Showing papers by "Chaitanya G. Joshi published in 2010"

Bacterial diversity in the rumen of Indian Surti buffalo (Bubalus bubalis), assessed by 16S rDNA analysis.

Abstract: Bacterial communities in buffalo rumen were characterized using a culture-independent approach for a pooled sample of rumen fluid from 3 adult Surti buffaloes. Buffalo rumen is likely to include species of various bacterial phyla, so 16S rDNA sequences were amplified and cloned from the sample. A total of 191 clones were sequenced and similarities to known 16S rDNA sequences were examined. About 62.82% sequences (120 clones) had >90% similarity to the 16S rDNA database sequences. Furthermore, about 34.03% of the sequences (65 clones) were 85–89% similar to 16S rDNA database sequences. For the remaining 3.14%, the similarity was lower than 85%. Phylogenetic analyses were also used to infer the makeup of bacterial communities in the rumen of Surti buffalo. As a result, we distinguished 42 operational taxonomic units (OTUs) based on unique 16S r DNA sequences: 19 OTUs affiliated to an unidentified group (45.23% of total OTUs), 11 OTUs of the phylum Firmicutes, also known as the low G+C group (26.19%), 7 OTUs of theCytophaga-Flexibacter-Bacteroides phylum (16.66%), 4 OTUs of Spirochaetes (9.52%), and 1 OTU of Actinobacteria (2.38%). These include 10 single-clone OTUs, so Good’s coverage (94.76%) of 16S rRNA libraries indicated that sequences identified in the libraries represent the majority of bacterial diversity present in rumen.

A modified enrichment protocol for adult caprine skeletal muscle stem cell.

Abstract: To establish an adequate model to study the proliferation and differentiation of adult caprine skeletal muscle in response to bioactive compounds, a pool of satellite cells (SC) was derived from the rectus abdominis muscle of adult goat. Skeletal muscle contains a population of adult stem cells, named as satellite cells that reside beneath the basal lamina of skeletal muscle fiber and other populations of cells. These SC are multipotent stem cells, since cells cultured in the presence of specific cell lineage inducing cocktails can differentiate into several types of mesenchymal lineage, such as osteocytes and adipocytes. In the present study, we have developed a modified protocol for isolating satellite cells (>90%) and examined their myogenic and contractile properties in vitro.

Pheno-genotypic characterization of Listeria monocytogenes from bovine clinical mastitis.

Abstract: The present study was carried out for pheno-genotypic characterization of L. monocytogenes (L. monocytogenes). A total of three isolates of L. monocytogenes were recovered from 85 mastitic milk samples (47 buffalos and 38 cows). Confirmation of the L. monocytogenes were based on biochemical tests followed by phenotypic characterization by hemolysis on sheep blood agar, the Christie Atkins Munch-Petersen (CAMP) test, phosphatidylinositol-specific phospholipase C (PIPLC) assay and phosphatidylcholine-specific phospholipase C (PIPLC) assay. The isolates were subjected to genotypic characterization with the help of PCR assay for five virulence associated genes namely, plcA, prfA, hlyA, actA and iap. The L. monocytogenes isolates were further subjected to multiplex-PCR based serotyping. All the three isolates of L. monocytogenes were hemolytic, CAMP positive, PI-PLC, PC-PLC positive, hlyA, pclA, actA, iap and prfA positive by PCR. All the three isolates of L. monocytogenes were serotyped as 4b.

Methanogenic diversity studies within the rumen of Surti buffaloes based on methyl coenzyme M reductase A (mcrA) genes point to Methanobacteriales.

Abstract: Methane emissions from ruminant livestock are considered to be one of the more potent forms of greenhouse gases contributing to global warming. Many strategies to reduce emissions are targeting the methanogens that inhabit the rumen, but such an approach can only be successful if it targets all the major groups of ruminant methanogens. Therefore, basic knowledge of the diversity of these microbes in breeds of buffalo is required. Therefore, the methanogenic community in the rumen of Surti buffaloes was analyzed by PCR amplification, cloning, and sequencing of methyl coenzyme M reductase (mcrA) gene. A total of 76 clones were identified, revealing 14 different sequences (phylotypes). All 14 sequences were similar to methanogens belonging to the order Methanobacteriales. Within Methanobacteriales, 12 clones (6 OTUs) were similar to Methanosphaera stadtmanae and the remaining 8 phylotypes (64 clones) were similar to unclassified Methanobacteriales. Overall, members of the Methanobacteriales dominated the mcrA clone library in the rumen of Surti buffalo. Further studies and effective strategies can be made to inhibit the growth of Methanobacteriales to reduce methane emission from the rumen which would help in preventing global warming.

Genetic diversity among Indian Gir, Deoni and Kankrej cattle breeds based on microsatellite markers

Abstract: D S Kale*, D N Rank, C G Joshi 1 , B R Yadav 2 , P G Koringa, K M Thakkar, T C Tolenkhomba 2 and J V Solanki Department of Animal Genetics and Breeding and Department of Animal Biotechnology College of Veterinary Sciences and Animal Husbandry, Anand Agricultural University, Anand 388 001, India Livestock Genome Analysis Laboratory, Dairy Cattle Breeding Division National Dairy Research Institute (NDRI), Karnal 132 001, India

Metagenomic analysis of poultry gut microbes

Abstract: A novel, unbiased approach of next generation sequencing i.e. Roche 454 GS-FLX technology was used to study metabiome of poultry gut microbes. Poultry gut microbiota is composed of approximately 1014 to 1015 microorganisms. We analyzed 6.3 million base pairs assembled to make 20927 sequences with average read length of 304.21 nucleotide. Nucleotide sequences were obtained from DNA of the caecal contains from five adult layer birds. Using MG-RAST, metabolic function analyses and phylogenetic analysis of identified genes were carried out. Metabolic analysis based on subsystem technology reveals significantly enriched maltose & maltodextrin utilization, methionine biosynthesis, mannose metabolism, cellulosome, peptidoglycan biosynthesis, and other carbohydrate metabolism with protein biosynthesis and virulence associated genes. The phylogenetic analysis was carried out using RDP, Silva LSU, Silva SSU, Green Gene and SEED based technology. Comprehensive results of sequence based phylogenetic analysis identified bacteroidetes/chlorobi group, firmicutes, proteobacteria, methanomicrobia, metazoa group, uncultured bacteria, salmonella typhimurium bacteriophase, streptococcus pneumoniae bacteriophase as well as other phages.

Genetic characterization of Malvi cattle using microsatellite markers

Abstract: Microsatellite markers offer great potential for genetic comparisons within and between populations. They are the best available molecular tools for characterization of breeds. Microsatellite markers (25) were used to screen 70 Malvi cattle. The mean number of alleles was 9.08 per microsatellite locus with the range of 4 (ILSTS011, ILSTS054) to 18 (CSSM663). The range in the allele size was from 88 bp (CSRM60) to 297 bp (ILSTS006). The microsatellite loci deviated significantly from Hardy-Weinberg equilibrium. Sign test and Wilcox on revealed no Bottleneck in recent past in Malvi population. The mean polymorphic information content was 0.71 ranging from 0.41 to 0.91. The overalls means for observed and expected heterozygosity were 0.54 and 0.75 respectively. The overall observed heterozygosity, PIC and Shannon index values were 0.54, 0.71 and 1.67 indicating high gene diversity. The markers used in the present study were highly informative.

Molecular basis of Post-surgical Peritoneal adhesions - An Overview

Abstract: Post surgical adhesion development remains a frequent occurrence and is often unrecognized by surgeons. Peritoneal adhesions are the leading cause of pelvic pain, bowel obstruction and infertility. The prevention of adhesion till date is speculative due to lack of understanding of mechanisms involved in adhesion development. Adhesions are proposed to the disorder of wound healing and imbalance between fibrinogenesis and fibrinolysis. The unprecedented advancement in Molecular Biology has led us to identify molecules involved in both wound healing and adhesion development. The role of these molecules in peritoneal biological functions is not well understood. Hypoxia is proposed to be major contributing factor for the development of adhesions. The major mechanisms behind adhesion development are increased fibrinogenesis, reduced fibrinolysis, increased Extra Cellular Matrix deposition, increased cytokine production, increased angiogenesis and reduced apoptosis. Better understanding of these events will make efficient management of adhesions possible.

Genetic polymorphism in the aldo-keto reductase family 1 member B1 (AKR1B1) gene of Murrah buffalo bulls (Bubalus bubalis).

Abstract: The present study was conducted to investigate the existence of polymorphism at the AKR1B1 locus by the PCR-RFLP method. The genotypes were confi rmed by checking specifi c, clearly distinguishable DNA band patterns resulting from digestion with the restriction enzyme Nde I. Among the 41 animals studied, 18 samples produced two bands of 463 bp and 333 bp referred as AA genotype, whereas 23 animals produced three bands of 796 bp, 463 bp and 333 bp referred as AG genotype. None of the animals revealed GG genotype. The allelic frequencies of ‘A’ and ‘G’ alleles were found to be 0.725 and 0.275, respectively. Association analysis revealed that none of the genotypes were associated with any semen quality traits studied. The current study is the fi rst report of DNA based genotyping of AKR1B1 gene of this indigenous buffalo breed of India.