Identification, Detection and Enumeration of Klebsiella based on Culture-Independent Real-Time PCR: “The Klebsiella: ‘An Emerging Environmental Threats’ in Riverine-Systems”

Klebsiella is one of the fecal coliform bacteria of family Enterobacteriaceae. It is an opportunistic pathogen which previously used as an indicator bacterium along with fecal coliforms namely Escherichia coli, Citrobacter, and Enterobacter, but is now known to cause various diseases in human beings. In present study we collected 126 samples from riverine-systems and confirmed the existence of Klebsiella using the conserved sequence of mdh gene with the application of SYBR green real-time PCR, which demonstrated very high specificity and stringency in detection strategy. The Klebsiella levels in surface water were 8.28 x102 ±17, 4.76 x103 ±122, 6.0 x103±178, 9.96 x103 ±315, 1.27 x104 ±463, 2.76 x104 ±1085, cfu/100ml; associated with hydrophytes were 5.80 x104 ±2320, 2.48x104±806, 2.86x104 ±963, 1.01x104 ±249, 2.62x104 ±830, 2.46x104 ±706 cfu/10g and in sediments were in the range of 1.89 x104 ±408, 2.46 x104 ±530, 3.20 x104 ±1004, 2.71 x104 ±777, 2.82 x104 ±693 and 8.88 x104 ±3214 cfu/10 g (p<0.05) at sampling site # 1, site # 2, site # 3, site # 4, site # 5 and, site # 6, respectively. This assay could be applied in the detection and monitoring of environmental samples to check waterborne diseases and Klebsiella caused outbreaks.