scholarly journals Isolation of Polyvalent Bacteriophages by Sequential Multiple-Host Approaches

2015 ◽  
Vol 82 (3) ◽  
pp. 808-815 ◽  
Author(s):  
Pingfeng Yu ◽  
Jacques Mathieu ◽  
Mengyan Li ◽  
Zhaoyi Dai ◽  
Pedro J. J. Alvarez
Keyword(s):  
Host Range ◽  
Pseudomonas Syringae ◽  
Burst Size ◽  
Broad Host Range ◽  
Ecological Significance ◽  
Content Type ◽  
Narrow Host Range ◽  
Isolation Methods ◽  
Culture Independent ◽  
K 12

ABSTRACTMany studies on phage biology are based on isolation methods that may inadvertently select for narrow-host-range phages. Consequently, broad-host-range phages, whose ecological significance is largely unexplored, are consistently overlooked. To enhance research on such polyvalent phages, we developed two sequential multihost isolation methods and tested both culture-dependent and culture-independent phage libraries for broad infectivity. Lytic phages isolated from activated sludge were capable of interspecies or even interorder infectivity without a significant reduction in the efficiency of plating (0.45 to 1.15). Two polyvalent phages (PX1 of thePodoviridaefamily and PEf1 of theSiphoviridaefamily) were characterized in terms of adsorption rate (3.54 × 10−10to 8.53 × 10−10ml/min), latent time (40 to 55 min), and burst size (45 to 99 PFU/cell), using different hosts. These phages were enriched with a nonpathogenic host (Pseudomonas putidaF1 orEscherichia coliK-12) and subsequently used to infect model problematic bacteria. By using a multiplicity of infection of 10 in bacterial challenge tests, >60% lethality was observed forPseudomonas aeruginosarelative to uninfected controls. The corresponding lethality forPseudomonas syringaewas ∼50%. Overall, this work suggests that polyvalent phages may be readily isolated from the environment by using different sequential hosts, and this approach should facilitate the study of their ecological significance as well as enable novel applications.

scholarly journals Identification of Bacteriophages for Biocontrol of the Kiwifruit Canker Phytopathogen Pseudomonas syringae pv. actinidiae

2014 ◽  
Vol 80 (7) ◽  
pp. 2216-2228 ◽  
Author(s):  
Rebekah A. Frampton ◽  
Corinda Taylor ◽  
Angela V. Holguín Moreno ◽  
Sandra B. Visnovsky ◽  
Nicola K. Petty ◽  
...  
Keyword(s):  
Host Range ◽  
Pseudomonas Syringae ◽  
Management Strategies ◽  
Pulse Field ◽  
Resistant Bacteria ◽  
Bacterial Canker ◽  
Content Type ◽  
Narrow Host Range ◽  
Pulse Field Gel Electrophoresis ◽  
Isolation And Characterization

ABSTRACTPseudomonas syringaepv. actinidiae is a reemerging pathogen which causes bacterial canker of kiwifruit (Actinidiasp.). Since 2008, a global outbreak ofP. syringaepv. actinidiae has occurred, and in 2010 this pathogen was detected in New Zealand. The economic impact and the development of resistance inP. syringaepv. actinidiae and other pathovars against antibiotics and copper sprays have led to a search for alternative management strategies. We isolated 275 phages, 258 of which were active againstP. syringaepv. actinidiae. Extensive host range testing onP. syringaepv. actinidiae, other pseudomonads, and bacteria isolated from kiwifruit orchards showed that most phages have a narrow host range. Twenty-four were analyzed by electron microscopy, pulse-field gel electrophoresis, and restriction digestion. Their suitability for biocontrol was tested by assessing stability and the absence of lysogeny and transduction. A detailed host range was performed, phage-resistant bacteria were isolated, and resistance to other phages was examined. The phages belonged to theCaudoviralesand were analyzed based on morphology and genome size, which showed them to be representatives ofMyoviridae,Podoviridae, andSiphoviridae. Twenty-oneMyoviridaemembers have similar morphologies and genome sizes yet differ in restriction patterns, host range, and resistance, indicating a closely related group. Nine of theseMyoviridaemembers were sequenced, and each was unique. The most closely related sequenced phages were a group infectingPseudomonas aeruginosaand characterized by phages JG004 and PAK_P1. In summary, this study reports the isolation and characterization ofP. syringaepv. actinidiae phages and provides a framework for the intelligent formulation of phage biocontrol agents against kiwifruit bacterial canker.

scholarly journals LamB, OmpC, and the Core Lipopolysaccharide of Escherichia coli K-12 Function as Receptors of Bacteriophage Bp7

2020 ◽  
Vol 94 (12) ◽  
Author(s):  
Peipei Chen ◽  
Huzhi Sun ◽  
Huiying Ren ◽  
Wenhua Liu ◽  
Guimei Li ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Host Range ◽  
Inner Core ◽  
Phage Infection ◽  
Broad Host Range ◽  
Content Type ◽  
E Coli ◽  
Phage Adsorption ◽  
Specific Receptors ◽  
K 12

ABSTRACT Bp7 is a T-even phage with a broad host range specific to Escherichia coli, including E. coli K-12. The receptor binding protein (RBP) of bacteriophages plays an important role in the phage adsorption process and determines phage host range, but the molecular mechanism involved in host recognition of phage Bp7 remains unknown. In this study, the interaction between phage Bp7 and E. coli K-12 was investigated. Based on homology alignment, amino acid sequence analysis, and a competitive assay, gp38, located at the tip of the long tail fiber, was identified as the RBP of phage Bp7. Using a combination of in vivo and in vitro approaches, including affinity chromatography, gene knockout mutagenesis, a phage plaque assay, and phage adsorption kinetics analysis, we identified the LamB and OmpC proteins on the surface of E. coli K-12 as specific receptors involved in the first step of reversible phage adsorption. Genomic analysis of the phage-resistant mutant strain E. coli K-12-R and complementation tests indicated that HepI of the inner core of polysaccharide acts as the second receptor recognized by phage Bp7 and is essential for successful phage infection. This observation provides an explanation of the broad host range of phage Bp7 and provides insight into phage-host interactions. IMPORTANCE The RBPs of T4-like phages are gp37 and gp38. The interaction between phage T4 RBP gp37 and its receptors has been clarified by many reports. However, the interaction between gp38 and its receptors during phage adsorption is still not completely understood. Here, we identified phage Bp7, which uses gp38 as an RBP, and provided a good model to study the phage-host interaction mechanisms in an enterobacteriophage. Our study revealed that gp38 of phage Bp7 recognizes the outer membrane proteins (OMPs) LamB and OmpC of E. coli K-12 as specific receptors and binds with them reversibly. HepI of the inner-core oligosaccharide is the second receptor and binds with phage Bp7 irreversibly to begin the infection process. Determining the interaction between the phage and its receptors will help elucidate the mechanisms of phage with a broad host range and help increase understanding of the phage infection mechanism based on gp38.

scholarly journals Silage Collected from Dairy Farms Harbors an Abundance of Listeriaphages with Considerable Host Range and Genome Size Diversity

2012 ◽  
Vol 78 (24) ◽  
pp. 8666-8675 ◽  
Author(s):  
Kitiya Vongkamjan ◽  
Andrea Moreno Switt ◽  
Henk C. den Bakker ◽  
Esther D. Fortes ◽  
Martin Wiedmann
Keyword(s):  
Host Range ◽  
Dairy Farm ◽  
Dairy Farms ◽  
Genomic Diversity ◽  
Broad Host Range ◽  
Content Type ◽  
Narrow Host Range ◽  
Food Borne ◽  
Size Diversity ◽  
Range Characterization

ABSTRACTSince the food-borne pathogenListeria monocytogenesis common in dairy farm environments, it is likely that phages infecting this bacterium (“listeriaphages”) are abundant on dairy farms. To better understand the ecology and diversity of listeriaphages on dairy farms and to develop a diverse phage collection for further studies, silage samples collected on two dairy farms were screened forL. monocytogenesand listeriaphages. While only 4.5% of silage samples tested positive forL. monocytogenes, 47.8% of samples were positive for listeriaphages, containing up to >1.5 × 104PFU/g. Host range characterization of the 114 phage isolates obtained, with a reference set of 13L. monocytogenesstrains representing the nine major serotypes and four lineages, revealed considerable host range diversity; phage isolates were classified into nine lysis groups. While one serotype 3c strain was not lysed by any phage isolates, serotype 4 strains were highly susceptible to phages and were lysed by 63.2 to 88.6% of phages tested. Overall, 12.3% of phage isolates showed a narrow host range (lysing 1 to 5 strains), while 28.9% of phages represented broad host range (lysing ≥11 strains). Genome sizes of the phage isolates were estimated to range from approximately 26 to 140 kb. The extensive host range and genomic diversity of phages observed here suggest an important role of phages in the ecology ofL. monocytogeneson dairy farms. In addition, the phage collection developed here has the potential to facilitate further development of phage-based biocontrol strategies (e.g., in silage) and other phage-based tools.

scholarly journals Cooccurrence of Broad- and Narrow-Host-Range Viruses Infecting the Bloom-Forming Toxic Cyanobacterium Microcystis aeruginosa

2019 ◽  
Vol 85 (18) ◽  
Author(s):  
Daichi Morimoto ◽  
Kento Tominaga ◽  
Yosuke Nishimura ◽  
Naohiro Yoshida ◽  
Shigeko Kimura ◽  
...  
Keyword(s):  
Host Range ◽  
Microcystis Aeruginosa ◽  
Current Knowledge ◽  
Broad Host Range ◽  
Antiviral Defense ◽  
Content Type ◽  
Narrow Host Range ◽  
Toxic Cyanobacterium ◽  
Antiviral Responses ◽  
Group Ii

ABSTRACT Viruses play important roles in regulating the abundance and composition of bacterial populations in aquatic ecosystems. The bloom-forming toxic cyanobacterium Microcystis aeruginosa is predicted to interact with diverse cyanoviruses, resulting in Microcystis population diversification. However, current knowledge of the genomes from these viruses and their infection programs is limited to those of Microcystis virus Ma-LMM01. Here, we performed a time series sampling at a small pond in Japan during a Microcystis bloom and then investigated the genomic information and transcriptional dynamics of Microcystis-interacting viruses using metagenomic and metatranscriptomic approaches. We identified 15 viral genomic fragments classified into three groups, groups I (including Ma-LMM01), II (high abundance and transcriptional activity), and III (new lineages). According to the phylogenetic distribution of Microcystis strains possessing spacers against each viral group, the group II-original viruses interacted with all three phylogenetically distinct Microcystis population types (phylotypes), whereas the groups I and III-original viruses interacted with only one or two phylotypes, indicating the cooccurrence of broad- (group II) and narrow (groups I and III)-host-range viruses in the bloom. These viral fragments showed the highest transcriptional levels during daytime regardless of their genomic differences. Interestingly, M. aeruginosa expressed antiviral defense genes in the environment, unlike what was seen with an Ma-LMM01 infection in a previous culture experiment. Given that broad-host-range viruses often induce antiviral responses within alternative hosts, our findings suggest that such antiviral responses might inhibit viral multiplication, mainly that of broad-host-range viruses like those in group II. IMPORTANCE The bloom-forming toxic cyanobacterium Microcystis aeruginosa is thought to have diversified its population through the interactions between host and viruses in antiviral defense systems. However, current knowledge of viral genomes and infection programs is limited to those of Microcystis virus Ma-LMM01, which was a narrow host range in which it can escape from the highly abundant host defense systems. Our metagenomic approaches unveiled the cooccurrence of narrow- and broad-host-range Microcystis viruses, which included fifteen viral genomic fragments from Microcystis blooms that were classified into three groups. Interestingly, Microcystis antiviral defense genes were expressed against viral infection in the environment, unlike what was seen in a culture experiment with Ma-LMM01. Given that viruses with a broad host range often induce antiviral responses within alternative hosts, our findings suggest that antiviral responses inhibit viral reproduction, especially that of broad-range viruses like those in group II. This paper augments our understanding of the interactions between M. aeruginosa and its viruses and fills an important knowledge gap.

scholarly journals Isolation of the Novel Phage PHB09 and Its Potential Use against the Plant Pathogen Pseudomonas syringae pv. actinidiae

Viruses ◽  
2021 ◽  
Vol 13 (11) ◽  
pp. 2275
Author(s):  
Yanxi Liu ◽  
Mengjiao Liu ◽  
Ran Hu ◽  
Jun Bai ◽  
Xiaoqing He ◽  
...  
Keyword(s):  
Host Range ◽  
Pseudomonas Syringae ◽  
Management Strategies ◽  
Lytic Bacteriophage ◽  
Bacterial Canker ◽  
Isolation And Identification ◽  
Narrow Host Range ◽  
Development Of Resistance ◽  
Wide Range ◽  
Complete Genome Sequence Analysis

Bacteriophages are viruses that specifically infect target bacteria. Recently, bacteriophages have been considered potential biological control agents for bacterial pathogens due to their host specificity. Pseudomonas syringae pv. actinidiae (Psa) is a reemerging pathogen that causes bacterial canker of kiwifruit (Actinidia sp.). The economic impact of this pest and the development of resistance to antibiotics and copper sprays in Psa and other pathovars have led to investigation of alternative management strategies. Phage therapy may be a useful alternative to conventional treatments for controlling Psa infections. Although the efficacy of bacteriophage φ6 was evaluated for the control of Psa, the characteristics of other DNA bacteriophages infecting Psa remain unclear. In this study, the PHB09 lytic bacteriophage specific to Psa was isolated from kiwifruit orchard soil. Extensive host range testing using Psa isolated from kiwifruit orchards and other Pseudomonas strains showed PHB09 has a narrow host range. It remained stable over a wide range of temperatures (4–50 °C) and pH values (pH 3–11) and maintained stability for 50 min under ultraviolet irradiation. Complete genome sequence analysis indicated PHB09 might belong to a new myovirus genus in Caudoviricetes. Its genome contains a total of 94,844 bp and 186 predicted genes associated with phage structure, packaging, host lysis, DNA manipulation, transcription, and additional functions. The isolation and identification of PHB09 enrich the research on Pseudomonas phages and provide a promising biocontrol agent against kiwifruit bacterial canker.

scholarly journals Evolution of IncA/CblaCMY-2-Carrying Plasmids by Acquisition of theblaNDM-1Carbapenemase Gene

2011 ◽  
Vol 56 (2) ◽  
pp. 783-786 ◽  
Author(s):  
Alessandra Carattoli ◽  
Laura Villa ◽  
Laurent Poirel ◽  
Rémy A. Bonnin ◽  
Patrice Nordmann
Keyword(s):  
Host Range ◽  
Large Scale ◽  
The United States ◽  
Broad Host Range ◽  
New Delhi ◽  
Yersinia Ruckeri ◽  
Content Type ◽  
Photobacterium Damselae ◽  
Environmental Bacteria ◽  
Broad Host Range Plasmids

ABSTRACTTheblaNDM-1gene has been reported to be often located on broad-host-range plasmids of the IncA/C type in clinical but also environmental bacteria recovered from the New Delhi, India, area. IncA/C-type plasmids are the main vehicles for the spread of the cephalosporinase geneblaCMY-2, frequently identified in the United States, Canada, and Europe. In this study, we completed the sequence of IncA/C plasmid pNDM-KN carrying theblaNDM-1gene, recovered from aKlebsiella pneumoniaeisolate from Kenya. This sequence was compared with those of three IncA/C-type reference plasmids fromEscherichia coli,Yersinia ruckeri, andPhotobacterium damselae. Comparative analysis showed that theblaNDM-1gene was located on a widely diffused plasmid scaffold known to be responsible for the spread ofblaCMY-2-like genes and consequently for resistance to broad-spectrum cephalosporins. Considering that IncA/C plasmids possess a broad host range, this scaffold might support a large-scale diffusion of theblaNDM-1gene among Gram-negative rods.

scholarly journals A Multispecies Cluster of VIM-1 Carbapenemase-Producing Enterobacterales Linked by a Novel, Highly Conjugative, and Broad-Host-Range IncA Plasmid Forebodes the Reemergence of VIM-1

2020 ◽  
Vol 64 (4) ◽  
Author(s):  
Gabriele Arcari ◽  
Federica Maria Di Lella ◽  
Giulia Bibbolino ◽  
Fabio Mengoni ◽  
Marzia Beccaccioli ◽  
...  
Keyword(s):  
Host Range ◽  
Bacterial Species ◽  
Klebsiella Oxytoca ◽  
Gene Cassette ◽  
University Hospital ◽  
Broad Host Range ◽  
Class 1 Integron ◽  
Content Type ◽  
Carbapenem Resistant ◽  
Class 1

ABSTRACT In this study, we investigated VIM-1-producing Escherichia coli, Klebsiella oxytoca, Klebsiella pneumoniae, Citrobacter freundii, and Enterobacter cloacae strains, isolated in 2019 during a period of active surveillance of carbapenem-resistant Enterobacterales in a large university hospital in Italy. VIM-1-producing strains colonized the gut of patients, with up to three different VIM-1-positive bacterial species isolated from a single rectal swab, but also caused bloodstream infection in one colonized patient. In the multispecies cluster, blaVIM-1 was identified in a 5-gene cassette class 1 integron, associated with several genetic determinants, including the blaSHV-12, qnrS1, and mph(A) genes, located on a highly conjugative and broad-host-range IncA plasmid. The characteristics and origin of this IncA plasmid were studied.

scholarly journals Construction of a Broad-Host-Range Tn7-Based Vector for Single-Copy PBAD-Controlled Gene Expression in Gram-Negative Bacteria

2012 ◽  
Vol 79 (2) ◽  
pp. 718-721 ◽  
Author(s):  
F. Heath Damron ◽  
Elizabeth S. McKenney ◽  
Herbert P. Schweizer ◽  
Joanna B. Goldberg
Keyword(s):  
Gene Expression ◽  
Pseudomonas Aeruginosa ◽  
Host Range ◽  
Single Copy ◽  
Broad Host Range ◽  
Gram Negative Bacteria ◽  
Gram Negative ◽  
Content Type ◽  
Delivery Vector ◽  
Inducible Gene

ABSTRACTWe describe a mini-Tn7-based broad-host-range expression cassette for arabinose-inducible gene expression from the PBADpromoter. This delivery vector, pTJ1, can integrate a single copy of a gene into the chromosome of Gram-negative bacteria for diverse genetic applications, of which several are discussed, usingPseudomonas aeruginosaas the model host.

scholarly journals The Sinorhizobium (Ensifer) fredii HH103 Nodulation Outer Protein NopI Is a Determinant for Efficient Nodulation of Soybean and Cowpea Plants

2016 ◽  
Vol 83 (5) ◽  
Author(s):  
Irene Jiménez-Guerrero ◽  
Francisco Pérez-Montaño ◽  
Carlos Medina ◽  
Francisco Javier Ollero ◽  
Francisco Javier López-Baena
Keyword(s):  
Adenylate Cyclase ◽  
Host Cell ◽  
Host Range ◽  
Pathogenic Bacteria ◽  
Defense Responses ◽  
Host Cells ◽  
Broad Host Range ◽  
Sinorhizobium Fredii ◽  
Symbiotic Efficiency ◽  
Content Type

ABSTRACT The type III secretion system (T3SS) is a specialized secretion apparatus that is commonly used by many plant and animal pathogenic bacteria to deliver proteins, termed effectors, to the interior of the host cells. These effectors suppress host defenses and interfere with signal transduction pathways to promote infection. Some rhizobial strains possess a functional T3SS, which is involved in the suppression of host defense responses, host range determination, and symbiotic efficiency. The analysis of the genome of the broad-host-range rhizobial strain Sinorhizobium fredii HH103 identified eight genes that code for putative T3SS effectors. Three of these effectors, NopL, NopP, and NopI, are Rhizobium specific. In this work, we demonstrate that NopI, whose amino acid sequence shows a certain similarity with NopP, is secreted through the S. fredii HH103 T3SS in response to flavonoids. We also determined that NopL can be considered an effector since it is directly secreted to the interior of the host cell as demonstrated by adenylate cyclase assays. Finally, the symbiotic phenotype of single, double, and triple nopI, nopL, and nopP mutants in soybean and cowpea was assayed, showing that NopI plays an important role in determining the number of nodules formed in both legumes and that the absence of both NopL and NopP is highly detrimental for symbiosis. IMPORTANCE The paper is focused on three Rhizobium-specific T3SS effectors of Sinorhizobium fredii HH103, NopL, NopP, and NopI. We demonstrate that S. fredii HH103 is able to secrete through the T3SS in response to flavonoids the nodulation outer protein NopI. Additionally, we determined that NopL can be considered an effector since it is secreted to the interior of the host cell as demonstrated by adenylate cyclase assays. Finally, nodulation assays of soybean and cowpea indicated that NopI is important for the determination of the number of nodules formed and that the absence of both NopL and NopP negatively affected nodulation.

scholarly journals Emergence and Dissemination of Enterobacteriaceae Isolates Producing CTX-M-1-Like Enzymes in Spain Are Associated with IncFII (CTX-M-15) and Broad-Host-Range (CTX-M-1, -3, and -32) Plasmids

2006 ◽  
Vol 51 (2) ◽  
pp. 796-799 ◽  
Author(s):  
Ângela Novais ◽  
Rafael Cantón ◽  
Raquel Moreira ◽  
Luísa Peixe ◽  
Fernando Baquero ◽  
...  
Keyword(s):  
Host Range ◽  
Broad Host Range ◽  
Narrow Host Range

ABSTRACT The spread of CTX-M-1-like enzymes in Spain is associated with particular plasmids of broad-host-range IncN (bla CTX-M-32, bla CTX-M-1), IncL/M (bla CTX-M-1), and IncA/C2 (bla CTX-M-3) or narrow-host-range IncFII (bla CTX-M-15). The identical genetic surroundings of bla CTX-M-32 and bla CTX-M-1 and their locations on related 40-kb IncN plasmids indicate the in vivo evolution of this element.

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