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Article: Effects of freshwater sponge Ephydatia fluviatilis on conjugative transfer of antimicrobial resistance in Enterococcus faecalis strains in aquatic environments

Cartwright, A / Arnscheidt, J / Conwell, M / Dooley, J.S.G / McGonigle, C / Naughton, P.J

Letters in applied microbiology. 2020 July, v. 71, no. 1

2020

Abstract: Filter feeding is a biotic process that brings waterborne bacteria in close contact with each other and may thus support the horizontal transfer of their antimicrobial resistance genes. This laboratory study investigated whether the freshwater sponge ... ...

Abstract	Filter feeding is a biotic process that brings waterborne bacteria in close contact with each other and may thus support the horizontal transfer of their antimicrobial resistance genes. This laboratory study investigated whether the freshwater sponge Ephydatia fluviatilis supported the transfer of vancomycin resistance between two Enterococcus faecalis strains that we previously demonstrated to exhibit pheromone responsive plasmid conjugation. Microcosm experiments exposed live and dead colonies of laboratory‐grown sponges to a vancomycin‐resistant donor strain and a rifampicin‐resistant recipient strain of Ent. faecalis. Enterococci with both resistance phenotypes were detected on double selection plates. In comparison to controls, abundance of these presumed transconjugants increased significantly in water from sponge microcosms. Homogenized suspensions of sponge cells also yielded presumed transconjugants; however, there was no significant difference between samples from live or dead sponges. Fluorescent in situ hybridization analysis of the sponge cell matrix using species‐specific probes revealed the presence of enterococci clusters with cells adjacent to each other. The results demonstrated that sponge colonies can support the horizontal transfer of antimicrobial resistance although the mechanism underlying this process, such as binding of the bacteria to the sponge collagen matrix, has yet to be fully elucidated.
Keywords	Enterococcus faecalis ; antibiotic resistance ; collagen ; fluorescence ; freshwater ; genetic conjugation ; hybridization ; pheromones ; vancomycin
Language	English
Dates of publication	2020-07
Size	p. 39-45.
Publishing place	John Wiley & Sons, Ltd
Document type	Article
Note	NAL-AP-2-clean ; JOURNAL ARTICLE
ZDB-ID	632584-1
ISSN	1472-765X ; 0266-8254
ISSN (online)	1472-765X
ISSN	0266-8254
DOI	10.1111/lam.13310
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Interspecies transfer of vancomycin, erythromycin and tetracycline resistance among Enterococcus species recovered from agrarian sources.

Conwell, M / Daniels, V / Naughton, P J / Dooley, J S G

BMC microbiology

2017 Volume 17, Issue 1, Page(s) 19

Abstract: Background: Enterococci are now well recognised for their ability to transfer antibiotic resistance and for their association with nosocomial infections, but less is known regarding their relevance in the wider environment. Enterococcus faecalis and ... ...

Abstract	Background: Enterococci are now well recognised for their ability to transfer antibiotic resistance and for their association with nosocomial infections, but less is known regarding their relevance in the wider environment. Enterococcus faecalis and Enterococcus faecium were isolated from a range of agrarian associated sources (low-flow water, septic tank, poultry litter, high flow water, slurry/soil) and were assessed for latent ability to transfer antimicrobial resistance. Results: The isolates were tested for phenotypic clumping in the presence of cell-free supernatant from other isolates. Some isolates were identified which demonstrated clumping, indicating that they possessed peptide sex pheromone conjugal machinery. All isolates were also tested for antibiotic resistance phenotypes using both disc diffusion and minimum inhibitory concentration (MIC) assays. These tests revealed that the enterococci demonstrated both phenotypic clumping and antibiotic resistance phenotypes. Based on these selection criteria, the isolates were identified as having the potential for horizontal gene transfer and were used to investigate the transfer of multiple antibiotic resistance phenotypes. Conjugal transfer of antibiotic resistance phenotypes was determined using a solid agar mating method followed by a standard antibiotic selection test resulting in different transfer patterns. An interspecies conjugal transfer of vancomycin resistance from E. faecalis to E. faecium was identified while the remaining reactions were within the same species. Transfer efficiencies ranging from 2 × 10 Conclusions: The conjugation reactions and selection conditions used in this study resulted in a variety of co-transferred resistance phenotypes suggesting the presence of different mobile elements in the set of natural isolates. This study highlights the potential for extensive horizontal gene transfer in a previously neglected reservoir for enterococci.
MeSH term(s)	Base Sequence ; Conjugation, Genetic/genetics ; Cross Infection ; DNA, Bacterial/genetics ; Drug Resistance, Multiple, Bacterial/genetics ; Enterococcus/drug effects ; Enterococcus/genetics ; Enterococcus/isolation & purification ; Enterococcus faecalis/drug effects ; Enterococcus faecalis/genetics ; Enterococcus faecalis/isolation & purification ; Enterococcus faecium/drug effects ; Enterococcus faecium/genetics ; Enterococcus faecium/isolation & purification ; Gene Transfer, Horizontal/genetics ; Genes, Bacterial/genetics ; Microbial Sensitivity Tests/methods ; Phenotype ; Sex Attractants ; Tetracycline Resistance/genetics ; Vancomycin Resistance/genetics ; Water Microbiology
Chemical Substances	DNA, Bacterial ; Sex Attractants
Language	English
Publishing date	2017-01-18
Publishing country	England
Document type	Journal Article
ISSN	1471-2180
ISSN (online)	1471-2180
DOI	10.1186/s12866-017-0928-3
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Effects of freshwater sponge Ephydatia fluviatilis on conjugative transfer of antimicrobial resistance in Enterococcus faecalis strains in aquatic environments.

Cartwright, A / Arnscheidt, J / Conwell, M / Dooley, J S G / McGonigle, C / Naughton, P J

Letters in applied microbiology

2020 Volume 71, Issue 1, Page(s) 39–45

Abstract	Filter feeding is a biotic process that brings waterborne bacteria in close contact with each other and may thus support the horizontal transfer of their antimicrobial resistance genes. This laboratory study investigated whether the freshwater sponge Ephydatia fluviatilis supported the transfer of vancomycin resistance between two Enterococcus faecalis strains that we previously demonstrated to exhibit pheromone responsive plasmid conjugation. Microcosm experiments exposed live and dead colonies of laboratory-grown sponges to a vancomycin-resistant donor strain and a rifampicin-resistant recipient strain of Ent. faecalis. Enterococci with both resistance phenotypes were detected on double selection plates. In comparison to controls, abundance of these presumed transconjugants increased significantly in water from sponge microcosms. Homogenized suspensions of sponge cells also yielded presumed transconjugants; however, there was no significant difference between samples from live or dead sponges. Fluorescent in situ hybridization analysis of the sponge cell matrix using species-specific probes revealed the presence of enterococci clusters with cells adjacent to each other. The results demonstrated that sponge colonies can support the horizontal transfer of antimicrobial resistance although the mechanism underlying this process, such as binding of the bacteria to the sponge collagen matrix, has yet to be fully elucidated.
MeSH term(s)	Animals ; Anti-Bacterial Agents/pharmacology ; Conjugation, Genetic/genetics ; Drug Resistance, Bacterial/genetics ; Enterococcus faecalis/drug effects ; Enterococcus faecalis/genetics ; Fresh Water ; In Situ Hybridization, Fluorescence ; Pheromones/pharmacology ; Plasmids/genetics ; Porifera/microbiology ; Vancomycin/pharmacology ; Vancomycin Resistance/genetics
Chemical Substances	Anti-Bacterial Agents ; Pheromones ; Vancomycin (6Q205EH1VU)
Language	English
Publishing date	2020-06-04
Publishing country	England
Document type	Journal Article
ZDB-ID	632584-1
ISSN	1472-765X ; 0266-8254
ISSN (online)	1472-765X
ISSN	0266-8254
DOI	10.1111/lam.13310
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Article: RT-TGGE as a guide for the successful isolation of phosphonoacetate degrading bacteria.

Panas, P / McMullan, G / Dooley, J S G

Journal of applied microbiology

2007 Volume 103, Issue 1, Page(s) 237–244

Abstract: Aims: Use of molecular techniques for the isolation of bacteria capable of phosphonoacetate mineralization as carbon, phosphorus and energy source.: Methods and results: RNA extracts obtained at three different stages of an enrichment selecting for ... ...

Abstract	Aims: Use of molecular techniques for the isolation of bacteria capable of phosphonoacetate mineralization as carbon, phosphorus and energy source. Methods and results: RNA extracts obtained at three different stages of an enrichment selecting for phosphonoacetate degrading bacteria were reverse transcribed using 16S rRNA-specific primers, amplified and analysed by temperature gradient gel electrophoresis (TGGE). This information was used to devise a strategy for the isolation of members of the enrichment that were otherwise difficult to obtain in pure culture. We were able to pull out, in total, four out of the six main microbial cultures that were detected by TGGE. Two of the isolates belonging to Mycobacterium and Agromyces genera were for the first time shown to grow in the presence of phosphonoacetate as sole carbon, phosphorus and energy source releasing almost equimolar levels of inorganic phosphate into the culture medium, and they were shown to exhibit phosphonoacetate hydrolase activity in vitro. Conclusions: The ubiquity of pseudomonad in degradation processes is more likely a consequence of our ignorance of bacterial requirements and physiology, rather than their possession of unique metabolic properties. Significance and impact of the study: RT-TGGE analysis can be used to guide the successful isolation of micro-organisms difficult to obtain by culture-dependent methods alone.
MeSH term(s)	Actinomycetales ; Alkaline Phosphatase ; Bacteria/genetics ; Bacteria/isolation & purification ; Bacteria/metabolism ; Biodegradation, Environmental ; Biodiversity ; DNA Fingerprinting ; Electrophoresis, Polyacrylamide Gel/methods ; Mycobacterium smegmatis/enzymology ; Phosphonoacetic Acid/metabolism ; Phosphoric Monoester Hydrolases/metabolism ; Phylogeny ; Polymerase Chain Reaction/methods ; RNA, Bacterial/genetics ; RNA, Ribosomal, 16S/genetics
Chemical Substances	RNA, Bacterial ; RNA, Ribosomal, 16S ; Alkaline Phosphatase (EC 3.1.3.1) ; Phosphoric Monoester Hydrolases (EC 3.1.3.2) ; phosphonoacetate hydrolase (EC 3.11.1.2) ; Phosphonoacetic Acid (N919E46723)
Language	English
Publishing date	2007-07
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1358023-1
ISSN	1365-2672 ; 1364-5072
ISSN (online)	1365-2672
ISSN	1364-5072
DOI	10.1111/j.1365-2672.2006.03213.x
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Article: Under the microscope: Arcobacter.

Snelling, W J / Matsuda, M / Moore, J E / Dooley, J S G

Letters in applied microbiology

2006 Volume 42, Issue 1, Page(s) 7–14

Abstract: This review describes characteristics of the genus Arcobacter. Unlike its close phenotypically related neighbour Campylobacter, Arcobacter is not currently a major public health concern, but is considered as an emerging human pathogen, and is of ... ...

Abstract	This review describes characteristics of the genus Arcobacter. Unlike its close phenotypically related neighbour Campylobacter, Arcobacter is not currently a major public health concern, but is considered as an emerging human pathogen, and is of significance towards animal health. This review focuses on the public health significance, culturing and typing, reservoirs, and antimicrobial studies of Arcobacter. Collectively, increasing knowledge in these areas will help to develop measures, which can be used to control this emerging pathogen.
MeSH term(s)	Animals ; Arcobacter/classification ; Arcobacter/isolation & purification ; Bacterial Typing Techniques/methods ; Campylobacter/classification ; Communicable Diseases, Emerging/microbiology ; Disease Reservoirs/microbiology ; Food Microbiology ; Gram-Negative Bacterial Infections/microbiology ; Humans ; Meat/microbiology ; Public Health ; Water Microbiology
Language	English
Publishing date	2006-01
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	632584-1
ISSN	1472-765X ; 0266-8254
ISSN (online)	1472-765X
ISSN	0266-8254
DOI	10.1111/j.1472-765X.2005.01841.x
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: RT-TGGE as a guide for the successful isolation of phosphonoacetate degrading bacteria

Panas, P / McMullan, G / Dooley, J.S.G

Journal of applied microbiology. 2007 July, v. 103, no. 1

2007

Abstract: Use of molecular techniques for the isolation of bacteria capable of phosphonoacetate mineralization as carbon, phosphorus and energy source. RNA extracts obtained at three different stages of an enrichment selecting for phosphonoacetate degrading ... ...

Abstract	Use of molecular techniques for the isolation of bacteria capable of phosphonoacetate mineralization as carbon, phosphorus and energy source. RNA extracts obtained at three different stages of an enrichment selecting for phosphonoacetate degrading bacteria were reverse transcribed using 16S rRNA-specific primers, amplified and analysed by temperature gradient gel electrophoresis (TGGE). This information was used to devise a strategy for the isolation of members of the enrichment that were otherwise difficult to obtain in pure culture. We were able to pull out, in total, four out of the six main microbial cultures that were detected by TGGE. Two of the isolates belonging to Mycobacterium and Agromyces genera were for the first time shown to grow in the presence of phosphonoacetate as sole carbon, phosphorus and energy source releasing almost equimolar levels of inorganic phosphate into the culture medium, and they were shown to exhibit phosphonoacetate hydrolase activity in vitro. The ubiquity of pseudomonad in degradation processes is more likely a consequence of our ignorance of bacterial requirements and physiology, rather than their possession of unique metabolic properties. RT-TGGE analysis can be used to guide the successful isolation of micro-organisms difficult to obtain by culture-dependent methods alone.
Keywords	Agromyces ; Mycobacterium ; Pseudomonas
Language	English
Dates of publication	2007-07
Size	p. 237-244.
Publisher	Blackwell Publishing Ltd
Publishing place	Oxford, UK
Document type	Article
ZDB-ID	1358023-1
ISSN	1364-5072
ISSN	1364-5072
DOI	10.1111/j.1365-2672.2006.03213.x
Database	NAL-Catalogue (AGRICOLA)

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Article: Polymerase chain reaction amplification: effect of dyes and other staining agents employed in clinical microbiology laboratories.

Hirayama, J / Hayashi, K / Goldsmith, C E / Coulter, W A / Millar, B C / Dooley, J S G / Matsuda, M / Moore, J E

British journal of biomedical science

2012 Volume 69, Issue 1, Page(s) 35–37

MeSH term(s)	Clinical Laboratory Techniques/methods ; Coloring Agents/adverse effects ; Coloring Agents/pharmacology ; Escherichia coli/drug effects ; Escherichia coli/genetics ; Escherichia coli/growth & development ; Escherichia coli/physiology ; Humans ; Microbiological Techniques/methods ; Nucleic Acids/drug effects ; Nucleic Acids/genetics ; Nucleic Acids/metabolism ; Polymerase Chain Reaction/methods ; RNA, Ribosomal, 16S/analysis ; RNA, Ribosomal, 16S/genetics ; Staining and Labeling/methods
Chemical Substances	Coloring Agents ; Nucleic Acids ; RNA, Ribosomal, 16S
Language	English
Publishing date	2012-05-03
Publishing country	England
Document type	Evaluation Studies ; Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1152119-3
ISSN	0967-4845
ISSN	0967-4845
DOI	10.1080/09674845.2012.11978243
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Article: Comparasion of five gene loci (rnpB, 16S rRNA, 16S-23S rRNA, sodA and dnaJ) to aid the molecular identification of viridans-group streptococci and pneumococci.

Maeda, Y / Goldsmith, C E / Coulter, W A / Mason, C / Dooley, J S G / Lowery, C J / Millar, B C / Moore, J E

British journal of biomedical science

2012 Volume 68, Issue 4, Page(s) 190–196

Abstract: Viridans-group streptococci (VGS) consist of several taxa which historically have been highly diverse. However, at times it may become necessary to have a reliable scheme for the identification of these organisms to the species level. The aim of this ... ...

Abstract	Viridans-group streptococci (VGS) consist of several taxa which historically have been highly diverse. However, at times it may become necessary to have a reliable scheme for the identification of these organisms to the species level. The aim of this study is to compare the ability of five gene loci, namely rnpB, 16S rRNA, 16S-23S rRNA, sodA and dnaJ, to speciate such organisms through a sequence typing-based approach. Reference organisms consisting of six VGS species were compared based on sequence typing, followed by comparison of 31 wild-type respiratory isolates, and showed that employment of sequence typing using the rnpB gene locus was the most specific and reliable. Therefore, the use of rnpB sequencing for the identification of VGS to species level is a reliable and feasible option, based on a single gene target.
MeSH term(s)	Genes, Bacterial/genetics ; Humans ; Phylogeny ; Pneumococcal Infections/diagnosis ; Pneumococcal Infections/microbiology ; RNA, Ribosomal, 16S/genetics ; RNA, Ribosomal, 23S/genetics ; Stomatitis/diagnosis ; Stomatitis/microbiology ; Streptococcal Infections/diagnosis ; Streptococcal Infections/microbiology ; Streptococcus pneumoniae/classification ; Streptococcus pneumoniae/genetics ; Streptococcus pneumoniae/isolation & purification ; Viridans Streptococci/classification ; Viridans Streptococci/genetics ; Viridans Streptococci/isolation & purification
Chemical Substances	RNA, Ribosomal, 16S ; RNA, Ribosomal, 23S
Language	English
Publishing date	2012-01-18
Publishing country	England
Document type	Comparative Study ; Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1152119-3
ISSN	0967-4845
ISSN	0967-4845
DOI	10.1080/09674845.2011.11730349
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Article: Campylobacter jejuni.

Snelling, W J / Matsuda, M / Moore, J E / Dooley, J S G

Letters in applied microbiology

2005 Volume 41, Issue 4, Page(s) 297–302

Abstract: This review describes characteristics of the family Campylobacteraceae and traits of Campylobacter jejuni. The review then focuses on the worldwide problem of C. jejuni antimicrobial resistance and mechanisms of pathogenesis and virulence. Unravelling ... ...

Abstract	This review describes characteristics of the family Campylobacteraceae and traits of Campylobacter jejuni. The review then focuses on the worldwide problem of C. jejuni antimicrobial resistance and mechanisms of pathogenesis and virulence. Unravelling these areas will help with the development of new therapeutic agents and ultimately decrease illness caused by this important human pathogen.
MeSH term(s)	Anti-Bacterial Agents/pharmacology ; Campylobacter Infections/microbiology ; Campylobacter jejuni/drug effects ; Campylobacter jejuni/pathogenicity ; Drug Resistance, Bacterial ; Humans ; Virulence
Chemical Substances	Anti-Bacterial Agents
Language	English
Publishing date	2005
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	632584-1
ISSN	1472-765X ; 0266-8254
ISSN (online)	1472-765X
ISSN	0266-8254
DOI	10.1111/j.1472-765X.2005.01788.x
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Article: Nucleic acid probes for the food industry

Dooley, J.S.G

Biotechnology advances. 1994. v. 12 (4)

1994

Keywords	DNA probes ; industrial applications ; food industry ; foodborne illness ; pathogens ; detection ; food analysis
Language	English
Size	p. 669-677.
Document type	Article
Note	In the special issue: Biotechnology and industry: Present and future / edited by C.R. Barnett, J.S.G. Dooley, A.P. McHale, and P.G. McKenna.
ZDB-ID	47165-3
ISSN	0734-9750
ISSN	0734-9750
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