Characterization, genome analysis and in vitro activity of a novel phage vB_EcoA_RDN8.1 active against multi-drug resistant and extensively drug-resistant biofilm-forming uropathogenic Escherichia coli isolates, India

Abstract

Aim:We aimed to study host range, stability, genome and antibiofilm activity of a novel phage vB_EcoA_RDN8.1 active against multi-drug resistant (MDR) and extensively drug-resistant (XDR) biofilm-forming uropathogenic Escherichia coli isolates. Methods and Results: A novel lytic phage vB_EcoA_RDN8.1 active against UPEC strains resistant to third-generation cephalosporins, fluoroquinolones, aminoglycosides, imipenem, beta-lactamase inhibitor combination and polymyxins was isolated from community raw sewage water of Chandigarh. It exhibited a clear plaque morphology and a burst size of 250. In the time-kill assay, the maximum amount of killing was achieved at MOI 1.0. vB_EcoA_RDN8.1 belongs to the family Autographiviridae, has a genome size of 39.5 kb with a GC content of 51.6%. It was stable over a wide range of temperatures and pH. It was able to inhibit biofilm formation which may be related to an endolysin encoded by ORF 19. Conclusions: The vB_EcoA_RDN8.1 is a novel lytic phage that has the potential for inclusion into phage cocktails being developed for the treatment of urinary tract infections (UTIs) caused by highly drug-resistant UPEC. Significance and Impact of the Study: We provide a detailed characterization of a novel lytic Escherichia phage with antibiofilm activity having a potential application against MDR and XDR UPEC causing UTIs.

Genome Analysis and Antibiofilm Activity of Phage 590B against Multidrug-Resistant and Extensively Drug-Resistant Uropathogenic Escherichia coli Isolates, India

“…It is challenging to eradicate bacteria present in the biofilms due to the antimicrobial-resistant phenotype that this structure confers. Additionally, persister cells found in biofilms exhibit a slowed metabolism, which raises the levels of AMR [10]. Lytic phages are emerging as promising agents to combat multi-drug-resistant and biofilm-associated infections [37].…”

“…Serotyping was performed at the Central Research Institute (CRI), Kasauli, India. Virulence genes of phage-sensitive bacterial isolates were conducted using PCR with previously described primers (Table S1) [10]. E. coli MTCC 739 obtained from Chandigarh was used as a biofilm-forming control strain.…”

“…The biofilm-forming capacity of the library of 50 UPEC strains was tested using the crystal violet assay [10,29,30]. The quantitative and qualitative antibiofilm activity of the phage 590B was estimated using a Foley silicone catheter and polystyrene coverslips models, respectively.…”

Vibrio Phage VMJ710 Can Prevent and Treat Disease Caused by Pathogenic MDR V. cholerae O1 in an Infant Mouse Model

“…V. cholerae O1 strains obtained from clinical cases of cholera and sewage water showed acquired non-susceptibility to at least one agent in three or more antimicrobial categories of the following classes of antibiotics: Third-generation cephalosporins, fluoroquinolones, aminoglycosides, tetracyclines, ampicillin, chloramphenicol, and cotrimoxazole were defined as MDR V. cholerae [11]. V. cholerae serotype O1 (strain ATCC 39315/El Tor Inaba N16961) was used to standardize the induction of cholera in mice.…”

Complete Genome Sequence of Salmonella Phage vB_SenA_SM5, Active against Multidrug-Resistant Salmonella enterica Serovar Typhi Isolates

“…A single plaque was selected and propagated twice as described by Valente et al ( 2 ). The phage preparation was concentrated by the addition of polyethylene glycol (10% PEG 8000), followed by ultracentrifugation at 55,000 rpm for 2 h ( 3 ). The final phage volume was resuspended in 1 mL of SM buffer (5.8 g/L NaCl, 100 mM MgSO 4 ·7H 2 O).…”