Journal of Environmental Biology

Bacterial diversity and antibiotic resistance in a wetland of

Lakhimpur-Kheri, Uttar Pradesh, India

Rupesh Kumar Mishra¹, Dev Kumar Verma¹, Ravindra¹, Manoj Kumar Yadav¹, Pravata Kumar Pradhan¹*, T. Raja Swaminathan² and Neeraj Sood¹

¹Fish Health Management Division, ICAR-National Bureau of Fish Genetic Resources, Lucknow-226 002, India

²Peninsular and Marine Fish Genetic Resources Division, NBFGR, CMFRI Campus, Kochi-682 018, India

*Corresponding Author E-mail: pradhanpk1@gmail.com

Key words

Antibiotic sensitivity,

Bactrial isolates,

16S rDNA sequencing,

Multiple antibiotic resistance,

Phylotypes

Publication Data

Paper received : 10.07.2015

Revised received : 21.11.2015

Re-revised received : 17.05.2016

Accepted : 26.07.2016

Abstract

Aim : Knowledge about the structure of microbial community is essential to assess the risks of exposure of farmed fish to pathogens and studying the antibiotic resistance. The present study was carried out to assess the bacterial diversity in a freshwater ecosystem.

Methodology : In the present study, microbial composition of a wetland in Lakhimpur-Kheri (N 27? 90.25' and E 80? 79.80'), Uttar Pradesh was assesesed employing amplified rDNA restriction analysis (ARDRA), using HaeIII enzyme as well as 16S rDNA gene sequencing. Furthermore, antibiotic resistance of the bacterial isolates was determined on the basis of multiple antibiotic resistance (MAR) index.  

Results : A total of one hundred and thirty five bacteria were isolated and the restriction analysis of amplified 16S rDNA gene of the above isolates, using HaeIII enzyme, indicated that isolated bacteria belonged to 33 single member phylotypes. Molecular characterization of the isolates revealed that these phylotypes belonged to 20 genera under 6 classes viz., Gammaproteobacteria (72.59%), Betaproteobacteria (4.44%), Flavobacteria (9.63%), Bacilli (7.41%) Actinobacteria (3.70%) and Sphingobacteria (2.22%). In the present study, four species i.e. Sphingobacterium kitahiroshimense, Yersinia nurmii, Pseudomonas koreensis and Vogesella alkaliphila have been reported for the first time from a wetland. The antibiotic sensitivity test indicated that most of the isolates were sensitive to norfloxacin followed by ofloxacin, whereas, majority of these bacteria were resistant to penicillin G followed by polymyxin B. Moreover, MAR indexing revealed that amongst the 33 single member phylotypes, 17 were multiple antibiotic resistant.

Interpretation : The bacterial diversity analysis of the wetland indicated bacterial population of aquatic, soil, animal and plant origin. Moreover, the study indicated that wetland can be a reservoir for multidrug resistant bacteria even when there is no application of antibiotics for aquaculture purposes. It can be assumed that the occurrence of antibiotic resistance and MAR among bacteria isolated from wetland could be due to flow of runoff from mainland. These results would help in strengthening our knowledge of bacterial ecology of the wetland ecosystem.

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