Juliane Bräunig

Virulence and resistance determinants of German Staphylococcus aureus ST398 isolates from nonhuman sources.

ABSTRACT A series of 100 Staphylococcus aureus isolates ascribed to sequence type 398 (ST398) and recovered from different sources (healthy carrier and diseased pigs, dust from pig farms, milk, and meat) in Germany were investigated for their virulence and antimicrobial resistance genetic background. Antimicrobial resistance was determined by the disk diffusion method. Virulence and resistance determinants (37 and 31 genes, respectively) were tested by PCR. Only two virulence profiles, including the accessory gene regulator agrI and three or four hemolysin-encoding genes, were detected. In contrast, 33 resistance profiles were distinguished (only 11 were shown by more than one isolate). Fifty-nine isolates were multiresistant (four or more antimicrobial classes), and 98 were methicillin resistant (mecA positive). All of the ST398 isolates showed resistance to tetracycline [encoded by tet(M) alone or together with tet(K) and/or tet(L)]. In addition, 98% were resistant to other antimicrobials, including macrolide-lincosamine-streptogramin B (70%, encoded by ermA, ermB, and ermC, alone or in combination), trimethoprim (65%, mostly due to dfrK and dfrG), kanamycin and gentamicin [29% and 14%, respectively, mainly related to aac(6′)-Ie-aph(2″)-Ia and/or ant(4′)-Ia but also to aph(3′)-IIIa], chloramphenicol (9%, fexA or cfr), quinupristin-dalfopristin (9%), ciprofloxacin (8%), and trimethoprim-sulfamethoxazole (4%). The heterogeneity of the resistance profiles underlines the ability of the ST398 clone to acquire multiple antimicrobial resistance genes. However, the virulence gene content of the tested isolates was low. Continuous surveillance is needed to clarify whether its pathogenicity potential for animals and humans will increase over time.

Prevalence of MRSA types in slaughter pigs in different German abattoirs.

To investigate the prevalence of types of meticillinresistant Staphylococcus aureus (MRSA) in slaughter pigs in German abattoirs, nasal swabs were collected from a total of 1026 pigs in five abattoirs after stunning in the course of two studies, and examined for MRSA. Study 1 included four abattoirs; study 2 was carried out in one large abattoir. Isolates were tested for antimicrobial susceptibility and characterised using spa-typing, multilocus sequence typing (MLST) and typing of the staphylococcal cassette chromosome, SCCmec. Overall, MRSA was isolated from 70·8 per cent of 520 samples in study 1 and from 49·0 per cent of 506 samples in study 2. The proportion of positive samples varied substantially between the abattoirs in study 1. Most isolates belonged to spa-types t011 and t034 and SCCmec types III and V. MLST of selected isolates revealed that they were all MLST ST398. Besides β-lactams, 100 per cent of the isolates were resistant to tetracycline, 80·5 per cent were resistant to erythromycin and 80·7 per cent were resistant to clindamycin. Less than 5 per cent of the isolates were resistant to other antimicrobials.

High Heterogeneity within Methicillin-Resistant Staphylococcus aureus ST398 Isolates, Defined by Cfr9I Macrorestriction-Pulsed-Field Gel Electrophoresis Profiles and spa and SCCmec Types

ABSTRACT During recent years, the animal-associated methicillin-resistant Staphylococcus aureus clone ST398 has extensively been studied. The DNA of these isolates turned out to be refractory to SmaI restriction, and consequently, SmaI is unsuitable for subtyping this clone by standard pulsed-field gel electrophoresis (PFGE). Very recently, ST398 DNA was shown to be digested by Cfr9I, a neoschizomer of SmaI. In the present study, we employed Cfr9I PFGE on 100 German and 5 Dutch ST398 isolates and compared their PFGE profiles, protein A gene variable repeat regions (spa types), and types of the staphylococcal cassette chromosome mec (SCCmec). The isolates (from healthy carrier pigs, clinical samples from pigs, dust from farms, milk, and meat) were assigned to 35 profiles, which were correlated to the SCCmec type. A dendrogram with the Cfr9I patterns assigned all profiles to two clusters. Cluster A grouped nearly all isolates with SCCmec type V, and cluster B comprised all SCCmec type IVa and V* (a type V variant first identified as III) carriers plus one isolate with SCCmec type V. Both clusters also grouped methicillin-susceptible S. aureus isolates. The association of the majority of isolates with SCCmec type V in one large cluster indicated the presence of a successful subclone within the clonal complex CC398 from pigs, which has diversified. In general, the combination of Cfr9I PFGE with spa and SCCmec typing demonstrated the heterogeneity of the series analyzed and can be further used for outbreak investigations and traceability studies of the MRSA ST398 emerging clone.

Investigations on the possible impact of a glyphosate-containing herbicide on ruminal metabolism and bacteria in vitro by means of the 'Rumen Simulation Technique'.

This study was performed in a well‐established in vitro model to investigate whether the application of a glyphosate‐containing herbicide might affect the bacterial communities and some biochemical parameters in a cows rumen.

Development of a Comparative Risk Ranking System for Agents Posing a Bioterrorism Threat to Human or Animal Populations

Various systems for prioritizing biological agents with respect to their applicability as biological weapons are available, ranging from qualitative to (semi)quantitative approaches. This research aimed at generating a generic risk ranking system applicable to human and animal pathogenic agents based on scientific information. Criteria were evaluated and clustered to create a criteria list. Considering availability of data, a number of 28 criteria separated by content were identified that can be classified in 11 thematic areas or categories. Relevant categories contributing to probability were historical aspects, accessibility, production efforts, and possible paths for dispersion. Categories associated with impact are dealing with containment measures, availability of diagnostics, preventive and treatment measures in human and animal populations, impact on society, human and veterinary public health, and economic and ecological consequences. To allow data-based scoring, each criterion was described by at least 1 measure that allows the assignment of values. These values constitute quantities, ranges, or facts that are as explicit and precise as possible. The consideration of minimum and maximum values that can occur due to natural variations and that are often described in the literature led to the development of minimum and maximum criteria and consequently category scores. Missing or incomplete data, and uncertainty resulting therefrom, were integrated into the scheme via a cautious (but not overcautious) approach. The visualization technique that was used allows the description and illustration of uncertainty on the level of probability and impact. The developed risk ranking system was evaluated by assessing the risk originating from the bioterrorism threat of the animal pathogen bluetongue virus, the human pathogen Enterohemorrhagic Escherichia coli O157:H7, the zoonotic Bacillus anthracis, and Botulinum neurotoxin.

Risikoabschätzung von laMRSA in der Lebensmittelkette - Molekulare Diagnostik als Werkzeug zur Aufklärung epidemiologischer Zusammenhänge

Methicillin resistente Staphylococcus aureus (MRSA) sind bei landwirtschaftlichen Nutztieren weit verbreitet. Personengruppen, die direkten Kontakt zu mit MRSA besiedelten landwirtschaftlichen Nutztieren haben, sind einem im Vergleich zur Normalbevolkerung erhohten Risiko einer Kolonisation und nachfolgenden Infektion ausgesetzt. Das von livestock associated (la-)MRSA in der Lebensmittelkette ausgehende Risiko einer Gesundheitsgefahrdung fur den Menschen wird insgesamt als gering eingeschatzt, ist momentan aber noch nicht exakt quantifizierbar. Uber Lebensmittel konnen breite Bevolkerungsschichten exponiert sein, nach derzeitigem Kenntnisstand ist die Konzentration der Erreger im Lebensmittel jedoch gering. Hinweise auf lebensmittelassoziierte menschliche Infektionen (z. B. durch kontaminiertes Rohfleisch) mit laMRSA liegen bisher nicht vor. Weitere Forschungs- und Untersuchungstatigkeiten sind notig, um die nach wie vor vorhandenen Wissenslucken, insbesondere zu den Ubertragungswegen und der Epidemiologie des Erregers, zu schliesen. Die molekulare Typisierung der laMRSA Isolate leistet hierbei einen unverzichtbaren Beitrag.