Characterizing the Biology of Lytic Bacteriophage vB_EaeM_φEap-3 Infecting Multidrug-Resistant Enterobacter aerogenes

Abstract

Carbapenem-resistant Enterobacter aerogenes strains are a major clinical problem because of the lack of effective alternative antibiotics. However, viruses that lyze bacteria, called bacteriophages, have potential therapeutic applications in the control of antibiotic-resistant bacteria. In the present study, a lytic bacteriophage specific for E. aerogenes isolates, designated vB_EaeM_φEap-3, was characterized. Based on transmission electron microscopy analysis, phage vB_EaeM_φEap-3 was classified as a member of the family Myoviridae (order, Caudovirales). Host range determination revealed that vB_EaeM_φEap-3 lyzed 18 of the 28 E. aerogenes strains tested, while a one-step growth curve showed a short latent period and a moderate burst size. The stability of vB_EaeM_φEap-3 at various temperatures and pH levels was also examined. Genomic sequencing and bioinformatics analysis revealed that vB_EaeM_φEap-3 has a 175,814-bp double-stranded DNA genome that does not contain any genes considered undesirable for the development of therapeutics (e.g., antibiotic resistance genes, toxin-encoding genes, integrase). The phage genome contained 278 putative protein-coding genes and one tRNA gene, tRNA-Met (AUG). Phylogenetic analysis based on large terminase subunit and major capsid protein sequences suggested that vB_EaeM_φEap-3 belongs to novel genus “Kp15 virus” within the T4-like virus subfamily. Based on host range, genomic, and physiological parameters, we propose that phage vB_EaeM_φEap-3 is a suitable candidate for phage therapy applications.

Phages from Ganges River curtail in vitro biofilms and planktonic growth of drug resistant Klebsiella pneumoniae in a zebrafish infection model

“…In addition, the phage was also tested against reference strains of E. coli (MG1655), Acenitobacter baumannii, Enterobacter cloacae, Pseudomonas aeruginosa and Enterococcus faecalis. Spot assay was done as reported earlier (Zhao et al 2019 ). 300 µl of fresh bacterial culture was added to soft agar, overlaid on nutrient agar plate and allowed to solidify.…”

Characteristics of a Series of Three Bacteriophages Infecting Salmonella enterica Strains

“…One-step growth experiments were performed as described previously, with some modifications [ 50 ]. In brief, S. enterica strains S. Typhimurium 12, S. Typhimurium 13, S. Enteritidis 64, and S. Enteritidis 1392 were grown at given temperatures (25, 37, or 42 °C) until reaching OD 600 = 0.2.…”

Combination Therapy of Phage vB_KpnM_P-KP2 and Gentamicin Combats Acute Pneumonia Caused by K47 Serotype Klebsiella pneumoniae

“…The genome of phage P-KP2 is a linear double-stranded DNA that is comprised of 172,138 bp with a G + C content of 41.9%, and the termini of the phage genome is located at 38,590 bp (Supplementary Figure 2). Comparative analysis of the complete nucleotide sequence indicates that several phages show high similarity to P-KP2, including Enterobacter phage phiEap-3 (Zhao et al, 2019), Klebsiella phage Matisse (Provasek et al, 2015), Klebsiella phage KP27, Klebsiella phage KP15, Klebsiella phage PMBT1 (Koberg et al, 2017), Klebsiella phage KOX10, Klebsiella phage Miro (Mijalis et al, 2015), Escherichia phage phT4A (Pereira et al, 2017), and Klebsiella phage KOX8 (Table 1), which can be classified as a member of pseudo-Teven myophages which belong to "KP15likevirus" genus. Multiple genome alignment confirmed that these phages have parallel gene functional modules (Figure 3).…”