Cross-sectional study on fecal carriage of Enterobacteriaceae with resistance to extended-spectrum cephalosporins in primary care patients

The aim of this study was to gain knowledge of the local epidemiology of extended-spectrum cephalosporin-resistant bacteria in primary care patients in a Swiss community. Fecal swabs were obtained from 291 primary care patients. Phenotyping and genotyping methods were used for further characterization of the isolates. Risk factors associated with carriage of ß-lactam-resistant strains were determined. Extended-spectrum cephalosporin-resistant Enterobacteriaceae were detected in 15 (5.2%) of the primary care patients. Thirteen isolates were CTX-M producers, one produced SHV-12, and three carried CMY-2. The pathogenic pandemic clone Escherichia coli ST131 was detected in 26.6% of the patients. Two patients (13.3%) carried two distinct strains simultaneously. There was a statistically significant risk of carriage of resistant strains for persons with a history of antibiotic therapy 4 months before sampling (p=0.05), markedly for therapy with ß-lactam (p=0.01). Age, gender, or history of hospitalization 4 months before sampling was not a risk factor for the acquisition of resistant bacteria in the analyzed patients. The relatively low prevalence of extended-spectrum cephalosporin-resistant strains in the community reflects the nationwide restrictive policy of antibiotic prescription as well as local implementation thereof. Nevertheless, our study shows that a potent antimicrobial resistance reservoir is present in primary care patients

Quinolone resistance mechanisms in Salmonella enterica serovars Hadar, Kentucky, Virchow, Schwarzengrund and S. enterica spp. enterica serovar 4,5,12:i:- isolated from humans in Switzerland: Identification of a novel qnrD variant, qnrD2

Human isolates of Salmonella enterica serovars Hadar, Kentucky, Virchow, Schwarzengrund, and the monophasic variant of S. Typhimurium, Salmonella enterica subsp. enterica serovar 4,5,12:i:- were examined for mutations within the quinolone resistance target genes gyrA, gyrB, parC, and parE and for plasmid-mediated resistance genes. Differences were observed among the serovars. A novel variant of qnrD, qnrD2, was detected in an S. Hadar isolate

Antimicrobial susceptibility of travel-related Salmonella enterica serovar Typhi isolates detected in Switzerland (2002-2013) and molecular characterization of quinolone resistant isolates

BACKGROUND: Typhoid fever is an acute, invasive, and potentially fatal systemic infection caused by Salmonella enterica subspecies enterica serotype Typhi (S. Typhi). Drug resistance to antimicrobials such as ciprofloxacin is emerging in developing countries, threatening the efficacy of treatment of patients in endemic regions as well as of travellers returning from these countries. METHODS: We compared the antimicrobial resistance profiles of 192 S. Typhi isolated from patients over a time span of twelve years. Susceptibility testing was done by the disk diffusion method. A representative selection of isolates (n = 41) was screened by PCR for mutations in the quinolone resistance-determining regions (QRDRs) of the gyrA and parC genes and all 192 isolates were screened for plasmid-mediated quinolone resistance (PMQR) genes. Multilocus sequence typing (MLST) was used to investigate the sequence type of isolates from patients with a known history of international travel. RESULTS: Resistance rates for nalidixic acid increased from 20 % to 66.7 % between 2002 and 2013. Resistance to ciprofloxacin was detected in 55.6 % of the isolates by 2013. Ciprofloxacin resistance was predominantly associated with the triple substitutions Ser83 → Phe and Asp87 → Asn in GyrA and Ser80 → Ile in ParC. The plasmid-mediated resistance gene qnrS1 was detected in two isolates. Sequence type ST1 was associated with the Indian subcontinent, while ST2 was distributed internationally. Multidrug resistance was noted for 11.5 % of the isolates. CONCLUSIONS: Fluoroquinolone resistant S. Typhi constitute a serious public health concern in endemic areas as well as in industrialized countries. Increased surveillance of global patterns of antimicrobial resistance is necessary and the control of resistant strains is of the utmost importance to maintain treatment options

Quinolone resistance mechanisms among extended-spectrum beta-lactamase (ESBL) producing Escherichia coli isolated from rivers and lakes in Switzerland

Sixty extended-spectrum β-lactamase (ESBL)-producing Escherichia coli isolated from rivers and lakes in Switzerland were screened for individual strains additionally exhibiting a reduced quinolone susceptibility phenotype. Totally, 42 such isolates were found and further characterized for their molecular (fluoro)quinolone resistance mechanisms. PCR and sequence analysis were performed to identify chromosomal mutations in the quinolone resistance-determining regions (QRDR) of gyrA, gyrB, parC and parE and to describe the occurrence of the following plasmid-mediated quinolone resistance genes: qepA, aac-6'-Ib-cr, qnrA, qnrB, qnrC, qnrD and qnrS. The contribution of efflux pumps to the resistance phenotype of selected strains was further determined by the broth microdilution method in the presence and absence of the efflux pump inhibitor phe-arg-β-naphthylamide (PAβN). Almost all strains, except two isolates, showed at least one mutation in the QRDR of gyrA. Ten strains showed only one mutation in gyrA, whereas thirty isolates exhibited up to four mutations in the QRDR of gyrA, parC and/or parE. No mutations were detected in gyrB. Most frequently the amino-acid substitution Ser83→Leu was detected in GyrA followed by Asp87→Asn in GyrA, Ser80→Ile in ParC, Glu84→Val in ParC and Ser458→Ala in ParE. Plasmid-mediated quinolone resistance mechanisms were found in twenty isolates bearing QnrS1 (4/20), AAC-6'-Ib-cr (15/20) and QepA (1/20) determinants, respectively. No qnrA, qnrB, qnrC and qnrD were found. In the presence of PAβN, the MICs of nalidixic acid were decreased 4- to 32-fold. (Fluoro) quinolone resistance is due to various mechanisms frequently associated with ESBL-production in E. coli from surface waters in Switzerland

Characteristics of extended-spectrum cephalosporin resistant Escherichia coli isolated from Swiss and imported poultry meat

A worrisome phenomenon is the progressive global spread of Enterobacteriaceae in poultry and chicken meat expressing plasmid-mediated enzymes that inactivate β-lactam antibiotics, suggesting that the food chain might play a role in the epidemiology and the transmission of extended-spectrum cephalosporin-resistant Enterobacteriaceae to humans. The aim of the present study was to further characterize 24 extended-spectrum cephalosporin-resistant Enterobacteriaceae isolated from domestic and imported poultry meat by antibiotic susceptibility testing, identification of the bla ESBL/bla pAmpC genes, conjugation mating experiments and determination of plasmid incompatibility types, multilocus sequence typing, and analysis of the Escherichia coli phylogenetic groups. On account of their resistance patterns, 21 of the total 24 isolates were classified as multidrug resistant. Eleven isolates carried a bla CMY-2 gene, whereas 13 isolates harbored a bla CTX-M-1 gene. All isolates harbored plasmids that were assigned to 8 of the 18 described plasmid incompatibility groups, the most frequent of which were IncI1, IncFIB, IncB/O, and IncFrepB. The bla ESBL/bla pAmpC genes were harbored mainly by transferable IncI1 and IncB/O plasmids. Multilocus sequence typing as well as E. coli phylogenetic group typing revealed a high heterogenicity even among different isolates of the same sample

MIC values of nalidixic acid in the presence and absence of Phe-Arg-β-naphthylamide in selected <i>E. coli</i> isolates.

<p>MIC values of nalidixic acid in the presence and absence of Phe-Arg-β-naphthylamide in selected <i>E. coli</i> isolates.</p

Plasmid-mediated quinolone resistance mechanisms, replicon types of transferred plasmids and quinolone resistance levels of the transconjugants of selected <i>E. coli</i> isolates.

<p><i>E. coli</i> HK225 Strep^RRif^R, recipient strain resistant to streptomycin and rifamycin.</p><p>TC_OW54E2, transconjugant receiving the plasmid from the donor strain OW54E2.</p

Quinolone resistance, amino acid substitutions in the QRDR of GyrA, ParC and ParE proteins in terms of amino acid positions and PMQR determinants in extended-spectrum β-lactamase (ESBL) producing <i>Escherichia coli</i> isolated from rivers and lakes in Switzerland.

a<p>) see Zurfluh et al. <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0095864#pone.0095864-Zurfluh1" target="_blank">[10]</a> for comparison.</p><p>AM, ampicillin; AMC, amoxicillin-clavulanic acid; CF, cephalothin; CTX, cefotaxime; CIP, ciprofloxacin; GM, gentamicin; TE, tetracycline; S, streptomycin; C, chloramphenicol; K, kanamycin; NA, nalidixic acid; SMZ, sulfamethoxazole; TMP, trimethoprim.</p><p>QRDR: quinolone resistance-determining regions; PMQR: plasmid-mediated quinolone resistance mechanisms.</p