Dataset Information

Isolation and Characterization of Lytic Bacteriophages Active against Clinical Strains of E. coli and Development of a Phage Antimicrobial Cocktail.

ABSTRACT: Pathogenic E. coli cause urinary tract, soft tissue and central nervous system infections, sepsis, etc. Lytic bacteriophages can be used to combat such infections. We investigated six lytic E. coli bacteriophages isolated from wastewater. Transmission electron microscopy and whole genome sequencing showed that the isolated bacteriophages are tailed phages of the Caudoviricetes class. One-step growth curves revealed that their latent period of reproduction is 20-30 min, and the average value of the burst size is 117-155. During co-cultivation with various E. coli strains, the phages completely suppressed bacterial host culture growth within the first 4 h at MOIs 10^-7 to 10^-3. The host range lysed by each bacteriophage varied from six to two bacterial strains out of nine used in the study. The cocktail formed from the isolated bacteriophages possessed the ability to completely suppress the growth of all the E. coli strains used in the study within 6 h and maintain its lytic activity for 8 months of storage. All the isolated bacteriophages may be useful in fighting pathogenic E. coli strains and in the development of phage cocktails with a long storage period and high efficiency in the treatment of bacterial infections.

SUBMITTER: Alexyuk P

PROVIDER: S-EPMC9697832 | biostudies-literature | 2022 Oct

REPOSITORIES: biostudies-literature

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Isolation and Characterization of Lytic Bacteriophages Active against Clinical Strains of E. coli and Development of a Phage Antimicrobial Cocktail.

Alexyuk Pavel P Bogoyavlenskiy Andrey A Alexyuk Madina M Akanova Kuralay K Moldakhanov Yergali Y Berezin Vladimir V

Viruses 20221028 11

Pathogenic E. coli cause urinary tract, soft tissue and central nervous system infections, sepsis, etc. Lytic bacteriophages can be used to combat such infections. We investigated six lytic E. coli bacteriophages isolated from wastewater. Transmission electron microscopy and whole genome sequencing showed that the isolated bacteriophages are tailed phages of the Caudoviricetes class. One-step growth curves revealed that their latent period of reproduction is 20-30 min, and t ...[more]

PMID: 36366479

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Project description:BackgroundEscherichia coli is a common fecal coliform, facultative aerobic, gram-negative bacterium. Pathogenic strains of such microbes have evolved to cause diarrhea, urinary tract infections, and septicemias. The emergence of antibiotic resistance urged the identification of an alternative strategy. The use of lytic bacteriophages against the control of pathogenic E. coli in clinics and different environmental setups (waste and drink water management) has become an alternative therapy to antibiotic therapy. Thus, this study aimed to isolate and characterize lytic bacteriophage from various sources in Addis Ababa, tested them against antimicrobial-resistant diarrheagenic E. coli strains and evaluated their therapeutic potential under in vitro conditions.MethodsA total of 14 samples were processed against six different diarrheagenic E. coli strains. The conventional culture and plaque analysis agar overlay method was used to recover lytic bacteriophage isolates. The phage isolates were characterized to determine their lytic effect, growth characteristics, host range activity, and stability under different temperature and pH conditions. Phage isolates were identified by scanning electron microscope (SEM), and molecular techniques (PCR).ResultsIn total, 17 phages were recovered from 84 tested plates. Of the 17 phage isolates, 11 (65%) were Myoviridae-like phages, and 6 (35%) phage isolates were Podoviridae and Siphoviridae by morphology and PCR identification. Based on the host range test, growth characteristics, and stability test 7 potent phages were selected. These phages demonstrated better growth characteristics, including short latent periods, highest burst sizes, and wider host ranges, as well as thermal stability and the ability to survive in a wide range of pH levels.ConclusionsThe promising effect of the phages isolated in this study against AMR pathogenic E. coli has raised the possibility of their use in the future treatment of E. coli infections.

Project description:Aim of studyThe objective of this study was isolation and morphological characterization of temperate bacteriophages obtained from M. haemolytica strains and evaluation of their lytic properties in vitro against M. haemolytica isolated from the respiratory tract of calves.Material and methodsThe material for the study consisted of the reference strain M. haemolytica serotype 1 (ATCC®) BAA-410™, reference serotypes A1, A2, A5, A6, A7, A9 and A11, and wild-type isolates of M. haemolytica. Bacteriophages were induced from an overnight bacterial starter culture of all examined M. haemolytica strains treated with mitomycin C. The lytic properties and host ranges were determined by plaque assays. The morphology of the bacteriophages was examined in negative-stained smears with 5% uranyl acetate solution using a transmission electron microscope. The genetic analysis of the bacteriophages was followed by restriction analysis of bacteriophage DNA. This was followed by analysis of genetic material by polymerase chain reaction (PCR).ResultsEight bacteriophages were obtained, like typical of the families Myoviridae, Siphoviridae and Podoviridae. Most of the bacteriophages exhibited lytic properties against the M. haemolytica strains. Restriction analysis revealed similarities to the P2-like phage obtained from the strain M. haemolytica BAA-410. The most similar profiles were observed in the case of bacteriophages φA1 and φA5. All of the bacteriophages obtained were characterized by the presence of additional fragments in the restriction profiles with respect to the P2-like reference phage. In the analysis of PCR products for the P2-like reference phage phi-MhaA1-PHL101 (DQ426904) and the phages of the M. haemolytica serotypes, a 734-bp phage PCR product was obtained. The primers were programmed in Primer-Blast software using the structure of the sequence DQ426904 of reference phage PHL101.ConclusionsThe results obtained indicate the need for further research aimed at isolating and characterizing bacteriophages, including sequence analysis of selected fragments. Moreover, standardization of methods for obtaining them in order to eliminate M. haemolytica bacteria involved in the etiopathogenesis of BRDC is essential.

Project description:Diabetic foot infection has become one of the most important public health concerns and is a growing problem. Pseudomonas aeruginosa is an important opportunistic multidrug-resistant bacterium in diabetic foot infections. In the absence of antibiotics active against MDR strains of P. aeruginosa, phage therapy becomes a key way to deal with P. aeruginosa infections. Out of 185 samples collected from diabetic foot ulcers, 50 (27.02%) isolates were identified as P. aeruginosa. The incidence increases with older ages, and males (n=34, 68%) predominated in all age groups. The tested isolates showed maximum susceptibility towards colistin (80%), imipenem (72%), amikacin (66%), and piperacillin/tazobactam (62%), while these isolates showed moderate susceptibility towards ceftazidime (58%), cefepime (52%) and gentamicin (46%). However, it showed complete resistance (100%) to ampicillin, cefaclor, and sulphamethoxazole/trimethoprim and highly resistance to clindamycin (90%) and amoxicillin/clavulanic acid (84%). Two bacteriophages (ϕPAE1 and ϕPAE2) isolated from sewage samples showed a broad host range against P. aeruginosaa clinical strains. ϕPAE2 infected 74% (37/50) and ϕPAE2 58% (29/50). Furthermore, both phages were host-specific, infecting only P. aeruginosa strains and could not infect other bacterial species in the cross-infectivity studies. Both phages were found to be relatively heat stable as over a period of 1 h, after exposure to a temperature range of 37-50°C, no significant loss in phage activity was observed. On the other hand, the lowest activity was observed at 70°C (39.15%) for ϕPAE1 whereas it was inactivated at 75°C while the lowest activity was observed at 75°C (38.01%) for ϕPAE2 whereas it was inactivated at 80°C. Isolated phages were able to survive and lyse host bacteria over a wide pH range. The optimum pH range for infection was from 6 to 8. Furthermore, ϕPAE1 lost its ability to lyses at pH 2, 3, 11 and 12, whereas; ϕPAE2 lost its infectivity at pH 2, 3 and 12. Chloroform was the most effective solvent that reduced the infectivity of ϕPAE1 and ϕPAE2 to 63.27% and 77.88%, respectively. On the other hand, petroleum ether showed the lowest effect on the infectivity of ϕPAE1 and ϕPAE2; it was reduced to 96.4% and 97.48%, respectively, followed by acetone and ethyl alcohol. The ability of P. aeruginosa phages to form plaques after different storage temperatures (4°C, 30°C, 37°C and 44°C) for a month was slightly affected. The storage of ϕPAE1 and ϕPAE2 at 4ºC showed the least effect on its infectivity, and the storage at 44ºC showed the highest reduction in its infectivity. Moreover, Phage counts were slightly decreased by increasing storage period and temperature.

Dataset Information

Isolation and Characterization of Lytic Bacteriophages Active against Clinical Strains of E. coli and Development of a Phage Antimicrobial Cocktail.

Publications

Isolation and Characterization of Lytic Bacteriophages Active against Clinical Strains of <i>E. coli</i> and Development of a Phage Antimicrobial Cocktail.

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