Aim: Phage therapy, as an effective and specific method in the treatment of multidrug-resistant (MDR) bacterial infections, has attracted the attention of many researchers. Methods and results: In this study, a double-stranded DNA phage with the ability of lysing some strains of MDR Klebsiella pneumoniae (vB_Kpn_3) was isolated from hospitals’ wastewater and then characterized morphologically and genetically. Transmission electron microscopy and genetic analyses have revealed that vB_Kpn_3 is a member of Siphoviridae family. One-step growth curve also showed a burst time of 35 min and a burst size of 31 PFU/ml. The genome of the phage is composed of 112,080 bp with 41.33% G + C content carrying 186 open reading frames. Conclusion: vB_Kpn_3 is a broad host range phage that infects MDR K. pneumoniae and some other species of Enterobacteriaceae such as Escherichia coli and Salmonella typhi. In addition, no antibiotic resistance and toxin genes were detected in its genome.
Background and Aim: This study aimed to isolate phages able to lyse some strains of multiple drug-resistant (MDR)-Klebsiella pneumoniae (named vB_Kp1 and vB_Kp2) and Enterobacter aerogenes (named vB_Ea1) from swage. Materials and Methods: Different K. pneumoniae and E. aerogenes strains were isolated from clinical specimens during January-September 2018 in three hospitals of Amol, Mazandaran, Iran. Kirby-Bauer disc diffusion method was used to determine the resistance profiles of these isolates using different antibiotics. The MDR strains were selected for investigating the effect of isolated phages from wastewater and hospital sewage. The presence of phage was confirmed by plaque formation. The isolated bacteriophages were enriched, concentrated, and stained. Afterwards, a transmitting electron microscope (TEM) was applied to observe the morphology of the bacteriophages. Phage identification tests, including host range and one-step growth, were performed. Results: The TEM analysis revealed that three phages had an icosahedral capsid and long contractile tail being classified as a member of the Myoviridae family. Phages were able to lyse 14 (56%) of the 25 MDR isolated bacterial strains. The onestep growth curve showed large bursts and short latent times. Conclusion:The formation of clear plaques demonstrates the high lyse power of phages. Therefore, they have good potential for further analysis for clinical use as a therapeutic agent in the future.
Background and Objectives: Multi-drug-resistant Enterobacter aerogenes is associated with various infectious diseases that cannot be easily treated by antibiotics. However, bacteriophages have potential therapeutic applications in the control of multi-drug-resistant bacteria. In this study, we aimed to isolate and characterize of a lytic bacteriophage that can lyse specifically the multi-drug-resistant (MDR) E. aerogenes. Materials and Methods: Lytic bacteriophage was isolated from Qaem hospital wastewater and characterized morphologically and genetically. Next-generation sequencing was used to complete genome analysis of the isolated bacteriophage. Results: Based on the transmission electron microscopy feature, the isolated bacteriophage (vB-Ea-5) belongs to the family Myoviridae. vB-Ea-5 had a latent period of 25 minutes, a burst size of 13 PFU/ml, and a burst time of 40 min. Genome sequencing revealed that vB-Ea-5 has a 135324 bp genome with 41.41% GC content. The vB-Ea-5 genome codes 212 ORFs 90 of which were categorized into several functional classes such as DNA replication and modification, transcriptional regulation, packaging, structural proteins, and a host lysis protein (Holin). No antibiotic resistance and toxin genes were detected in the genome. SDS-PAGE of vB-Ea-5 proteins exhibited three major and four minor bands with a molecular weight ranging from 18 to 50 kD. Conclusion: Our study suggests vB-Ea-5 as a potential candidate for phage therapy against MDR E. aerogenes infections.
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