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Welfare recipients in Germany are allowed to take up supplementary jobs while receiving welfare. In the present study, we use the German Panel Study "Labour Market and Social Security" (PASS) for the years 2006–2014 to analyze the impact of these supplementary jobs on the chances of welfare exit. Dynamic multinomial logit models controlling for unobserved heterogeneity and endogenous initial conditions reveal that full‐time employed men and women are more likely to exit welfare into employment than their non‐employed counterparts. For supplementary part‐time jobs, however, we find no or (only in some specifications for men) much smaller stepping stone effects. [ABSTRACT FROM AUTHOR]

Little is known about zoonotic pathogens and their antimicrobial resistance in South American camelids (SAC) in Germany including Clostridioides (C.) difficile. The aim of this study was to investigate prevalence, molecular characteristics and antimicrobial resistance of C. difficile in SAC. Composite SAC faecal samples were collected in 43 husbandries in Central Germany and cultured for C. difficile. Toxinotyping and ribotyping was done by PCR. Whole genome sequencing was performed with Illumina® Miseq™. The genomes were screened for antimicrobial resistance determinants. Genetic relatedness of the isolates was investigated using core genome multi locus sequence typing (cgMLST) and single nucleotide polymorphism analysis. Antimicrobial susceptibility testing was done using the Etest® method. Eight C. difficile isolates were recovered from seven farms. The isolates belonged to different PCR ribotypes. All isolates were toxinogenic. cgMLST revealed a cluster containing isolates recovered from different farms. Seven isolates showed similar resistance gene patterns. Different phenotypic resistance patterns were found. Agreement between phenotypic and genotypic resistance was identified only in some cases. Consequently, SAC may act as a reservoir for C. difficile. Thus, SAC may pose a risk regarding zoonotic transmission of toxinogenic, potentially human-pathogenic and resistant C. difficile isolates. [ABSTRACT FROM AUTHOR]

Listeria (L.) monocytogenes is a foodborne pathogen that can cause disease, mainly in elderly, pregnant or immunocompromised persons through consumption of contaminated food, including pork products. It is widespread in the environment and can also be found in asymptomatic carrier animals, for example, in different tissues of pigs. To learn more about their nature, 16 Listeria spp. isolates found in tonsils and intestinal content of pigs and 13 isolates from the slaughterhouse environment were characterized using next-generation sequencing (NGS). A wide distribution of clonal complexes was observed in pigs, as well as in the pork production chain, suggesting multiple sources of entry. Hypervirulent clones were found in pig tonsils, showing the potential risk of pigs as source of isolates causing human disease. The presence of closely related isolates along the production chain suggests a cross-contamination in the slaughterhouse or recontamination from the same source, strengthening the importance of efficient cleaning and disinfection procedures. The phenotypical antimicrobial resistance status of L. monocytogenes isolates was examined via broth microdilution and revealed a low resistance level. Nevertheless, genotypical resistance data suggested multiple resistances in some non-pathogenic L. innocua isolates from pig samples, which might pose a risk of spreading resistances to pathogenic species. [ABSTRACT FROM AUTHOR]

Staphylococcus aureus can be a harmless coloniser, but it can also cause severe infections in humans, livestock and wildlife. Regarding the latter, only few studies have been performed and knowledge on virulence factors is insufficient. The aim of the present study was to study S. aureus isolates from deceased wild beavers (Castor fiber). Seventeen isolates from eleven beavers, found in Germany and Austria, were investigated. Antimicrobial and biocide susceptibility tests were performed. Isolates were characterised using S. aureus-specific DNA microarrays, spa typing and whole-genome sequencing. From two isolates, prophages were induced by mitomycin C and studied by transmission electron microscopy. Four isolates belonged to clonal complex (CC) 8, CC12, and CC398. Twelve isolates belonged to CC1956 and one isolate was CC49. The CC49 and CC1956 isolates carried distinct lukF/S genes related to the Panton-Valentine leukocidin (PVL) from human isolates of S. aureus. These genes were located on related, but not identical, Siphovirus prophages. The beavers, from which those isolates originated, suffered from abscesses, purulent organ lesions and necrotising pneumonia, i.e., clinical manifestations resembling symptoms of severe PVL-associated disease in humans. It might thus be assumed that the "Beaver Leukocidin (BVL, lukF/S-BV)"-positive strains are beaver-specific pathogens, and further studies on their clinical role as well as on a possible transmissibility to other species, including humans, are warranted. [ABSTRACT FROM AUTHOR]

Background: Extended-spectrum-β-lactamase-producing, carbapenemase-producing, and colistin-resistant Enterobacterales (ESBL-E, CPE, and Col-E) are multiresistant pathogens that are increasingly being encountered in both human and veterinary medicine. In this review, we discuss the frequency, sources, and significance of the zoonotic transmission of these pathogens between animals and human beings. Methods: This review is based on pertinent publications retrieved by a selective literature search. Findings for Germany are presented in the global context. Results: ESBL-E are common in Germany in both animals and human beings, with a 6-10% colonization rate in the general human population. A major source of ESBL-E is human-to-human transmission, partly through travel. Some colonizations are of zoonotic origin (i.e., brought about by contact with animals or animal-derived food products); in the Netherlands, more than 20% of cases are thought to be of this type. CPE infections, on the other hand, are rare in Germany in both animals and human beings. Their main source in human beings is nosocomial transmission. Col-E, which bear mcr resistance genes, have been described in Germany mainly in food-producing animals and their meat. No representative data are available on Col-E in human beings in Germany; in Europe, the prevalence of colonization is less than 2%, with long-distance travel as a risk factor. The relevance of animals as a source of Col-E for human beings is not yet entirely clear. Conclusion: Livestock farming and animal contact affect human colonization with the multiresistant Gram-negative pathogens CPE, ESBL-E and Col-E to differing extents. Improved prevention will require the joint efforts of human and veterinary medicine. [ABSTRACT FROM AUTHOR]

The aim of this study was to investigate the presence of β-lactamase producing or fluoroquinolone-resistant members of the family Enterobacteriaceae in European mouflons (Ovis orientalis musimon). The mouflon samples originated from nasal and perineal swabs and/or organ samples in cases of a suspected infection. Only one of the 32 mouflons was tested positive for the presence of Enterobacteriaceae that displayed either an ESBL/AmpC phenotype or were resistant to ciprofloxacin. The positively tested swab originated from a sample of the jejunal mucosa of a four-year old female mouflon. Two different colony morphotypes were identified as Escherichia coli and Klebsiella pneumoniae. These isolates were phenotypically and genotypically characterized in detail by a polyphasic approach. Both isolates were multi-drug resistant. The E. coli isolate belonged to the phylogenetic group B1 and sequence type (ST) 744 and harboured the β-lactamase genes blaCTX-M-15 and blaOXA-1. The K. pneumoniae, identified as ST11, harboured the β-lactamase genes blaSHV-11, blaOXA-1, and blaDHA-1 as well as the plasmid-mediated quinolone resistance (PMQR) gene qnrB55. The present study demonstrates that wild animals can acquire human-derived resistance determinants and such findings may indicate environmental pollution with resistance determinants from other sources. [ABSTRACT FROM AUTHOR]

Objectives: This study aimed to compare ESBL-producing Escherichia coli causing infections in humans with infecting or commensal isolates from animals and isolates from food of animal origin in terms of the strain types, the ESBL gene present and the plasmids that carry the respective ESBL genes. Methods: A collection of 353 ESBL-positive E. coli isolates from the UK, the Netherlands and Germany were studied by MLST and ESBL genes were identified. Characterization of ESBL gene-carrying plasmids was performed using PCR-based replicon typing. Moreover, IncI1-Iγ and IncN plasmids were characterized by plasmid MLST. Results: The ESBL-producing E. coli represented 158 different STs with ST131, ST10 and ST88 being the most common. Overall, blaCTX-M-1 was the most frequently detected ESBL gene, followed by blaCTX-M-15, which was the most common ESBL gene in the human isolates. The most common plasmid replicon type overall was IncI1-Iγ followed by multiple IncF replicons. Conclusions: ESBL genes were present in a wide variety of E. coli STs. IncI1-Iγ plasmids that carried the blaCTX-M-1 gene were widely disseminated amongst STs in isolates from animals and humans, whereas other plasmids and STs appeared to be more restricted to isolates from specific hosts. [ABSTRACT FROM AUTHOR]

Background Enteric Redmouth Disease (ERM), caused by Yersinia ruckeri, is one of the most important infectious diseases in rainbow trout (Oncorhynchus mykiss) aquaculture in Europe. More recently, non-motile vaccine resistant isolates appear to have evolved and are causing disease problems throughout Europe, including Germany. The aim of this study was to analyse the variation of biochemical and molecular characteristics of Y. ruckeri isolates collected in north west Germany as a basis for strain differentiation. The isolates originated mainly from rainbow trout and were characterised by biochemical profiling, 16S rDNA sequencing, repetitive sequence-based PCRs, including (GTG)5-PCR, BOX-PCR, ERIC-PCR and REPPCR, and pulsed-field gel electrophoresis (PFGE). Results In total, 83 isolates were characterised, including 48 isolates collected during a field study in north west Germany. All isolates were confirmed as Y. ruckeri by the API 20E system. Five isolates were additionally confirmed as Y. ruckeri by Y. ruckeri-specific PCR and 16S rDNA sequencing. Only 17 isolates hydrolyzed Tween 80/20. Sixty-six isolates (79.5%) were nonmotile. Two different patterns were obtained by REP-PCR, five patterns by ERIC-PCR, four patterns by (GTG)5-PCR and three patterns by BOX-PCR. NotI-directed PFGE resulted in 17 patterns that differed from each other by 25-29 fragments. Isolates from the field study clustered together as PFGE type C. According to the results of API 20E, repetitive sequencebased PCRs and PFGE, these isolates could be subdivided into 27 different groups. Conclusions The detailed molecular and phenotypic characterisation scheme developed in this study could be used to help trace the dissemination of Y. ruckeri isolates, and thus may represent part of improved disease monitoring plans in the future. [ABSTRACT FROM AUTHOR]

Objectives: Fifty-four methicillin-resistant Staphylococcus aureus (MRSA) ST398 isolates from unrelated diseased swine collected all over Germany were comparatively investigated for their antimicrobial resistance and virulence properties, and for their genomic relatedness. [ABSTRACT FROM PUBLISHER]

Plasmids are mobile genetic elements, contributing to the spread of resistance determinants by horizontal gene transfer. Plasmid-mediated quinolone resistances (PMQRs) are important determinants able to decrease the antimicrobial susceptibility of bacteria against fluoroquinolones and quinolones. The PMQR gene qnrS1, especially, is broadly present in the livestock and food sector. Thus, it is of interest to understand the characteristics of plasmids able to carry and disseminate this determinant and therewith contribute to the resistance development against this class of high-priority, critically important antimicrobials. Therefore, we investigated all commensal Escherichia (E.) coli isolates, with reduced susceptibility to quinolones, recovered during the annual zoonosis monitoring 2017 in the pork and beef production chain in Germany (n = 2799). Through short-read whole-genome sequencing and bioinformatics analysis, the composition of the plasmids and factors involved in their occurrence were determined. We analysed the presence and structures of predominant plasmids carrying the PMQR qnrS1. This gene was most frequently located on IncX plasmids. Although the E. coli harbouring these IncX plasmids were highly diverse in their sequence types as well as their phenotypic resistance profiles, the IncX plasmids-carrying the qnrS1 gene were rather conserved. Thus, we only detected three distinct IncX plasmids carrying qnrS1 in the investigated isolates. The IncX plasmids were assigned either to IncX1 or to IncX3. All qnrS1-carrying IncX plasmids further harboured a β-lactamase gene (bla). In addition, all investigated IncX plasmids were transmissible. Overall, we found highly heterogenic E. coli harbouring conserved IncX plasmids as vehicles for the most prevalent qnr gene qnrS1. These IncX plasmids may play an important role in the dissemination of those two resistance determinants and their presence, transfer and co-selection properties require a deeper understanding for a thorough risk assessment. [ABSTRACT FROM AUTHOR]

Fluoroquinolones are the highest priority, critically important antimicrobial agents. Resistance development can occur via different mechanisms, with plasmid-mediated quinolone resistance (PMQR) being prevalent in the livestock and food area. Especially, qnr genes, commonly located on mobile genetic elements, are major drivers for the spread of resistance determinants against fluoroquinolones. We investigated the prevalence and characteristics of qnr-positive Escherichia (E.) coli obtained from different monitoring programs in Germany in 2017. Furthermore, we aimed to evaluate commonalities of qnr-carrying plasmids in E. coli. We found qnr to be broadly spread over different livestock and food matrices, and to be present in various sequence types. The qnr-positive isolates were predominantly detected within selectively isolated ESBL (extended spectrum beta-lactamase)-producing E. coli, leading to a frequent association with other resistance genes, especially cephalosporin determinants. Furthermore, we found that qnr correlates with the presence of genes involved in resistance development against quaternary ammonium compounds (qac). The detection of additional point mutations in many isolates within the chromosomal QRDR region led to even higher MIC values against fluoroquinolones for the investigated E. coli. All of these attributes should be carefully taken into account in the risk assessment of qnr-carrying E. coli from livestock and food. [ABSTRACT FROM AUTHOR]

The surveillance of antimicrobial resistance among humans and food-producing animals is important to monitor the zoonotic transmission of multidrug-resistant bacteria (MDRB). We assessed the prevalence of four MDRB within the meat production chain, including extended-spectrum β-lactamase (ESBL)-producing, carbapenemase-producing Enterobacterales (CPE) and colistin-resistant Enterobacterales (Col-E), as well as vancomycin-resistant enterococci (VRE). In total, 505 samples from four stages of meat production, i.e., slaughterhouses, meat-processing plants, fresh food products and the urban environment, were collected in northwestern Germany in 2018/2019 and screened for the presence of MDRB using both culture-based and PCR-based techniques. We detected genes encoding for carbapenemases in 9–56% (blaOXA-48, blaKPC, blaNDM, blaVIM) and colistin resistance-encoding mcr genes in 9–26% of the samples from all stages. Culture-based analysis found CPE and VRE only in environmental samples (11% and 7%, respectively), but Col-E and ESBL-producers in 1–7% and 12–46% of samples from all stages, respectively. Overall, our results showed that ESBL-producers and mcr-carrying Col-E were common in food-producing animals at slaughterhouses, in meat-processing plants and in food items at retail, while CPE and VRE were only found in the environment. The discrepancy between detected carbapenemase genes and isolated CPE emphasizes the need for more sensitive detection methods for CPE monitoring. [ABSTRACT FROM AUTHOR]

The putative virulence and antimicrobial resistance gene contents of extended spectrum β-lactamase (ESBL)-positive E. coli (n=629) isolated between 2005 and 2009 from humans, animals and animal food products in Germany, The Netherlands and the UK were compared using a microarray approach to test the suitability of this approach with regard to determining their similarities. A selection of isolates (n=313) were also analysed by multilocus sequence typing (MLST). Isolates harbouring blaCTX-M-group-1 dominated (66%, n=418) and originated from both animals and cases of human infections in all three countries; 23% (n=144) of all isolates contained both blaCTX-M-group-1 and blaOXA-1-like genes, predominantly from humans (n=127) and UK cattle (n=15). The antimicrobial resistance and virulence gene profiles of this collection of isolates were highly diverse. A substantial number of human isolates (32%, n=87) did not share more than 40% similarity (based on the Jaccard coefficient) with animal isolates. A further 43% of human isolates from the three countries (n=117) were at least 40% similar to each other and to five isolates from UK cattle and one each from Dutch chicken meat and a German dog; the members of this group usually harboured genes such as mph(A), mrx, aac(6’)-Ib, catB3, blaOXA-1-like and blaCTX-M-group-1. forty-four per cent of the MLST-typed isolates in this group belonged to ST131 (n=18) and 22% to ST405 (n=9), all from humans. Among animal isolates subjected to MLST (n=258), only 1.2% (n=3) were more than 70% similar to human isolates in gene profiles and shared the same MLST clonal complex with the corresponding human isolates. The results suggest that minimising human-to-human transmission is essential to control the spread of ESBL-positive E. coli in humans. [ABSTRACT FROM AUTHOR]

Highlights • P. multocida isolates with high sulfonamide/trimethoprim MICs were detected in 3/29 farms. • These P. multocida isolates were related in their ApaI and SmaI PFGE patterns. • All of them harboured a plasmid of 6050 bp that was closely related to a plasmid of A. pleuropneumoniae. • The plasmid carried intact sul2 and dfrA14 genes, the latter was integrated into a strA gene. Abstract Pasteurella multocida is an important respiratory tract pathogen in intensive livestock farming, especially in pigs. Antimicrobial agents are frequently used to combat infections caused by this pathogen. In a study on antimicrobial resistance among respiratory tract pathogens of pigs from 30 German pig-producing farms, P. multocida isolates (n = 9) with high minimal inhibitory concentration (MIC) values of 16/304 mg/L (n = 2), 32/608 mg/L (n = 3) or ≥64/1216 mg/L (n = 4) for trimethoprim/sulfamethoxazole (1:19) and of ≥512 mg/L (n = 9) for trimethoprim (TMP) were detected in three of these farms. The genetic relatedness of the isolates was investigated via capsule-specific PCR and macrorestriction analyses with Apa I and Sma I. Pulsed-field gel electrophoresis revealed indistinguishable restriction patterns per farm, with slight differences between the three farms. All isolates represented capsular type A. Four representative isolates, that were subjected to whole genome sequencing, shared the multi-locus sequence type (ST) 3. Their plasmids were transformed into E. coli TOP10 with subsequent selection on TMP-containing agar plates. Antimicrobial susceptibility testing and plasmid analysis of the transformants confirmed that they were resistant to sulfonamides and trimethoprim and carried only a single small plasmid. This plasmid was completely sequenced and revealed a size of 6050 bp. Sequence analyses identified the presence of a resistance gene cluster comprising the genes sul2-ΔstrA-dfrA14-ΔstrA-ΔstrB. Further analysis identified a dfrA14 gene cassette being integrated into the strA reading frame. Neither the gene dfrA14 nor this gene cluster have been detected before in P. multocida. [ABSTRACT FROM AUTHOR]

The aim of this study was to identify extended-spectrum β-lactamase (ESBL)-producing Escherichia coli collected from diseased food-producing animals in Germany. A total of 6849 E . coli isolates, collected from diseased cattle, pigs and poultry in the German national monitoring program GE RM -Vet (2008–2014), were characterized by antimicrobial susceptibility testing and screened for the ESBL phenotype. ESBL genes were identified by PCR and sequencing. The isolates were further characterized by PCR-based phylotyping. The 419/6849 (6.1%) ESBL-producers identified included 324/2896 (11.2%) isolates from cattle, 75/1562 (4.8%) from pigs and 20/2391 (0.8%) from poultry. The ESBL genes detected were: bla CTX-M-1 (69.9%), bla CTX-M-15 (13.6%), bla CTX-M-14 (11.7%), bla TEM-52 (1.9%), bla SHV-12 (1.4%), bla CTX-M-3 (1.0%), and bla CTX-M-2 (0.5%). The phylogroup A was the dominant phylogroup (57.0%) followed by phylogroups D (23.4%), B1 (17.9%), and B2 (1.7%). Bovine isolates belonged predominantly to the phylogroups A and D, whereas the porcine and avian isolates mainly belonged to A and B1. The majority of the ESBL-producing isolates found in each phylogroup were from animals suffering from gastrointestinal infections. In 399/419 isolates (95.2%), additional resistance to non-β-lactam antibiotics was seen. Multidrug-resistance [resistance to aminoglycosides, fluoro(quinolones), sulphonamides, tetracyclines, and trimethoprim] was seen in 369/419 (88.1%) isolates, which may facilitate the co-selection of ESBL genes, when located on the same mobile genetic element as the others resistance genes, and may compromise the therapeutic options. [ABSTRACT FROM AUTHOR]

During a survey of fresh chicken and turkey meat as well as chicken and turkey meat products for the presence of methicillin-resistant Staphylococcus aureus (MRSA) isolates in Germany, 32 (37.2%) of 86 samples were MRSA positive. Twenty-eight of these MRSA isolates belonged to clonal complex 398 (CC398), which is widespread among food-producing animals. These CC398 isolates carried SCCmec elements of type IV or V and exhibited spa type t011, t034, t899, t2346 or t6574 and either the known dru types dt2b, dt6j, dt10a, dt10q, dt11a, dt11v, and dt11ab or the novel dru types dt6m, dt10as, and dt10at. In addition, two MRSA sequence type 9 (ST9) isolates with a type IV SCCmec cassette, spa type t1430, and dru type dt10a as well as single MRSA ST5 and ST1791 isolates with a type III SCCmec cassette, spa type t002, and dru type dt9v were identified. All but two isolates were classified as multiresistant. A wide variety of resistance phenotypes and genotypes were detected. All isolates were negative for the major virulence factors, such as Panton-Valentine leukocidin, toxic shock syndrome toxin 1, or exfoliative toxins. In contrast to the MRSA CC398 isolates, the four ST9, ST5, or ST1791 isolates harbored the egc gene cluster for enterotoxin G, I, M, N, O, and U genes. Although the relevance of contamination of fresh poultry meat or poultry products with MRSA is currently unclear, the presence of multiresistant and, in part, enterotoxigenic MRSA emphasizes the need for further studies to elucidate possible health hazards for consumers. [ABSTRACT FROM AUTHOR]

Background: Extended-spectrum cephalosporins (ESCs) are third and fourth generation cephalosporin antimicrobials used in humans and animals to treat infections due to multidrug-resistant (MDR) bacteria. Resistance to ESCs (ESC-R) in Enterobacterales is predominantly due to the production of extended-spectrum β-lactamases (ESBLs) and plasmid-mediated AmpC β-lactamases (AmpCs). The dynamics of ESBLs and AmpCs are changing across countries and host species, the result of global transmission of ESC-R genes. Plasmids are known to play a key role in this dissemination, but the relative importance of different types of plasmids is not fully understood., Methods: In this study, Escherichia coli with the major ESC-R genes bla CTX-M-1 , bla CTX-M-15 , bla CTX-M-14 (ESBLs) and bla CMY-2 (AmpC), were selected from diverse host species and other sources across Canada, France and Germany, collected between 2003 and 2017. To examine in detail the vehicles of transmission of the ESC-R genes, long- and short-read sequences were generated to obtain complete contiguous chromosome and plasmid sequences (n = 192 ESC-R E. coli). The types, gene composition and genetic relatedness of these plasmids were investigated, along with association with isolate year, source and geographical origin, and put in context with publicly available plasmid sequences., Findings: We identified five epidemic resistance plasmid subtypes with distinct genetic properties that are associated with the global dissemination of ESC-R genes across multiple E. coli lineages and host species. The IncI1 pST3 bla CTX-M-1 plasmid subtype was found in more diverse sources than the other main plasmid subtypes, whereas IncI1 pST12 bla CMY-2 was more frequent in Canadian and German human and chicken isolates. Clonal expansion also contributed to the dissemination of the IncI1 pST12 bla CMY-2 plasmid in ST131 and ST117 E. coli harbouring this plasmid. The IncI1 pST2 bla CMY-2 subtype was predominant in isolates from humans in France, while the IncF F31:A4:B1 bla CTX-M-15 and F2:A-:B- bla CTX-M-14 plasmid subtypes were frequent in human and cattle isolates across multiple countries. Beyond their epidemic nature with respect to ESC-R genes, in our collection almost all IncI1 pST3 bla CTX-M-1 and IncF F31:A4:B1 bla CTX-M-15 epidemic plasmids also carried multiple antimicrobial resistance (AMR) genes conferring resistance to other antimicrobial classes. Finally, we found genetic signatures in the regions surrounding specific ESC-R genes, identifying the predominant mechanisms of ESC-R gene movement, and using publicly available databases, we identified these epidemic plasmids from widespread bacterial species, host species, countries and continents., Interpretation: We provide evidence that epidemic resistance plasmid subtypes contribute to the global dissemination of ESC-R genes, and in addition, some of these epidemic plasmids confer resistance to multiple other antimicrobial classes. The success of these plasmids suggests that they may have a fitness advantage over other plasmid types and subtypes. Identification and understanding of the vehicles of AMR transmission are crucial to develop and target strategies and interventions to reduce the spread of AMR., Funding: This project was supported by the Joint Programming Initiative on Antimicrobial Resistance (JPIAMR), through the Medical Research Council (MRC, MR/R000948/1), the Canadian Institutes of Health Research (CFC-150770), and the Genomics Research and Development Initiative (Government of Canada), the German Federal Ministry of Education and Research (BMBF) grant no. 01KI1709, the French Agency for food environmental and occupational health & safety (Anses), and the French National Reference Center (CNR) for antimicrobial resistance. Support was also provided by the Biotechnology and Biological Sciences Research Council (BBSRC) through the BBSRC Institute Strategic Programme Microbes in the Food ChainBB/R012504/1 and its constituent project BBS/E/F/000PR10348 (Theme 1, Epidemiology and Evolution of Pathogens in the Food Chain)., Competing Interests: Declaration of interests AEM is an inventor on International Patent Application No. PCT/GB2023/050906 filed in the name of Quadram Institute Bioscience–Determination and quantification of the microbial communities and antimicrobial resistance genes on food. All other authors declare no competing interests., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

One Health refers to a concept that links human, animal, and environmental health. In Germany, there is extensive data on antibiotic resistance (AMR) and multidrug-resistant (micro)organisms (MDRO) in human and veterinary medicine, as well as from studies in various environmental compartments (soil, water, wastewater). All these activities are conducted according to different specifications and standards, which makes it difficult to compare data. A focus on AMR and MDRO of human therapeutic importance is helpful to provide some guidance. Most data are available across sectors on methicillin-resistant Staphylococcus aureus (MRSA) and multiresistant Enterobacterales such as Escherichia coli and Klebsiella pneumoniae. Here, the trends of resistance are heterogeneous. Antibiotic use leads to MRE selection, which is well documented. Success in minimizing antibiotic use has also been demonstrated in recent years in several sectors and could be correlated with success in containing AMR and MDRO (e.g., decrease in MRSA in human medicine). Sector-specific measures to reduce the burden of MDRO and AMR are also necessary, as not all resistance problems are linked to other sectors. Carbapenem resistance is still rare, but most apparent in human pathogens. Colistin resistance occurs in different sectors but shows different mechanisms in each. Resistance to antibiotics of last resort such as linezolid is rare in Germany, but shows a specific One Health correlation. Efforts to harmonize methods, for example in the field of antimicrobial susceptibility testing and genome-based pathogen and AMR surveillance, are an important first step towards a better comparability of the different data collections., (© 2023. The Author(s).)

Methicillin-resistant Staphylococcus aureus (MRSA) of sequence types ST398 and ST9 are dominant lineages among livestock in Europe and Asia, respectively. Although both STs were commonly found as colonizers of the skin and the mucosal membranes, MRSA ST398, rather than MRSA ST9, has been reported to cause infections in humans and animals. Herein, we comparatively analyzed the genomic characteristics, fitness and virulence of MRSA ST398 and ST9 isolated from pigs in both China (CHN) and Germany (GER) to explore the factors that lead to differences in their epidemics and pathogenicity. We observed that the CHN-MRSA ST9 and the GER-MRSA ST9 have evolved independently, whereas the CHN-MRSA ST398 and GER-MRSA ST398 had close evolutionary relationships. Resistance to antimicrobial agents commonly used in livestock, the enhanced ability of biofilm formation, and the resistance to desiccation contribute to the success of the dominant clones of CHN-MRSA ST9 and GER-MRSA ST398, and the vwb νSaα gene on the genomic island might in part contribute to their colonization fitness in pigs. All MRSA ST398 strains revealed more diverse genome structures, higher tolerance to acids and high osmotic pressure, and greater competitive fitness in co-culture experiments. Notably, we identified and characterized a novel hysA νSaβ gene, which was located on the genomic island νSaβ of MRSA ST398 but was absent in MRSA ST9. The enhanced pathogenicity of the MRSA ST398 strains due to hysA νSaβ might in part explain why MRSA ST398 strains are more likely to cause infections.

Background: Resistance to 3rd-generation cephalosporins in Escherichia coli is mostly mediated by extended-spectrum beta-lactamases (ESBLs) or AmpC beta-lactamases. Besides overexpression of the species-specific chromosomal ampC gene, acquisition of plasmid-encoded ampC genes, e.g. bla CMY-2 , has been described worldwide in E. coli from humans and animals. To investigate a possible transmission of bla CMY-2 along the food production chain, we conducted a next-generation sequencing (NGS)-based analysis of 164 CMY-2-producing E. coli isolates from humans, livestock animals and foodstuff from Germany., Results: The data of the 164 sequenced isolates revealed 59 different sequence types (STs); the most prevalent ones were ST38 (n = 19), ST131 (n = 16) and ST117 (n = 13). Two STs were present in all reservoirs: ST131 (human n = 8; food n = 2; animal n = 6) and ST38 (human n = 3; animal n = 9; food n = 7). All but one CMY-2-producing ST131 isolates belonged to the clade B (fimH22) that differed substantially from the worldwide dominant CTX-M-15-producing clonal lineage ST131-O25b clade C (fimH30). Plasmid replicon types IncI1 (n = 61) and IncK (n = 72) were identified for the majority of bla CMY-2 -carrying plasmids. Plasmid sequence comparisons showed a remarkable sequence identity, especially for IncK plasmids. Associations of replicon types and distinct STs were shown for IncK and ST57, ST429 and ST38 as well as for IncI1 and ST58. Additional β-lactamase genes (bla TEM , bla CTX-M , bla OXA , bla SHV ) were detected in 50% of the isolates, and twelve E. coli from chicken and retail chicken meat carried the colistin resistance gene mcr-1., Conclusion: We found isolates of distinct E. coli clonal lineages (ST131 and ST38) in all three reservoirs. However, a direct clonal relationship of isolates from food animals and humans was only noticeable for a few cases. The CMY-2-producing E. coli-ST131 represents a clonal lineage different from the CTX-M-15-producing ST131-O25b cluster. Apart from the ST-driven spread, plasmid-mediated spread, especially via IncI1 and IncK plasmids, likely plays an important role for emergence and transmission of bla CMY-2 between animals and humans.

Extended-spectrum β-lactamase (ESBL)-producing Escherichia coli isolates have been increasingly reported in different reservoirs. The aims of this study were to investigate the presence of ESBL-producing E. coli in fresh vegetables and to characterise their ESBL gene-carrying plasmids. Among the 245 samples from vegetables investigated during 2011-2013, seven putative ESBL-producing E. coli (salad n = 2, sprouts n = 5) were found. They were subjected to ESBL phenotypic confirmatory tests, detection/sequencing of ESBL genes, antimicrobial susceptibility testing (AST), phylotyping, XbaI-macrorestriction analysis, multilocus sequence typing and transformation. Transformants were characterised by AST, S1-nuclease PFGE, replicon typing, conjugation and investigated for co-located antimicrobial resistance genes. Two ESBL gene-carrying plasmids were sequenced using a HiSeq 2500 system. The seven isolates were confirmed as ESBL producers, displayed unrelated XbaI-patterns and unique sequence types (STs) and belonged to the phylogroups A, B1 or D. The ESBL genes were located on plasmids. Two plasmids carrying bla CTX-M-14 genes (incompatibility group IncK or IncHI2) were seen in isolates from salad (ST973) and sprout (ST527). Two bla CTX-M-15 - (IncFIB; non-typeable) and the IncN bla CTX-M-65 - and IncHI2 bla CTX-M-125 -carrying plasmids were found in isolates from sprouts (ST410, ST847, ST10, ST542). All plasmids were conjugative, except for the IncFIA-FIB bla CTX-M-2 -carrying plasmid. Sequence analysis of two plasmids identified the ESBL genes in close location to other resistance genes: sulfonamide resistance gene sul2, streptomycin resistance genes strA and strB, the plasmid-mediated quinolone resistance gene qnrS1 and bla TEM-1 (sul2-strA-strB-IS66-bla TEM-1 -tnpR-ΔtnpA-ISEcp1-bla CTX-M-15 -Δorf477-ΔtnpA-qnrS1) or the fosfomycin resistance gene fosA3 (ΔISEcp1-bla CTX-M-125 -ΔIS903B-fosA3). These observations underline the importance of vegetables as reservoirs for multidrug resistant ESBL-producing E. coli., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Extended-spectrum β-lactamase (ESBL)-producing isolates have been increasingly reported during recent years. The aims of this study were to characterize ESBL-producing Escherichia coli from bovine mastitis as well as their ESBL gene-carrying plasmids. A culture collection of E. coli isolated from bovine quarter milk samples (2009-2013), was screened for ESBL production using ESBL selective agar plates. Putative ESBL producers (n=16) were investigated by phenotypic confirmatory tests and were characterized by the detection/sequencing of ESBL genes, XbaI macrorestriction analysis, multilocus sequence typing (MLST), phylotyping and antimicrobial susceptibility testing. ESBL gene-carrying plasmids were investigated by transfer experiments, PCRs for the detection of co-located antimicrobial resistance genes, PCR-based replicon typing and S1-nuclease pulsed-field gel electrophoresis. Twelve ESBL-producing isolates were found. They showed eleven different XbaI patterns and were distributed among eight MLST types [ST10 (n=3), ST117 (n=2), ST361 (n=1), ST362 (n=1), ST540 (n=1), ST1431 (n=2), ST1508 (n=1), and the novel ST5447 (n=1)] and the phylogenetic groups A (n=6), B1 (n=2), B2 (n=1) and D (n=3). ESBL genes bla CTX-M-1 (n=5), bla CTX-M-2 (n=2), bla CTX-M-14 and bla CTX-M-15 (n=4) were found on conjugative plasmids (35-225kb) of diverse incompatibility groups (e.g. IncF, IncI1 or HI2+P). Co-located resistance to sulfonamides, tetracycline, trimethoprim, and chloramphenicol/florfenicol was detected on five ESBL gene-carrying plasmids, but seven plasmids conferred solely resistance to β-lactam antibiotics. The presence of additional resistance genes on the ESBL gene-carrying plasmids suggests that co-selection of ESBL genes may occur even in the absence of β-lactam antibiotics., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Multidrug-resistant Escherichia coli encoding CTX-M-type extended-spectrum β-lactamases (ESBLs) are isolated in increasing numbers from humans, companion animals and livestock, raising concern regarding the exchange and spread of isolates in these populations. In this study, whole-genome sequencing of CTX-M-15-producing E. coli isolates recently sampled from humans, companion animals, livestock and farm environments was performed. In total, 26 different sequence types (STs) were detected, of which ST410 was the most frequent and was the only ST present in all populations studied. Five clades (designated A-E) were detected within the ST410 isolates. In particular, isolates of clade B were present in all four populations and had core genomes that differed by less than 70 single nucleotide polymorphisms (SNPs). Isolates of clades B and C were also clonally marked, exhibiting identical chromosomal insertions of blaCTX-M-15 at distinct loci. These data provide strong evidence for the clonal dissemination of specific clades of CTX-M-15-producing E. coli ST410 in human and animal populations., (Copyright © 2016 Elsevier B.V. and International Society of Chemotherapy. All rights reserved.)

Enteric redmouth disease (ERM), caused by Yersinia ruckeri, is among the most important infectious diseases in rainbow trout Oncorhynchus mykiss aquaculture in Europe. Our aim was to analyse the persistence of Y. ruckeri strains in trout farms in northwest Germany and their dissemination between farms based on a detailed molecular and phenotypical characterisation scheme. The data on identification and characterisation of Y. ruckeri strains and examining the distribution of these strains in the field could serve as a basis for preventive disease monitoring plans. During the observation period from June 2011 until June 2012, we collected 48 Y. ruckeri isolates from 12 different rainbow trout hatcheries. In total, 44 (91.7%) of the isolates were non-motile; in particular, all isolates recovered during the sampling period in winter and early spring were non-motile. In several trout farms, characteristic farm-specific Y. ruckeri isolates from particular typing groups were isolated throughout the year, while in other farms, which had a trading relationship between each other, ERM outbreaks were caused by Y. ruckeri from the same typing group. Our data indicate that in some farms, the causative Y. ruckeri strains persisted in the respective trout farm. The presence of Y. ruckeri from the same typing group in farms with a trading relationship indicates a dissemination of the infection between the farms.

Methicillin-resistant Staphylococcus aureus (MRSA) causes colonization and infection both in animals and humans. In Germany, cases of MRSA colonization among humans, w+hich affect 0.5-1.5% of the general population and 1.0-2.5% of patients at hospital admission, are still mostly associated with previous healthcare contact and defined epidemic clonal lineages. However, MRSA is also distributed in livestock production in Germany, mostly without causing infections in the animals. These MRSA predominantly belong to the clonal complex (CC) 398, but also to CC9 and CC97. Zoonotic transmission of MRSA CC398 from livestock to humans occurs predominantly in people with occupational livestock contact. Spread of MRSA CC398 to household members of these persons is also frequently observed, but dissemination in the general population is limited so far However, especially in areas with intensive livestock husbandry, about 20-38% of MRSA CC398 cases among humans cannot be epidemiologically linked to direct livestock contact, indicating other transmission pathways. MRSA CC398 currently causes about 2% of all human MRSA infections (wound infections, pneumonia, sepsis) in Germany, but up to 10% in regions characterized by a high density of livestock-farming. The burden of MRSA in companion animals was demonstrated to range between 3.6-9.4% within wound samples obtained from dogs, cats and horses, respectively. In contrast to livestock and horses, MRSA distributed in pet animals are mostly associated with MRSA clonal lineages that are also prevalent in human healthcare facilities. Overall, zoonotic exchange of MRSA between humans and animals has relevant impact on the epidemiology of MRSA in Germany.

Extended-spectrum-beta-lactamase (ESBL) and plasmid-encoded cephamycinase (pAmpC) producing Escherichia (E.) coli in livestock farms have recently been matter of growing scientific and public concern. This article summarises selected European studies which focus on the prevalence and risk factors associated with the presence of such resistant E. coli isolates in livestock farms. Due to the different methodologies used in these studies, they cannot be compared directly; nonetheless, the overall prevalence found is very high. The prevalence found in broiler farms was higher than 40% and the individual animal prevalence was ca. 30%. The prevalence was more variable in pigs, with reports of pig farms showing prevalence of 1 to 80% and reports of individual animal prevalence of 15 to 100% In studies on cattle farms the production type as well as the age of animals had an influence on the number of positive samples. The highest prevalence was found with calves after birth and in the first weeks, whereas with older cattle the numbers of positive samples were considerably lower. Samples taken from dairy cows were positive more often after calving than before calving. According to the livestock species different risk factors may be assessed for the occurrence of ESBL/pAmpC-producing E. coli isolates. In some studies an association between the occurrence of ESBL-producing E. coli and factors like the use of antimicrobial agents or management factors, as the duration of the fattening period and the acquisition of animals from different origins, were identified. At the moment, there is a lack of systematic and standardised transnational epidemiological investigations on the occurrence of ESBL/pAmpC-producing E. coli in livestock. To control the further spread of ESBL/pAmpC-producing E. coli and the effectiveness of preventive measures, comprehensive monitoring and surveillance systems with harmonised methods are essential. Modern typing methods, in particular the sequence-based methods, can provide more information to clarify transmission pathways.

The putative virulence and antimicrobial resistance gene contents of extended spectrum β-lactamase (ESBL)-positive E. coli (n=629) isolated between 2005 and 2009 from humans, animals and animal food products in Germany, The Netherlands and the UK were compared using a microarray approach to test the suitability of this approach with regard to determining their similarities. A selection of isolates (n=313) were also analysed by multilocus sequence typing (MLST). Isolates harbouring bla(CTX-M-group-1) dominated (66%, n=418) and originated from both animals and cases of human infections in all three countries; 23% (n=144) of all isolates contained both bla(CTX-M-group-1) and bla(OXA-1-like) genes, predominantly from humans (n=127) and UK cattle (n=15). The antimicrobial resistance and virulence gene profiles of this collection of isolates were highly diverse. A substantial number of human isolates (32%, n=87) did not share more than 40% similarity (based on the Jaccard coefficient) with animal isolates. A further 43% of human isolates from the three countries (n=117) were at least 40% similar to each other and to five isolates from UK cattle and one each from Dutch chicken meat and a German dog; the members of this group usually harboured genes such as mph(A), mrx, aac(6')-Ib, catB3, bla(OXA-1-like) and bla(CTX-M-group-1). forty-four per cent of the MLST-typed isolates in this group belonged to ST131 (n=18) and 22% to ST405 (n=9), all from humans. Among animal isolates subjected to MLST (n=258), only 1.2% (n=3) were more than 70% similar to human isolates in gene profiles and shared the same MLST clonal complex with the corresponding human isolates. The results suggest that minimising human-to-human transmission is essential to control the spread of ESBL-positive E. coli in humans.

Objectives: The aims of this study were (i) to detect extended-spectrum β-lactamase (ESBL) genes among 1378 Escherichia coli isolates from defined disease conditions of companion and farm animals and (ii) to determine the localization and organization of ESBL genes., Methods: E. coli isolates from the German resistance monitoring programme GERM-Vet were included in the study. Plasmids were transferred by conjugation or transformation and typed by PCR-based replicon typing. ESBL genes were detected by PCR; the complete ESBL genes and their flanking regions were sequenced by primer walking. Phylogenetic grouping and multilocus sequence typing (MLST) were performed for all ESBL-producing E. coli isolates., Results: Of the 27 ESBL-producing E. coli isolates detected, 22 carried blaCTX-M-1 genes on IncN (n = 16), IncF (n = 3), IncI1 (n = 2) or multireplicon (n = 1) plasmids. A blaCTX-M-3 gene was located on an IncN plasmid and a blaCTX-M-15 gene was located on an IncF plasmid. A multireplicon plasmid and an IncHI1 plasmid harboured blaCTX-M-2. A blaTEM-52c gene was identified within Tn2 on an IncI1 plasmid. The blaCTX-M genes located within the same or related genetic contexts showed differences due to the integration of insertion sequences. Various MLST types were detected, with ST10 (n = 7), ST167 (n = 4) and ST100 (n = 3) being the most common., Conclusions: This study showed that the blaCTX-M-1 gene is the predominant ESBL gene among E. coli isolates from diseased animals in Germany and a considerable structural heterogeneity was found in the regions flanking the blaCTX-M-1 gene. Insertion sequences, transposons and recombination events are likely to be involved in alterations of the ESBL gene regions.

Background: Decompressive hemicraniectomy reduces mortality after space-occupying MCA infarction. Data on the general public's opinion toward interventions that can save lives but leave the survivors impaired are lacking., Methods: In this population-based epidemiological study in a German city, we surveyed 312 adults in a telephone interview. Here, we presented a scenario of a space-occupying MCA infarct. We evaluated probands' attitude toward decompressive surgery in general, and toward outcome scenarios according to Rankin scale (RS) definitions., Results: 312 persons (157 women, 52 ± 20 years) were interviewed. 58 persons had difficulty comprehending the proposed scenario, most of them being of advanced age (79 ± 5 years). From the remaining 254 responders 5 (2%) persons favoured surgical intervention, 149 (58%) were undecided, and 100 (39%) were opposed to surgery. The number of individuals opting for surgery rose in scenarios with a better outcome: If very severe impairment was anticipated (RS 5), only 3 (1%) persons favored surgery. With severe (RS 4), moderate (RS3), and slight impairment (RS2) the numbers were at 16 (6%), 60 (24%), and 161 (63%), respectively. We found no association with age, sex, religion, education, self-estimated health status, or marital status., Conclusions: Explaining complex medical situations to laypersons poses a major problem, particularly to those of old age. Only a minority favors life-saving medical interventions if survival is associated with deficits of unpredictable degree. The majority of persons does not favor intervention even if only moderate impairment is anticipated. Decompressive surgery may in fact be against the values of many individuals.

The correct assessment of mastitis pathogens for their susceptibility/resistance to cefoperazone is currently hampered by the lack of harmonized test conditions and interpretive criteria. The aim of this study was to provide a proposal for clinical breakpoints of cefoperazone which are applicable to Staphylococcus aureus, coagulase-negative staphylococci, Escherichia coli, Streptococcus agalactiae, Streptococcus dysgalactiae and Streptococcus uberis from cases of bovine mastitis and better reflect the situation in the bovine udder than breakpoints adopted from human medicine. For this, pharmacological data and clinical efficacy data of the documents submitted for approval of cefoperazone have been revisited. In addition, 1086 bacterial pathogens of the aforementioned six species/groups collected in Germany and in the USA during recent years were tested in parallel for their cefoperazone MICs and the zone diameters using a 75 μg disk. Subsequently, MICs were plotted against zone diameters. Based on the pharmacological data, the clinical efficacy and the microbiological data, a proposal was made for veterinary-specific breakpoints which classify members of the aforementioned species/groups as (a) susceptible to cefoperazone when their MIC is ≤ 2 μg/ml and their zone diameters are ≥ 27 mm (staphylococci or E. coli) or ≥ 21 mm (streptococci), (b) intermediate when their MIC is 4 μg/ml and their zone diameters are 22-26 mm (staphylococci or E. coli) or 16-20mm (streptococci), and (c) resistant when their MIC is ≥ 8 μg/ml and their zone diameters are ≤ 21 mm (staphylococci or E. coli) or ≤ 15 mm (streptococci)., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Objectives: Twenty-five MRSA ST398 isolates from cases of bovine clinical mastitis and two isolates from farm personnel collected from 17 dairy farms in Germany were investigated for genetic relatedness, antimicrobial resistance and virulence properties., Methods: Genomic relationships were determined by ApaI PFGE, spa typing, SCCmec typing and dru typing. Antimicrobial resistance phenotypes were determined by broth microdilution. Resistance and virulence genes were detected via a diagnostic DNA microarray and specific PCRs., Results: Nine major ApaI PFGE patterns were detected. Three spa types (t011, t034 and t2576) and two SCCmec types (IV and V) were identified. Five different dru types were seen with dt11a being predominant. All isolates were negative for Panton-Valentine leucocidin, enterotoxin and exfoliative toxin genes. Ten resistance patterns were observed with 11 (40.7%) isolates being resistant to only beta-lactam antibiotics and tetracyclines. Several resistance genes were detected: blaZ (penicillin resistance); tet(M), tet(K) and tet(L) (tetracycline resistance); erm(A), erm(B), erm(C) and erm(T) (macrolide/lincosamide/streptogramin B resistance); aacA-aphD, aphA3, aadD and spc (aminoglycoside or aminocyclitol resistance); fexA (phenicol resistance); dfrK (trimethoprim resistance); and vga(A) and vga(C) (pleuromutilin/lincosamide/streptogramin A resistance). The two human isolates were indistinguishable in their genotypic and phenotypic characteristics from the mastitis isolates of the same farm., Conclusions: As previously described for ST398 from swine, isolates of this sequence type from cases of bovine mastitis also demonstrated a high degree of variability when ApaI PFGE profiles and other genotypic and phenotypic characteristics were compared. A uniform virulence gene pattern appeared to be conserved between ST398 isolates from both animal species.

Objectives: Three florfenicol-resistant Pasteurella multocida isolates from Germany, two from swine and one from a calf, were investigated for the genetics and transferability of florfenicol resistance., Methods: The isolates were investigated for susceptibility to antimicrobial agents and plasmid content. Florfenicol resistance plasmids carrying the gene floR were identified by transformation and PCR. Plasmids were mapped, and a novel plasmid type was sequenced completely. PFGE served to determine the clonality of the isolates., Results: In one porcine and the bovine P. multocida isolate, florfenicol resistance was associated with the plasmid pCCK381 previously described in a bovine P. multocida isolate from the UK. The remaining porcine isolate harboured a new type of floR-carrying plasmid, the 10 226 bp plasmid pCCK1900. Complete sequence analysis identified an RSF1010-like plasmid backbone with the mobilization genes mobA, mobB and mobC, the plasmid replication genes repA, repB and repC, the sulphonamide resistance gene sul2 and the streptomycin resistance genes strA and strB. The floR gene area was integrated into a region downstream of strB, which exhibited homology to the floR flanking regions found in various bacteria. PFGE revealed that the floR-carrying P. multocida strains from Germany were unrelated and also different from the UK strain., Conclusions: After the UK and France, floR-mediated florfenicol resistance has now also been identified in target bacteria from Germany. PFGE data and the analysis of plasmids strongly suggested that the spread of florfenicol resistance is due to the horizontal transfer of plasmids rather than the clonal dissemination of a resistant P. multocida isolate.

Objectives: In the BfT-GermVet monitoring study, 417 Escherichia coli isolates collected during 2004-06 in Germany from various disease conditions of pigs (n = 87), horses (n = 102) or cats/dogs (n = 228) were investigated for their susceptibility to 24 antimicrobial agents. This study dealt with the identification of integron-associated resistance genes among these isolates., Methods: Class 1 and class 2 integrons were detected by PCR. The variable parts of the integrons were cloned and sequenced. Transformation and conjugation experiments were conducted to confirm a plasmid location of the integrons., Results: Class 1 and/or class 2 integrons, alone or in different combinations, were detected in 79 of the 417 E. coli isolates. Four trimethoprim resistance genes (dfrA1/12/14/17), five streptomycin/spectinomycin resistance genes (aadA1/2/4/5/6), two streptothricin resistance genes (estX, sat2), one gentamicin/tobramycin/kanamycin resistance gene (aadB) and one chloramphenicol resistance gene (catB3) were detected. Seven different cassette arrangements were identified within class 1 integrons: aadA1 (21 isolates), dfrA1 + aadA1 (18 isolates), dfrA17 + aadA5 (9 isolates), dfrA12 + orfF + aadA2 (8 isolates), aadB + aadA1 (1 isolate), dfrA14 + recombined aadA6 (1 isolate) and dfrA1 + catB3 + aadA4 (1 isolate). Three different cassette arrangements in class 2 integrons, dfrA1 + sat2 + aadA1 (24 isolates), estX + sat2 + aadA1 (6 isolates) and estX + sat2 + DeltaaadA1 (1 isolate), were identified. The plasmid location of class 1 and/or class 2 integrons was confirmed in 37 isolates., Conclusions: Class 1 and/or class 2 integrons carrying resistance gene cassettes were detected in 18.9% of the isolates tested. This molecular analysis complements the phenotypic susceptibility testing conducted in the BfT-GermVet monitoring study and helps to explain the persistence of resistance genes even without direct selective pressure.

Objectives: During recent years, methicillin-resistant Staphylococcus aureus (MRSA) strains from animals have become the focus of various studies. In the present study, coagulase-positive staphylococci obtained from pigs, dogs and cats suffering from acute infections, which had been collected in the BfT-GermVet monitoring programme in Germany and phenotypically identified as oxacillin-resistant, were characterized., Methods: The staphylococci were comparatively investigated for their resistance phenotypes and genotypes. Resistance genes were identified by PCR. MRSA strains were further characterized by SmaI macrorestriction analysis and spa typing to assess their genomic relationships., Results: Among the 248 strains tested, 7 strains (5 porcine S. aureus and 2 canine Staphylococcus pseudintermedius) carried the resistance gene mecA. Gentamicin resistance was based on the presence of the gene aacA/aphD while three different tetracycline resistance genes, tet(K), tet(L) and tet(M), alone or in combinations, were detected. The single macrolide/lincosamide-resistant strain carried an erm(A) gene. All MRSA strains proved to be non-typeable by SmaI macrorestriction analysis and exhibited the spa types t011 (four strains) or t034 (one strain)., Conclusions: Based on their spa types and their non-respondence to SmaI digestion, all five porcine MRSA strains resembled MRSA strains of multilocus sequence type ST398, previously detected among pigs in neighbouring countries such as The Netherlands or Denmark. The results of this study indicate that such strains are also involved in defined disease conditions of pigs from various parts of Germany.

A total of 120 isolates of Klebsiella spp. and Proteus spp. collected from horses and small animals (dogs and cats) were screened for their susceptibility to 24 different antimicrobial agents. Klebsiella spp. were included from infections of the genital tract (GT) of horses (36 isolates) and the urinary/genital tract (UGT) from dogs and cats (17 isolates), while Proteus spp. were from small animal (dogs and cats) infections of the UGT (37 strains) and the skin (incl. ear/mouth) (30 isolates). In Klebsiella spp. resistance appeared most frequently to ampicillin (53-67%), sulfamethoxazole (19-29%) and potentiated sulfonamides (trimethoprim/sulfamethoxazole 1/19 combination) (19-24%). A further 29% of enrofloxacin resistant Klebsiella isolates were observed for the UGT of small animals. From the GT of horses for this antimicrobial agent there was no isolate detected with a comparably high minimum inhibitory concentration (MIC) value. In Proteus spp. highest percentages of resistance occurred against tetracycline (90-92%). Due to drug efflux proteins, high MIC values against this antimicrobial agent have been frequently reported in literature. In Proteus spp. relevant resistance percentages also occurred for potentiated sulfonamides (27-37%), sulfamethoxazole (24-37%) and chloramphenicol (24-37%).

To determine the current status of antimicrobial susceptibility of bacterial pathogens from animals in Germany, the Bff-GermVet monitoring program was initiated as a complementary program to the German national monitoring program GERM-Vet conducted by the Federal Office of Consumer Protection and Food Safety (BVL). In the Bff-GermVet program, a total of 1,626 bacterial strains, obtained during a 27-month period (01/2004-03/2006) from 31 indications, was screened for susceptibility against 22 antimicrobial agents and two combinations of antimicrobial agents. Selected bacteria were additionally tested for their susceptibility against additional three combinations of antimicrobial agents and the corresponding single substances. This paper describes the overall aims and the structure of the program with particular reference to the sampling strategy, the methodology for susceptibility testing and the interpretive criteria used for evaluation of the results.

During the BfT-GermVet monitoring program, Pseudomonas (P) aeruginosa from dogs and cats (n = 99) as well as Arcanobacterium (A.) pyogenes from cattle and swine (n = 90) were examined for their antimicrobial susceptibility. In general, P. aeruginosa is known to be resistant against many antimicrobial agents whereas A. pyogenes is thought to be susceptible to most agents in-vitro. However, representative and actual minimum inhibitory concentration (MIC) values are missing for both veterinary pathogens. In the present study, MIC values were determined and categorized according to the recommendations given in the Clinical and Laboratory Standards Institute (CLSI) documents M31-A2 and M31-S1. For susceptibility testing of A. pyogenes, the CLSI methodology was slightly modified. Specific breakpoints were not available for most of the antimicrobial agents tested. P. aeruginosa isolates from infections of the skin, ear and mouth as well as the urinary and genital tract of dogs and cats were either resistant or exhibited high MIC values to most antimicrobial agents tested. However, gentamicin resistant isolates were observed in only 27% and 11% (intermediate isolates 29% and 39%), respectively. For the same bacterium/host animal/organ system combinations, enrofloxacin resistance was detected in only 24% and 11% of the isolates (intermediate isolates 49% and 61%). For A. pyogenes, resistance was most prevalent against tetracycline (33%-56%, bovine and porcine isolates) and sulfonamides (26%-40%, bovine isolates).

A total of 248 coagulase-positive and coagulase-variable staphylococci from two indications of swine (infections of the urinary/genital tract including strains from the mastitis metritis agalactia syndrome as well as infections of the skin) as well as two indications of dogs/cats (respiratory tract infections and infections of skin/ear/mouth) were investigated for their susceptibility to numerous antimicrobial agents. Regardless of the animal origin and indication, the most frequently detected resistance properties were resistances against penicillin G (53-77%) and ampicillin (42-75%), tetracycline (33-52%) as well as erythromycin (13-27%). Oxacillin-resistant staphylococci were rarely detected.

A total of 500 streptococci from two indications of swine (beta-haemolytic streptococci from infections of the urinary/genital tract including strains from the mastitis metritis agalactia syndrome as well as S. suis from infections of the central nervous system and the musculoskeletal system), two indications of horses (S. equi from respiratory tract infections and beta-haemolytic streptococci from infections of the genital tract), as well as three indications of dogs and cats (beta-haemolytic streptococci from infections of the respiratory tract, the urinary/genital tract, and skin/ear/mouth) were investigated for their susceptibility to antimicrobial agents. Regardless of the animal origin and indication, the most frequently detected resistance properties were resistances against sulfamethoxazole (20-78%), tetracycline (17-93%) as well as gentamicin (14-79%). Resistance to penicillins or cephalosporins was very rarely detected - if at all.

A total of 417 isolates of Escherichia coli collected from five animal species/organ system combinations from swine [urinary/genital tract (UGT) incl. mastitis metritis agalactia syndrome], horses [genital tract (GT)] and dogs/cats [respiratory tract (RT), UGT and gastrointestinal tract (GIT)] were analysed quantitatively for their susceptibility against different antimicrobial agents by determination of minimum inhibitory concentrations. Regardless of which animal species the strains originated from, resistance appeared most frequently against sulfamethoxazole (18-59%), tetracycline (14-54 %), and ampicillin (14-39%). High percentages of intermediate isolates were observed for cephalothin (39-46 %). In general, low prevalences of resistance were detected for amoxicillin/clavulanic acid (1-4%), gentamicin (1-9%), and cefazolin (0-11%). Generally speaking, the antimicrobial resistance situation among E. coli isolates from horses and small animals is relatively good.

A total of 92 canine/feline Pasteurella multocida strains form respiratory tract infections or infections of skin/ear/mouth as well as 42 canine/feline Bordetella bronchiseptica strains from respiratory tract infections were investigated for their susceptibility to antimicrobial agents. While the P. multocida strains were susceptible to all antimicrobial agents tested - except sulfonamides -, a considerable number of the B. bronchiseptica strains was resistant or exhibited high MIC values against a number of antimicrobial agents including penicillin G, oxacillin, cefazolin, ceftiofur, cefquinome, sulfamethoxazole, and trimethoprim/sulfamethoxazole.

In this study, erythromycin- and/or clindamycin-resistant isolates among 248 coagulase-positive and coagulase-variable staphylococci and 500 streptococci, collected all over Germany during 2004-2006 in the resistance monitoring program BfT-GermVet, were investigated for their genetic basis of macrolide and/or lincosamide resistance. Staphylococci were sampled from various disease conditions of dogs/cats or pigs, whereas streptococci were from dogs/cats, pigs or horses. Resistant staphylococci were further identified biochemically to species and subspecies level and tested for the resistance genes erm(A), erm(B), erm(C), erm(TR), msr(A), msr(D), mef(A), mph(C), lnu(A), lnu(B) and lnu(C). The methylase genes erm(A), erm(B) and erm(C) were detected in staphylococci, alone or in different combinations. The erm(B) gene was the predominant gene in Staphylococcus intermedius and streptococci. The efflux gene msr(A) and the genes mph(C) and lnu(A) coding for inactivating enzymes were detected in single staphylococcal isolates. The efflux genes mef(A) and msr(D) were detected in three streptococci, in one of them together with the erm(B) gene. The lnu(B) gene was detected in seven porcine streptococcal isolates with reduced susceptibility to clindamycin. These data confirm that high-level resistance to erythromycin and clindamycin in staphylococci and streptococci was mainly due to rRNA methylases. The lnu(B) gene was detected for the first time in streptococci of animal origin.

In accordance with NCCLS guideline M31-A2, the DVG working group "antimicrobial resistance" developed a standard operating procedure (SOP) for the determination of the minimal inhibitory concentration (MIC) of antimicrobial agents by broth microdilution. This SOP was evaluated for its fitness for use in a national interlaboratory test. A total of 32 participating laboratories tested five strains (including two internationally accepted reference strains and three field strains representing in total three different bacterial species) three times at a one week interval each, using uniform microtitre plates. In 31 of the 32 laboratories more than 80% of MIC determinations performed yielded values in the expected range. In total 94.0% of the results were reproducible, with a lesser deviation of 4.0% from the expected values for laboratories performing MIC determination as a matter of routine (46.9%), compared to 7.9% for laboratories without such routine (53.1%). Comparing the consistency of results on the basis of the tested strains, a higher reproducibility of the results was observed for reference strains (96.1%) than for field strains (92.6%). In particular results obtained for the Streptococcus uberis field strain were afflicted with a higher error ratio (98 deviations from the expected values). Among the tested antimicrobial agents, a higher variability of results was recorded only for gentamicin with 16.7% divergent MIC determinations (mean value 6.0%). The high reproducibility of the results confirmed by this interlaboratory study underlines the robustness of the developed SOP as well as broth microdilutions as the method of choice for MIC determina tion.

MICs for 349 Bordetella bronchiseptica isolates from respiratory tract infections of swine were determined by broth microdilution. The lowest MIC at which 90% of isolates tested are inhibited (MIC90) was that of tetracycline and enrofloxacin (0.5 microg/ml), whereas the highest MIC90s were those of tilmicosin and cephalothin (32 microg/ml) as well as streptomycin (256 microg/ml).

Two layouts for microtitre plates, which should serve for in-vitro susceptibility testing in routine diagnostics, have been set up by the working group "Antibiotic resistance" of the German Society for Veterinary Medicine. One of these layouts was designed for the testing of bacteria from cases of mastitis and the other for bacteria from infections in large food-producing animals. The choice of the antimicrobial agents and their concentrations to be included in these layouts were based on (1) the bacteria frequently associated with the respective diseases/animals, (2) the antimicrobial agents licensed for therapeutic use in these diseases/animals, (3) the currently available breakpoints, and (4) cross-resistances between the antimicrobial agerts so far known to occur in the respective bacteria.

Objectives: Prior to the renewal of spectinomycin licensing for veterinary uses in Germany, 154 Pasteurella multocida and 148 Mannheimia haemolytica strains from respiratory tract infections in cattle were investigated for their MICs of spectinomycin and other antimicrobial agents. The data obtained should serve as a baseline from which to judge the future development of resistance. Moreover, the in vitro activity of spectinomycin in comparison with other antimicrobials should be assessed., Methods: MIC determination for all 302 strains was performed by the broth dilution method and evaluated according to NCCLS standards. MIC(50) and MIC(90) values were calculated. Strains resistant to spectinomycin were subjected to PCR assays for genes known to mediate spectinomycin resistance in Gram-negative and Gram-positive bacteria., Results: With the exception of resistance to sulfamethoxazole in P. multocida and M. haemolytica, and resistance to ampicillin in M. haemolytica, an overall low level of resistance was detected. A total of 93.5% of the P. multocida and 98.6% of the M. haemolytica strains were susceptible to spectinomycin, with MIC(90)s of 32 mg/L. PCR analysis showed that none of the spectinomycin-resistant strains carried any of the aadA gene subtypes, nor the genes spc or aad(9)., Conclusions: Prior to the renewal of spectinomycin, only a small number of spectinomycin-resistant strains was detected among bovine P. multocida and M. haemolytica. The genes responsible for spectinomycin resistance in these strains seemed to be different from those so far known to occur in other Gram-negative and Gram-positive bacteria.