About
Summary
Myself Santhosh K.S., from India. I completed my PhD in May 2015 with specialization in Microbiology from Karnataka Veterinary, Animal and Fisheries science university, Bidar, India. For my PhD research, I worked on the "Antibiotic resistance associated with gram negative bacteria from the environment and its impact". In my study Gram negative bacteria were isolated from different sources, identified and characterized. Isolates having higher percentage of antibiotic resistances to multidrugs were sequenced. The sequence data were analysed using various bioinformatic tools. Antibiotic resistant determinants were identified and the mode of transformation of resistance i.e., transduction, transformation and conjugation between bacteria were studied. The bacteriophages were isolated and were analysed for the resistance determinants. These phages were also used in studying the transfer of resistance by phage mediated transduction. Also, mutations in genes responsible for quinolone resistance were analyzed in the study. Over all I became proficient in microbiology, molecular microbiology, bioinformatics.
After my PhD I worked in few projects that helped me in gaining knowledge on real time PCR, cloning and expression of proteins, protein purification, vaccine development, development of siRNA as a vaccine candidate, isolation of food borne and shrimp pathogens and their specific bacteriophages, applications of bacteriophages as a biocontrol of pathogens in food industry and also as a therapeutic agents to control diseases in aquatic animals. Detection of pathogens using antibody based flow-through assay. Quantitative real time expression analysis of novel immune genes and antimicrobial peptides from Eukaryotes and their protein characterization. Handling small animals like mice, rabbit etc.
Positions
Research Associate Jan 2015 -
project “Capture and removal of ammonia from waste water using Archaea” funded by National Fund for Basic and Strategic Research in Agriculture (NFBSRA).
Theme:
a) To identify archaeal members in the oxic, suboxic and anoxic stages of seafood processing effluent treatment plant and in aquaculture pond sediments that show deterioration.
b) To study optimum physiological parameters for mass production of the identified archaeal strains.
c) To enrich archaeal strains and apply them for efficient effluent treatment.
d) To harvest the archaeal biomass after effluent treatment for use as biofertiliser.
PhD student Sep 2011 - Dec 2014
In my study Gram negative bacteria were isolated from different sources, identified and characterized. Isolates having higher percentage of antibiotic resistances to multidrugs were sequenced. The sequence data were analysed using various bioinformatic tools. Antibiotic resistant determinants were identified and the mode of transformation of resistance i.e., transduction, transformation and conjugation between bacteria were studied. The bacteriophages were isolated and were analysed for the resistance determinants. These phages were also used in studying the transfer of resistance by phage mediated transduction. Also, mutations in genes responsible for quinolone resistance were analyzed in the study. Over all I became proficient in microbiology, molecular microbiology, bioinformatics.
Education
College of Fisheries, Mangalore 2011 - 2015
Field of study: Fisheries Microbiology
Degree: PhD
Doctoral research (2011-2015): During my PhD, I worked on the topic “Antibiotic resistance associated with Gram negative bacteria from the environment and its impact’’ to understand the susceptibility of antimicrobial drugs against the major pathogenic Gram negative bacteria isolated from the environment under the joint supervision of Prof. Indrani Karunasagar, Director (R & D), NITTE University, Mangalore and Dr. M.N. Venugopal, Professor and Head, Dept. of Microbiology, College of Fisheries, Mangalore. The topic of my doctoral research is very relevant and deals with one of the most challenging problems of modern world, i.e. the development and spread of antibiotic resistance among bacteria of human health significance.
Objective-1: My research work was well planned and suitable standard protocols were used for isolation, identification and confirmation of Gram negative bacteria from different environment sources. Antibiotic resistance pattern was drawn for isolated bacteria by examining their susceptibility to major group of antimicrobial drugs which were frequently used.
Objective-2: The genotypic determinants responsible for the resistance in bacterial isolates were characterized by nucleic acid based molecular techniques - polymerase chain reaction (PCR) and agents of transfer such as plasmids, class1 integrons and phages accountable for resistance were studied. Mutation analysis- The selective evolutionary pressure of antimicrobials on bacterial population makes the previously sensitive isolate resistant. This kind of acquired antibiotic resistance is caused due to mutation (change in nucleotide) in bacterial genes. In this study, quinolone resistant isolates were analyzed for point mutation and results showed two reported single point mutations and one novel point mutation.
Objective-3: Antibiotic resistance is naturally transmissible between bacteria through genes by different modes involving genetic recombination mechanisms such as transformation, transduction or conjugation. Horizontal transmission of antibiotic resistance gene between bacteria was performed experimentally. In present study, the different mode of resistance transfer was examined. Transformation – the naked DNA (plasmid – resistance gene carrier (tetB)) extracted from multi-drug resistant isolate (EC10) was transferred into sensitive isolate (E. coli K12). Conjugation - is a mode of genetic recombination that involves the transfer of genetic element between two living bacterial cells with direct physical contact. The resistant isolate (carrying tetA gene in their plasmid) selected for conjugation with E. coli K12 cell and result revealed the transfer of resistance gene. Transduction – where the resistance genes are transferred from one bacterium to another by a phage. In the current study, the transfer of resistance gene (blaCTX) to isolate (EC153) through phage (p13) was experimentally demonstrated the transduction mode of gene transfer between bacteria.
College of Fisheries, Mangalore 2009 - 2011
Field of study: Fisheries Microbiology
Degree: MFSc
During my master degree program I worked on the dissertation topic entitled “Nucleic acid based identification of Nodavirus and Iridovirus associated with wild caught and cultured marine finfish”. Two years Master degree program with research work has endowed me with the expertise needed to deal with problems encountered in research and the course work has strengthened my theoretical knowledge as well.
Colleg of Fisherries, Mangalore 2005 - 2009
Field of study: Fisheries sciences
Degree: BSc
Skills
Molecular Biology: Isolation of DNA, RNA, RT-PCR, Real Time PCR (q-PCR), SDS-PAGE, 2D Gel-Electrophoresis, RFLP, RAPD, Microarray analysis.
Microbiology: Isolation and identification of bacteria (classical and molecular methods), bacterial culture techniques and preparation of competent bacterial cells and maintenance of glycerol stock cultures. Antibiotic profiling, detection of resistance determinants, mode of resistance transfer in in-vitro condition using different molecular techniques- transformation, conjugation and transduction.
Protein: Experience in the cloning, expression and purification of proteins (in prokaryotic system). Perform affinity chromatography, preparation of sub-cellular fractions of bacteria, Western blot analyses, polyclonal antibody production and Enzyme-Linked Immuno Sorbent Assay (ELISA).
Professional interests
Molecular biology, Microbiology, Biotechnology, Cell biology, Genetics and engeneering
CV
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