Abstract

ISOLATION AND CHARACTERIZATION OF BACTERIOPHAGES AGAINST CARBAPENEM RESISTANT ENTEROBACTERIACEAE FROM SEWAGE

Swarnalatha B.*

Abstract

Antibiotic resistant „superbugs‟ have become one of the world‟s most important public health concerns. The increase in Carbapenemresistant enterobacteriaceae[CRE] poses a challenge in administration of effective treatment for the most common infections. In this study, carbapenem-resistant Klebsiella pneumoniae was isolated from raw sewage samples and identified by standard methods such as modified Hodge test and biochemical testing. Bacteriophages against this pathogen were isolated by double layer method. The bacteriophage was observed as plaques which were later purified and the potent phage was concentrated by PEG precipitation. The isolated and purified phage was characterized for structural, genomic and growth properties. Transmission electron microscopy revealed that the phage belonged to the family Myoviridae. The bacteriophage also showed a narrow host range. One step growth kinetics showed that the latent period was 20 minutes and the burst size was found to be 9-11 virions/cell. The bacteriophage was viable at pH range of 4-9 and a temperature up to 600c. Genomic characters were studied by using enzyme technique and dsDNA genome was detected. The objective of this study was to isolate and characterize a bacteriophage which may exhibit properties indicative of potential use in phage therapy and to add new insights of finding alternatives for treatment of infections caused by carbapenemresistant enterobacteriaceae.

Keywords: Bacteriophage, Klebsiella Pneumoniae, Phage Therapy, Carbapenem- Resistant Enterobacteriaceae.