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Riehen, Switzerland

Steinmann I.C.,University of Zurich | Pfluger V.,Mabritec AG | Schaffner F.,University of Zurich | Mathis A.,University of Zurich | Kaufmann C.,University of Zurich
Parasitology | Year: 2013

Matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) was evaluated for the rapid identification of ceratopogonid larvae. Optimal sample preparation as evaluated with laboratory-reared biting midges Culicoides nubeculosus was the homogenization of gut-less larvae in 10% formic acid, and analysis of 0·2 mg/ml crude protein homogenate mixed with SA matrix at a ratio of 1:1·5. Using 5 larvae each of 4 ceratopogonid species (C. nubeculosus, C. obsoletus, C. decor, and Dasyhelea sp.) and of 2 culicid species (Aedes aegypti, Ae. japonicus), biomarker mass sets between 27 and 33 masses were determined. In a validation study, 67 larvae belonging to the target species were correctly identified by automated database-based identification (91%) or manual full comparison (9%). Four specimens of non-target species did not yield identification. As anticipated for holometabolous insects, the biomarker mass sets of adults cannot be used for the identification of larvae, and vice versa, because they share only very few similar masses as shown for C. nubeculosus, C. obsoletus, and Ae. japonicus. Thus, protein profiling by MALDI-TOF as a quick, inexpensive and accurate alternative tool is applicable to identify insect larvae of vector species collected in the field. © Cambridge University Press 2012.

Stephan R.,University of Zurich | Joutsen S.,University of Helsinki | Hofer E.,University of Zurich | Sade E.,University of Helsinki | And 3 more authors.
European Journal of Clinical Microbiology and Infectious Diseases | Year: 2013

Yersinia enterocolitica biotype 1A strains are frequently isolated from the environment, foods, and animals, and also from humans with yersiniosis. There are controversial reports on the pathogenicity of biotype 1A strains. In this study, 811 fecal samples from asymptomatic humans from Switzerland were studied for the presence of Y. enterocolitica. Nine (1.1 %) of the 811 samples were positive for Y. enterocolitica 1A. These strains were compared with 12 Y. enterocolitica 1A strains from Swiss patients with diarrhea isolated in the same year. Almost all (20/21) Y. enterocolitica 1A strains carried the ystB gene, seven strains carried the hreP gene, and none carried the ail, ystA, myfA, yadA, or virF genes. Most (17/21) Y. enterocolitica 1A strains belonged to two major clusters, A and B, by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Strains of cluster B were only isolated from humans with diarrhea; however, ystB and hreP genes were detected in strains from both clinical and non-clinical samples and from strains of clusters A and B. Using ribotyping, six restriction patterns among biotype 1A strains were obtained with HindIII enzyme. The most common ribotype (RT I) was found in strains isolated from humans with and without diarrhea. All biotype 1A strains had a unique NotI profile by pulsed-field gel electrophoresis (PFGE), showing a very high genetic diversity. In this study, Y. enterocolitica 1A strains from clinical and non-clinical samples could not be clearly differentiated from each other. More research is needed in order to prove that biotype 1A strains are a primary cause for human yersiniosis and not only a secondary finding. © 2013 Springer-Verlag Berlin Heidelberg.

Hahn D.,Texas State University | Mirza B.,Texas State University | Benagli C.,Cantonal Institute of Microbiology | Vogel G.,Mabritec AG | And 2 more authors.
Systematic and Applied Microbiology | Year: 2011

Matrix-assisted laser desorption ionization-time-of-flight (MALDI-TOF) mass spectrometry (MS) was evaluated as a technique to characterize strains of the nitrogen-fixing actinomycete Frankia. MALDI-TOF MS reliably distinguished 37 isolates within the genus Frankia and assigned them to their respective host infection groups, i.e., the Alnus/. Casuarina and the Elaeagnus host infection groups. The assignment of individual strains to sub-groups within the respective host infection groups was consistent with classification based on comparative sequence analysis of nifH gene fragments, confirming the usefulness of MALDI-TOF MS as a rapid and reliable tool for the characterization of Frankia strains. © 2010 Elsevier GmbH.

Moser A.,University of Zurich | Stephan R.,University of Zurich | Ziegler D.,Mabritec AG | Johler S.,University of Zurich
Schweizer Archiv fur Tierheilkunde | Year: 2013

Coagulase-negative staphylococci (CNS) are the predominant cause of bovine intra-mammary infections. They can lead to chronic infections and were reported to signifi cantly increase milk somatic cell counts. The goal of our study was to determine the species distribution and antimicrobial susceptibility profi les of CNS in bovine mastitis milk samples in Switzerland. Between March 2011 and February 2012, a total of 120 CNS were isolated from mastitis milk samples from 117 different animals at 77 farms. The isolates were identifi ed by matrix-assisted laser desorption ionization - time of fl ight mass spectrometry (MALDI-TOF MS) and subsequently tested for sensitivity to various antibiotic agents by disk diffusion. Antimicrobial agents were selected mainly based on their relevance to the treatment of bovine mastitis in Switzerland. MALDI-TOF MS assigned the 120 isolates to 12 different staphylococcal species - S. chromogenes (33 %), S. xylosus (28 %), S. sciuri (13 %), S. haemolyticus (9 %), S. epidermidis (4 %), S. simulans (4 %), S. warneri (3 %), S. equorum (2 %), S. hyicus (2 %), S. cohnii (1 %), S. succinus (1 %), and S. fl euretti (1 %). Resistance rates in CNS were high, with 39% of isolates exhibiting resistance to ampicillin and penicillin, 6% of isolates being resistant to amoxicillin with clavulanic acid, ceftiofur, cephalothin, and cefoxitin, and 5 % being resistant to erythromycin. In rare cases resistance to gentamicin (2 %), kanamycin (2 %), and kanamycin- cefalexin (1 %) was detected. © 2013 Verlag Hans Huber, Hogrefe AG, Bern.

Schaffner F.,University of Zurich | Kaufmann C.,University of Zurich | Pfluger V.,Mabritec AG | Mathis A.,University of Zurich
Parasites and Vectors | Year: 2014

Background: Invasive aedine mosquito species have become a major issue in many parts of the world as most of them are recognised vectors or potentially involved in transmission of pathogens. Surveillance of these mosquitoes (e.g. Ae. aegypti, Yellow fever mosquito, Aedes albopictus, Asian tiger mosquito) is mainly done by collecting eggs using ovitraps and by identification of the larvae hatched in the laboratory. In order to replace this challenging and laborious procedure, we have evaluated matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) for easy and rapid species identification. Methods. Individual protein profiles were generated using five eggs each of nine aedine species (Ae. aegypti, Ae. albopictus, Ae. atropalpus, Ae. cretinus, Ae. geniculatus, Ae. japonicus, Ae. koreicus, Ae. phoeniciae, Ae. triseriatus) from various geographical origins, and species-specific biomarker mass sets could be generated. A blinded validation using our reference data base for automated egg identification was performed. In addition, pools of 10 aedine eggs (132 two-species and 18 three-species pools) in different ratios were evaluated. Results: Specific biomarker mass sets comprising 18 marker masses could be generated for eggs of nine container-inhabiting aedine species, including all the major invasive and indigenous species of Europe and North America. Two additional masses shared by all investigated aedine species are used as internal calibrators. Identification of single eggs was highly accurate (100% specificity, 98.75% sensitivity), and this method is also of value for the identification of species in pools of ten eggs. When mixing two or three species, all were identified in all pools in at least 2 or 1 of the 4 loaded replicates, respectively, if the "lesser abundant" species in the pool accounted for three or more eggs. Conclusions: MALDI-TOF MS, which is widely applied for routine identification of microorganisms in clinical microbiology laboratories, is also suited for robust, low-cost and high throughput identification of mosquito vectors in surveillance programmes. This tool can further be developed to include a wide spectrum of arthropods but also other Metazoa for which surveillance is required, and might become the method of choice for their centralised identification via online platforms. © 2014 Schaffner et al.; licensee BioMed Central Ltd.

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