Isolation, Molecular Characterization, and Therapeutic Evaluation of Bacteriophages Targeting Shiga Toxin-Producing Escherichia coli (STEC)

Hillary  Debbarma; Moon Moon Sathpathy; Ambika Arun; Abhishek; Bablu Kumar

doi:10.56093/ijans.v95i12.170345

Authors

Hillary Debbarma
Moon Moon Sathpathy
Ambika Arun
Abhishek
Bablu Kumar

https://doi.org/10.56093/ijans.v95i12.170345

Keywords:

STEC phage, Phage Therapy, AMR

Abstract

Bacteriophages offer a promising alternative for controlling pathogenic drug-resistant bacteria. Shiga toxin-producing Escherichia coli (STEC), a significant foodborne pathogen with zoonotic potential, poses health risks through contaminated food sources. This study aimed to isolate phages against STEC from twenty-three sewage samples, including those from poultry, swine, cattle farms, and hospitals sources. A total number of twenty-two STEC isolates were biochemically and molecularly characterized. Molecular detection of Shiga toxin producing genes revealed that 86.36% (19/22) had the stx1 gene and 50% (11/22) had the eae gene. None had stx2 or a combination of stx1 and stx2 genes. Thirteen lytic bacteriophages were isolated against the indicator strain (ATCC 2469), with twelve showing lytic activity against STEC isolates. In order to determine the lytic spectrum, these twelve phages were tested on 66 E. coli isolates (22 STEC and 44 non-STEC). Phages (n=5) showing highest lysis i.e. >=22.72% (15/66), were morphologically characterized via Transmission Electron Microscopy (TEM), followed by stability testing across various range of temperature and pH. Three phages belonged to Podoviridae and two belonged to Siphoviridae families. The phages were found stable at a wide range of temperature (-20°C to 37°C) and pH (4 to 9), with optimal stability observed at 4°C and pH 7.5. Phage Ecp10, exhibiting the highest lytic range (33.33%, 22/66), was tested in spiked milk samples at different MOIs. The highest reduction in bacterial count was recorded at MOI 0.1 after 6 hours of incubation. This study indicates that Ecp10 can be a promising decontaminating agent in milk and a potential treatment for STEC infections in animals and humans.

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Author Biographies

Hillary Debbarma

M.V.Sc Scholar, Division of Bacteriology & Mycology, ICAR-IVRI, Bareilly, 243122, India
Moon Moon Sathpathy

Ph.D. Scholar, Division of Bacteriology & Mycology, ICAR-IVRI, Bareilly, 243122, India
Bablu Kumar

Assistant Professor
Deptt Of Vety Microbiology
Bihar Veterinary College
Patna
800014

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