Folia Microbiologica

, Volume 64, Issue 6, pp 875–887 | Cite as

Genetic diversity and multiple antibiotic resistance index study of bacterial pathogen, Klebsiella pneumoniae strains isolated from diseased Indian major carps

  • A. Das
  • B. K. BeheraEmail author
  • S. Acharya
  • P. Paria
  • H. J. Chakraborty
  • P. K. Parida
  • B. K. Das
Original Article


Intensive fish farming systems have led to increase in disease incidence, due to higher stocking density, high organic matter levels, and poor quality of the aquatic environment. Diseased fish samples showing hemorrhages and reddish lesions were collected from different freshwater fish farms located at three different districts of West Bengal, India (Burdwan, North 24 Parganas, and Nadia). The present study was conducted to evaluate the genetic diversity of ten different Klebsiella pneumoniae strains isolated from different infected freshwater fish samples based on 16S rRNA gene sequence analysis. Primarily, Klebsiella-specific media was used for the isolation and characterization of Klebsiella pneumoniae. Further, through a biochemical test, all the strains were confirmed as K. pneumoniae. PCR analysis of 16S–23S internal transcribed spacer (PCR ribotyping) was carried out to study the species variation within different Klebsiella pneumoniae isolates. For all the isolates, a conserved PCR ribotype pattern was observed while differing from other bacterial species. Phylogenetic study showed the high degree of homology with diverse source of other strains. The multiple antibiotic resistance (MAR) values of the present study for the isolates were found to be 0.468. MAR value above 0.2 indicates that the source of isolation was highly contaminated with antibiotics. Based on the 16S rRNA gene sequence analysis, the present study revealed the genetic diversity of Klebsiella pneumoniae isolated from the different diseased fish farms of West Bengal. All the strains were found to be hypermucoviscous and multidrug-resistant, thus making it pathogenic towards the host organisms. Further, the study revealed a high prevalence of K. pneumoniae in aquaculture farms, representing a risk towards successful aquaculture.


Klebsiella pneumoniae Genetic diversity Carp fish Antibiotic resistance index India 



The authors are thankful to Dr. Joykrushna Jena, Deputy Director General (Fishery Science), Indian Council of Agricultural Research, New Delhi, for his support and guidance. The authors are also thankful to Mr. Asim Kumar Jana for laboratory assistance.

Funding information

This work was funded by the National Fisheries Development Board (NFDB) (G/Nat. Surveillance/2013 dated 16.08.2013), Hyderabad, under the “National Surveillance Program for Aquatic Animal Diseases (NSPAAD)” Project.

Compliance with ethical standards

Ethical approval

The fish samples for this study were collected and handled in accordance with the ICAR-Central Inland Fisheries Research Institute (CIFRI), Kolkata, India ethical guidelines, and regulations. Further, the experimental protocols for this study were approved by the Institute Research Committee of ICAR-CIFRI (Project Code: OXXO3049).

Conflict of interest

The authors have declared no conflict of interest.

Supplementary material

12223_2019_701_MOESM1_ESM.docx (185 kb)
ESM 1 (DOCX 184 kb)


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Copyright information

© Institute of Microbiology, Academy of Sciences of the Czech Republic, v.v.i. 2019

Authors and Affiliations

  1. 1.ICAR-Central Inland Fisheries Research InstituteKolkataIndia
  2. 2.Department of ZoologyVidyasagar UniversityMedinipurIndia

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