Characterization of Enterobacter phage vB_EcRAM-01, a new Pseudotevenvirus against Enterobacter cloacae, isolated in an urban river in Panama

Abstract

The Enterobacter cloacae complex, a prominent bacterium responsible worldwide for most bloodstream infections in the hospital environment, has shown broad-spectrum antibiotic resistance, including carbapenems. Therefore, bacteriophages have again attracted the attention of the science and medical community as an alternative to control Multidrug resistant bacteria. In this study, water samples from Rio Abajo River, in Panama City, Panama, were collected, for phage isolation, purification, characterization and propagation against the E. cloacae complex. As result, a phage produced clear and round plaque-forming units indicating a lytic phage was isolated. Further analyses concluded that this phage is stable at temperatures between 25?C and 50?C, it remains infective in a pH range between 7 to 11, with high sensitivity to Ultraviolet light. Remarkedly, it exhibits a narrow host specificity only infecting E. cloacae. Whole genome sequencing revealed that is a myovirus with a genome size of 178,477 bp, a G-C content of 45.8%, and containing approximately 294 genes. Among them, protein-encoding genes involved in morphology, inactivation, adsorption to cells, DNA injection and lytic enzymes were identified. Additionally, the genome contained two tRNA sequences. Genes that encode holins and endolysins, typical of lytic bacteriophages, were also present. A whole-genome sequencing analysis indicated that, according to the genus demarcation criteria, this phage belongs to a novel species within the Family Straboviridae, called genus Pseudotevenvirus.