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Birsa Agricultural University, Ranchi
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ThesisItem Open Access GENETIC ARCHITECTURE OF “T&D” AND ITS CROSSES WITH EXOTIC PIGS(Birsa Agricultural University, Kanke, Ranchi, Jharkhand, 2010) Himkar, Himanshu Kumar; Singh, S. K.1. It was concluded that performance of crosses of T&D with Russian charmukha was superior than T&D × T&D followed by T&D × Tamworth and T&D × Hampshire. 2. Growth rate of suckling piglets during pre-weaning period decreased with the increase of litter size at birth of dam. 3. There was decrease in mortality rate with the advancement of ages. Majority of mortality was noticed during first two weeks of pre-weaning period. 4. The medium to high estimates of heritability for growth trait indicated that there is a good scope for increasing weight at different ages through selection. 5. The positive and significant phenotypic correlations among body weights at various ages would form an excellent basis for selection at an earlier age. 6. The high and positive genetic correlations among body weights at various ages would form an excellent basis for selection at an earlier age.ThesisItem Open Access IMMUNO RESPONSIVENESS: A TOOL FOR SELECTION IN SWINE(Birsa Agricultural University, Kanke, Ranchi, Jharkhand, 2010) KUJUR, TRIPTI VIJAYA; Singh "Dron", D.K.On the basis of observed findings, the present study could be concluded as follows: I. Classification of piglets and sows population into two lines viz. High and Low responder to Sheep Red Cells (SRBCs). II. High Responder Piglets (HRP) to SRBCs could be selected for better growth with 12th week of age and also better survivability up to weaning age i.e. 56th day of age. III. Selection of sows on the basis of its response to SRBCs could be effective for altering its reproductive traits like litter size and farrowing interval. IV. Improvement in Humoral Immune status of piglets could be possible by exploiting additive genetic variances. 122 V. Selection of piglets could be based on its response to SRBCs followed by breeding programme by involving Hampshire and Tamworth genes in combination, if exotic germplasm are to be incorporated for better humoral immune status of progenies and its positive impact on growth up to 12th week of age and survivability during pre-weaning period.ThesisItem Open Access MOLECULAR CHARACTERIZATION OF GOAT GENETIC RESOURCES OF WEST BENGAL AND JHARKHAND(Birsa Agricultural University, Kanke, Ranchi, Jharkhand, 2010) KUMARI, NANDANI; SINGH, L. B.Two goat breeds, Black Bengal and Black Bengal type goats of Jharkhand were characterized by using RAPD markers in the present investigation. DNA was extracted from 50 blood samples each for Black Bengal and Jharkhand Black i.e. total of 100 examples. Goat is an important asset for the livestock breeders as well as the poor farmers. It is the earliest ruminant domesticated around 9000 to 7000 B.C. Black Bengal goat found in the entire area of Jharkhand (referred to as Jharkhand Black in my thesis for simplicity) have similarities as well as differences with the original Black Bengal goats of West Bengal, yet it has no identity of its own. Genomic DNA was isolated and purified from white blood cells using Proteinase K digestion and standard phenol: chloroform extraction method as per the standard protocol described by Sambrook et al (1989). Agarose gel electrophoresis for checking the quality of isolated genomic DNA samples was done by diluting the DNA samples in ratio of 1:10. This was followed by Polymerase chain reaction. And PCR technique was used to amplify the chosen marker. For this, optimization of PCR technique was followed by Agarose gel electrophoresis of PCR amplified product of DNA. The amplified product were statistically analyzed to measure the between and within population diversity. The genetic diversity within and between population was analyzed as the observed and expected number of alleles and Shannon’s information Index using popgene software . Ewan’s Watterson test was performed to test the neutrality for RAPD markers ;the statistics F (sum of square of allelic frequency ) and limit (upper and lower) at 95% confidence region for the test were calculated using the algorithm by mainly using 100 simulated samples and implemented in popgene software package. In order to quantify the percentage of molecular variance due to differences among the difference among different populations and significance was tested by a non-random permutation approach using AMOVA programme included in Arleqin software package. Migration rate which showed the gene flow per generation was also found out. A dendrogram by UPGMA method was constructed. The result can be summarized as follows- *Gene frequency:-Gene frequency in Black Bengal ranged from 0.125 to 0.729 for allele 0 and from 0.271 to 0.875 for allele 1.Similarly in case of Jharkhand Black , Gene frequency ranged from 0.146 to 0.625 for allele0 and from 0.375 to 0.854 for allele 1. *Polymorphic information content:-Polymorphic information content (PIC) or expected heterozygosity scores varied from 0.219 to 0.486 with overall mean 0.411 in Black Bengal and from 0.278 to 0.496 with overall mean 0.413 in Jharkhand Black goat. *Mean observed and effective number of alleles:-Mean observed number of alleles was 2 in both Jharkhand Black and Black Bengal with mean effective number of alleles was 1.6991 for Black Bengal and 1.6935 for Jharkhand Black. *Nei’s gene diversity value (h):-Nei’s gene diversity value was 0.3750 for Black Bengal and 0.4022 for Jharkhand Black. *Shannon’s Information Index:-Shannon’s Information Index was 0.6792 for Black Bengal and 0.5898 for Jharkhand Black *Gene flow:-Gene flow (Nm) value was 25.68 , Hs(Mean sample gene diversity) between two population was 0.412and Ht (Total gene diversity )in total sample was 0.420. *Genetic Identity and Genetic Distance:-Nei’s genetic identity was found to be 0.9727 and genetic distance was 0.0276. * Evan’s Watterson’s test of neutrality:-Not even a single locus showed the F value beyond the standardized range of U95 and L95 at 95% confidence level and so all the locus were neutral to selection pressure when these were taken separately but when combined together the two locus SIGMA06-4 AND SIGMA10-1 were not found neutral to selection pressure. *Dendrogram:-The clustering between the breeds was not very sharp with their intermingling at a few places .This showed the dilution between the gene pools of Black Bengal and Jharkhand Black. *AMOVA:-The analysis of molecular variance (AMOVA) showed 1.69983 % among population variation and 98.30017 %between population variation. A significant amount of differentiation among the two breeds and high level of gene flow between Jharkhand black & Black Bengal was observed. As Jharkhand black Goat also shows significant gene diversity, it should be given a separate identity. The result was crucial for in situ conservation and on the basis of this result, it can be recommended that within breed diversity is actively maintained to enable these extensively unmanaged stockists to adapt to further demands and conditions and there is ample scope for further improvement in its productively through appropriate breeding strategies. The results presented added important information on the puzzle of goat genetic diversity and conservation in India where it is of crucial economic relevance to increasingly marginalized rural communities. Therefore, it can be recommended that within-breed diversity is actively maintained to enable these extensively unmanaged stocks to adapt to future demands and conditions and there is ample scope for further improvement in its productivity through appropriate breeding strategies. On an whole it was concluded that Jharkhand black shows considerable amount of similarities and few of dissimilarities and so Jharkhand black may be a strain or the derivative of Black Bengal and should be given a separate identity considering the importance of gene conservation amidst high gene flow due to geographical closeness.ThesisItem Open Access EVALUATION OF SOME GENOTYPES TO DEVELOP A BREED FOR RURAL POULTRY FARMING(Birsa Agricultural University, Kanke, Ranchi, Jharkhand, 2010) KUMAR, BASANT; Singh Dron, D. K.On the basis of findings of the present study, the following conclusions were drawn: i) Red Cornish genotype is better than Dulham Red and two synthetic strains but both synthetic strains are better than Dulham Red. ii) Humoral immune response is negatively associated with growth traits, hence it could be utilized for developing selection and breeding strategies. iii) Humoral immune response had no impact on egg quality traits but it varied due to genetic groups. iv) Red Cornish and both synthetic strains could be utilized to develop a suitable breed for rural poultry farming in the agro-climatic conditions of Jharkhand.ThesisItem Open Access MHC II GENE POLYMORPHISM WITH IMMUNE RESPONSE AND TOLERANCE TO HAEMONCHUS SPECIES IN CHOTANAGPURI SHEEP(Birsa Agricultural University, Kanke, Ranchi, Jharkhand, 2011) MURMU, AKHILESHWAR; Singh “Dron”, D. K.The present study was carried out on Chotanagpuri sheep which are moderately resistant to nematode (Haemonchous contortus). Animals are being maintained under Mega Sheep Seed Project at RVC, BAU. In this investigation, sheep were screened for their Humoral Immune Response against Goat RBC to develop divergent lines by truncation selection on the basis of the antibody response against Goat Red Blood Cells (RBC). Base population of divergent lines were selected on the basis of HA titres and egg per gram. Sheep of both high and low lines were evaluated for MHC class II gene polymorphism. Haemonchus contortus load of ewes upto 1200 EPG had no adverse effect on birth weight and subsequent growth of lambs till yearling age indicating that Chotanagpuri sheep are tolerant to it a greater extent. Humoral Antibody Response of Chotanagpuri ewes against Goat Red Blood Cells (GRBC) did not affect birth weight and subsequent growth of their lambs till yearling age significantly, though the lambs of high responder dams had better growth rate than those of low responder ones. Lambs of comparatively heavier ewes had better growth rate during pre- and postweaning periods, though birth weight of lambs did not vary significantly due to variation in body weight of their dams at lambing. Lambs born heavier continue to weigh comparatively more than others revealed that the selection of birth weight for improvement in weaning as well marketing weight at 40 to 48 weeks of age would be effective. Heritability estimates of body weight from 4 week to 36 week of age were high in magnitude (0.64±0.35 to0.75±0.37).Heritability estimates of birth weight and weights at 48th week were low in magnitude. Heritability estimates of antibody response were also low in magnitude. The phenotypic relationship of antibody response of dams with growth of their lambs were erratic and non significant. Phenotypic relationship among body weights at different stages of growth were positive and significant (P≤0.01). Antibody response of dams had positive and medium to high genetic relationship with growth of their lambs from 4 to 36-week of age. The genetic association among body weights at different stages of growth were positive and high in magnitude in general. DRA gene sequence subjected to amplification in PCR with sheep genome gives amplified product though did not showed polymorphism in SSCP (Single stranded confirmation polymorphism) analysis.ThesisItem Open Access CHROMOSOMAL PROFILE OF BLACK BENGAL GOAT OF JHARKHAND(Birsa Agricultural University, Kanke, Ranchi, Jharkhand, 2011) MANDAL, BANANI; SINGH, L. B.Black Bengal goat was characterised cytogenetically by using morphometric measurement of chromosomes through karyotyping in the present investigation. A total of 20 animals (10 male and 10 females) were utilized for this study. (Instructional Farm Small Ruminants, Ranchi Veterinary College, Birsa Agricultural University, Ranchi). Blood samples were collected and short-term lymphocyte culture method was followed as given by Moorehead et al. with slight modification. Slides were prepared by dropping cell suspension on to the slide and checked under a microscope to determine the good quality metaphase spreads. Slides were stained with 2% Giemsa stain. The scanning of the slides was done at 10 X magnification and photos were taken under 100 X magnification. The relative lengths of chromosomes were measured as percentage of the total genome length. Since the data was not following normal distribution the data were transformed to arc sin values. The protocol for goat lymphocyte culture was standardized. The best culture result was found at 38.50C. Fetal bovine serum was used at the rate of 12 ml for 40 ml of culture media. 150 μl of PHA (1mg/ml) provided the best result in culturing goat lymphocyte. The concentration of RPMI1640 (dried medium), L-glutamine and antibiotics (Penicillin and Streptomycin) were kept constant as per established protocol. 20μl of colchicines (10μg/ml) was used per culture to arrest the lymphocyte in metaphase stage. A total of 500 metaphase spreads of Black Bengal goats were screened. Out of them, the best 200 metaphase spreads (10 spreads per animal) were examined for karyotyping in the present investigation. The normal diploid (2n) chromosome complement of Black Bengal goat was found to be 60 having 29 pair of autosome and 1 pair of sex chromosome All the chromosome excluding Y-chromosome were acrocentric. The Ychromosome was found to be smallest one in Black Bengal goat. It appeared dot like structure and seemed to be sub-metacentric. X-chromosome was longest in both sexes. Length of X chromosome (3.14±0.05μm) in male goat was significantly higher than that of female goat (3.03±0.06 μm). Smallest chromosome was Y in male goat and was found to be 0.79±0.02μm. Length of autosome in male goat varied from 0.96±0.01 to 2.78±0.04μm. Length of autosome pairs in female goat varies from 0.96±0.02 to 2.86±0.06μm. Overall length of autosome in Black Bengal goat, pooled over sexes ranged from 0.96±0.02 to 2.78±0.04μm. Most of the autosome pairs varied significantly in length while length of some of the autosome pair did not differ significantly from its adjacent chromosome pairs in male, female and overall in Black Bengal goat. Length of chromosome pair 27 and 28 of male goat was found significantly (p0.01) higher than that of the female goat. The length of other autosome pairs did not differ significantly. The relative length of autosome ranged from 1.79±0.02 to 5.19±0.03 percent in male goat and 1.78±0.03 to 5.25±0.04 percent in female goat. The X-chromosome which was the longest among all, contributed 5.95±0.05 percent in male and 5.57±0.05 percent in female with overall 5.76±0.05 percent to total genome in Black Bengal goat. The contribution of Y-chromosome to total genome in male goat was only 1.47±0.03 percent. The relative length of chromosome pair 21 & 22 and 24 & 25 in autosome did not differ significantly to each other in male goat. Similarly in female goat chromosome pair 9 & 10, 20 & 21 and 23 & 24 did not differ significantly to each other. When data were pooled over sex, the relative length of autosome in Black Bengal goat differed significantly (p0.01) among themselves. Relative length of chromosome pair 3, 19, 23, 27, 28 and X were significantly (p0.01) higher in male than female goat of Black Bengal breed. While, relative lengths of chromosome pair 5, 6, 8, 9, 10, 11, 12, 13 and 16 were significantly (p0.01) lower in male than that of female Black Bengal goat. The arm ratio, centromeric index and morphological index have not been calculated as acrocentric nature of all the chromosome in Black Bengal goat. The idiogram for male, female and pooled of Black Bengal breed were constructed based on absolute and transformed relative lengths of chromosome.ThesisItem Open Access GENETIC ARCHITECTURE OF BLACK BENGAL GOAT IN CONTEXT TO ITS FECUNDITY(Birsa Agricultural University, Kanke, Ranchi, Jharkhand, 2013) KUMAR, VIVEK; Singh, L. B.The present study was carried out on Black Bengal goats which is prolific meat purpose breed of India. Animals were maintained at Instructional Small Ruminant Farm, Ranchi Veterinary College, B.A.U., Ranchi. In this investigation, goat population was screened for polymorphism of FecB gene. A study of association of different variants with different reproductive traits and absolute body weights at different stages of growth was also done. With the primers having forward and reverse base sequence as 5'-CCAGAGGACAATAGCAAAGCAAA-3' and 5'-CAAGATGTTTTC ATGCCTCATCAACAGGTC-3', PCR products obtained were subjected to polyacrylamide gel electrophoresis for the detection of Single Strand Conformational Polymorphism (SSCP). As a result, three different SSCP variants were found which were designated as AA, AB and BB. The highest genotype frequency was observed for AB (0.38), which was followed by BB (0.33) and AA (0.29). Least-square analysis of variance showed significant (P<0.01) effect of genotype on litter size at birth. The result revealed that highest litter size at birth was found to be 02.06±00.08 for genotype BB, which differed significantly (P<0.01) from that of genotype AA (01.14±00.08) and genotype AB (01.27±00.07). Further, it was found that genotype had significant (P<0.01) effect on litter size at weaning. The highest litter size at weaning was found to be 01.81±00.06 for genotype BB, which differed significantly from that of genotype AA (01.00±00.08) and genotype AB (01.12±00.06). There was non-significant effect of genotype on age at first kidding, service period, kidding interval and gestation period. Moreover, there is significant (P<0.01) effect of genotype on body weight at birth. The least square mean of body weights at birth for genotype BB (01.17±00.03 kg) was significantly lower than that of genotype AA (01.43±00.03 kg) and genotype AB (01.36±00.02 kg). The different genotype were non-significantly associated with absolute body weight at 4-week, 8-week, 12-week, 24-week, 36-week and 48-week. Nucleotide sequences of the allelic variants were also analyzed. The DNA sequence showing polymorphism observed were used to identify SNPs. Principal SNP was found at 78 position of gene sequence, which shows transition of Adenine (AA genotype) to Guanine (BB genotype). On the basis of this investigation, following conclusion were observed: 1. The PCR-SSCP analysis of FecB gene revealed the polymorphic pattern of genotypes in Black Bengal Goat. It suggests that flow of FecB mutation was present in Black Bengal Goat. 2. There was significant association of different polymorphic variants of FecB gene with the litter size at birth and at weaning and body weight at birth.ThesisItem Open Access GENETIC ARCHITECTURE OF CHHOTANAGPURI SHEEP IN CONTEXT TO ITS FECUNDITY(Birsa Agricultural University, Kanke, Ranchi, Jharkhand, 2013) ORAON, THANESH; Singh “Dron”, D. K.The present study was conducted to study polymorphism of FecB genes and ascertain genotypes of Chhotanagpuri lambs on the basis of alleles of FecB gene with objectives to estimate the effects of genotypes and some non- genetic factors on growth in terms of absolute body weights of lambs from birth to 52-week of age as well as variation in growth rate in terms of average daily gain in weight during different periods of growth. Attempts were also made to study relationship of ewe’s weight at lambing with growth of their lambs as well as among body weights at different stages of growth from birth to 52-week of age. Regression equations with their coefficient of determination were also developed to predict body weight at later stages of life on the basis of earlier weights. The findings of this study are being summarized as follows: (1) Six single strand conformation polymorphism variants of FecB gene were obtained at Locus-1 and Locus-2. (2) AT Locus-1, the genotype frequency of AA was highest (0.424) followed by AB (0.326) and BB (0.250) .revealed that the frequency of A and B alleles were 0.587 and 0.413, respectively. (3) Locus-2, the genotype frequency of CD was higher (0.402) followed by CC (0.381) and DD (0.217), indicating that the frequency C and D were 0.587 and 0.418, respectively. (4) Genotypes at Locus-1 had significant effect on body weights of lambs from 8th week of age to 52nd week of age, though the variations in birth weight and weight at 4th week due to it was not significant. (5) Lambs with BB genotype at Locus-1 had better growth rate up to 52-wk of age than those with AA and AB genotypes. (6) Growth of lambs remains unchanged due to genotypes at Locus 2, though lambs of CC genotype had better growth than CD and DD genotypes. (7) Body weights of lambs from 4th week onwards varied significantly due to dam’s weight at lambing. (8) Lambs of heavier dams continued to weigh more than those of lighter dams till 52- wk of age. (9) Birth weight of lambs had significant effect on their growth up to 52-week of age. Lambs born with higher birth weight at grow faster rate than those born having lower birth weight particularly during pre-weaning period i.e up to 12-week of age. (10) Average Daily Gain (ADG) in body weight of lambs was maximum (81.64±3.83 g/day) irrespective of genotypes. (11) ADG declined with the advancement in age. (12) Lambs with BB genotype at Locus had better growth rate in terms of ADG than those of AA and AB at this locus (13) Lambs with higher birth weight had better ADG particularly during pre-weaning period. (14) Dam’s weight at lambing had positive and significant phenotypic association with body weight of their lambs up to 52 – week of age. (15) Phenotypic association among body weights at various stages of growth were positive and significant, though the magnitude of relationship declined with the increase in gap between two weights taken. Regression study revealed that the body weight at growth could be predicted with moderately higher accuracy on the basis of preceding weight The following conclusions have been drawn: (1) Chhotanagpuri lambs with BB genotypes should be selected for better growth. (2) Breeding program should be designed to increase the frequency of ‘B’ allele among individuals in a flock of Chhotanagpuri sheep. (3) Lambs with high birth weight should be cared properly particularly after weaning at 12-wk of age for better post-weaning growth. (4) Keeping in view the positive association of dam’s weight at lambing on growth of their lambs up to 52-week of age the ewes particularly during advance stage of pregnancy should be fed properly and cared properly for improving birth weight of lambs and its growth. (5) Lambs could be selected on the basis of weight at at birth and 12-wk of age for better growth and more meat yield. (6) Polymorphism of FecB gene should be studied with a large number of primers to derive valid conclusion in context of fecundity.