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.Methicillin-resistant Staphylococcus aureus (MRSA) of sequence type 398 (ST398) has gained particular attention during recent years because of its association with pigs and its ability to colonize pig farmers and other people in close contact with pigs (7,12,47). The MRSA isolates of ST398 usually lack important virulence determinants that are typical in other community and hospital MRSA isolates. The majority of the ST398 isolates analyzed so far carry only hemolysin-encoding genes (13,21,31,32), although a small number of cases in which the isolates carried the bicomponent leukotoxin PantonValentine (lukPV genes) (43, 49) or staphylococcal enterotoxins (SEs, se genes) (21, 26) have also been reported. Genes for other toxins, like exfoliatins (ET, et genes), leukotoxins, and toxic shock syndrome toxin (TSST-1, tst gene) have not been found yet in ST398 isolates (13,21,31,32,44). The regulation of the expression of most extracellular virulence factors in S. aureus is under the control of a two-component signaling system called the accessory gene regulator (agr), which is polymorphic and divided into four distinct genetic groups (I to IV). A correlation exists between some agr groups and certain pathotypes and clonal complexes (CCs) (48), and CC398 seem to be associated with agr group I (ag...

To investigate the prevalence of types of meticillin-resistant 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 beta-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.

Decisions on food safety involve consideration of a wide range of concerns including the public health impact of foodborne illness, the economic importance of the agricultural sector and the food industry, and the effectiveness and efficiency of interventions. To support such decisions, we propose an integrated scientific approach combining veterinary and medical epidemiology, risk assessment for the farm-to-fork food chain as well as agricultural and health economy. Scientific advice is relevant in all stages of the policy cycle: to assess the magnitude of the food safety problem, to define the priorities for action, to establish the causes for the problem, to choose between different control options, to define targets along the food chain and to measure success.

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.

Aims: 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 cow's rumen. Methods and Results: The test item was applied in two concentrations (high and low) for 5 days. In a second trial, fermentation vessels were inoculated with Clostridium sporogenes before the high dose was applied. Effluents were analysed by biochemical, microbiological and genetic methods. A marginal increase in short-chain fatty acid production and a reduction in NH 3 -N were observed. There were minor and rather equivocal changes in the composition of ruminal bacteria but no indications of a shift towards a more frequent abundance of pathogenic Clostridia species. Clostridium sporogenes counts declined consistently. Conclusions: No adverse effects of the herbicide on ruminal metabolism or composition of the bacterial communities could be detected. In particular, there was no evidence of a suspected stimulation of Clostridia growth. Significance and Impact of the Study: Antibiotic activity of glyphosate resulting in microbial imbalances has been postulated. In this exploratory study, however, intraruminal application of concentrations reflecting potential exposure of dairy cows or beef cattle did not exhibit significant effects on bacterial communities in a complex in vitro system. The low number of replicates (n = 3/dose) may leave some uncertainty.