Agniswar Sarkar

Identification and Typing of Aeromonas Hydrophila through 16S rDNA-PCR Fingerprinting

16S rRNA is a conserved biomolecule within a cell. Sequencing data was analyzed and examined by sequencing the corresponding 16s rDNA regions of isolates from different sources. Specific culture media and different biochemical tests primarily confirmed isolates as Aeromonas hydrophila; those were designed and tested following PCR assays and identified by 16S rDNA gene sequence analysis. To automate the method few online and offline computational tools have been used. The method has proven useful for identification of Aeromonas species. Our results emphasize the need to take into account the intragenomic diversity of the 16S rRNA gene. Different software and programme used in this study are freely available online in different website.

Introduction to Establish the Comparative Analysis of 16S rRNA Gene Sequences with amoA and nxrA for Nitrifying Bacteria Isolated from East Kolkata Wetland: an International Ramsar Site

The nitrogen cycle is most complex and very important for the life on earth. Nitrifying bacteria which carried out nitrogen cycling process play a vital role in water quality control, thus creating a genomic fingerprint database for surveillance and monitoring of genetic variability of nitrifying bacteria is very important and also to establish a Biosecurity protocol for the Bheries located in different areas of West Bengal. The currently evolutionary relationships and the natural diversity of Ammonia Oxidizing Bacteria (AOB) and Nitrite Oxidizing Bacteria (NOB) is mainly based on comparative sequence analyses of their genes encoding the 16S rRNA and the active site polypeptide of the ammonia monooxygenase (AmoA) and Nitrite oxidoreductase (NxrA) in the East Kolkata Wetland. This study extended significantly the 16S rRNA and amoA databases for AOB, nxrA databases for NOB. Therefore, either of the functional markers (amoA or nxrA) can be used to trace ammonia oxidizers or structural marker (16S rDNA) can be used to trace specific species in environmental studies. These techniques included the use of 16S rDNA genes to characterize natural AOB and NOB populations and to analyze their taxonomic and phylogenetic features. The current perception of AOB and NOB phylogeny established by comparative 16S rRNA sequence analysis could be confirmed independently by exploiting the gene amoA and nxrA genes and proved as an alternative phylogenetic marker. The aim of our work is to investigate the potential of the ammonia monooxygenase subunit A gene (amoA) and Nitrite oxidoreductase (nxrA) gene as functional markers for AOBs and NOBs along with 16S rDNA sequence analysis in EKW. For ecological surveillance, genes like amoA and nxrA specific for the nitrifying bacteria under present research work could be a more reliable tool.

Isolation, characterization and protein profiling of lead resistant bacteria.

Aims: To isolate lead resistant bacteria from industrial effluent and characterize them by biochemical tests, effect of physical and chemicals factors on their growth and protein profiling of the isolates on with or without metal stress. Study Design: Cross-sectional study of related research articles and papers. Place and Duration of Study: Samples were collected from effluent water of Indian Iron and Steel Company (IISCO), Burnpur, West Bengal, India and studied at The Department of Biotechnology, The University of Burdwan. West Bengal, India, between September 2010 and June 2011. Methodology: Isolation of the isolates was done by pour plating the diluted effluent water onto the metal (Lead acetate) containing agar plates. The basic characterization of the isolates was done on the basis of morphological, physical, biochemical and antibiogram tests. Quantification of metal absorption was determined by Atomic absorption Spectrophotometric analysis. Protein expressions of the isolates on metal stress were studied by SDS-PAGE. Research Article British Microbiology Research Journal, 4(1): 116-131, 2014 117 Results: Two bacterial isolates namely PbB1 and PbB2 were isolated; they showed resistance to lead up to 6mM Lead (Lead acetate) and also to different antibiotics. They are able to ferment different sugars even in presence of low concentration of metals. Their protein profiling by SDS-PAGE shows different expressions of proteins upon metal stress. Conclusion: The isolates can be utilized to bioremediate lead from contaminated environment and further study of molecular mechanisms underlying lead accumulation process can be studied.

Development of Molecular Identification of Nitrifying Bacteria in Water Bodies of East Kolkata Wetland, West Bengal

Nitrifying bacteria plays a major role in converting the waste water to valuable renewable resources for the society. Ammonia is the toxic excretory product of most aquatic organisms and nitrite formed by the oxidation of ammonia is also toxic. Ammonia oxidizing bacteria converts ammonia to nitrite and Nitrite oxidizing bacteria convert nitrite to nitrate. Among them, Nitrosomonas sp. and Nitrobacter sp. has been the most widely studied organisms. An advanced molecular technique made it possible to explore the nitrifying bacteria in the environment and to enhance our knowledge of its functioning. In view of this it would be of prime importance for rapid detection of these strains/ isolates using molecular techniques. The present study was undertaken to develop and validate 16S rDNA sequence analysis of Nitrosomonas sp. and Nitrobacter sp. collected from different bheries in East Kolkata Wetland, West Bengal.