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1.
Viruses ; 14(2)2022 01 19.
Artigo em Inglês | MEDLINE | ID: mdl-35215775

RESUMO

The number of sequenced Acinetobacter phage genomes in the International Nucleotide Sequence Database Collaboration has increased significantly in recent years, from 37 in 2017 to a total of 139 as of January 2021 with genome sizes ranging from 31 to 378 kb. Here, we explored the genetic diversity of the Acinetobacter phages using comparative genomics approaches that included assessment of nucleotide similarity, shared gene content, single gene phylogeny, and the network-based classification tool vConTACT2. Phages infecting Acinetobacter sp. are genetically diverse and can be grouped into 8 clusters (subfamilies) and 46 sub-clusters (genera), of which 8 represent genomic singletons (additional genera). We propose the creation of five new subfamilies and suggest a reorganisation of the genus Obolenskvirus. These results provide an updated view of the viruses infecting Acinetobacter species, providing insights into their diversity.


Assuntos
Acinetobacter/virologia , Bacteriófagos/genética , Variação Genética , Myoviridae/genética , Bacteriófagos/classificação , Bacteriófagos/isolamento & purificação , Sequência de Bases , Genoma Viral , Genômica , Myoviridae/classificação , Myoviridae/isolamento & purificação , Filogenia
2.
Viruses ; 14(2)2022 01 25.
Artigo em Inglês | MEDLINE | ID: mdl-35215834

RESUMO

Salmonella Typhimurium, a foodborne pathogen, is a major concern for food safety. Its MDR serovars of animal origin pose a serious threat to the human population. Phage therapy can be an alternative for the treatment of such MDR Salmonella serovars. In this study, we report on detailed genome analyses of a novel Salmonella phage (Salmonella-Phage-SSBI34) and evaluate its therapeutic potential. The phage was evaluated for latent time, burst size, host range, and bacterial growth reduction in liquid cultures. The phage stability was examined at various pH levels and temperatures. The genome analysis (141.095 Kb) indicated that its nucleotide sequence is novel, as it exhibited only 1-7% DNA coverage. The phage genome features 44% GC content, and 234 putative open reading frames were predicted. The genome was predicted to encode for 28 structural proteins and 40 enzymes related to nucleotide metabolism, DNA modification, and protein synthesis. Further, the genome features 11 tRNA genes for 10 different amino acids, indicating alternate codon usage, and hosts a unique hydrolase for bacterial lysis. This study provides new insights into the subfamily Vequintavirinae, of which SSBI34 may represent a new genus.


Assuntos
Myoviridae/genética , Fagos de Salmonella/genética , Salmonella typhimurium/virologia , Animais , Bacteriólise , Agentes de Controle Biológico , Genoma Viral , Especificidade de Hospedeiro , Myoviridae/classificação , Myoviridae/isolamento & purificação , Myoviridae/fisiologia , Fases de Leitura Aberta , Terapia por Fagos , Filogenia , Aves Domésticas/microbiologia , Infecções por Salmonella/terapia , Fagos de Salmonella/classificação , Fagos de Salmonella/isolamento & purificação , Fagos de Salmonella/fisiologia , Salmonella typhimurium/isolamento & purificação
3.
Viruses ; 14(2)2022 01 29.
Artigo em Inglês | MEDLINE | ID: mdl-35215879

RESUMO

Salmonella and Escherichia coli (E. coli) food contamination could lead to serious foodborne diseases. The gradual increase in the incidence of foodborne disease invokes new and efficient methods to limit food pathogenic microorganism contamination. In this study, a polyvalent broad-spectrum Escherichia phage named Tequatrovirus EP01 was isolated from pig farm sewage. It could lyse both Salmonella Enteritidis (S. Enteritidis) and E. coli and exhibited broad host range. EP01 possessed a short latent period (10 min), a large burst size (80 PFU/cell), and moderate pH stability (4-10) and appropriate thermal tolerance (30-80 °C). Electron microscopy and genome sequence revealed that EP01 belonged to T4-like viruses genus, Myoviridae family. EP01 harbored 12 CDSs associated with receptor-binding proteins and lacked virulence genes and drug resistance genes. We tested the inhibitory effect of EP01 on S. Enteritidis, E. coli O157:H7, E. coli O114:K90 (B90), and E. coli O142:K86 (B) in liquid broth medium (LB). EP01 could significantly reduce the counts of all tested strains compared with phage-free groups. We further examined the effectiveness of EP01 in controlling bacterial contamination in two kinds of foods (meat and milk) contaminated with S. Enteritidis, E. coli O157:H7, E. coli O114:K90 (B90), and E. coli O142:K86 (B), respectively. EP01 significantly reduced the viable counts of all the tested bacteria (2.18-6.55 log10 CFU/sample, p < 0.05). A significant reduction of 6.55 log10 CFU/cm2 (p < 0.001) in bacterial counts on the surface of meat was observed with EP01 treatment. Addition of EP01 at MOI of 1 decreased the counts of bacteria by 4.3 log10 CFU/mL (p < 0.001) in milk. Generally, the inhibitory effect exhibited more stable at 4 °C than that at 28 °C, whereas the opposite results were observed in milk. The antibacterial effects were better at MOI of 1 than that at MOI of 0.001. These results suggests that phage EP01-based method is a promising strategy of controlling Salmonella and Escherichia coli pathogens to limit microbial food contamination.


Assuntos
Escherichia coli/virologia , Contaminação de Alimentos/prevenção & controle , Myoviridae/fisiologia , Salmonella enteritidis/virologia , Animais , Bacteriólise , Escherichia coli/crescimento & desenvolvimento , Microbiologia de Alimentos , Genoma Viral , Especificidade de Hospedeiro , Carne/microbiologia , Leite/microbiologia , Myoviridae/classificação , Myoviridae/genética , Myoviridae/isolamento & purificação , Filogenia , Salmonella enteritidis/crescimento & desenvolvimento , Esgotos/virologia , Suínos
4.
Sci Rep ; 12(1): 495, 2022 01 11.
Artigo em Inglês | MEDLINE | ID: mdl-35017610

RESUMO

Among food preservation methods, bacteriophage treatment can be a viable alternative method to overcome the drawbacks of traditional approaches. Bacteriophages are naturally occurring viruses that are highly specific to their hosts and have the capability to lyse bacterial cells, making them useful as biopreservation agents. This study aims to characterize and determine the application of bacteriophage isolated from Indonesian traditional Ready-to-Eat (RTE) food to control Enterotoxigenic Escherichia coli (ETEC) population in various foods. Phage DW-EC isolated from Indonesian traditional RTE food called dawet with ETEC as its host showed a positive result by the formation of plaques (clear zone) in the bacterial host lawn. Transmission electron microscopy (TEM) results also showed that DW-EC can be suspected to belong to the Myoviridae family. Molecular characterization and bioinformatic analysis showed that DW-EC exhibited characteristics as promising biocontrol agents in food samples. Genes related to the lytic cycle, such as lysozyme and tail fiber assembly protein, were annotated. There were also no signs of lysogenic genes among the annotation results. The resulting PHACTS data also indicated that DW-EC was leaning toward being exclusively lytic. DW-EC significantly reduced the ETEC population (P ≤ 0.05) in various food samples after two different incubation times (1 day and 6 days) in chicken meat (80.93%; 87.29%), fish meat (63.78%; 87.89%), cucumber (61.42%; 71.88%), tomato (56.24%; 74.51%), and lettuce (46.88%; 43.38%).


Assuntos
Bacteriófagos/isolamento & purificação , Bacteriófagos/fisiologia , Escherichia coli Enterotoxigênica/virologia , Conservação de Alimentos/métodos , Myoviridae/isolamento & purificação , Myoviridae/fisiologia , Animais , Bacteriófagos/classificação , Bacteriófagos/genética , Galinhas , Escherichia coli Enterotoxigênica/fisiologia , Fast Foods/virologia , Peixes , Contaminação de Alimentos/prevenção & controle , Carne/microbiologia , Myoviridae/classificação , Myoviridae/genética , Verduras/microbiologia , Proteínas Virais/genética , Proteínas Virais/metabolismo
5.
Viruses ; 14(1)2022 01 08.
Artigo em Inglês | MEDLINE | ID: mdl-35062312

RESUMO

Viruses are far more abundant than cellular microorganisms in the marine ecosystem. However, very few viruses have so far been isolated from marine sediments, especially hydrothermal vent sediments, hindering the understanding of the biology and ecological functions of these tiny organisms. Here, we report the isolation and characterization of a temperate bacteriophage, named PVJ1, which infects Psychrobacillus from a hydrothermal vent field in Okinawa Trough. PVJ1 belongs to the Myoviridae family of the order Caudovirales. The tailed phage possesses a 53,187 bp linear dsDNA genome, with 84 ORFs encoding structural proteins, genome replication, host lysis, etc. in a modular pattern. The phage genome is integrated into the host chromosome near the 3'-end of deoD, a gene encoding purine nucleoside phosphorylase (PNP). The phage integration does not appear to disrupt the function of PNP. The phage DNA is packaged by the headful mechanism. Release of PVJ1 from the host cell was drastically enhanced by treatment with mitomycin C. Phages encoding an MCP sharing significant similarity (≥70% identical amino acids) with that of PVJ1 are widespread in diverse environments, including marine and freshwater sediments, soils, artificial ecosystems, and animal intestines, and primarily infect Firmicutes. These results are valuable to the understanding of the lifestyle and host interactions of bacterial viruses at the bottom of the ocean.


Assuntos
Bacillaceae/virologia , Bacteriófagos/classificação , Bacteriófagos/isolamento & purificação , Sedimentos Geológicos/virologia , Filogenia , Animais , Bacteriófagos/genética , Caudovirales/genética , Caudovirales/isolamento & purificação , DNA Viral/isolamento & purificação , Ecossistema , Genoma Viral , Myoviridae/genética , Myoviridae/isolamento & purificação , Fases de Leitura Aberta
6.
Int J Mol Sci ; 22(22)2021 Nov 18.
Artigo em Inglês | MEDLINE | ID: mdl-34830335

RESUMO

The food industry is still searching for novel solutions to effectively ensure the microbiological safety of food, especially fresh and minimally processed food products. Nowadays, the use of bacteriophages as potential biological control agents in microbiological food safety and preservation is a promising strategy. The aim of the study was the isolation and comprehensive characterization of novel bacteriophages with lytic activity against saprophytic bacterial microflora of minimally processed plant-based food products, such as mixed leaf salads. From 43 phages isolated from municipal sewage, four phages, namely Enterobacter phage KKP 3263, Citrobacter phage KKP 3664, Enterobacter phage KKP 3262, and Serratia phage KKP 3264 have lytic activity against Enterobacter ludwigii KKP 3083, Citrobacter freundii KKP 3655, Enterobacter cloacae KKP 3082, and Serratia fonticola KKP 3084 bacterial strains, respectively. Transmission electron microscopy (TEM) and whole-genome sequencing (WGS) identified Enterobacter phage KKP 3263 as an Autographiviridae, and Citrobacter phage KKP 3664, Enterobacter phage KKP 3262, and Serratia phage KKP 3264 as members of the Myoviridae family. Genome sequencing revealed that these phages have linear double-stranded DNA (dsDNA) with sizes of 39,418 bp (KKP 3263), 61,608 bp (KKP 3664), 84,075 bp (KKP 3262), and 148,182 bp (KKP 3264). No antibiotic resistance genes, virulence factors, integrase, recombinase, or repressors, which are the main markers of lysogenic viruses, were annotated in phage genomes. Serratia phage KKP 3264 showed the greatest growth inhibition of Serratia fonticola KKP 3084 strain. The use of MOI 1.0 caused an almost 5-fold decrease in the value of the specific growth rate coefficient. The phages retained their lytic activity in a wide range of temperatures (from -20 °C to 50 °C) and active acidity values (pH from 4 to 11). All phages retained at least 70% of lytic activity at 60 °C. At 80 °C, no lytic activity against tested bacterial strains was observed. Serratia phage KKP 3264 was the most resistant to chemical factors, by maintaining high lytic activity across a broader range of pH from 3 to 11. The results indicated that these phages could be a potential biological control agent against saprophytic bacterial microflora of minimally processed plant-based food products.


Assuntos
Bacteriófagos/genética , Citrobacter freundii/virologia , Enterobacter cloacae/virologia , Inocuidade dos Alimentos/métodos , Genoma Viral , Myoviridae/genética , Serratia/virologia , Bacteriólise/fisiologia , Bacteriófagos/classificação , Bacteriófagos/isolamento & purificação , Agentes de Controle Biológico/classificação , Agentes de Controle Biológico/isolamento & purificação , DNA Viral/genética , Microbiologia de Alimentos/métodos , Embalagem de Alimentos/métodos , Conservação de Alimentos/métodos , Humanos , Myoviridae/classificação , Myoviridae/isolamento & purificação , Filogenia , Esgotos/virologia , Verduras/microbiologia
7.
Microbiol Spectr ; 9(2): e0046321, 2021 10 31.
Artigo em Inglês | MEDLINE | ID: mdl-34643440

RESUMO

Alteromonas is a ubiquitous, abundant, copiotrophic and phytoplankton-associated marine member of the Gammaproteobacteria with a range extending from tropical waters to polar regions and including hadal zones. Here, we describe a novel Alteromonas phage, ZP6, that was isolated from surface coastal waters of Qingdao, China. ZP6 contains a linear, double-stranded, 38,080-bp DNA molecule with 50.1% G+C content and 47 putative open reading frames (ORFs). Three auxiliary metabolic genes were identified, encoding metal-dependent phosphohydrolase, diaminopurine synthetase, and nucleotide pyrophosphohydrolase. The first two ORFs facilitate the replacement of adenine (A) by diaminopurine (Z) in phage genomes and help phages to evade attack from host restriction enzymes. The nucleotide pyrophosphohydrolase enables the host cells to stop programmed cell death and improves the survival rate of the host in a nutrient-depleted environment. Phylogenetic analysis based on the amino acid sequences of whole genomes and comparative genomic analysis revealed that ZP6 is most closely related to Enhodamvirus but with low similarity (shared genes, <30%, and average nucleotide sequence identity, <65%); it is distinct from other bacteriophages. Together, these results suggest that ZP6 could represent a novel viral genus, here named Mareflavirus. Combining its ability to infect Alteromonas, its harboring of a diaminopurine genome-biosynthetic system, and its representativeness of an understudied viral group, ZP6 could be an important and novel model system for marine virus research. IMPORTANCEAlteromonas is an important symbiotic bacterium of phytoplankton, but research on its bacteriophages is still at an elementary level. Our isolation and genome characterization of a novel Alteromonas podovirus, ZP6, identified a new viral genus of podovirus, namely, Mareflavirus. The ZP6 genome, with a diaminopurine genome-biosynthetic system, is different from those of other isolated Alteromonas phages and will bring new impetus to the development of virus classification and provide important insights into novel viral sequences from metagenomic data sets.


Assuntos
Alteromonas/virologia , Bacteriófagos/genética , Bacteriófagos/isolamento & purificação , Genoma Viral , Myoviridae/genética , Myoviridae/isolamento & purificação , Bacteriófagos/classificação , Bacteriófagos/crescimento & desenvolvimento , China , Myoviridae/classificação , Fases de Leitura Aberta , Filogenia , Água do Mar/virologia
8.
Sci Rep ; 11(1): 19393, 2021 09 29.
Artigo em Inglês | MEDLINE | ID: mdl-34588479

RESUMO

In recent years, the use of bacteriophages (or 'phages') against multidrug-resistant (MDR) bacteria including Pseudomonas aeruginosa has drawn considerable attention, globally. In this work, we report the isolation and detailed characterization of a highly lytic Pseudomonasphage DRL-P1 isolated from wastewater. Under TEM, DRL-P1 appeared as a member of the phage family Myoviridae. DRL-P1 featured rapid adsorption (~ 5 min), short-latency (~ 30 min), and large burst size (~ 100 PFU per infected cell). DRL-P1 can withstand a wide temperature range (4 °C to 40 °C) and pH (5.0 to 10.0) conditions. The 66,243 bp DRL-P1 genome (MN564818) encodes at least 93 ORFs, of which 36 were functionally annotated based on homology with similar phage proteins available in the databases. Comparative analyses of related genomes suggest an independent evolutionary history and discrete taxonomic position of DRL-P1 within genus Pbunavirus. No toxin or antibiotic resistance genes was identified. DRL-P1 is tolerant to lyophilization and encapsulation techniques and retained lytic activity even after 18 months of storage. We also demonstrated decontaminating potentials of DRL-P1 in vitro, on an artificially contaminated cover-slip model. To the best of our knowledge, this is the first Pbunavirus to be reported from India. Our study suggests DRL-P1 as a potential candidate for various applications.


Assuntos
Myoviridae , Fagos de Pseudomonas , Pseudomonas aeruginosa/virologia , Águas Residuárias , DNA Viral , Farmacorresistência Bacteriana Múltipla/genética , Genoma Viral , Myoviridae/classificação , Myoviridae/isolamento & purificação , Myoviridae/fisiologia , Fagos de Pseudomonas/classificação , Fagos de Pseudomonas/isolamento & purificação , Fagos de Pseudomonas/fisiologia , Águas Residuárias/microbiologia , Águas Residuárias/virologia
9.
Viruses ; 13(8)2021 07 21.
Artigo em Inglês | MEDLINE | ID: mdl-34452284

RESUMO

Kosakonia cowanii (syn. Enterobacter cowanii) is a highly competitive bacterium that lives with plant, insect, fish, bird, and human organisms. It is pathogenic on some plants and an opportunistic pathogen of human. Nine novel viruses that lyse plant pathogenic strains and/or human strains of K. cowanii were isolated, sequenced, and characterized. Kc166A is a novel kayfunavirus, Kc261 is a novel bonnellvirus, and Kc318 is a new cronosvirus (all Autographiviridae). Kc237 is a new sortsnevirus, but Kc166B and Kc283 are members of new genera within Podoviridae. Kc304 is a new winklervirus, and Kc263 and Kc305 are new myoviruses. The viruses differ in host specificity, plaque phenotype, and lysis kinetics. Some of them should be suitable also as pathogen control agents.


Assuntos
Bacteriólise , Bacteriófagos/fisiologia , Caudovirales/fisiologia , Enterobacteriaceae/virologia , Folhas de Planta/microbiologia , Bacteriófagos/classificação , Bacteriófagos/genética , Bacteriófagos/isolamento & purificação , Caudovirales/classificação , Caudovirales/genética , Caudovirales/isolamento & purificação , Enterobacteriaceae/fisiologia , Genoma Viral , Especificidade de Hospedeiro , Humanos , Myoviridae/classificação , Myoviridae/genética , Myoviridae/isolamento & purificação , Myoviridae/fisiologia , Filogenia , Doenças das Plantas/microbiologia , Microbiologia do Solo , /microbiologia
10.
Science ; 373(6554)2021 07 30.
Artigo em Inglês | MEDLINE | ID: mdl-34326207

RESUMO

Bacteriophage predation selects for diverse antiphage systems that frequently cluster on mobilizable defense islands in bacterial genomes. However, molecular insight into the reciprocal dynamics of phage-bacterial adaptations in nature is lacking, particularly in clinical contexts where there is need to inform phage therapy efforts and to understand how phages drive pathogen evolution. Using time-shift experiments, we uncovered fluctuations in Vibrio cholerae's resistance to phages in clinical samples. We mapped phage resistance determinants to SXT integrative and conjugative elements (ICEs), which notoriously also confer antibiotic resistance. We found that SXT ICEs, which are widespread in γ-proteobacteria, invariably encode phage defense systems localized to a single hotspot of genetic exchange. We identified mechanisms that allow phage to counter SXT-mediated defense in clinical samples, and document the selection of a novel phage-encoded defense inhibitor. Phage infection stimulates high-frequency SXT ICE conjugation, leading to the concurrent dissemination of phage and antibiotic resistances.


Assuntos
Farmacorresistência Bacteriana/genética , Sequências Repetitivas Dispersas , Myoviridae/fisiologia , Vibrio cholerae/efeitos dos fármacos , Vibrio cholerae/virologia , Bacteriólise , Cólera/microbiologia , Conjugação Genética , Epigênese Genética , Fezes/microbiologia , Fezes/virologia , Gammaproteobacteria/genética , Gammaproteobacteria/virologia , Genes Bacterianos , Genes Virais , Genoma Bacteriano , Genoma Viral , Especificidade de Hospedeiro , Humanos , Interações Microbianas , Myoviridae/genética , Myoviridae/isolamento & purificação , Vibrio cholerae/genética , Vibrio cholerae/isolamento & purificação , Proteínas Virais/genética , Proteínas Virais/metabolismo
11.
Biomol Concepts ; 12(1): 85-93, 2021 Jul 02.
Artigo em Inglês | MEDLINE | ID: mdl-34218551

RESUMO

Bacillus cereus is considered to be an important food poisoning agent causing diarrhea and vomiting. In this study, the occurrence of B. cereus bacteriophages in Thai fermented soybean products (Thua Nao) was studied using five B. cereus sensu lato indicator strains (four B. cereus strains and one B. thuringiensis strain). In a total of 26 Thua Nao samples, there were only two bacteriophages namely BaceFT01 and BaceCM02 exhibiting lytic activity against B. cereus. Morphological analysis revealed that these two bacteriophages belonged to the Myoviridae. Both phages were specific to B. cereus and not able to lyse other tested bacteria including B. licheniformis and B. subtilis. The two phages were able to survive in a pH range between 5 and 12. However, both phages were inactive either by treatment of 50°C for 2 h or exposure of UV for 2 h. It should be noted that both phages were chloroform-insensitive, however. This is the first report describing the presence of bacteriophages in Thua Nao products. The characterization of these two phages is expected to be useful in the food industry for an alternative strategy including the potential use of the phages as a biocontrol candidate against foodborne pathogenic bacteria.


Assuntos
Fagos Bacilares/isolamento & purificação , Bacillus cereus/virologia , Fermentação , Myoviridae/isolamento & purificação , Fagos Bacilares/fisiologia , Microbiologia de Alimentos , Humanos , Myoviridae/classificação , Myoviridae/genética , Myoviridae/crescimento & desenvolvimento , Tailândia
12.
Genes (Basel) ; 12(7)2021 06 29.
Artigo em Inglês | MEDLINE | ID: mdl-34209474

RESUMO

Marine viruses are widely distributed and influence matter and energy transformation in ecosystems by modulating hosts' metabolism. The hadal trenches represent the deepest marine habitat on Earth, for which the viral communities and related biogeochemical functions are least explored and poorly understood. Here, using the sediment samples (8720 m below sea level) collected from the New Britain Trench (NBT), we investigated the viral community, diversity, and genetic potentials in the hadal sediment habitat for the first time by deep shotgun metagenomic sequencing. We found the NBT sediment viral community was dominated by Siphoviridae, Myoviridae, Podoviridae, Mimiviridae, and Phycodnaviridae, which belong to the dsDNA viruses. However, the large majority of them remained uncharacterized. We found the hadal sediment virome had some common components by comparing the hadal sediment viruses with those of hadal aquatic habitats and those of bathypelagic and terrestrial habitats. It was also distinctive in community structure and had many novel viral clusters not associated with the other habitual virome included in our analyses. Further phylogenetic analysis on its Caudovirales showed novel diversities, including new clades specially evolved in the hadal sediment habitat. Annotation of the NBT sediment viruses indicated the viruses might influence microbial hydrocarbon biodegradation and carbon and sulfur cycling via metabolic augmentation through auxiliary metabolic genes (AMGs). Our study filled in the knowledge gaps on the virome of the hadal sediment habitats and provided insight into the evolution and the potential metabolic functions of the hadal sediment virome.


Assuntos
Ecossistema , Sedimentos Geológicos/virologia , Metagenômica , Vírus/isolamento & purificação , Humanos , Metagenoma/genética , Mimiviridae/genética , Mimiviridae/isolamento & purificação , Myoviridae/genética , Myoviridae/isolamento & purificação , Phycodnaviridae/genética , Phycodnaviridae/isolamento & purificação , Filogenia , Podoviridae/genética , Podoviridae/isolamento & purificação , Siphoviridae/genética , Siphoviridae/isolamento & purificação , Vírus/classificação , Vírus/genética
13.
J Virol ; 95(19): e0239120, 2021 09 09.
Artigo em Inglês | MEDLINE | ID: mdl-34287047

RESUMO

The majority of previously described Staphylococcus aureus bacteriophages belong to three major groups, namely, P68-like podophages, Twort-like or K-like myophages, and a more diverse group of temperate siphophages. Here, we present the following three novel S. aureus "jumbo" phages: MarsHill, Madawaska, and Machias. These phages were isolated from swine production environments in the United States and represent a novel clade of S. aureus myophage. The average genome size for these phages is ∼269 kb with each genome encoding ∼263 predicted protein-coding genes. Phage genome organization and content are similar to those of known jumbo phages of Bacillus sp., including AR9 and vB_BpuM-BpSp. All three phages possess genes encoding complete virion and nonvirion RNA polymerases, multiple homing endonucleases, and a retron-like reverse transcriptase. Like AR9, all of these phages are presumed to have uracil-substituted DNA which interferes with DNA sequencing. These phages are also able to transduce host plasmids, which is significant as these phages were found circulating in swine production environments and can also infect human S. aureus isolates. IMPORTANCE This study describes the comparative genomics of the following three novel S. aureus jumbo phages: MarsHill, Madawaska, and Machias. These three S. aureus myophages represent an emerging class of S. aureus phage. These genomes contain abundant introns which show a pattern consistent with repeated acquisition rather than vertical inheritance, suggesting intron acquisition and loss are active processes in the evolution of these phages. These phages have presumably hypermodified DNA which inhibits sequencing by several different common platforms. Therefore, these phages also represent potential genomic diversity that has been missed due to the limitations of standard sequencing techniques. In particular, such hypermodified genomes may be missed by metagenomic studies due to their resistance to standard sequencing techniques. Phage MarsHill was found to be able to transduce host DNA at levels comparable to that found for other transducing S. aureus phages, making it a potential vector for horizontal gene transfer in the environment.


Assuntos
Genoma Viral , Myoviridae/genética , Fagos de Staphylococcus/genética , Staphylococcus aureus/virologia , Animais , DNA Viral/genética , RNA Polimerases Dirigidas por DNA/genética , Genômica , Íntrons , Myoviridae/isolamento & purificação , Myoviridae/fisiologia , Myoviridae/ultraestrutura , Análise de Sequência de DNA , Fagos de Staphylococcus/isolamento & purificação , Fagos de Staphylococcus/fisiologia , Fagos de Staphylococcus/ultraestrutura , Suínos , Transdução Genética , Proteínas Virais/genética
14.
Arch Virol ; 166(9): 2597-2602, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34117533

RESUMO

This work describes the characterization and genome annotation of the newly isolated lytic phage vB_SsoM_Z31 (referred to as Z31), isolated from wastewater samples collected in Dalian, China. Transmission electron microscopy revealed that phage Z31 belongs to the family Myoviridae, order Caudovirales. This phage specifically infects Shigella sonnei, Shigella dysenteriae, and Escherichia coli. The genome of the phage Z31 is an 89,355-bp-long dsDNA molecule with a G+C content of 38.87%. It was predicted to contain 133 ORFs and encode 24 tRNAs. No homologs of virulence factor genes or antimicrobial resistance genes were found in this phage. Based on the results of nucleotide sequence alignment and phylogenetic analysis, phage Z31 was assigned to the genus Felixounavirus, subfamily Ounavirinae.


Assuntos
Myoviridae/classificação , Myoviridae/genética , Myoviridae/isolamento & purificação , Shigella sonnei/virologia , Bacteriófagos/genética , Bacteriófagos/isolamento & purificação , Composição de Bases , China , DNA Viral/genética , Farmacorresistência Bacteriana Múltipla , Escherichia coli/virologia , Genoma Viral , Especificidade de Hospedeiro , Fases de Leitura Aberta , Filogenia , Análise de Sequência de DNA , Shigella dysenteriae/virologia , Águas Residuárias/virologia
15.
APMIS ; 129(8): 461-469, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-33950561

RESUMO

Enterobacter cloacae (E. cloacae) is an emerging nosocomial pathogen that had acquired antibiotic resistance against multiple classes of antibiotics. The current study was aimed to isolate and characterize lytic bacteriophage against E. cloacae. The bacteriophage EBP was isolated from a sewage water sample using E. cloacae as a host strain by double-layer agar technique. EBP was found stabile at a wide range of temperatures (25, 37, 60, and 80°C) and pH (5, 6, 7, 8, and 9) with antibacterial activity up to 24 h of infection. The latent period of EBP was 20 min with a burst size of 252 phages per cell. It showed a narrow host range and infected 12/21 (57%) isolates of E. cloacae tested. It has helical symmetry with a head size of 105 and 120 nm long tail with contractile sheath. The EBP has 179.1 kb long double-stranded DNA genome with 44.8% GC content. Majority of identified ORFs (187/281) were encoding putative proteins with unknown function. Necessary replication enzymes, structural proteins, and lytic enzymes were detected in the genome of EBP. Phylogenetic analysis revealed that EBP closely resembles with Coronobacter phage vB_CsaM_IeN, vB_CsaM_IeE, vB_CsaM_IeB, and Citrobacter phage Margaery. Based on electron microscopy and molecular characterization, EBP was classified as a Myoviridae phage.


Assuntos
Bacteriófagos/isolamento & purificação , Genoma Viral , Myoviridae/isolamento & purificação , Bacteriófagos/classificação , Bacteriófagos/genética , Bacteriófagos/fisiologia , Enterobacter cloacae/virologia , Tamanho do Genoma , Especificidade de Hospedeiro , Myoviridae/classificação , Myoviridae/genética , Myoviridae/fisiologia , Filogenia , Esgotos/virologia
16.
J Microbiol Biotechnol ; 31(5): 710-716, 2021 May 28.
Artigo em Inglês | MEDLINE | ID: mdl-33782222

RESUMO

A risk analysis of Shiga toxin (Stx)-encoding bacteriophage was carried out by confirming the transduction phage to non-Stx-producing Escherichia coli (STEC) and subsequent expression of the Shiga toxin genes. The virulence factor stx1 was identified in five phages, and both stx1 and stx2 were found in four phages from a total of 19 phage isolates with seven non-O157 STEC strains. The four phages, designated as φNOEC41, φNOEC46, φNOEC47, and φNOEC49, belonged morphologically to the Myoviridae family. The stabilities of these phages to temperature, pH, ethanol, and NaClO were high with some variabilities among the phages. The infection of five non-STEC strains by nine Stx-encoding phages occurred at a rate of approximately 40%. Non-STEC strains were transduced by Stx-encoding phage to become lysogenic strains, and seven convertant strains had stx1 and/or stx2 genes. Only the stx1 gene was transferred to the receptor strains without any deletion. Gene expression of a convertant having both stx1 and stx2 genes was confirmed to be up to 32 times higher for Stx1 in 6% NaCl osmotic media and twice for Stx2 in 4% NaCl media, compared with expression in low-salt environments. Therefore, a new risk might arise from the transfer of pathogenic genes from Stx-encoding phages to otherwise harmless hosts. Without adequate sterilization of food exposed to various environments, there is a possibility that the toxicity of the phages might increase.


Assuntos
Colífagos/genética , Escherichia coli/genética , Toxina Shiga/genética , Transdução Genética , Colífagos/classificação , Colífagos/isolamento & purificação , Colífagos/fisiologia , Escherichia coli/crescimento & desenvolvimento , Escherichia coli/metabolismo , Escherichia coli/virologia , Expressão Gênica , Lisogenia , Myoviridae/classificação , Myoviridae/genética , Myoviridae/isolamento & purificação , Toxina Shiga/metabolismo , Fatores de Virulência/genética , Fatores de Virulência/metabolismo
17.
Viruses ; 13(2)2021 02 20.
Artigo em Inglês | MEDLINE | ID: mdl-33672748

RESUMO

Here we present the characterization of a Francisella bacteriophage (vB_FhiM_KIRK) including the morphology, the genome sequence and the induction of the prophage. The prophage sequence (FhaGI-1) has previously been identified in F. hispaniensis strain 3523. UV radiation induced the prophage to assemble phage particles consisting of an icosahedral head (~52 nm in diameter), a tail of up to 97 nm in length and a mean width of 9 nm. The double stranded genome of vB_FhiM_KIRK contains 51 open reading frames and is 34,259 bp in length. The genotypic and phylogenetic analysis indicated that this phage seems to belong to the Myoviridae family of bacteriophages. Under the conditions tested here, host cell (Francisella hispaniensis 3523) lysis activity of KIRK was very low, and the phage particles seem to be defective for infecting new bacterial cells. Nevertheless, recombinant KIRK DNA was able to integrate site-specifically into the genome of different Francisella species after DNA transformation.


Assuntos
Bacteriófagos/genética , Francisella/virologia , Myoviridae/genética , Bacteriófagos/classificação , Bacteriófagos/isolamento & purificação , Bacteriófagos/ultraestrutura , Genoma Viral , Myoviridae/classificação , Myoviridae/isolamento & purificação , Myoviridae/ultraestrutura , Fases de Leitura Aberta , Filogenia , Proteínas Virais/genética
18.
BMC Microbiol ; 21(1): 57, 2021 02 20.
Artigo em Inglês | MEDLINE | ID: mdl-33607940

RESUMO

BACKGROUND: Staphylococcus aureus is the causative agent of chronic mastitis, and can form a biofilm that is difficult to completely remove once formed. Disinfectants are effective against S. aureus, but their activity is easily affected by environmental factors and they are corrosive to equipment and chemically toxic to livestock and humans. Therefore, we investigated the potential utility of a bacteriophage as a narrow-spectrum disinfectant against biofilms formed by S. aureus. In this study, we isolated and characterized bacteriophage vB_SauM_SDQ (abbreviated to SDQ) to determine its efficacy in removing S. aureus biofilms. RESULTS: SDQ belongs to the family Myoviridae and consists of a hexagonal head, long neck, and short tail. This phage can sterilize a 109 CFU/mL culture of S. aureus in 12 h and multiply itself 1000-fold in that time. Biofilms formed on polystyrene, milk, and mammary-gland tissue were significantly reduced after SDQ treatment. Fluorescence microscopy and scanning electron microscopy showed that SDQ destroyed the biofilm structure. Moreover, the titer of SDQ remained relatively high after the lysis of the bacteria and the removal of the biofilm, exerting a continuous bacteriostatic effect. SDQ also retained its full activity under conditions that mimic common environments, i.e., in the presence of nonionic detergents, tap water, or organic materials. A nonionic detergent (Triton X-100) enhanced the removal of biofilm by SDQ. CONCLUSIONS: Our results suggest that SDQ, a specific lytic S. aureus phage, can be used to control biofilm infections. SDQ maintains its full activity in the presence of nonionic detergents, tap water, metal chelators, and organic materials, and can be used in combination with detergents. We propose this phage as a narrow-spectrum disinfectant against S. aureus, to augment or supplement the use of broad-spectrum disinfectants in the prevention and control of the mastitis and dairy industry contamination caused by S. aureus.


Assuntos
Biofilmes , Mastite/veterinária , Myoviridae/isolamento & purificação , Infecções Estafilocócicas/prevenção & controle , Fagos de Staphylococcus/isolamento & purificação , Staphylococcus aureus/virologia , Animais , Bovinos , Indústria de Laticínios , Desinfetantes , Feminino , Mastite/microbiologia , Mastite/prevenção & controle , Mastite/terapia , Microscopia Eletrônica de Varredura , Myoviridae/genética , Myoviridae/fisiologia , Terapia por Fagos , Esgotos/microbiologia , Infecções Estafilocócicas/microbiologia , Infecções Estafilocócicas/terapia , Fagos de Staphylococcus/genética , Fagos de Staphylococcus/fisiologia , Staphylococcus aureus/fisiologia
19.
Arch Virol ; 166(4): 1263-1265, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33585960

RESUMO

Xanthomonas oryzae pv. oryzae is a bacterial pathogen that gives rise to diseases in rice all over the world. A bacteriophage infecting this bacterium was isolated from rice fields in China. Here, we report the complete genome sequence of this phage, which has a linear dsDNA genome of 309,023 bp and a G + C content of 42.43%. It contains 401 open reading frames and encodes 28 tRNAs. It belongs to the family Myoviridae and has a broad host range, making it a possible candidate for phage therapy.


Assuntos
Bacteriófagos/genética , Genoma Viral/genética , Xanthomonas/virologia , Bacteriófagos/classificação , Bacteriófagos/isolamento & purificação , Composição de Bases , Sequência de Bases , DNA Viral/genética , Especificidade de Hospedeiro , Myoviridae/classificação , Myoviridae/genética , Myoviridae/isolamento & purificação , Fases de Leitura Aberta , Oryza/microbiologia , Doenças das Plantas/microbiologia , RNA de Transferência/genética , Análise de Sequência de DNA , Homologia de Sequência , Microbiologia do Solo
20.
Arch Microbiol ; 203(4): 1345-1356, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33386871

RESUMO

This work describes the characterization and genome annotation of Salmonella phage vB_SalM_8-19 (referred to as 8-19) isolated from sewage samples collected in a pig farm in Jilin, China. This phage was capable of infecting 60% Salmonella strains in our lab stock. The genome of phage 8-19 is composed of linear double-stranded DNA that is 52,648 bp in length with a G + C content of 46.02%; containing 74 ORFs and no tRNA genes. In October 2019, phylogenetic analyses indicated that phage 8-19 might belong to a novel cluster among the other similar phages which have not been specifically classified within some new genus in family Myoviridae. Recently, the International Committee on Taxonomy of Viruses (ICTV) defined phage 8-19 and its related phages as genus Rosemountvirus, family Myoviridae. This new genus, known as Rosemountvirus, is rarely reported in the literature.


Assuntos
Genoma Viral , Myoviridae/genética , Fagos de Salmonella/genética , Animais , Composição de Bases , China , Myoviridae/classificação , Myoviridae/isolamento & purificação , Fases de Leitura Aberta , Filogenia , Fagos de Salmonella/classificação , Fagos de Salmonella/isolamento & purificação , Esgotos/virologia , Suínos
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