616 (CSSM31) to 0.9136 (BM827) with a mean of 0.8171. The FIS values were found positive and ranged from 0.8201 (OARHH64) to 1.000 (BM1314, BM6506, BM8125, HUJ616, OARCP20, OARFCB128, OARJMP8). The mean FIS value across the twenty four loci observed was 0.9257. The χ2 test revealed that all the twenty four loci deviated significantly from Hardy- Weinberg Equilibrium.ThesisItem Open Access ASSESSMENT OF GENETIC DIVERSITY IN ONGOLE AND PUNGANUR CATTLE BREEDS THROUGH MICROSATELLITE TYPING(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2013-04) ASHA, UPPUTURI; Sakunthala Devi, K(MAJOR); Rajkumar, U; Harikrishna, ChABSTRACT: Twenty dinucleotide microsatellite markers viz ETHOLO, ILSTSOOS, TGLA122, INRA035, INRA063, HEL001, TGLA 126, INRAOOS, ILSTS006, INRA032, ETH225, CSRM060, BMI 824, TGLA053, INRAO37, ETHO03, TGLA227, MM012, HAUT024 and HAUT027 were used for Assessment of genetic diversity in Ongole and Punganur cattle breeds through Microsatellite typing. The mean quantity of DNA was 2.96 pdml in Ongole and 3.16 pg/ml in Punganur cattle. Mean optical absorbance ratio (2601280nm) was 1.78 in both Ongole and Punganur cattle indicating good quality of genomic DNA. Out of the total 216 alleles detected, 98 alleles were specific to Ongole and 75 alleles were specific to Punganur. Mean number of alleles obtained at each locus varied from 1 to 12 in Ongole and 1 to 13 in Punganur cattle. The overall mean effective number of alleles was found to be 4.09 in Ongole and 3.41 in Punganur cattle. The overall mean expected heterozygosity and observed heterozygosity were 0.65 and 0.33 in Ongole and 0.55 and 0.3 1 in Punganur breed, respectively. The overall mean PIC values observed were 0.62 and 0.53 in Ongole and Punganur breeds respectively. The mean inbreeding coefficient (Fls) obtained in the present study was 0.456 for Ongole and 0.434 in Punganur breed. F-statistics revealed moderate inbreeding within the breeds and existence of moderate relationship between the Ongole and Punganur cattle. In Ongole population, loci ETHO10, TGLA122, INRA035, INRA063, HEL001, ILSTS006, TGLA126, ETH225, CSRM60, TGLA053, INRA037, ETH003, MM012, HAUT027 and HAUT024 were deviated significantly from the equilibrium frequency (I'c0.01). In Punganur breed, except the locus TGLA126, ETH225 and CSRMO60, all other loci deviated significantly (Ps0.01) from the Hardy-Weinberg equilibrium.ThesisItem Open Access ANALYSIS OF POLYMORPHISM AT THE TNF-α AND PGR LOCI AND THEIR ASSOCIATION WITH REPRODUCTIVE TRAITS IN ONGOLE CATTLE(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2018-01) NAGA SHIVA KUMAR, KONAPALLI; MURALIDHAR, M(MAJOR); SUDHAKAR, K; VENKATA NAIDU, GABSTRACT: The present study was aimed at understanding genetic polymorphism at two loci namely Tumour Necrosis Factor-α (TNF-) and Progesterone receptor (PGR) genes in Ongole cattle using PCR-RFLP method. Two Single Nucleotide Polymorphisms (SNPs) were analysed, one for each locus. Primers were designed to amplify the targeted SNP of both the genes. DNA samples from a total of 150 Ongole cattle breed were used in the present study. The amplicons of TNF-α were subjected to RFLP with RsaI restriction enzyme and the PGR amplicons with DraIII enzyme. Based on RFLP profile obtained, the genotypic and allelic frequencies were determined for the targeted SNP. In the present study the PGR gene was monomorphic in all the samples. The TNF-α gene was polymorphic, only two genotypes G/G and G/A were observed. No individual in the present study was homozygous for the allele A at this locus. The G/G genotype is predominant in Ongole population with a frequency of 0.74 followed by G/A genotype with 0.36. The major allele in this locus in Ongole population is G and the minor allele is A with a frequency of 0.13. The Ongole population at this SNP locus is under equilibrium in the present study. Association analysis with the phenotypes that indicate reproductive performance parameters namely, calving interval, service period and dry period with the available data indicated no significant association between the genotype and any of these traits. Further studies with more information are required to understand precise role of the SNP in gene regulation and functionThesisItem Open Access GENETIC DIVERGENCE OF VIZIANAGARAM AND MACHERLA BROWN SHEEP WITH NELLORE SHEEP: A STUDY(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2018-01) TEJASWANI, VENNELA; VINOO, R(MAJOR); SUDHAKAR, K; VENKATA SESHAIAH, ChABSTRACT : In India, most of the small and marginal farmers depend on sheep for their livelihood for which reason, they are considered as an important small ruminant. Several sheep breeds like Nellore and Deccani have been reared in different places of Andhra Pradesh for the past few decades and showing marginal gain to the shepherd community. Apart from these two characterized breeds, there are two other local genetic groups viz. Vizianagaram and Macherla Brown sheep reared by local shepherds. The Vizianagaram sheep are distributed in the North coastal region of Andhra Pradesh whereas Macherla Brown sheep are distributed in the villages adjacent to Krishna river in Guntur, Prakasam and Krishna districts. These two genetic groups of sheep are yet to be characterized and the genetic distances with other breeds is to be established. Phenotypic and production performances indicate that these two genetic groups are different from Nellore sheep. The objective of the present research is to study the genetic variation in the three genetic groups of Andhra Pradesh and their phylogenetic status using mitochondrial DNA control region. Blood samples were collected from six Macherla and Vizianagaram sheep respectively. Genomic DNA was isolated and 885 bp mitochondrial control region was amplified and sequenced. Metadata of Nellore and other Indian breeds were obtained from NCBI database and total of 341 sequences were aligned and trimmed to have a final length of 721 bp. The measures like polymorphism, divergence, gene flow and genetic differentiation were estimated using DnaSP software. A phylogenetic tree was constructed using Neighbour joining method by MEGA software. The tree topology was tested using 1000 bootstrap replications. The results of polymorphism and divergence analysis disclosed the existence of polymorphism in three genetic groups of Andhra Pradesh and these genetic groups were distinct. The findings of gene flow and genetic differentiation estimates concluded the presence of high differentiation and moderate sub-structuring of the three genetic groups in Andhra Pradesh similar to other sheep breeds in India. The phylogenetic analysis indicated that, all the haplotypes of the two genetic groups of sheep (Vizianagaram and Macherla) under present study are in lineage A along with majority of the other Indian sheep breeds. In Nellore sheep, out of 28 haplotypes, 26 were grouped in lineage A and 2 were grouped in lineage B. This separation could be due to the existing hypothesis that there may be two independent domestication events in India.ThesisItem Open Access IDENTIFICATION OF BOVINE TUBERCULOSIS ASSOCIATED PUTATIVE SNP IN ONGOLE CATTLE AND MURRAH BUFFALO(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2018-01) SUSHMA DEVI, POSIMSETTI; SUDHAKAR, K(MAJOR); VINOO, R; SUBRAMANYAM, K.VABSTRACT: Bovine Tuberculosis is a chronic infectious respiratory disease caused by Mycobacterium bovis. The conventional preventive and control measures are not so effective in combating the disease, hence alternate strategies like selective breeding of animals resistant to the disease by exploiting the natural variation of the genes underlying the disease resistance trait to the disease could be considered. The present study is aimed at identifying those genes and SNPs to be associated with bTB susceptibility / resistance and exploiting the genetic variations in the gene regions by using PCR-RFLP method. Two genes (CCL4 and RNF185) were identified to be promising candidate genes in association with bTB and their SNPs that create restriction enzyme sites were considered for the present study. DNA was isolated from blood samples of 128 Ongole cattle and 155 Murrah buffaloes by modified high salt method. Two sets of primers were designed so as to capture the SNP regions of the genes. The PCR products were subjected to RFLP with BtsαI and TseI enzymes for CCL4 and RNF185 genes respectively. In the population under study, both the genes were found to be monomorphic. The fixation of allele ‘A’ is revealed in CCL4 gene with the absence of ‘T’ allele while in RNF185 gene, ‘A’ allele is fixed with absence of ‘G’ allele. Both the loci were found to be monomorphic in both cattle and buffalo.ThesisItem Open Access AMPLIFICATION, SEQUENCING AND BIOINFORMATIC CHARACTERIZATION OF TRANSMEMBRANE PROTEIN 95 GENE (TMEM95) IN MURRAH BUFFALO (Bubalus bubalis)(2017-01) SHIREESHA, SAMPATHIRAO; SUDHAKAR, K(MAJOR); VINOO, R; VENKATA SESHAIAH, ChABSTRACT: TMEM95 gene is known to be involved in idiopathic male subfertility in cattle. It is required for the integrity of sperm plasma membrane and thus has a role in fertilization. The present study was aimed at bioinformatic characterization of the Transmembrane protein 95 (TMEM95) gene in Murrah buffalo. The TMEM95 gene was amplified and sequenced in Murrah. Analysis with CodonCode Aligner revealed three heterozygous positions in Murrah sequence at positions 1284 (T and G), 1460 (C and A) and 1897 (G and A) with respect to the 2631 bp reference fragment from the Hereford cattle genome assembly. A 2 bp deletion (at 937 bp) was observed in Murrah buffalo which is causing frame shift mutation. The isoforms were predicted using GeneWise, a comparative gene annotation tool and found that the 2bp deletion resulted in the truncation of isoforms 1, 3 and 4 and were unlikely to form. The transmembrane topology and signal peptide were predicted using Phobius program, and it was observed that the isoforms 1, 2 and 3 doesn’t have any transmembrane domains but the isoform 5 has two transmembrane domains. Analysis of isoforms with MotifScan predicted the presence of Casein kinase II phosphorylation site, which has an important role in sperm morphology. Leucine zipper pattern, N-myristoylation site, protein kinase C phosphorylation site, CHRD domain profile, N-glycosylation site and HIT zinc finger motifs were also predicted. Divergence analysis was carried out for the TMEM95 region across different mammalian species and less divergence was observed among cattle, bison, yak and mithun. Buffalo and sheep are moderately divergent from cattle. Based on the sequence analysis and functional prediction, it was concluded that the TMEM95 gene in Murrah buffalo is likely to have function in male fertility. However there may be some species-specific differences with respect to their function between the two speciesThesisItem Open Access A STUDY ON GENETIC POLYMORPHISM OF MYOSTATIN (GDF8) GENE IN NELLORE AND MACHERLA BROWN SHEEP(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2017-01) VENKATA PRANEETH, D; Vinoo, R(MAJOR); Muralidhar, M; Jagadeeswara Rao, SABSTRACT : The primary objective in small animal production is to have higher meat production. The present study is aimed at understanding natural variation of GDF8 locus in Nellore and Macherla Brown sheep genetic groups by using PCR-RFLP. DNA was isolated from 100 sheep blood samples using a modified high salt method. Two sets of primers were designed to amplify regions of exon 3 and intron 1 of GDF8 gene. A total of 100 blood samples from the three sheep populations were used for amplification of exon 3 and 65 samples were used for amplification of intron 1. The PCR products were subjected to RFLP with HaeIII restriction enzyme for exon 3 and HpyCH4V restriction enzyme for intron 1. Based on different genotypes obtained the genotypic and allelic frequencies were determined. In the present study exon 3 is monomorphic in all the samples and hence, further analysis could not be performed on this locus. The intron 1 region was polymorphic representing HH, Hh, hh genotypes. The heterozygosity values for intron 1 region were 0.48, 0.49 and 0.47 for Nellore Jodipi, Nellore Brown and Macherla Brown genetic groups respectively. The PIC values for Nellore Jodipi, Nellore Brown and Macherla Brown are 0.37, 0.37 and 0.36 respectively, suggesting considerable amount of variation exist in these populations. The allelic frequencies of Nellore Jodipi, Nellore Brown and Macherla Brown were 0.59 and 0.41, 0.56 and 0.44, 0.62 and 0.38 for H and h alleles, respectively. Diversity estimates (FIS) were negative for the three populations indicating that there is no differentiation among the three populations. The test for Hardy Weinberg Equilibrium indicated that the three populations are departing from the equilibrium assumptions. No significant (P>0.05) association of a genotype with body weights at different ages was observed in the present study in both the genetic groups of Nellore Jodipi and Nellore Brown. A new PCR-RFLP marker is designed for intron 1 in this study which is found to be polymorphic and useful in population studies.ThesisItem Open Access STUDIES ON GENETIC VARIABILITY IN NELLORE BROWN SHEEP USING MICROSATELLITE MARKERS(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2015) SHEELA MANJARI JONNA KUTTI, JONNA KUTTI; PUNYA KUMARI, B(MAJOR)ABSTRACT: Nellore sheep is the tallest mutton breed of India and its home tract is Nellore and Prakasam districts of Andhra Pradesh, but widely distributed throughout the state of Andhra Pradesh and parts of Telangana. Three varieties are distinguished phenotypically based on coat colour as Palla (completely white), Jodipi (white with black patches) and Dora/Brown (completely brown). Nellore Brown sheep are predominantly present in Rayalseema region (Kadapa, Kurnool, Anantapur districts) of Andhra Pradesh. This breed is well known for its disease resistance and thrives well in harsh climatic conditions of Rayalseema. The assessment of genetic variability is important to monitor the gene flow in populations, for conservation of breed and also to determine the level of inbreeding in the population. The present study was undertaken to evaluate the genetic variability present in Nellore Brown sheep population by using twenty four microsatellite markers from the panel of markers recommended by ISAG-FAO for the genetic diversity studies in sheep. A total of fifty blood samples were collected at random from unrelated Nellore Brown animals maintained by farmers in its breeding tract. Phenol-chloroform method was used for isolation of DNA from blood samples. The quality and quantity of isolated DNA was checked using gel electrophoresis and Nanodrop spectrophotometer, respectively. The PCR products were resolved using agarose gel electrophoresis. The allele size for each locus was scored manually from the electrophoresed gels. The allele frequencies, observed number of alleles (Na), effective number of alleles (Ne), observed (Ho) and expected (He) heterozygosity, within population inbreeding estimates (FIS) were computed using POPGENE version 1.32 software. A total of 216 alleles were identified across all the twenty four microsatellite loci studied. The number of alleles at each locus varied from a minimum of four (HUJ616) to a maximum of thirteen (OarCP49) with a mean of 9.0 alleles across all loci. Allele size ranged from a minimum of 75 bp (OarCP49) to a maximum of 297 bp (HSC), while allele frequencies ranged from 0.0102 to 0.3980. The most frequent allele number was nine. The number of effective alleles ranged from 3.8757 (HUJ616) to 9.6232 (OarCP49). The mean number of effective alleles with an overall mean of 6.78 ± 1.79 alleles. The observed heterozygosity ranged from 0.0408 (BM1314) to 0.1429 (OarHH35, OarHH41) with a mean value of 0.0985 ± 0.025, whereas, the expected heterozygosity ranged from 0.7420 (HUJ616) to 0.8961(OarCP49) with a mean value of 0.8409 ± 0.046. All the twenty four microsatellite loci (100 percent) were found to be highly polymorphic and the PIC values ranged from 0.6945 (HUJ616) to 0.8961 (OarCP49) with a mean PIC value of 0.8240 ± 0.061. The inbreeding estimates obtained in this study were all positive and varied from 0.8318 (OarHH35) to 0.9534 (BM1314). The mean FIS value of 0.8825 ± 0.03 indicated the deficiency of heterozygotes. The Chi-square test revealed that all the twenty four loci were showing significant deviation from Hardy-Weinberg Equilibrium.