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Article ; Online: Reconciling the Potentially Irreconcilable? Genotypic and Phenotypic Amoxicillin-Clavulanate Resistance in

2020 Volume 64, Issue 6

Abstract: Resistance to amoxicillin-clavulanate, a widely used beta-lactam/beta-lactamase inhibitor combination antibiotic, is rising globally, and yet susceptibility testing remains challenging. To test whether whole-genome sequencing (WGS) could provide a more ... ...

Abstract	Resistance to amoxicillin-clavulanate, a widely used beta-lactam/beta-lactamase inhibitor combination antibiotic, is rising globally, and yet susceptibility testing remains challenging. To test whether whole-genome sequencing (WGS) could provide a more reliable assessment of susceptibility than traditional methods, we predicted resistance from WGS for 976
MeSH term(s)	Amoxicillin-Potassium Clavulanate Combination/pharmacology ; Anti-Bacterial Agents/pharmacology ; Anti-Bacterial Agents/therapeutic use ; Clavulanic Acid/pharmacology ; Escherichia coli/genetics ; Microbial Sensitivity Tests ; Phenotype ; United Kingdom ; beta-Lactamases/genetics
Chemical Substances	Anti-Bacterial Agents ; Clavulanic Acid (23521W1S24) ; Amoxicillin-Potassium Clavulanate Combination (74469-00-4) ; beta-Lactamases (EC 3.5.2.6)
Language	English
Publishing date	2020-05-21
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	217602-6
ISSN	1098-6596 ; 0066-4804
ISSN (online)	1098-6596
ISSN	0066-4804
DOI	10.1128/AAC.02026-19
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Article ; Online: Cefotaxime resistant Escherichia coli collected from a healthy volunteer; characterisation and the effect of plasmid loss.

Kirchner, Miranda / Abuoun, Manal / Mafura, Muriel / Bagnall, Mary / Hunt, Theresa / Thomas, Christopher / Weile, Jan / Anjum, Muna F

PloS one

2013 Volume 8, Issue 12, Page(s) e84142

Abstract: In this study 6 CTX-M positive E. coli isolates collected during a clinical study examining the effect of antibiotic use in a human trial were analysed. The aim of the study was to analyse these isolates and assess the effect of full or partial loss of ... ...

Abstract	In this study 6 CTX-M positive E. coli isolates collected during a clinical study examining the effect of antibiotic use in a human trial were analysed. The aim of the study was to analyse these isolates and assess the effect of full or partial loss of plasmid genes on bacterial fitness and pathogenicity. A DNA array was utilised to assess resistance and virulence gene carriage. Plasmids were characterised by PCR-based replicon typing and addiction system multiplex PCR. A phenotypic array and insect virulence model were utilised to assess the effect of plasmid-loss in E. coli of a large multi-resistance plasmid. All six E. coli carrying bla CTX-M-14 were detected from a single participant and were identical by pulse field gel electrophoresis and MLST. Plasmid profiling and arrays indicated absence of a large multi-drug resistance (MDR) F-replicon plasmid carrying blaTEM, aadA4, strA, strB, dfrA17/19, sul1, and tetB from one isolate. Although this isolate partially retained the plasmid it showed altered fitness characteristics e.g. inability to respire in presence of antiseptics, similar to a plasmid-cured strain. However, unlike the plasmid-cured or plasmid harbouring strains, the survival rate for Galleria mellonella infected by the former strain was approximately 5-times lower, indicating other possible changes accompanying partial plasmid loss. In conclusion, our results demonstrated that an apparently healthy individual can harbour bla CTX-M-14 E. coli strains. In one such strain, isolated from the same individual, partial absence of a large MDR plasmid resulted in altered fitness and virulence characteristics, which may have implications in the ability of this strain to infect and any subsequent treatment.
MeSH term(s)	Anti-Bacterial Agents/pharmacology ; Cefotaxime/pharmacology ; Drug Resistance, Bacterial/genetics ; Escherichia coli/drug effects ; Escherichia coli/genetics ; Escherichia coli/isolation & purification ; Escherichia coli/physiology ; Genotype ; Healthy Volunteers ; Humans ; Phenotype ; Plasmids/genetics ; beta-Lactamases/genetics
Chemical Substances	Anti-Bacterial Agents ; beta-lactamase CTX-2 (EC 3.5.2.-) ; beta-Lactamases (EC 3.5.2.6) ; Cefotaxime (N2GI8B1GK7)
Language	English
Publishing date	2013-12-27
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	1932-6203
ISSN (online)	1932-6203
DOI	10.1371/journal.pone.0084142
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Article ; Online: The role of Campylobacter jejuni cytolethal distending toxin in gastroenteritis: toxin detection, antibody production, and clinical outcome.

Mortensen, Ninell P / Schiellerup, Peter / Boisen, Nadia / Klein, Bjarke M / Locht, Henning / Abuoun, Manal / Newell, Diane / Krogfelt, Karen A

APMIS : acta pathologica, microbiologica, et immunologica Scandinavica

2011 Volume 119, Issue 9, Page(s) 626–634

Abstract: The role of Campylobacter jejuni cytolethal distending toxin (CDT) on clinical outcome after gastroenteritis was investigated. Clinical data, blood serum samples, and Campylobacter spp. isolated, from each of 30 patients were collected over a period of 6 ...

Abstract	The role of Campylobacter jejuni cytolethal distending toxin (CDT) on clinical outcome after gastroenteritis was investigated. Clinical data, blood serum samples, and Campylobacter spp. isolated, from each of 30 patients were collected over a period of 6 months. The CDT encoding genes, cdtABC, characterized by PCR, revealed that all but one of the C. jejuni strains had the wild-type sequence. Sequencing of cdtABC from this strain showed two major deletions. From all of the strains, CDT titers were determined, and toxin neutralizing antibodies were documented using an in vitro assay. Three of the thirty clinical isolates, including the one with the mutant cdtABC coding genes, did not have a detectable CDT activity. Analyzing the relationship between CDT titer, serum neutralization of CDT, and the clinical outcome showed that campylobacteriosis caused by CDT-negative strains was clinically indistinguishable from that of patients infected with an isolate that produced high levels of CDT. These results suggest that CDT does not solely determine severity of infection and clinical outcome.
MeSH term(s)	Adolescent ; Antibodies, Bacterial/blood ; Antibody Formation ; Bacterial Toxins/genetics ; Bacterial Toxins/toxicity ; Campylobacter Infections/microbiology ; Campylobacter Infections/pathology ; Campylobacter jejuni/growth & development ; Campylobacter jejuni/isolation & purification ; Campylobacter jejuni/metabolism ; Campylobacter jejuni/pathogenicity ; Gastroenteritis/microbiology ; Gene Expression Regulation, Bacterial ; Genes, Bacterial ; HeLa Cells ; Humans ; Treatment Outcome
Chemical Substances	Antibodies, Bacterial ; Bacterial Toxins ; cytolethal distending toxin
Language	English
Publishing date	2011-06-21
Publishing country	Denmark
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	93340-5
ISSN	1600-0463 ; 0903-4641
ISSN (online)	1600-0463
ISSN	0903-4641
DOI	10.1111/j.1600-0463.2011.02781.x
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Article: Campylobacter colonization of the chicken induces a proinflammatory response in mucosal tissues.

Smith, Christopher K / Abuoun, Manal / Cawthraw, Shaun A / Humphrey, Tom J / Rothwell, Lisa / Kaiser, Pete / Barrow, Paul A / Jones, Michael A

FEMS immunology and medical microbiology

2008 Volume 54, Issue 1, Page(s) 114–121

Abstract: Campylobacter jejuni is a major cause of human inflammatory enteritis, but colonizes the gastrointestinal tract of poultry to a high level in a commensal manner. In vitro, C. jejuni induces the production of cytokines from both human and avian-model ... ...

Abstract	Campylobacter jejuni is a major cause of human inflammatory enteritis, but colonizes the gastrointestinal tract of poultry to a high level in a commensal manner. In vitro, C. jejuni induces the production of cytokines from both human and avian-model epithelial cell and macrophage infections. This suggests that, in vivo, Campylobacter could induce proinflammatory signals in both hosts. We investigated whether a proinflammatory cytokine response can be measured in both day-of-hatch and 2-week-old Light Sussex chickens during infection with C. jejuni. A significant induction of proinflammatory chemokine transcript was observed in birds of both ages, compared with levels in mock-infected controls. This correlated with an influx of heterophils but was not associated with any pathology. These results suggest that in poultry there may be a controlled inflammatory process during colonization.
MeSH term(s)	Animals ; Campylobacter Infections/immunology ; Campylobacter Infections/microbiology ; Campylobacter Infections/veterinary ; Campylobacter jejuni/growth & development ; Campylobacter jejuni/immunology ; Campylobacter jejuni/isolation & purification ; Cecum/immunology ; Cecum/microbiology ; Chickens ; Colony Count, Microbial ; Cytokines/metabolism ; Ileum/immunology ; Ileum/microbiology ; Intestinal Mucosa/immunology ; Intestinal Mucosa/microbiology ; Poultry Diseases/immunology ; Poultry Diseases/microbiology ; Specific Pathogen-Free Organisms
Chemical Substances	Cytokines
Language	English
Publishing date	2008-10
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1143208-1
ISSN	1574-695X ; 0928-8244
ISSN (online)	1574-695X
ISSN	0928-8244
DOI	10.1111/j.1574-695X.2008.00458.x
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Article ; Online: A novel virulence strategy for Pseudomonas aeruginosa mediated by an autotransporter with arginine-specific aminopeptidase activity.

Luckett, Jeni C A / Darch, Owen / Watters, Chase / Abuoun, Manal / Wright, Victoria / Paredes-Osses, Esteban / Ward, Jenny / Goto, Hana / Heeb, Stephan / Pommier, Stéphanie / Rumbaugh, Kendra P / Cámara, Miguel / Hardie, Kim R

PLoS pathogens

2012 Volume 8, Issue 8, Page(s) e1002854

Abstract: The opportunistic human pathogen, Pseudomonas aeruginosa, is a major cause of infections in chronic wounds, burns and the lungs of cystic fibrosis patients. The P. aeruginosa genome encodes at least three proteins exhibiting the characteristic three ... ...

Abstract	The opportunistic human pathogen, Pseudomonas aeruginosa, is a major cause of infections in chronic wounds, burns and the lungs of cystic fibrosis patients. The P. aeruginosa genome encodes at least three proteins exhibiting the characteristic three domain structure of autotransporters, but much remains to be understood about the functions of these three proteins and their role in pathogenicity. Autotransporters are the largest family of secreted proteins in Gram-negative bacteria, and those characterised are virulence factors. Here, we demonstrate that the PA0328 autotransporter is a cell-surface tethered, arginine-specific aminopeptidase, and have defined its active site by site directed mutagenesis. Hence, we have assigned PA0328 with the name AaaA, for arginine-specific autotransporter of P. aeruginosa. We show that AaaA provides a fitness advantage in environments where the sole source of nitrogen is peptides with an aminoterminal arginine, and that this could be important for establishing an infection, as the lack of AaaA led to attenuation in a mouse chronic wound infection which correlated with lower levels of the cytokines TNFα, IL-1α, KC and COX-2. Consequently AaaA is an important virulence factor playing a significant role in the successful establishment of P. aeruginosa infections.
MeSH term(s)	Aminopeptidases/genetics ; Aminopeptidases/metabolism ; Animals ; Bacterial Proteins/genetics ; Bacterial Proteins/metabolism ; Carrier Proteins/genetics ; Carrier Proteins/metabolism ; Chronic Disease ; Cytokines/genetics ; Cytokines/metabolism ; Disease Models, Animal ; Humans ; Mice ; Mutagenesis, Site-Directed ; Peptides/metabolism ; Pseudomonas Infections/enzymology ; Pseudomonas Infections/genetics ; Pseudomonas aeruginosa/enzymology ; Pseudomonas aeruginosa/genetics ; Pseudomonas aeruginosa/pathogenicity ; Virulence Factors/genetics ; Virulence Factors/metabolism ; Wound Infection/enzymology ; Wound Infection/genetics ; Wound Infection/microbiology
Chemical Substances	Bacterial Proteins ; Carrier Proteins ; Cytokines ; Peptides ; Virulence Factors ; Aminopeptidases (EC 3.4.11.-) ; aminopeptidase B (EC 3.4.11.6)
Language	English
Publishing date	2012-08-23
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2205412-1
ISSN	1553-7374 ; 1553-7366
ISSN (online)	1553-7374
ISSN	1553-7366
DOI	10.1371/journal.ppat.1002854
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Article ; Online: Differential phenotypic and genotypic characteristics of qnrS1-harboring plasmids carried by hospital and community commensal enterobacteria.

Vien, Le Thi Minh / Abuoun, Manal / Morrison, Victoria / Thomson, Nicholas / Campbell, James I / Woodward, Martin J / Van Vinh Chau, Nguyen / Farrar, Jeremy / Schultsz, Constance / Baker, Stephen

Antimicrobial agents and chemotherapy

2011 Volume 55, Issue 4, Page(s) 1798–1802

Abstract: The qnrS1 gene induces reduced susceptibility to fluoroquinolones in enterobacteria. We investigated the structure, antimicrobial susceptibility phenotype, and antimicrobial resistance gene characteristics of qnrS1 plasmids from hospitalized patients and ...

Abstract	The qnrS1 gene induces reduced susceptibility to fluoroquinolones in enterobacteria. We investigated the structure, antimicrobial susceptibility phenotype, and antimicrobial resistance gene characteristics of qnrS1 plasmids from hospitalized patients and community controls in southern Vietnam. We found that the antimicrobial susceptibilities, resistance gene characteristics, and plasmid structures of qnrS1 plasmids from the hospital differed from those from the community. Our data imply that the characteristics of the two plasmid groups are indicative of distinct selective pressures in the differing environments.
MeSH term(s)	Anti-Bacterial Agents/pharmacology ; Bacterial Proteins/genetics ; Bacterial Proteins/physiology ; Drug Resistance, Multiple, Bacterial/genetics ; Enterobacteriaceae/drug effects ; Enterobacteriaceae/genetics ; Fluoroquinolones/pharmacology ; Genotype ; Microbial Sensitivity Tests ; Plasmids/genetics
Chemical Substances	Anti-Bacterial Agents ; Bacterial Proteins ; Fluoroquinolones
Language	English
Publishing date	2011-01-31
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	217602-6
ISSN	1098-6596 ; 0066-4804
ISSN (online)	1098-6596
ISSN	0066-4804
DOI	10.1128/AAC.01200-10
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Article: Epidemic multidrug-resistant (MDR-AmpC) Salmonella enterica serovar Newport strains contain three phage regions and a MDR resistance plasmid.

Wu, Guanghui / Abuoun, Manal / Hackl, Evelyn / La Ragione, Roberto M / Fookes, Maria / Fenner, Jackie / Pan, Zhensheng / Wenzl, Peter / Anjum, Muna F / Woodward, Martin J

Environmental microbiology reports

2010 Volume 2, Issue 2, Page(s) 228–235

Abstract: Multidrug-resistant (MDR-AmpC) Salmonella enterica serovar Newport has caused serious disease in animals and humans in North America, whereas in the UK S. enterica serovar Newport is not associated with severe disease and usually sensitive to antibiotics; ...

Abstract	Multidrug-resistant (MDR-AmpC) Salmonella enterica serovar Newport has caused serious disease in animals and humans in North America, whereas in the UK S. enterica serovar Newport is not associated with severe disease and usually sensitive to antibiotics; MDR S. Newport (not AmpC) strains have only been isolated from poultry. We found that UK poultry strains belonged to MLST type ST166 and were distinct from cattle isolates for being able to utilize D-tagotose and when compared by pulsed-field gel electrophoresis (PFGE), comparative genomic hybridization (CGH) and diversity arrays technology (DArT). Cattle strains belonged to the ST45 complex differing from ST166 at all seven loci. PFGE showed that 19 out of 27 cattle isolates were more than 85% similar to each other and some UK and US strains were indistinguishable. Both CGH and DArT identified genes (including phage-related ones) that were uniquely present in the US isolates and two such genes identified by DArT showed sequence similarities with the pertussis-like (artAB) toxin. This work demonstrates that MDR-AmpC S. Newport from the USA are genetically closely related to pan-susceptible strains from the UK, but contained three extra phage regions and a MDR plasmid.
Language	English
Publishing date	2010-04
Publishing country	United States
Document type	Journal Article
ISSN	1758-2229
ISSN	1758-2229
DOI	10.1111/j.1758-2229.2009.00095.x
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Article: Cytolethal distending toxin (CDT)-negative Campylobacter jejuni strains and anti-CDT neutralizing antibodies are induced during human infection but not during colonization in chickens.

Abuoun, Manal / Manning, Georgina / Cawthraw, Shaun A / Ridley, Anne / Ahmed, If H / Wassenaar, Trudy M / Newell, Diane G

Infection and immunity

2005 Volume 73, Issue 5, Page(s) 3053–3062

Abstract: The cytolethal distending toxin (CDT) of Campylobacter jejuni was detectable, using an in vitro assay, in most but not all of 24 strains tested. The reason for the absence of toxin activity in these naturally occurring CDT-negative C. jejuni strains was ... ...

Abstract	The cytolethal distending toxin (CDT) of Campylobacter jejuni was detectable, using an in vitro assay, in most but not all of 24 strains tested. The reason for the absence of toxin activity in these naturally occurring CDT-negative C. jejuni strains was then investigated at the genetic level. CDT is encoded by three highly conserved genes, cdtA, -B, and -C. In the CDT-negative strains, two types of mutation were identified. The CDT activities of C. jejuni strains possessing both types of mutation were successfully complemented with the functional genes of C. jejuni 11168. The first type of mutation comprised a 667-bp deletion across cdtA and cdtB and considerable degeneration in the remainder of the cdt locus. Using a PCR technique to screen for this deletion, this mutation occurred in fewer than 3% of 147 human, veterinary, and environmental strains tested. The second type of mutation involved at least four nonsynonymous nucleotide changes, but only the replacement of proline with serine at CdtB position 95 was considered important for CDT activity. This was confirmed by site-directed mutagenesis. This type of mutation also occurred in fewer than 3% of strains as determined using a LightCycler biprobe assay. The detection of two CDT-negative clinical isolates raised questions about the role of CDT in some cases of human campylobacteriosis. To determine if anti-CDT antibodies are produced in human infection, a toxin neutralization assay was developed and validated using rabbit antisera. Pooled human sera from infected patients neutralized the toxin, indicating expression and immunogenicity during infection. However, no neutralizing antibodies were detected in colonized chickens despite the expression of CDT in the avian gut as indicated by reverse transcription-PCR.
MeSH term(s)	Amino Acid Sequence ; Animals ; Antibodies, Bacterial/blood ; Bacterial Toxins/genetics ; Bacterial Toxins/immunology ; Campylobacter Infections/immunology ; Campylobacter Infections/microbiology ; Campylobacter jejuni/immunology ; Chickens ; Genetic Complementation Test ; HeLa Cells ; Humans ; Molecular Sequence Data ; Mutagenesis, Site-Directed ; Mutation ; Neutralization Tests ; Poultry Diseases/immunology ; Poultry Diseases/microbiology ; Species Specificity
Chemical Substances	Antibodies, Bacterial ; Bacterial Toxins ; cytolethal distending toxin
Language	English
Publishing date	2005-01-25
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	218698-6
ISSN	1098-5522 ; 0019-9567
ISSN (online)	1098-5522
ISSN	0019-9567
DOI	10.1128/IAI.73.5.3053-3062.2005
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