Browsing by Author "Bankar, P. S."
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ThesisItem Open Access Genetic Analysis of Production and Reproduction Traits in Malpura Sheep(MAFSU, 39703) Bankar, P. S.; Sirothia, A. R.The present study was undertaken in Malpura sheep for genetic analysis of production and reproduction traits. The data consisting 1727 Malpura sheep spread over from 1996 to 2007, maintained at CSWRI, Avikanagar, Rajasthan were used to estimate LSMs, genetic parameters and sire evaluation. The traits studied were BWT, 3WT, 6WT, 9WT, 12WT, ADG1, ADG2, ADG3, GFY1, GFY2, GFY3, AFS, WTFS and AFL. The data were subjected to LSMLMW and MIXMDL packages of Harvey (1990) and derivative free-REML package of Meyer (1998), taking sex, year, season as fixed effects; sire as a random effect; ewe weight and age at shearing as covariates. The least squares means (±S.E.) for above mentioned traits were 3.01±0.01, 13.62±0.07, 20.69±0.11, 23.57±0.12, 26.89±0.12, 118±0.76, 77.01±0.65, 38.11±0.82, 0.586±0.004, 0.371±0.01, 0.381±0.0 (kg.), 645±7.49 days, 26.32±0.13kg and 729.15±4.57 days, respectively. LSMs for all traits were in range. Fixed effects sex and year had significant (p<0.01) effect on almost all traits studied as compared to season. The males were heavier at all ages and produced more wool than female. The sire effect was significant for almost all traits. The estimates of heritability for growth and wool traits were highest for 6WT (0.254±0.07) and GFY-3(0.305±0.05). Model 2 and model 8 had higher estimates of heritability than Univariate DF-REML procedure. The genetic, phenotypic and environmental correlations among the growth traits under study were observed positive and from moderate to very high, except ADG 2 and ADG3. The ranking of sires was done by estimating their breeding values under all methods. All the rank correlations were significant. It can be stated from the present study that the 6WT and GFY1 can be continued for selection of animals. Also, the BLUP and DF-REML reveals better genetic potential of animals than earlier models, hence can be employed for ranking sires for growth and wool traits.ThesisItem Open Access Growth Hormone Gene Polymorphism and Its Association with Growth Performance in Deccani Sheep(MAFSU, Nagpur, 2022-01-17) Bhavekar, A. P.; Bankar, P. S.The present research was conducted with the objectives to study the genetic polymorphism of Growth hormone gene using PCR-RFLP as well as to investigate the association of polymorphic genotypes with growth performance in Deccani sheep in the Molecular Genetics Laboratory, Department of Animal Genetics and Breeding, PGIVAS, Akola. In the present study, growth performance (body weight) and important morphometric traits (Body length, height at wither, chest girth and Paunch girth) were recorded in 63 Deccani sheep in Yavatmal district, belonging to both sexes and different age groups. The growth performance and morphometric traits in 32 lambs were systematically recorded from birth to 3 months of age, at 15 days intervals each. The body weight and respective morphometric traits in randomly selected 31unrelated ewe and ram belonging to 6, 9 and 12 months of age were recorded once during a visit to herds. To screen the polymorphism of GH gene (422 bp), total 32 Deccani lambs from different herds and sexes were considered. The DNA was collected from the blood using blood genomic DNA isolation kit. The reported primer of GH gene (422 bp) with Forward (5’CTCTGCCTGCCCTGGACT3’) and Reverse primer: (5’ GGAGAAGCAGAAGGCAACC 3’) was used to amplify the selected locus. After the digestion of HaeIII restriction enzyme, the samples executed three different genotypes, which were assigned on the basis of bands, screened through gel electrophoresis. The present study revealed polymorphic pattern for GH gene i.e. distinct restriction conformational patterns indicating a total of three genotypes AA (366, 56 bp), AB (422,366 and 56 bp) and BB (422 bp) were observed. The highest genotypic frequency (0.72) was noted for AB genotype as compared to BB (0.16) and AA (0.12) genotypes with a gene frequency of A and B allele as 0.48 and 0.52, respectively for GH gene in studied Deccani sheep. The findings indicated that the B allele was more frequent than the A allele among the studied Deccani population for the GHgene locus (422 bp). The data on growth performance traits in Deccani lambs viz. body weight and morphometric traits measured periodically was subjected for Least Squares Mean estimation. The overall least squares mean for body weight (kg) at birth, 1 month (30 days), 2 months (60 days), 3 months (90 days) in Deccani lambs were 2.72±0.67, 9.86±0.49, 14.19±0.57 and 19.32±0.63 kg, respectively. The mean values observed for body weight (kg) at 6, 9 and 12 months of age were 27.38±0.44, 32.04±1.00, and 37.89±1.80 kg, respectively. The present investigation revealed, body length at birth, 1, 2, 3, 6, 9 and 12 months of age months estimates (cm) were 32.72±1.05, 42.86±0.97, 51.71±1.48, 60.41±1.6, 65.1±1.28, 65.3±1.61 and 78.06±0.58 cm respectively. Similarly, the estimates (cm) for height at wither were 38.86±0.93, 48.24±1.28, 56.30±1.52, 64.30±1.58, 61.8±1.04, 62.9±0.69, and 74.16±0.77, respectively; the estimates (cm) for chest girth were 40.11±1.06, 47.62±1.13, 54.59±1.27, 62.61±1.20 74.4±1.35, 74.6±0.87 and 78.91±1.51, respectively and the estimates (cm) for paunch girth were 38.54±0.88, 45.72±0.98, 53.11±1.01, 59.90±1.04, 78.95±2.93, 78.3±1.39 and 78.19±1.17, respectively. The effect of sex, herd and genotype were investigated as sources of variation. The effect of sex was highly significant (p<0.01) source of variation for body weight at birth only. The effect of herd was found non significant source of variation at all ages except for body length at one month of age (p<0.05) in Deccani lambs. However, the effect of sex at other stages as well as effect of herd and genotypes at all stages were found as non significant sources of variation for body weight and morphometric traits in studied population of Deccani lambs. In present investigation, the body weight and morphometric traits studied in both sexes at different ages were indicating that Deccani sheep as mutton purpose breed. The present study suggested that, there was requirement for assessment of other loci and genes for the association of growth traits in Deccani sheep.ThesisItem Open Access Studies on Phenotypic Characterization and HSP-70 Gene Polymorphism in Khamgaon Cattle(MAFSU, NAGPUR, 2023-04-03) Chavhan Rahul Rupesh; Bankar, P. S.The present study was undertaken to record the phenotypic traits of Khamgaon cattle in Buldhana district alongwith the assessment of socioeconomic status and husbandry practices of stakeholders as well as detection of genetic polymorphism of HSP-70 gene using PCR RFRLP technique. Total 251 local cattle in the Buldhana district of the Maharashtra state, locally known as “Khamgaon cattle”, were surveyed using preformed questionnaire based on ICAR-NBAGR. It was found that, Khamgaon cattle were of medium sized with prominent head and tanned brown coloured coat (64.94%) colour as well as light brown coloured muzzle (98.40%), eyelid (99.60%), hoof (90.03%) and tail switch (93.62%) respectively. The horns were prominently brown (99.60%) and straight (100%) with inward orientation (56.97%). The udder was medium (58.65%) and bowl shaped (93.26%) with cylindrical shaped teats with pointed tips (96.15%), respectively. The brisket was small. Khamgaon cattle were having medium (58.65%) and bowl (93.26%) shaped udder; cylindrical (96.15%) shaped teat and round shaped teat tips. Total 109 owners, belonging to villages of Khamgaon, Shegaon and Nandura tehsils of Buldhana district, were personally interviewed by author. The Khamgaon cattle were primarily reared by farmer’s communities belonging to OBC caste (60.55%). The higher proportion of medium (36.69%) to large (36.69%) family size was involved in cattle farming. The average family size was estimated as 6.9 in studied population. Amongst the surveyed cattle owners, 84.40 respondents were having agriculture as main business, however, 15.59 % were labour by occupation. Total 65.22% cattle were literate. The majority of Khamgaon cattle owners (83.48%) were having small land holder i.e. upto 6 acres of land. Most of the Khamgaon cattle farmers were low to medium level of income (i.e. from Rs. 56,000 to 1,00,000/-). Khamgaon cattle were mainly reared for draft purpose (36.39 %) as compared to milk (32.15%) and breeding purpose (31.44%). Farmers were more inclined for natural service than artificial insemination (91.74%). The cattle were allowed for grazing whole day (98.16%) and housed during night (55.04%) and were kept in Open and Kachcha type of cattle shed, which was a part of residence of farmer. The specific region of HSP-70 gene i.e. 169 bp of exon 3 region was amplified with a synthetic primer (Forward primer-5’ACATGGCAAGGTGGAGATCA- 3’ and Reverse primer- 5’GAACTTGCGTCCGATCAGTC-3’) to detect the genetic polymorphism. The amplified DNA samples for HSP-70 gene (169bp) through PCR, were subjected to digestion with restriction endonuclease BsaAI at 37oC for 3 hours. It was found that, monomorphic pattern for AA genotype was observed for all the studied samples when the HSP-70 gene (169bp) digested with BsaAI enzyme. All the samples exhibited uncut fragment (lacking cutting site) indicating band size as 169 bp, which was assigned as AA genotype. The gene and genotype frequency was 1 for ‘A allele’ and AA genotype respectively, indicating its fixation in studied local Khamgaon cattle population. The distinct phenotypic features observed in studied population of Khamgaon cattle was not only verified with the earlier reports but also exhibited its phenotypic divergent from adjoining cattle populations and admitted its existence in the breeding tract. Hence, recognition of this lesser known germplasm at National level is proposed.ThesisItem Open Access Studies on Phenotypic Morphometric Traits and HSP-70 Gene Polymorphism in Local Donkeys of Akola(MAFSU, NAGPUR, 2023-04-03) Shinde Shubhangi Sanjay; Bankar, P. S.This study documented the morphometric, phenotypic traits and breeding practices of local donkeys in Akola district, Maharashtra. Total 86 local donkeys were subjected for recording and socioeconomic profile of 96 donkey farmers was also surveyed using preformed questionnaire. Additionally, genetic polymorphism of the HSP-70 gene in donkeys were investigated. 50 unrelated Kaikadi donkeys were analyzed for genetic polymorphism using PCR RFLP of the HSP-70 gene (169 bp) with BsaAI restriction enzyme. The donkeys were of medium sized with grey coat colour prominently. Phenotypic characteristics of the donkeys, such as black eyes, eyelids and hooves, a light grey muzzle, and dark brown tail switch were also noticed. Nasal bone was concave and straight. The overall mean values for various morphometric traits (cm) recorded in adult local donkeys (n=86) belonging to both sexes exhibited mean values for Body length (BL), Height at wither (WH), Height at croup (HC), Height at knee (HN), Height at hock (HH), Arm length (AL), Dis. bet. withers to croup (WC) and Dis. bet. croup and tail head (CT), Chest girth (CG), Paunch girth (PG), Tail length (TL), Face length (FL), Face width (FW), Dis. bet. eyes (ED), Ear length (EL), Ear width (EW) and Body weight (BWT) were 73.43±0.32 cm, 91.20±0.58 cm, 93.44 ±0.60 cm, 31.39±0.14 cm, 35.19 ±0.14 cm, 84.52 ±0.61 cm, 55.11 ±0.48 cm, 17.94±0.08 cm, 97.31±0.54 cm, 101.29 ±0.54 cm, 51.90 ±0.32 cm, 41.73±0.35 cm, 19.24 ±0.27 cm, 9.94 ±0.11 cm, 21.00 ±0.17 cm, 10.03 ±0.16 cm and 58.66 ±0.90 kg, resp. The Least square analysis of variances exhibited effect of non-genetic factors viz. age, sex and location etc. on various morphometric traits. Kaikadi donkeys were primarily reared by Kaikadi community, belonging to NT category with medium family size, mostly illiterate and landless and labour. All the donkey farmers surveyed were below poverty line. The families kept donkeys as their traditional business for draft or pack purposes. All the donkey farmers preferred for Natural breeding using their own jenny and no specific criteria for selection of male for breeding. The donkeys were kept in open (61.46%) and Kaccha (97.79%) shed, which located near residence (92.71%). The PCR-RFLP of HSP-70 gene (169 bp) using synthetic primer Forward primer-5-ACATGGCAAGGTGGAGATCA-3 and Reverse primer-5’-GAACTTGCGTCCGATCAGTC-3’) and digesion with BsaAI restriction enzyme exhibited uncut fragment of 169 bp i.e. monomorphic AA genotype for all the studied samples. Thus, the frequency of observed genotypes AA, AB and BB was estimated as 1, 0 and 0 respectively and complete absence ‘B’ allele and fixation of allele A in population in studied local donkey of Akola district. The study found that the Kaikadi donkey population in Akola district was of small size and has unique physical characteristics compared to other donkey populations in India. The HSP-70 gene locus studied was found to have a fixed AA genotype in the donkey population.ThesisItem Open Access Study of Polymorphism in a-Lactalbumin and b-Lactoglobulin Genes in Marathwadi Buffalo(MAFSU, Nagpur, 2016) Joshi, A. S.; Bankar, P. S.The present research work was conducted to study the polymorphism of -Lactalbumin and -Lactoglobulin genes (whey protein genes) in Marathwadi buffaloes using PCR-RFLP technique. The experiment material was comprised of (n=30) blood samples and (n=9) milk samples of Marathwadi buffaloes, collected from the Marathwadi Buffalo Farm, College of Veterinary and Animal Sciences, Udgir during June-August, 2016. The research work was conducted at Molecular Genetics Laboratory of Department of Animal Genetics and Breeding, Post Graduate Institute of Veterinary and Animal Sciences, Akola. DNA was isolated from blood using standard Phenol-Chloroform extraction method followed by checking of its quality and quantity by Spectrophotometer and subsequently by electrophoresis method. A 309 bp fragment of -Lactalbumin gene amplified using forward (5’ TTG GTT TTA CTG GCC TCT CTT GTC ATC 3’) and reverse (5’ TGA ATT ATG GGA CAA AGC AAA ATA GCA G 3’) primer with MspI restriction enzyme yielded only one type of restriction pattern (220 bp and 89 bp) in all 30 Marathwadi buffaloes and accordingly monomorphic genotype was identified as BB. A 247 bp fragment of -Lactoglobulin (LGB) gene amplified using forward (5’ TGT GCT GGA CAC CGA CTA CAA AAA 3’) and reverse (5’ GCT CCC GGT ATA TGA CCA CCC TCT 3’) primer with HaeIII restriction enzyme yielded only one type of restriction pattern (148, 99 and 74 bp) in all 30 Marathwadi buffaloes and accordingly monomorphic genotype was identified as AB. The estimates of protein, fat and lactose contents were recorded thrice at monthly interval from 9 unrelated lactating buffaloes belonging to Marathwadi Buffalo Farm, College of Veterinary and Animal Sciences, Udgir using Lactoscan (Milk analyzer) during June to August 2016. The mean values for protein, fat and Lactose content in Marathwadi buffalo ranged from 3.8 percent to 4.3 percent, 3.9 percent to 5.8 percent and 3.7 percent to 4.2 percent respectively. The total yields for protein, fat and Lactose content in Marathwadi buffalo estimated by Test Interval Method ranged from 19.66 kg to 30.52 kg, 21.99 kg to 31.11 kg and 18.68 kg to 28.65 kg, respectively. The monomorphic pattern revealed by screening of -Lactalbumin gene (BB) and -Lactoglobulin gene (AB) in small population of Marathwadi buffaloes pointed out the need of genotyping whole or larger population of ‘true to breed type’ Marathwadi buffaloes from its breeding tract so as to identify possible genotypes of whey protein genes. The monomorphic genotype AB observed for -LG gene in Marathwadi buffaloes is suggestive of presence of AA and BB genotypes in population of Marathwadi buffaloes. Screening larger population of Marathwadi buffaloes for finding polymorphism of candidate milk genes like whey protein genes is prerequisite to explore their association with economically important milk production traits will provide a robust platform for marker assisted selection.ThesisItem Open Access Study of Polymorphism in Candidate Milk Protein Genes and Their Association with Milk Traits in Gaolao Breed of Cattle(MAFSU, Nagpur, 2020-11-25) Bankar, P. S.; Kuralkar, S. V.The present research was undertaken to study the genetic polymorphism of candidate milk protein genes in Gaolao cattle with the objectives to investigate the polymorphism in - casein, -casein, - lactalbumin and -lactoglobulin genes, to detect their relationship with milk traits and to sequence their known polymorphism. Blood samples and milk yield data was collected from total 83 unrelated lactating Gaolao cattle belonging to Bull Mother Farm, Pohara Dist. Amravati (n=60) and from breeding tract i.e. villages of Wardha district (n=23) during 2015-2017. The research work was carried out in the Molecular Genetic Laboratory, Department of Animal Genetics and Breeding, Post Graduate Institute of Veterinary and Animal Sciences, Akola. The milk yield (kg) was estimated by the Test Interval Method following the guidelines of International Committee for Animal Recording. The milk composition traits viz. Fat%, Protein %, Lactose% and SNF % estimated using milk analyzer were subjected for daily yields with adjustment of milking interval and stage of lactation. The polymorphism in - casein gene was detected for loci 121 bp, 251 bp, 495 bp and 357 bp; in -casein for loci 379 bp (Exon III to V), 453 bp (Exon IV), 350 bp (Exon IV) and synthetic primer of 350 bp; in -lactalbumin for loci 309 bp, 166 bp, 268 bp (Exon III) and 133 bp and in -lactoglobulin genes for loci 247 bp (Exon IV), 854 bp, 262 bp (Exon IV) and 119 bp, respectively. All the loci were subjected for PCR-RFLP method, except locus 357 bp in -casein gene, 350 bp synthetic primer in –casein gene, for locus 133 bp in -lactalbumin gene and for locus 119 bp in -lactoglobulin gene, which were screened by PCR SSCP method following silver staining technique. All the studied loci exhibited polymorphism except, the locus 495 bp (Exon VII) of ß-casein gene showed monomorphic pattern (BB genotype). Amongst 15 polymorphic loci, 121 bp of ß-casein and 854 bp of ßlactoglobulin gene loci showed AA and AB genotypes only. However, rest of other loci exhibited AA, AB and BB genotypes. The higher gene frequency for A allele was observed for loci 357 bp (0.55) of ß-casein, 379 bp (0.93) and 453 bp (0.87) of -casein, 166 bp (0.88) of -lactalbumin gene, 854 bp (0.91), 262 bp (0.73) and 119 bp (0.60) of ß-lactoglobulin gene. However, higher gene frequency for B allele was observed for loci 121bp (0.96), 495 bp (1.0) and 251 bp (0.87) of ß-casein, 350 bp (0.64) and 350 bp synthetic (0.67) of - casein, 309 bp (0.72), 268 bp (0.85) and 133 bp (0.72) of -lactalbumin gene, 247 bp (0.71) of ß-lactoglobulin gene Mean values for Total Milk yield (kg), Total Fat yield (%), Total Protein yield (%), Total Lactose yield (%) and Total SNF yield (%) were 312.47 ± 85.52, 4.14± 1.15, 3.21±0 .86, 4.99±1.35 and 9.30± 2.47, respectively in Gaolao cattle. All the loci exhibited effect of genotype as a non significant source of variation in Gaolao cattle, except locus i.e. 251 bp of - casein gene which exhibited significant influence of genotype (A2A2) on all the studied milk traits except TFY. On the other hand, the effect of genotype could not be estimated for locus 495 bp of -casein gene, due to its monomorphic pattern. The effect of location and season of calving was also found non-significant with respect to milk traits under study. However, the effect of parity was found significant with TMY, TPY and TLY for all the loci as well as with TFY for locus 453 bp of -casein gene and with TSNFY for loci -casein 495 bp of -casein gene and 379 bp of -casein gene respectively. The representative polymorphic amplicons of 11 loci of studied milk protein genes, belonging to different genotypes detected by PCR-RFLP method only were outsourced for DNA sequencing for confirmation of restriction sites and thereby the genotypes observed, with the help of Mega X and Chromas softwares. Restriction sites were confirmed for loci 121 bp and 251 bp of ß-casein gene, 379 bp, 453 bp and 350 bp of -casein gene, 309 bp and 166 bp of -lactalbumin gene and 247 bp, 854 bp and 262 bp of ßlactoglobulin gene. However, the cutting site and genotype could not get confirmed for locus 268 bp of -lactalbumin gene due to poor quality of sequencing. Thus, it can be concluded from present study that, polymorphic nature of studied loci can be utilized as baseline information for future genomic selection database for Gaolao cattle. The predominance of B allele for loci 121 bp and 357 bp and A2 allele for locus 251 bp in ß-casein gene was indicating preponderance and thereby confirming the uniqueness of Gaolao breed as Bos indicus germplasm for studied loci. The estimates of TMY, TFY, TPY, TLY and TSNFY when compared with other recognized cattle breeds of Maharashtra, were supporting the classification of Gaolao as draught purpose breed. Non specific trends or genotypes observed were suggestive of differential expression of respective allele and or variation in sample size under different genotypes of each locus. The automated DNA sequencing confirmed the restriction sites for studied polymorphic loci of - casein, - casein, -lactalbumin and -lactoglobulin genes identified by PCR-RFLP method in Gaolao cattle.