Isolation and Characterization of Shewanella Phage Thanatos Infecting and Lysing Shewanella oneidensis and Promoting Nascent Biofilm Formation

Abstract

Species of the genus Shewanella are widespread in nature in various habitats, however, little is known about phages affecting Shewanella sp. Here, we report the isolation of phages from diverse freshwater environments that infect and lyse strains of Shewanella oneidensis and other Shewanella sp. Sequence analysis and microscopic imaging strongly indicate that these phages form a so far unclassified genus, now named Shewanella phage Thanatos, which can be positioned within the subfamily of Tevenvirinae (Duplodnaviria; Heunggongvirae; Uroviricota; Caudoviricetes; Caudovirales; Myoviridae; Tevenvirinae). We characterized one member of this group in more detail using S. oneidensis MR-1 as a host. Shewanella phage Thanatos-1 possesses a prolate icosahedral capsule of about 110 nm in height and 70 nm in width and a tail of about 95 nm in length. The dsDNA genome exhibits a GC content of about 34.5%, has a size of 160.6 kbp and encodes about 206 proteins (92 with an annotated putative function) and two tRNAs. Out of those 206, MS analyses identified about 155 phage proteins in PEG-precipitated samples of infected cells. Phage attachment likely requires the outer lipopolysaccharide of S. oneidensis, narrowing the phage's host range. Under the applied conditions, about 20 novel phage particles per cell were produced after a latent period of approximately 40 min, which are stable at a pH range from 4 to 12 and resist temperatures up to 55 • C for at least 24 h. Addition of Thanatos to S. oneidensis results in partial dissolution of established biofilms, however, early exposure of planktonic cells to Thanatos significantly enhances biofilm formation. Taken together, we identified a novel genus of Myophages affecting S. oneidensis communities in different ways.

Characterization of Three Novel Virulent Aeromonas Phages Provides Insights into the Diversity of the Autographiviridae Family

“…Among these subclusters, there is also another example of inter-family connections between phages. This includes autographiviruses (10 Okabevirinae phages and 9 phages yet unassigned to any subfamily but located within the Okabevirinae clique) and six siphoviruses infecting Stenotrophomonas (BUCT603, MW934263.1) and Xanthomonas hosts (phiL7 [ 107 ], CP1 [ 108 ], OP1 [ 109 ], Xp10 [ 110 ], and Xop411 [ 110 ]). Similarities are shared through the entire replication module ( Figure S15 ), that is, their DNA primases, DNA helicases, DNA polymerases, DUF669-containing proteins, 5′-3′ exonucleases, DNA endonucleases VII, ribonuclease H-like domain-containing proteins RNA polymerases, and lysozymes ( Figure S17 ).…”

A rolling circle-replicating plasmid as an Inovirus phage satellite

“…In a study aimed at the isolation and identification of phages infecting Shewanella oneidensis MR-1 21 , water and sediment samples of limnic environments were sampled. One of the enrichments from a fresh water sediment yielded a phage isolate characterized by small turbid plaques ( Fig.…”

Identifying the components of theShewanellaphage LambdaSo lysis system

“…To determine the latent period and burst size of LambdaSo, a one-step growth experiment was performed (43)(44)(45). Briefly, 20 ml of cell culture of S. oneidensis ΔLambdaSo ΔMuSo1 ΔMuSo2 was prepared in LB medium and incubated at RT.…”