Isolation and characterisation of novel phages infecting Lactobacillus plantarum and proposal of a new genus, “Silenusvirus”
Ifigeneia KyrkouAlexander Byth CarstensLea Ellegaard‐JensenWitold KotAthanasios ZervasAmaru Miranda DjurhuusHorst NeveCharles M. A. P. FranzMartin HansenLars Hestbjerg Hansen
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Abstract Bacteria of Lactobacillus sp. are very useful to humans. However, the biology and genomic diversity of their (bacterio)phage enemies remains understudied. Knowledge on Lactobacillus phage diversity should broaden to develop efficient phage control strategies. To this end, organic waste samples were screened for phages against two wine-related Lactobacillus plantarum strains. Isolates were shotgun sequenced and compared against the phage database and each other by phylogenetics and comparative genomics. The new isolates had only three distant relatives from the database, but displayed a high overall degree of genomic similarity amongst them. The latter allowed for the use of one isolate as a representative to conduct transmission electron microscopy and structural protein sequencing, and to study phage adsorption and growth kinetics. The microscopy and proteomics tests confirmed the observed diversity of the new isolates and supported their classification to the family Siphoviridae and the proposal of the new phage genus “Silenusvirus”.Keywords:
Siphoviridae
Phage therapy
Isolation
Siphoviridae
Myoviridae
Prophage
Podoviridae
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Abstract Bacteriophage P8625 is a lytic bacteriophage that infects the verrucomicrobial strain IMCC8625, a marine bacterium affiliated with Verrucomicrobia subdivision 4. Both the bacteriophage and the host bacterial strain were isolated from surface seawater samples collected off the east coast of Korea. The phage particle has an icosahedral capsid with a diameter of ~47 nm and a long tail of ~75 nm in length, showing the distinctive morphology of the Siphoviridae family. The complete genome sequence of phage P8625 is 32,894 bp long with 51.0 % G + C content. This is the first report of the complete genome sequence of a lytic phage that infects the Verrucomicrobia , for which the name “verrucophage” is proposed.
Siphoviridae
Lytic cycle
Verrucomicrobia
Phage therapy
Strain (injury)
Myoviridae
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Siphoviridae
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From the hospital waste-water, a novel bacteriophage was isolated and characterized. According to characterization properties, this bacteriophage belongs to the Siphoviridae family, the maximum bacteriophage titer was recorded at 37°C and a pH of 7.2, had a 44,789 bp linear double-strand DNA genome, and within the genome sequence, there are 61 genes, all of which are encoded into proteins. Although this bacteriophage does not have any virulence factors or antimicrobial resistance genes and had specific lytic activity against some antimicrobial resistance S. aureus clinical isolates.
Siphoviridae
Lytic cycle
Myoviridae
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Vibrio parahaemolyticus is a foodborne pathogen that is frequently isolated from a variety of seafood. To control this pathogenic Vibrio spp., the implementation of bacteriophages in aquaculture and food industries have shown a promising alternative to antibiotics. In this study, six bacteriophages isolated from the seafood samples demonstrated a narrow host range specificity that infecting only the V. parahaemolyticus strains. Morphological analysis revealed that bacteriophages Vp33, Vp22, Vp21, and Vp02 belong to the Podoviridae family, while bacteriophages Vp08 and Vp11 were categorized into the Siphoviridae family. All bacteriophages were composed of DNA genome and showed distinctive restriction fragment length polymorphism. The optimal MOI for bacteriophage propagation was determined to be 0.001 to 1. One-step growth curve revealed that the latent period ranged from 10 to 20 min, and the burst size of bacteriophage was approximately 17 to 51 PFU/cell. The influence of temperature and pH levels on the stability of bacteriophages showed that all bacteriophages were optimally stable over a wide range of temperatures and pH levels. In vitro lytic activity of all bacteriophages demonstrated to have a significant effect against V. parahaemolyticus . Besides, the application of a bacteriophage cocktail instead of a single bacteriophage suspension was observed to have a better efficiency to control the growth of V . parahaemolyticus . Results from this study provided a basic understanding of the physiological and biological properties of the isolated bacteriophages before it can be readily used as a biocontrol agent against the growth of V. parahaemolyticus .
Lytic cycle
Siphoviridae
Podoviridae
Myoviridae
Phage therapy
Coliphage
Lysogenic cycle
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Phage therapy
Lytic cycle
Lysogenic cycle
Lysin
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Siphoviridae
Lysogen
Myoviridae
Podoviridae
Shiga-like toxin
Prophage
Lambda phage
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Objective To isolate bacteriophages from raw sewage and to investigate the biological properties of bacteriophages using 14 strains of Enterobacter cloacae as host cells,and to identify its structure.Methods Bacteriophages were isolated from sewage with double-agar and identified by plaque method.After purification and negative-staining of bacteriophages,the morphologic features of bacteriophages were examined by electron microscope,the bacteriophage genome was extracted for electrophoresis;the lytic spectrum of host was carried out for specificity and cracking range of the Enterobacter cloacae.Results Bacteriophage ФEc53 was isolated and identified,electron micrograph showed that the head of bacteriophage was spherical,60 nm in diameter and it had a tail,about 130 nm long.Electrophoresis of phage DNA revealed that the size of genome was about 40 000 bp and contained lots of restriction sites.The lytic spectrum trial showed it had a relatively narrow host range.Conclusion The isolated Enterobacter cloacae(named ФEc53) bacteriophage belongs to Caudovirales,Siphoviridae.According to its biologic features,bacteriophage ФEc53 has strong lytic and high specificity.
Siphoviridae
Lytic cycle
Enterobacter cloacae
Podoviridae
Phage therapy
Myoviridae
Temperateness
Agarose gel electrophoresis
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From the hospital wastewater, a novel bacteriophage was isolated and characterized. According to characterization properties, this bacteriophage belongs to the Siphoviridae family, the maximum bacteriophage titer was recorded at 37°C and a pH of 7.2, had a 44,789 bp linear double-strand DNA genome, and within the genome sequence, there are 61 genes, all of which are encoded into proteins. Although this bacteriophage does not have any virulence factors or antimicrobial resistance genes and had specific lytic activity against some antimicrobial resistance S. aureus clinical isolates.
Lytic cycle
Siphoviridae
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The resistance to antibiotics in clinically important bacteria is one of the major global health concerns. Phage therapy could be one reliable alternative therapeutic strategy to combat these superbugs. In this study, we assessed host range of a novel bacteriophage, JHP, and characterized for its potential use in phage therapy. The bacteriophage demonstrated infectivity over a broad range of genera including multidrug resistant clinical isolates of Pseudomonas aeruginosa, members of family Enterobacteracae, and other important human pathogens. The antibacterial activity was highest at pH 7, and at temperature of 37 °C. The phage lytic activity gradually decreased till 60 °C and showed no activity when temperature was further raised. The bacteriophage could safely be stored at 4 °C or -20 °C. The latent period of the bacteriophage was 25 min and showed a burst size of 433 virions per cell. The size of JHP genome was approximately 30 kb. Family, Siphoviridae was assigned to JHP based on its icosahedral head with non-contractile tail. The diameter of JHP head and tail length was found 115 and 152 nm, respectively. To sum up, the broad spectrum Siphoviridae phage JHP is an ingenious candidate for phage therapy.
Siphoviridae
Lytic cycle
Phage therapy
Infectivity
Myoviridae
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