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Isolation and Characterization of a Myoviridae MJ1 Bacteriophage Against Multi-Drug Resistant Escherichia coli 3.

ABSTRACT: BACKGROUND:Antibiotic resistance in Escherichia coli, a member of the Enterobacteriaceae, is of particular concern because it is the most common (Gram-negative) pathogen causing nosocomial and community infections. Researchers are now considering the use of phages for the control of various antibiotic-resistant bacterial infections. OBJECTIVES:The purpose of this study was to isolate and characterize a novel pathogenic/lytic phage that targets multi-drug resistant (MDR) E. coli 3, and to investigate its effectiveness at lysing this bacterium. MATERIALS AND METHODS:A clinical strain of E. coli 3 was identified based on its 16S rRNA sequencing and its antibiotic resistance profile was determined by the disc diffusion method. A bacteriophage was isolated from wastewater and its various characteristics, such as host range, heat tolerance, pH stability, one step growth, total protein content, and genome size, were determined. The antibacterial property of the phage was determined against log-phase bacterial planktonic cells at 37°C. RESULTS:The bacteriophage, designated MJ1, was isolated by testing against a clinical MDR E. coli 3 strain. The MJ1 phage showed a wide range of heat and pH stability. The phage morphology, determined by transmission electron microscopy, revealed a structure comprised of a head (108 ± 0.2 nm long by 128 ± 0.5 nm wide) and a contractile tail (123 ± 0.5 nm long by 15 - 26 nm wide). These features placed the MJ1 phage in the family Myoviridae and the order Caudovirales. Eleven structural proteins (17 to 200 kDa) for this phage were detected by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). A double stranded DNA, approximately 32 kb, in size was detected for this phage on agarose gels. The phage efficacy against E. coli 3 planktonic cells was also investigated. The MJ1 phage demonstrated a very good capability to reduce the numbers of E. coli 3 planktonic cells, as determined by a change in the bacterial growth (an optical density decrease at 600 nm from 0.40 to 0.12). CONCLUSIONS:MJ1 phage has much potential for use in phage therapy and other applications.

SUBMITTER: Jamal M

PROVIDER: S-EPMC4744325 | biostudies-literature | 2015 Nov

REPOSITORIES: biostudies-literature

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Isolation and Characterization of a Myoviridae MJ1 Bacteriophage Against Multi-Drug Resistant Escherichia coli 3.

Jamal Muhsin M Hussain Tahir T Rajanna Das Chythanya C Andleeb Saadia S

Jundishapur journal of microbiology 20151121 11

<h4>Background</h4>Antibiotic resistance in Escherichia coli, a member of the Enterobacteriaceae, is of particular concern because it is the most common (Gram-negative) pathogen causing nosocomial and community infections. Researchers are now considering the use of phages for the control of various antibiotic-resistant bacterial infections.<h4>Objectives</h4>The purpose of this study was to isolate and characterize a novel pathogenic/lytic phage that targets multi-drug resistant (MDR) E. coli 3, ...[more]

PMID: 26865936

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Project description:Atypical enteropathogenic E. coli (aEPEC) is a group of diarrhoeagenic Escherichia coli with high diversity of serogroups, which lack the bundle-forming pili (BFP) and genes encoding for shiga toxins. The aim of this study was to isolate, identify and determine virulence and antibiotic resistance profiles of aEPEC O177 strains from cattle feces. A total of 780 samples were collected from beef and dairy cattle and analyzed for the presence of E. coli O177. One thousand two hundred and seventy-two (1272) presumptive isolates were obtained and 915 were confirmed as E. coli species. Three hundred and seventy-six isolates were positively confirmed as E. coli O177 through amplification of rmlB and wzy gene sequences using multiplex PCR. None of these isolates harbored bfpA gene. A larger proportion (12.74%) of the isolates harbored hlyA gene while 11.20, 9.07, 7.25, 2.60, and 0.63% possessed stx 2, stx 1 , eaeA, stx 2a , and stx 2d , respectively. Most of E. coli O177 isolates carried stx 2/hlyA (9.74%). Furthermore, 7.40% of the isolates harbored stx 1/stx 2 while 7.09% possessed stx 1/stx 2/hlyA genes. Only one isolate harbored stx 1/stx 2/hly/eaeA/stx 2a /stx 2d while 5.11% of the isolates harbored all the four major virulence genes stx 1/stx 2/hlyA/eaeA, simultaneously. Further analysis revealed that the isolates displayed varied antimicrobial resistance to erythromycin (63.84%), ampicillin (21.54%), tetracycline (13.37%), streptomycin (17.01%), kanamycin (2.42%), chloramphenicol (1.97%), and norfloxacin (1.40%). Moreover, 20.7% of the isolates exhibited different phenotypic multi-drug resistance patterns. All 73 isolates harbored at least one antimicrobial resistance gene. The aadA, streA, streB, erm, and tetA resistance genes were detected separately and/or concurrently. In conclusion, our findings indicate that environmental isolates of aEPEC O177 strains obtained from cattle in South Africa harbored virulence and antimicrobial resistance gene determinants similar to those reported in other shiga-toxin producing E. coli strains and suggest that these determinants may contribute to the virulence of the isolates.

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Dataset Information

Isolation and Characterization of a Myoviridae MJ1 Bacteriophage Against Multi-Drug Resistant Escherichia coli 3.

Publications

Isolation and Characterization of a Myoviridae MJ1 Bacteriophage Against Multi-Drug Resistant Escherichia coli 3.

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