Marco Maria D'Andrea1, Domenico Frezza2, Elena Romano2, Pasquale Marmo2, Lucia Henrici De Angelis3, Nicoletta Perini2, Maria Cristina Thaller2, Gustavo Di Lallo4. 1. Department of Medical Biotechnologies, University of Siena, Siena, Italy; Department of Biology, University of Rome "Tor Vergata", Rome, Italy. 2. Department of Biology, University of Rome "Tor Vergata", Rome, Italy. 3. Department of Medical Biotechnologies, University of Siena, Siena, Italy. 4. Department of Biology, University of Rome "Tor Vergata", Rome, Italy. Electronic address: dilallo@uniroma2.it.
Abstract
OBJECTIVES: The aim of this study is to characterize a new bacteriophage able to infect Enterococcus faecalis, and to evaluate its ability to disrupt biofilm. METHODS: The vB_EfaH_EF1TV (EF1TV) host-range was determined by spot test and efficiency of plating using a collection of 15E. faecalis clinical strains. The phage genome was sequenced with a next generation sequencing approach. Anti-biofilm activity was tested by crystal violet method and confocal laser scanning microscopy. Phage-resistant mutants were selected and sequenced to investigate receptors exploited by phage for infection. RESULTS: EF1TV is a newly discoveredE. faecalis phage which belongs to the Herelleviridae family. EF1TV, whose genome is 98% identical to φEF24C, is characterized by a linear dsDNA genome of 143,507 bp with direct terminal repeats of 1,911 bp. The phage is able to infect E. faecalis and shows also the ability to degrade biofilm produced by strains of this species. The results were confirmed by confocal laser scanning microscopy analyzing the biofilm reduction in the same optical field before and after phage infection. CONCLUSIONS: The EF1TV phage shows promising features such as an obligatory lytic nature, an anti-biofilm activity and the absence of integration-related proteins, antibiotic resistance determinants and virulence factors, and therefore could be a promising tool for therapeutic applications.
OBJECTIVES: The aim of this study is to characterize a new bacteriophage able to infect Enterococcus faecalis, and to evaluate its ability to disrupt biofilm. METHODS: The vB_EfaH_EF1TV (EF1TV) host-range was determined by spot test and efficiency of plating using a collection of 15E. faecalis clinical strains. The phage genome was sequenced with a next generation sequencing approach. Anti-biofilm activity was tested by crystal violet method and confocal laser scanning microscopy. Phage-resistant mutants were selected and sequenced to investigate receptors exploited by phage for infection. RESULTS: EF1TV is a newly discoveredE. faecalis phage which belongs to the Herelleviridae family. EF1TV, whose genome is 98% identical to φEF24C, is characterized by a linear dsDNA genome of 143,507 bp with direct terminal repeats of 1,911 bp. The phage is able to infect E. faecalis and shows also the ability to degrade biofilm produced by strains of this species. The results were confirmed by confocal laser scanning microscopy analyzing the biofilm reduction in the same optical field before and after phage infection. CONCLUSIONS: The EF1TV phage shows promising features such as an obligatory lytic nature, an anti-biofilm activity and the absence of integration-related proteins, antibiotic resistance determinants and virulence factors, and therefore could be a promising tool for therapeutic applications.
Authors: Colin Buttimer; Tom Sutton; Joan Colom; Ellen Murray; Pedro H Bettio; Linda Smith; Andrei S Bolocan; Andrey Shkoporov; Akihiko Oka; Bo Liu; Jeremy W Herzog; R Balfour Sartor; Lorraine A Draper; R Paul Ross; Colin Hill Journal: Front Microbiol Date: 2022-07-22 Impact factor: 6.064