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Volume: 1; Issue: 4, Nov-Dec, 2013
Isolation and identification of pathogenic microbes from tomato puree and their delineation of distinctness by molecular techniques
R.K. Garg, N. Batav, N. Silawat and R.K. Singh
The present investigation is on isolation and identification of pathogenic microbes of tomato puree. On the basis of results of the gram's staining, growth on selective and differential media and biochemical analysis of sample S-1 and S-2 of tomato puree, total 12 pathogenic species i.e., Proteus mirabilis, Aeromonas hydrophila, Klebsiella sp., Vibrio furnissii, Vibrio sp., Erwinia herbicola, Vibrio furnissii, Serratia marcescens, Vibrio furnissii, Proteus sp., Pseudomonas sp. and Klebsiella sp. were recorded. Pathogenic Vibrio sp., Klebsiella sp., Serratia sp., and Pseudomonas sp. were seen in unsterilized sample S-2 and negligible in sterilized sample S-1. The amplification of genomic DNA from the seven bacterial species with the four primers showed the species have different RAPD profiles. The bands obtained ranged in size from 100-3000bp. DNA fingerprints were obtained using 04 random primers and gave 151 bands of which 71 are polymorphic, 64 monomorphic and 16 are unique bands representing 47.01%, 42.38% and 10.59% respectively. RAPD analysis successfully identified and 04 RAPD-PCR markers accurately distinguish between these seven species of bacteria. The technique is reliable, reproducible and relatively easy to use with these species. The result reported in this work is the first step in the genetic characterization of the one bacterial species, further step will include the use of RAPD in studies on population genetics.
Keywords: Microbes, pathogens, biochemical analyses, PCR, molecular diversity, microbial diversity.