Yersinia enterocolitica strains associated with human infections in Switzerland 2001-2010

Yersinia enterocolitica infections are common in humans. However, very scarce data are available on the different biotypes and virulence factors of human strains, which has proved to be problematic to assess the clinical significance of the isolated strains. In this study, the presence of the ail gene and distribution of different bio- and serotypes among human Y. enterocolitica strains and their possible relation to the genotype and antimicrobial resistance were studied. In total, 128 Y. enterocolitica strains isolated from human clinical samples in Switzerland during 2001-2010 were characterised. Most (75 out of 128) of the Y. enterocolitica strains belonged to biotypes 2, 3 or 4 and carried the ail gene. One of the 51 strains that belonged to biotype 1A was also ail positive. Most of the ail-positive strains belonged to bioserotype 4/O:3 (47 out of 76) followed by 2/O:9 (22 out of 76). Strains of bioserotype 4/O:3 were dominant among patients between 20 and 40years old and strains of biotype 1A dominate in patients over 40years. Strains belonging to biotypes 2, 3 and 4, which all carried the ail gene, exhibited a high homogeneity with PFGE typing. Y. enterocolitica 2/O:5,27 and 2/O:9 strains showed resistance to amoxicillin/clavulanic acid and cefoxitin, but Y. enterocolitica 4/O:3 strains did no

Antimicrobial resistance patterns and genotypes of Salmonella enterica serovar Hadar strains associated with human infections in Switzerland, 2005-2010

Salmonella Hadar ranks in the top ten serovars reported from humans in Switzerland. In this study, all 64 S. Hadar strains isolated from different patients from 2005 to 2010 in Switzerland were characterized by (i) assessing phenotypic antimicrobial resistance profiles using the disk diffusion method and (ii) by genotyping using pulsed-field gel electrophoresis (PFGE) in order to evaluate the relationship of the strains. The annual incidences varied between 0·32/100000 in 2005 (highest incidence) and 0·065/100000 in 2007 (lowest incidence). In total 71·8% of the isolates were resistant to nalidixic acid. Although 40·6% of the strains were resistant to the β-lactam antibiotic ampicillin, they remained susceptible to the third-generation cephalosporin cefotaxime. Genotyping revealed a primary cluster consisting of 42 strains, sharing a similarity of >92%, with a subcluster of 18 strains with indistinguishable patterns. Resistance profiles allowed further differentiation within this subcluster providing a link of two strains to an outbreak in Spai

Pathogenic Yersinia enterocolitica O:3 isolated from a hunted wild alpine ibex

Occurrence of Yersinia spp. in wild ruminants was studied and the strains were characterized to get more information on the epidemiology of enteropathogenic Yersinia in the wildlife. In total, faecal samples of 77 red deer, 60 chamois, 55 roe deer and 27 alpine ibex were collected during 3 months of the hunting season in 2011. The most frequently identified species was Y. enterocolitica found in 13%, 10%, 4% and 2% of roe deer, red deer, alpine ibex and chamois, respectively. Interestingly, one Y. enterocolitica O:3 strain, isolated from an alpine ibex, carried the important virulence genes located on the virulence plasmid (yadA and virF) and in the chromosome (ail, hreP, myfA and ystA). Most of the Y. enterocolitica strains belonged to biotype 1A of which 14 were ystB positive. Further studies are needed to clarify the importance of alpine ibex as a reservoir of pathogenic Y. enterocolitic

Function impairing mutations in blaZ and blaR genes of penicillin susceptible Staphylococcus aureus strains isolated from bovine mastitis

Molecular based approaches have gained increasing importance in routine mastitis diagnostics for typing and antibiotic resistance testing of Staphylococcus aureus. Out of 78 S. aureus strains isolated from bovine mastitis milk 10 of them harbored blaZ, blaI and blaR genes. Although 5 strains were phenotypically resistant to penicillin, the other 5 (all belonging the clonal complex 8) were penicillin susceptible. PCR amplification confirmed the presence of the blaZ, blaR and blaI genes in all 5 strains. Sequencing of these genes uncovered a 29 base deletion within the blaZ gene in all these strains that causes a translational frame shift, which is predicted to induce abrogation of BlaZ expression. Additionally single nucleotide insertions and deletions were detected in blaR of 3 strains. These insertions cause translation reading frame shifts and premature stop codons that are predicted to induce expression of truncated BlaR proteins. Using the genetically altered blaZ genes detected as targets, a real-time PCR system for detecting CC8 associated blaZ positive S. aureus strains that still remain susceptible to penicillin was developed. Such strains are part of detection challenges that must be considered in routine application of genotypic resistance testing of bovine mastitis S. aureus

Fecal carriage of extended-spectrum beta-lactamase-producing Enterobacteriaceae in swine and cattle at slaughter in Switzerland

During the past decade, extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae have become a matter of great concern in human medicine. ESBL-producing strains are found in the community, not just in hospital-associated patients, which raises a question about possible reservoirs. Recent studies describe the occurrence of ESBL-producing Enterobacteriaceae in meat, fish, and raw milk; therefore, the impact of food animals as reservoirs for and disseminators of such strains into the food production chain must be assessed. In this pilot study, fecal samples of 59 pigs and 64 cattle were investigated to determine the occurrence of ESBL-producing Enterobacteriaceae in farm animals at slaughter in Switzerland. Presumptive-positive colonies on Brilliance ESBL agar were subjected to identification and antibiotic susceptibility testing including the disc diffusion method and E-test ESBL strips. As many as 15.2% of the porcine and 17.1% of the bovine samples, predominantly from calves, yielded ESBL producers. Of the 21 isolated strains, 20 were Escherichia coli, and one was Citrobacter youngae. PCR analysis revealed that 18 strains including C. youngae produced CTX-M group 1 ESBLs, and three strains carried genes encoding for CTX-M group 9 enzymes. In addition, eight isolates were PCR positive for TEM beta-lactamase, but no bla(SHV) genes were detected. Pulsed-field gel electrophoresis showed a high genetic diversity within the strains. The relatively high rates of occurrence of ESBLproducing strains in food animals and the high genetic diversity among these strains indicate that there is an established reservoir of these organisms in farm animals. Further studies are necessary to assess future trends

Evaluation of seven different commercially available real-time PCR assays for detection of shiga toxin 1 and 2 gene subtypes

Following the recent outbreak of Shiga toxin-producing Escherichia coli (STEC) O104:H4 infection in Germany, the demand for fast detection of STEC has again increased. Various real-time PCR-based methods enabling detection of Shiga toxin genes (stx) have been developed and can be used for applications in food microbiology. The present study was conducted to evaluate the reliability of seven commercially available real-time PCR systems for detection of stx1 and stx2 subtypes. For this purpose, pure cultures of 18 STEC strains harboring all known stx1 and/or stx2 subtypes were tested. Only one of the seven real-time PCR systems detected all known stx1 and stx2 subtypes. Six systems failed to detect the stx2f subtype. One system missed stx2 subtypes reported in association with severe human disease. Because the presence of certain stx genes (subtypes) is considered an important indicator of STEC virulence, systems differentiating between the stx1 and stx2 gene groups provide added value. Reliable and fast detection of stx genes is of major importance for both diagnostic laboratories and the food industry

Methicillin-resistant Staphylococcus lentus strains isolated from chicken carcasses and employees of a poultry abattoir

Methicillin-resistant coagulase-negative staphylococci (MR-CNS) are increasingly reported in animals and humans as colonizing organisms and as opportunistic pathogens. Within a collection of MR-CNS isolates from livestock, chicken carcasses, bulk tank milk, minced meat, and contact persons, matrix-assisted laser desorption ionization-time of flight mass spectrometry identified 37 isolates as S. lentus. All 37 methicillin-resistant S. lentus strains originated from either chicken carcasses (30 strains) or employees working in a poultry abattoir (seven strains). To assess the phenotypic antibiotic resistance to selected antibiotics (ciprofloxacin, clindamycin, erythromycin, gentamicin, rifampin, sulfamethoxazole/trimethoprime, tetracycline, vancomycin), the disk diffusion method was used and none of the strains was resistant to gentamicin, rifampin, or vancomycin. Pulsed-field gel electrophoresis (SmaI) was applied to evaluate the genotypic relationship of the 37 methicillin-resistant S. lentus strains. With a cut-off level of 80 % similarity, 30 (81.1 %) strains were grouped into only two clusters. Overall, the seven human strains showed between 85 % and 100 % similarity to the closest related chicken isolates. Our results suggest potential transmission of methicillin-resistant S. lentus between slaughtered chickens and abattoir personnel

Evaluation of three commercially available real-time PCR based systems for detection of Cronobacter species

In the last few years, various PCR based methods have been developed that enable detection of Cronobacter spp. to the genus and species level. Moreover, several real-time PCR based systems for detection of Cronobacter spp. are available, however, comparative evaluation studies are not available. The current study represents a comparative evaluation of three commercial diagnostic systems, namely the BAX® System PCR Assay Enterobacter sakazakii (DuPont, Qualicon, Wilmington, USA), the Assurance GDS™ Enterobacter sakazakii (BioControl, Bellvue, USA) and the foodproof® Enterobacter sakazakii Detection Kit (Biotecon Diagnostics, Potsdam, Germany) for the rapid identification of Cronobacter spp. Twenty-one target and non-target strains were included in the study and results were compared for specificity and convenience in performance. A specificity of 100% was observed for two of the three real time PCR systems tested, namely the Assurance GDS™ Enterobacter sakazakii and the foodproof® Enterobacter sakazakii Detection Kit for pure cultures as well as artificially contaminated powdered infant formula (PIF) samples. Copyright © 2011 Elsevier B.V. All rights reserved

Occurrence of Vibrio spp. in fish and shellfish collected from the Swiss market

The genus Vibrio includes gram-negative bacteria that inhabit estuarine ecosystems. V. cholerae, V. parahaemolyticus, and V. vulnificus pose a considerable public health threat as agents of sporadic and epidemic foodborne infections associated with the consumption of raw or undercooked contaminated fish or shellfish. In this study, we analyzed 138 fish and shellfish samples collected from the Swiss market (fish fillets [n = 102], bivalves [n = 34], and squid [n = 2]). Microbiological analysis was done according to International Organization for Standardization method 21872-1/21872-2:2007, using thiosulfate citrate bile sucrose agar and chromID Vibrio agar as selective agar. Presumptive-positive colonies on thiosulfate citrate bile sucrose agar or chromID Vibrio agar were picked and were identified by the API 20E and species-specific PCR systems. V. cholerae isolates were tested further by PCR for the presence of the cholera toxin A subunit gene (ctxA). V. parahaemolyticus isolates were tested by PCR for genes encoding for thermostable direct hemolysin (tdh) and TDH-related hemolysin (trh). V. cholerae was isolated from three samples and V. parahaemolyticus from eight samples. None of these strains harbored species-specific virulence factors. Further, V. alginolyticus was isolated from 40 samples, and V. fluvialis was isolated from 1 sample. Our study provides, for the first time, data for the assessment of exposure to Vibrio spp. in raw fish and bivalves consumed in Switzerland

Salmonella enterica serotype Kentucky associated with human infections in Switzerland: genotype and resistance trends 2004-2009

Salmonella serotype Kentucky emerged since 2002 and now ranks among the top ten serovars isolated from humans in Europe, and 8th to 10th in Switzerland. A total of 106 strains isolated from different patients from 2004 through 2009 in Switzerland were further characterized by (i) assessing phenotypic antibiotic resistance profiles using the disk diffusion method and (ii) by genotyping using pulsed-field gel electrophoresis (PFGE) after macrorestriction with XbaI in order to evaluate strain relationships and trends. In Switzerland, there is evidence for an outbreak in 2006 as the annual incidence almost doubled. A total of 30 strains (28%) were resistant or showed intermediate resistance to one to three antimicrobials and 48 strains (45%) displayed resistance to more than three antibiotics. We found a high prevalence (67%) of nalidixic acid resistance, and 58% were resistant to ciprofloxacin. One strain was a producer of extended-spectrum β-lactamase (ESBL). PFGE discriminated four clusters (similarity coefficient cut off at 80%). The resistance situation among the strains isolated from 2004 to 2009 in Switzerland is discussed and shown to coincide with findings in other European countries. Based on genetic subtyping, a so far undetected outbreak is likely to have occurred in Switzerland in 2006. Finally, our data identified travelling to Northern Africa as a risk factor for S. Kentucky infections