scholarly journals Bacteriophage GH15: Developing A Novel Weapon Against MRSA

2019 ◽  
Vol 11 (1) ◽  
Author(s):  
Adam Card
Keyword(s):  
Host Range ◽  
Home Health Care ◽  
Therapeutic Agent ◽  
Bacterial Species ◽  
Alternative Methods ◽  
Broad Host Range ◽  
Antibiotic Resistant ◽  
Humoral Immune ◽  
Humoral Immune Responses ◽  
Antibiotic Resistant Bacterium

In recent years, numerous bacterial species have developed antibiotic resistance due to the overuse of antibiotics in the home, health care setting, and in agriculture. Alternative methods of treatment, including phage therapy (PT), have been proposed as solutions to this problem. PT is showing promise as an alternative method of treatment against the bacteria methicillin-resistant Staphylococcus aureus (MRSA). MRSA is a virulent and antibiotic resistant bacterium capable of causing infections of the skin, respiratory system, and various other body systems. In this research proposal, we propose investigating the use of the Staphylococcal bacteriophage (phage) GH15 as a therapeutic agent against MRSA infections due to its broad host range, its lack of bacterial virulence genes, and its strong ability to lyse various strains of MRSA. Specifically, we propose to evaluate the tail fibre genes of GH15 contributing to the phage’s host range, in addition to the ability of the phage to induce antiphage humoral immune responses in human cells, in the interest of exploring GH15 as a therapeutic agent for use in PT, specifically against MRSA. 

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 Fate of Antibiotic Resistant Pseudomonas putida and Broad Host Range Plasmid in Natural Soil Microcosms

2019 ◽  
Vol 10 ◽  
Author(s):  
Xiao-Ting Fan ◽  
Hu Li ◽  
Qing-Lin Chen ◽  
Yu-Sen Zhang ◽  
Jun Ye ◽  
...  
Keyword(s):  
Pseudomonas Putida ◽  
Host Range ◽  
Natural Soil ◽  
Broad Host Range ◽  
Antibiotic Resistant ◽  
Soil Microcosms ◽  
Broad Host Range Plasmid

scholarly journals Large scale changes in host methylation patterns induced by IncA/C plasmid transformation inVibrio cholerae

2018 ◽  
Author(s):  
Ruibai Wang ◽  
Kanglin Wan
Keyword(s):  
Drug Resistance ◽  
Host Range ◽  
Vibrio Cholerae ◽  
Horizontal Transfer ◽  
Large Scale ◽  
Epigenetic Modification ◽  
Bacterial Species ◽  
Multi Drug Resistance ◽  
Broad Host Range ◽  
Methylation Patterns

AbstractDNA methylation is a central epigenetic modification and has diverse biological functions in eukaryotic and prokaryotic organisms alike. The IncA/C plasmid genomes are approximately 150kb in length and harbour three methylase genes, two of which demonstrate cytosine specificity. Transformation of theVibrio choleraestrain C6706 with the IncA/C plasmid pVC211 resulted in a significant relabelling of the methylation patterns on the host chromosomes. The new methylation patterns induced by transformation with IncA/C plasmid were accepted by the restriction enzymes of the host’s restriction modification (RM) system. These data uncover a novel mechanism by which plasmids can be compatible with a host’s RM system and suggest a possible reason that plasmids of the IncA/C family are broad-host-range.Author summaryAntibiotic resistance of bacteria is a growing serious problem worldwidely and the horizontal transfer of multi-drug resistance genes mediated by plasmids within and between species of bacteria is the main reason. In the researches of multi-drug resistance ofVibrio cholerae, I have isolated several IncA/C plasmids. What impressed me most is their ability to accumulate the resistant genes. Moreover, they can transfer with high frequency and are stable in several bacterial species. There are at least three Tra regions on the IncA/C plasmid which containing components of the Type 4 Secretion System and are important for conjugative transfer of plasmids. So the horizontal transfer ability of IncA/C plasmids is reasonable. There are three methylase genes on the small genome of IncA/C plasmids, which demonstrate cytosine specificity and are seldom in bacteria. Their modification target and roles are interesting. Here, we analysed the methylation profiles of the hostV. choeraeinduced by the plasmid pVC211 and found that they were completely changed. In addition to replicons, this may be a novel mechanism that plasmid cross the barrier of the host’s RM system and become broad-host range. Changing the activity of methylase in IncA/C plasmids may be a new way to affect the stability of IncA/C plasmids to eliminate these multidrug-resistant plasmids from bacteria.

scholarly journals Broad-Host-Range Plasmids for Constitutive and Inducible Gene Expression in the Absence of Antibiotic Selection

2019 ◽  
Vol 8 (36) ◽  
Author(s):  
Lindsey Burbank ◽  
Wei Wei
Keyword(s):  
Gene Expression ◽  
Host Range ◽  
Gene Function ◽  
Bacterial Species ◽  
Limited Range ◽  
Broad Host Range ◽  
Bacterial Gene ◽  
Antibiotic Selection ◽  
Broad Host Range Plasmids ◽  
Inducible Gene

Plasmid vectors are a valuable research tool for characterizing bacterial gene function, but there is a limited range of plasmids that are functional in nonmodel bacterial species. Described here is a set of broad-host-range plasmids modified for stability in the absence of antibiotic selection and for gene expression manipulation.

scholarly journals Burkholderia thailandensis E125 Harbors a Temperate Bacteriophage Specific for Burkholderia mallei

2002 ◽  
Vol 184 (14) ◽  
pp. 4003-4017 ◽  
Author(s):  
Donald E. Woods ◽  
Jeffrey A. Jeddeloh ◽  
David L. Fritz ◽  
David DeShazer
Keyword(s):  
Host Range ◽  
Bacterial Species ◽  
Attachment Site ◽  
Broad Host Range ◽  
Burkholderia Mallei ◽  
Burkholderia Thailandensis ◽  
O Antigen ◽  
Broad Host Range Plasmid ◽  
Transmission Electron ◽  
Cytosine Methyltransferase

ABSTRACT Burkholderia thailandensis is a nonpathogenic gram-negative bacillus that is closely related to Burkholderia mallei and Burkholderia pseudomallei. We found that B. thailandensis E125 spontaneously produced a bacteriophage, termed φE125, which formed turbid plaques in top agar containing B. mallei ATCC 23344. We examined the host range of φE125 and found that it formed plaques on B. mallei but not on any other bacterial species tested, including B. thailandensis and B. pseudomallei. Examination of the bacteriophage by transmission electron microscopy revealed an isometric head and a long noncontractile tail. B. mallei NCTC 120 and B. mallei DB110795 were resistant to infection with φE125 and did not produce lipopolysaccharide (LPS) O antigen due to IS407A insertions in wbiE and wbiG, respectively. wbiE was provided in trans on a broad-host-range plasmid to B. mallei NCTC 120, and it restored LPS O-antigen production and susceptibility to φE125. The 53,373-bp φE125 genome contained 70 genes, an IS3 family insertion sequence (ISBt3), and an attachment site (attP) encompassing the 3′ end of a proline tRNA (UGG) gene. While the overall genetic organization of the φE125 genome was similar to λ-like bacteriophages and prophages, it also possessed a novel cluster of putative replication and lysogeny genes. The φE125 genome encoded an adenine and a cytosine methyltransferase, and purified bacteriophage DNA contained both N6-methyladenine and N4-methylcytosine. The results presented here demonstrate that φE125 is a new member of the λ supergroup of Siphoviridae that may be useful as a diagnostic tool for B. mallei.

scholarly journals Genetic Modification of Sodalis Species by DNA Transduction

mSphere ◽  
2021 ◽  
Vol 6 (1) ◽  
Author(s):  
Chelsea M. Keller ◽  
Christopher G. Kendra ◽  
Roberto E. Bruna ◽  
David Craft ◽  
Mauricio H. Pontes
Keyword(s):  
Host Range ◽  
Genetic Study ◽  
Genetic Manipulation ◽  
Bacterial Species ◽  
Broad Host Range ◽  
Insect Host ◽  
Exogenous Dna ◽  
Content Type ◽  
Sodalis Glossinidius ◽  
Genetically Manipulated

ABSTRACT Bacteriophages (phages) are ubiquitous in nature. These viruses play a number of central roles in microbial ecology and evolution by, for instance, promoting horizontal gene transfer (HGT) among bacterial species. The ability of phages to mediate HGT through transduction has been widely exploited as an experimental tool for the genetic study of bacteria. As such, bacteriophage P1 represents a prototypical generalized transducing phage with a broad host range that has been extensively employed in the genetic manipulation of Escherichia coli and a number of other model bacterial species. Here we demonstrate that P1 is capable of infecting, lysogenizing, and promoting transduction in members of the bacterial genus Sodalis, including the maternally inherited insect endosymbiont Sodalis glossinidius. While establishing new tools for the genetic study of these bacterial species, our results suggest that P1 may be used to deliver DNA to many Gram-negative endosymbionts in their insect host, thereby circumventing a culturing requirement to genetically manipulate these organisms. IMPORTANCE A large number of economically important insects maintain intimate associations with maternally inherited endosymbiotic bacteria. Due to the inherent nature of these associations, insect endosymbionts cannot be usually isolated in pure culture or genetically manipulated. Here we use a broad-host-range bacteriophage to deliver exogenous DNA to an insect endosymbiont and a closely related free-living species. Our results suggest that broad-host-range bacteriophages can be used to genetically alter insect endosymbionts in their insect host and, as a result, bypass a culturing requirement to genetically alter these bacteria.

scholarly journals Characterization of novel bacteriophage phiC119 capable of lysing multidrug-resistant Shiga toxin-producingEscherichia coliO157:H7

PeerJ ◽  
2016 ◽  
Vol 4 ◽  
pp. e2423 ◽  
Author(s):  
Luis Amarillas ◽  
Cristóbal Chaidez ◽  
Arturo González-Robles ◽  
Yadira Lugo-Melchor ◽  
Josefina León-Félix
Keyword(s):  
Host Range ◽  
Shiga Toxin ◽  
Pathogenic Bacteria ◽  
Genetic Material ◽  
Multidrug Resistant ◽  
Broad Host Range ◽  
Antibiotic Resistant ◽  
E Coli ◽  
Tract Infections

BackgroundShiga toxin-producingEscherichia coli(STEC) is one of the most common and widely distributed foodborne pathogens that has been frequently implicated in gastrointestinal and urinary tract infections. Moreover, high rates of multiple antibiotic-resistantE. colistrains have been reported worldwide. Due to the emergence of antibiotic-resistant strains, bacteriophages are considered an attractive alternative to biocontrol pathogenic bacteria. Characterization is a preliminary step towards designing a phage for biocontrol.MethodsIn this study, we describe the characterization of a bacteriophage designated phiC119, which can infect and lyse several multidrug-resistant STEC strains and someSalmonellastrains. The phage genome was screened to detect thestx-genes using PCR, morphological analysis, host range was determined, and genome sequencing were carried out, as well as an analysis of the cohesive ends and identification of the type of genetic material through enzymatic digestion of the genome.ResultsAnalysis of the bacteriophage particles by transmission electron microscopy showed that it had an icosahedral head and a long tail, characteristic of the familySiphoviridae. The phage exhibits broad host range against multidrug-resistant and highly virulentE. coliisolates. One-step growth experiments revealed that the phiC119 phage presented a large burst size (210 PFU/cell) and a latent period of 20 min. Based on genomic analysis, the phage contains a linear double-stranded DNA genome with a size of 47,319 bp. The phage encodes 75 putative proteins, but lysogeny and virulence genes were not found in the phiC119 genome.ConclusionThese results suggest that phage phiC119 may be a good biological control agent. However, further studies are required to ensure its control of STEC and to confirm the safety of phage use.

scholarly journals DNA transduction in Sodalis species: implications for the genetic modification of uncultured endosymbionts of insects

2020 ◽  
Author(s):  
Chelsea M. Keller ◽  
Christopher G. Kendra ◽  
Roberto E. Bruna ◽  
David Craft ◽  
Mauricio H. Pontes
Keyword(s):  
Host Range ◽  
Genetic Study ◽  
Genetic Manipulation ◽  
Bacterial Species ◽  
Broad Host Range ◽  
Insect Host ◽  
Exogenous Dna ◽  
Bacteriophage P1 ◽  
Sodalis Glossinidius ◽  
Genetically Manipulated

AbstractBacteriophages (phages) are ubiquitous in nature. These viruses play a number of central roles in microbial ecology and evolution by, for instance, promoting horizontal gene transfer (HGT) among bacterial species. The ability of phages to mediate HGT through transduction has been widely exploited as an experimental tool for the genetic study of bacteria. As such, bacteriophage P1 represents a prototypical generalized transducing phage with a broad host range that has been extensively employed in the genetic manipulation of Escherichia coli and a number of other model bacterial species. Here we demonstrate that P1 is capable of infecting, lysogenizing and promoting transduction in members of the bacterial genus Sodalis, including the maternally inherited insect endosymbiont Sodalis glossinidius. While establishing new tools for the genetic study of these bacterial species, our results suggest that P1 may be used to deliver DNA to many Gram negative endosymbionts in their insect host, thereby circumventing a culturing requirement to genetically manipulate these organisms.SummaryA large number of economically important insects maintain intimate associations with maternally inherited endosymbiotic bacteria. Due to the inherit nature of these associations, insect endosymbionts cannot be usually isolated in pure culture nor genetically manipulated. Here we use a broad-host range bacteriophage to deliver exogenous DNA to an insect endosymbiont and a closely related free-living species. Our results suggest that broad host range bacteriophages can be used to genetically alter insect endosymbionts in their insect host and, as a result, bypass a culturing requirement to genetically alter these bacteria.

scholarly journals Molecular and Functional Characterization ofSalmonella enterica Serovar Typhimurium poxAGene: Effect on Attenuation of Virulence and Protection

1998 ◽  
Vol 66 (12) ◽  
pp. 5599-5606 ◽  
Author(s):  
Koné Kaniga ◽  
Melissa S. Compton ◽  
Roy Curtiss ◽  
Preeti Sundaram
Keyword(s):  
Bacterial Species ◽  
Expression System ◽  
Functional Characterization ◽  
Wild Type ◽  
Mesenteric Lymph Nodes ◽  
Reading Frame ◽  
Humoral Immune ◽  
Humoral Immune Responses ◽  
Serovar Typhimurium ◽  
Dna Fragment

ABSTRACT Salmonella enterica poxA mutants exhibit a pleiotropic phenotype, including reduced pyruvate oxidase activity; reduced growth rate; and hypersensitivity to the herbicide sulfometuron methyl, α-ketobutyrate, and amino acid analogs. These mutants also failed to grow in the presence of the host antimicrobial peptide, protamine. In this study, PoxA− mutants of S. entericaserovar Typhimurium (S. typhimurium) were found to be 10,000-fold attenuated in orally inoculated BALB/c mice and 1,000-fold attenuated in intraperitoneally inoculated BALB/c mice, compared to wild-type S. typhimurium UK-1. In addition,poxA mutants were found to be capable of colonizing the spleen, mesenteric lymph nodes, and Peyer’s patches; to induce strong humoral immune responses; and to protect mice against a lethal wild-type Salmonella challenge. A 2-kb DNA fragment was isolated from wild-type S. typhimurium UK-1 based on its ability to complement an isogenic poxA mutant. The nucleotide sequence of this DNA fragment revealed an open reading frame of 325 amino acids capable of encoding a polypeptide of 36.8 kDa that was confirmed in the bacteriophage T7 expression system. Comparison of the translated sequence to the available databases indicated high homology to a family of lysyl-tRNA synthetases. Our results indicate that a mutation of poxA has an attenuating effect onSalmonella virulence. Further, poxA mutants are immunogenic and could be useful in designing live vaccines with a variety of bacterial species. To our knowledge, this is the first report on the effect of poxA mutation on bacterial virulence.

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