Dynamin A, Myosin IB and Abp1 couple phagosome maturation to F-actin binding.

The role of actin, class I myosins and dynamin in endocytic uptake processes is well characterized, but their role during endo-phagosomal membrane trafficking and maturation is less clear. In Dictyostelium, knockout of myosin IB (myoB) leads to a defect in membrane protein recycling from endosomes back to the plasma membrane. Here, we show that actin plays a central role in the morphology and function of the endocytic pathway. Indeed, latrunculin B (LatB) induces endosome tubulation, a phenotype also observed in dynamin A (dymA)-null cells. Knockout of dymA impairs phagosome acidification, whereas knockout of myoB delays reneutralization, a phenotype mimicked by a low dose of LatB. As a read out for actin-dependent processes during maturation, we monitored the capacity of purified phagosomes to bind F-actin in vitro, and correlated this with the presence of actin-binding and membrane-trafficking proteins. Phagosomes isolated from myoB-null cells showed an increased binding to F-actin, especially late phagosomes. In contrast, early phagosomes from dymA-null cells showed reduced binding to F-actin while late phagosomes were unaffected. We provide evidence that Abp1 is the main F-actin-binding protein in this assay and is central for the interplay between DymA and MyoB during phagosome maturation.

Subgrouping of ESBL-producing Escherichia coli from animal and human sources: an approach to quantify the distribution of ESBL types between different reservoirs.

Escherichia (E.) coli producing extended-spectrum beta-lactamases (ESBLs) are an increasing problem for public health. The success of ESBLs may be due to spread of ESBL-producing bacterial clones, transfer of ESBL gene-carrying plasmids or exchange of ESBL encoding genes on mobile elements. This makes it difficult to identify transmission routes and sources for ESBL-producing bacteria. The objectives of this study were to compare the distribution of genotypic and phenotypic properties of E. coli isolates from different animal and human sources collected in studies in the scope of the national research project RESET. ESBL-producing E. coli from two longitudinal and four cross-sectional studies in broiler, swine and cattle farms, a cross-sectional and a case-control study in humans and diagnostic isolates from humans and animals were used. In the RESET consortium, all laboratories followed harmonized methodologies for antimicrobial susceptibility testing, confirmation of the ESBL phenotype, specific PCR assays for the detection of bla(TEM), bla(CTX), and bla(SHV) genes and sequence analysis of the complete ESBL gene as well as a multiplex PCR for the detection of the four major phylogenetic groups of E. coli. Most ESBL genes were found in both, human and non-human populations but quantitative differences for distinct ESBL-types were detectable. The enzymes CTX-M-1 (63.3% of all animal isolates, 29.3% of all human isolates), CTX-M-15 (17.7% vs. 48.0%) and CTX-M-14 (5.3% vs. 8.7%) were the most common ones. More than 70% of the animal isolates and more than 50% of the human isolates contained the broadly distributed ESBL genes bla(CTX-M-1), bla(CTX-M-15), or the combinations bla(SHV-12)+bla(TEM) or bla(CTX-M-1)+bla(TEM). While the majority of animal isolates carried bla(CTX-M-1) (37.5%) or the combination bla(CTX-M-1)+bla(TEM) (25.8%), this was the case for only 16.7% and 12.6%, respectively, of the human isolates. In contrast, 28.2% of the human isolates carried bla(CTX-M-15) compared to 10.8% of the animal isolates. When grouping data by ESBL types and phylogroups bla(CTX-M-1) genes, mostly combined with phylogroup A or B1, were detected frequently in all settings. In contrast, bla(CTX-M-15) genes common in human and animal populations were mainly combined with phylogroup A, but not with the more virulent phylogroup B2 with the exception of companion animals, where a few isolates were detectable. When E. coli subtype definition included ESBL types, phylogenetic grouping and antimicrobial susceptibility data, the proportion of isolates allocated to common clusters was markedly reduced. Nevertheless, relevant proportions of same subtypes were detected in isolates from the human and livestock and companion animal populations included in this study, suggesting exchange of bacteria or bacterial genes between these populations or a common reservoir. In addition, these results clearly showed that there is some similarity between ESBL genes, and bacterial properties in isolates from the different populations. Finally, our current approach provides good insight into common and population-specific clusters, which can be used as a basis for the selection of ESBL-producing isolates from interesting clusters for further detailed characterizations, e.g. by whole genome sequencing.

Zoonoses research in the German National Cohort : feasibility of parallel sampling of pets and owners.

Cats and dogs live in more than 20 % of German households and the contact between these pets and their owners can be very close. Therefore, a transmission of zoonotic pathogens may occur. To investigate whether zoonotic research questions can be examined in the context of population-based studies like the German National Cohort (GNC), two studies on different study populations were conducted as part of the feasibility tests of the GNC. The aim of the first study was to quantify the actual exposure of participants of the GNC to cats and dogs. In the second study summarised here the feasibility of the sampling of cats and dogs by their owners was tested. To quantify the exposure of participants of the GNC to cats and dogs 744 study participants of the Pretests of the GNC were asked whether they had contact with animals. Currently 10 % have a dog and 14 % have a cat in their household. These figures confirm that a large proportion of the German population has contact with pets and that there is a need for further zoonoses research. To establish the collection of biological samples from cats and dogs in the context of large-scale population-based studies feasible methods are needed. Therefore, a study was conducted to test whether pet owners can take samples from their cats and dogs and whether the quality of these samples is comparable to samples taken by a qualified veterinarian. A total of 82 dog and 18 cat owners were recruited in two veterinary practices in Hannover and the Clinic for Small Animals at the University of Veterinary Medicine in Hannover. Sampling instructions and sample material for nasal and buccal swabs, faecal samples and, in the case of cat owners, a brush for fur samples, were given to the pet owners. The pet owners were asked to take the samples from their pets at home and to send the samples by surface mail. Swab samples were cultured and bacterial growth was quantified independent of bacterial species. The growth of Gram-positive and Gram-negative bacteria from samples taken by the veterinarian and the pet owners were compared. For Gram-positive bacteria the agreement of laboratory results was 71 % for nasal swabs and 78 % for oral swabs while for Gram-negative bacteria the agreement of laboratory results was 55 % for nasal swabs and 87 % for oral swabs. In conclusion it has been shown that participants of the GNC are exposed to cats and dogs and that the sampling of cats and dogs by their owners is a feasible method which can be a useful tool for zoonoses research in population-based studies.

Evaluation of a questionnaire to assess selected infectious diseases and their risk factors : findings of a multicenter study.

BACKGROUND/OBJECTIVES: The risk to die from an infectious disease in Germany has been continuously decreasing over the last century. Since infections are, however, not only causes of death but risk factors for diseases like cardiovascular diseases, it is essential to monitor and analyze their prevalence and frequency, especially in consideration of the increased life expectancy. To gain more knowledge about infectious diseases as risk factors and their implications on the condition and change of the immune status, the German National Cohort (GNC), a population-based prospective cohort study, will recruit 200,000 subjects between 2014 and 2017. In Pretest 1, a feasibility study for the GNC, we evaluated a self-administered and self-report questionnaire on infectious diseases and on the use of health care facilities (hereinafter called "ID Screen") for feasibility and validity. METHODS: From August-November 2011, 435 participants between the ages of 20-69 completed the ID Screen. All subjects had been recruited via a random sample from the local residents' registration offices by 4 of the 18 participating study centers. The questionnaire encompasses 77 variables in six sections assessing items such as 12-month prevalence of infections, cumulative prevalence of infectious diseases, visit of health care facilities and vaccination. The feasibility was amongst others evaluated by assessing the completeness and comprehensiveness of the questionnaire. To assess the questionnaires ability to measure "immune status" and "susceptibility to infections", multivariate analysis was used. RESULTS: The overall practicability was good and most items were well understood, demonstrated by < 2/33 missing questions per questionnaire and only three variables: vaccination for influenza and pneumococci and infection with chickenpox had a frequency > 5 % of missing values. However, direct comparison of the items 12-month prevalence and lifetime prevalence of nephritis/pyelitis showed poor agreement and thereby poor understanding by 80 % of the participants, illustrating the necessity for a clear, lay person appropriate description of rare diseases to increase comprehensibility. The questionnaire will be used to support the assessment of immune dysfunction and frequency of infection. An analysis of these constructs in an exploratory factor analysis revealed limited applicability due to low interitem correlation (Cronbach's α < 0.5). This is corroborated by the extraction of more than one factor with a Kaiser-Meyer-Olkin measure of 0.6 instead of a unidimensional latent construct for "immune status". CONCLUSION: All in all, the ID Screen is a good and reliable tool to measure infectious diseases as risk factors and outcome in general, but requires a better translation of infection specific terms into lay person terms. For the assessment of the overall immune status, the tool has strong limitations. Vaccinations status should also rather be assessed based on vaccination certificates than on participants' recall.

Carriage of Extended Spectrum Beta Lactamase-Producing Escherichia coli: Prevalence and Factors Associated with Fecal Colonization of Dogs from a Pet Clinic in Lower Saxony, Germany.

Extended spectrum beta-lactamase (ESBL)-producing Escherichia coli are an emerging problem in veterinary and human medicine. Our study concentrated on the estimation of the prevalence and factors associated with the carriage of ESBL-producing E. coli in dogs who visited a veterinary clinic in northern Germany in 2017. For this reason, 1000 patients (healthy and sick dogs) were tested, resulting in 1000 samples originating from rectal swabs. Additional data were collected using a self-reported questionnaire that was completed by the dog owner. Factors associated with ESBL carriage were considered for further modeling if p < 0.05 using a two-sided Fisher test. Using a backward elimination procedure, the variables for the final multivariable logistic regression model were identified. In total, 8.9% of the dogs tested were positive for carriage of ESBL-producing E. coli. Seven factors were associated with the colonization of dogs with ESBL-E. coli within the multivariable model, namely husbandry system (p = 0.0019, OR = 3.00; 95% CI: 1.50-6.00), contact with puppies (p = 0.0044, OR = 2.43; 95% CI: 1.32-4.46), feeding of raw meat (p = 0.011, OR = 2.28; 95% CI: 1.21-4.31), food residues (p = 0.0151, OR = 2.31; 95% CI: 1.18-4.53) and food supplements (p = 0.0487, OR = 0.426; 95% CI: 0.18-0.96), and antibiotic treatments of dogs (p = 0.0005, OR = 3.030; 95% CI: 1.62-5.68) or owners (p = 0.041, OR = 2.74; 95% CI: 1.04-7.19) prior to the study. These factors refer to the animals themselves as well as to the owners and their habits or medical treatments. Although the causality and direction of transmission from owners to their dogs cannot be proven, the factor of antibiotic treatment of the owner is clearly associated with the dog's status.

Pandemic management: Analysis of availability and relevance of surveillance indicators by COVID-Task-Forces in the German federal state of Lower Saxony.

Background: Locally, the introduction of measures during times of a pandemic emergency is embodied in a pandemic containment plan created by the Robert Koch Institute in 2017. In addition to central indicators such as incidence rates and number of deaths, various indicators are used at the local level to assess the pandemic situation. So far, there hasn't been analyses of the availability and perceived relevance of the surveillance indicators used to manage the SARS-CoV-2 pandemic by the local German pandemic task forces. Aim: This study examined whether local decision-makers had access to surveillance-related indicators in a way that they could be used to make informed decisions in response to the pandemic situation. Methods: A cross sectional study was conducted, using an online questionnaire developed by experts of The Public Health Agency of Lower Saxony and The University Medical Center Göttingen (UMG). All local COVID-19 task forces of the German state of Lower-Saxony were enrolled in the study. Findings: The surveillance indicators assessed by survey respondents as most available and relevant are included under the German Infection Protection Act (IfSG). In contrast, the indicators that are not bound by the IfSG have a significantly lower availability and an inconsistent assessment of relevance. Conclusion: Against the background of efficiency, it seems central to be able to reliably provide the highly weighted surveillance indicators. Nevertheless, the relevance assessment gap between the indicators embedded in the IfSG and the ones that are not may be explained by cognitive processes such as anchoring bias. The collection and use of indicators to assess the pandemic situation and to evaluate measures should be the subject of continuous multidisciplinary discussions.

Veterinary drug administration in German veal calves: An exploratory study on retrospective data.

The aim of this cross-sectional study was to characterise the usage of antibiotic and non-antibiotic drugs with a withdrawal period in German veal calves in more detail. In Germany, the documentation of the usage of veterinary medicinal products in food producing animals is mandatory. In the German monitoring system antibiotic use in calves under the age of eight months is recorded irrespective of the production type and only some basic measures (50 % and 75 % percentiles of the entire distributions of the treatment frequencies) are published regularly. Within this study 57 farms were included contributing data of 169 veal calf groups with a total of 91,196 individual calves. To assess the drug use the treatment frequency (TF) was calculated. Most treatments were applied in the first weeks after arrival at the farm against respiratory and gastrointestinal disorders, accounting for 65.2 % and 28.6 % of the TF, respectively. Antibiotics account for 91 % of the TF. The antibiotics used most frequently were tetracyclines (35.6 %), beta-lactams (21.9 %), macrolides (12.7 %), sulphonamides (6.0 %) and trimethoprimes (5.3 %). Tetracyclines and polypeptides were administered as group treatments in more than half of the recorded applications. The number of antibiotic group treatments decreased considerably from the first to the second half of the fattening period. Logistic regression analyses revealed no statistically significant association between TF and groups size or mortality. Nevertheless, the results indicate a negative associated between TF and mortality. Concerning non-antibiotics mainly iron compounds, arylpropionic acids, mucolytics and avermectines were applied, accounting for about 5 % of the total TF. The present study provides basic data on antibiotic and non-antibiotic use in German veal calf production.

Diversity in prevalence and characteristics of ESBL/pAmpC producing E. coli in food in Germany.

The spread of extended-spectrum ß-lactamases (ESBLs) in Escherichia coli is a major public health issue and ESBL-producing bacteria are frequently reported in livestock. For the assessment of the role of the foodborne transmission pathway in Germany, detailed data on the prevalence and characteristics of isolates of food origin are necessary. The objective of this study was to describe the prevalence of cefotaxime resistant E. coli as well as ESBL/pAmpC-producing E. coli and their characteristics in foods in Germany. Out of 2256 food samples, the highest prevalence of cefotaxime resistant E. coli was observed in chicken meat (74.9%), followed by turkey meat (40.1%). Prevalence in beef, pork and minced meat was considerably lower (4.2-15.3%). Whereas 18.0% of the raw milk samples, collected at farm level were positive, this was true only for few cheese samples (1.3%). In one out of 399 vegetable samples a cefotaxime-resistant E. coli was isolated. ESBL resistance genes of the CTX-M-group (10.1% of all samples) were most frequently detected, followed by genes of the pAmpC (2.6%), SHV (2.0%) and TEM (0.8%) families. Distribution of ESBL/AmpC-encoding E. coli resistance genes and E. coli phylogroups was significantly different between the chicken related food samples and all other food items. Our study results reflect that consumers might get exposed to ESBL/pAmpC-producing E. coli through several food chains. These results together with those collected at primary production and in the human population in other studies will allow more detailed analysis of the foodborne pathways, considering transmission from livestock populations to food at retail and to consumers in Germany.

Investigation of potential risk factors for the occurrence of Escherichia coli isolates from German fattening pig farms harbouring the mcr-1 colistin-resistance gene.

Within the scope of a cross-sectional investigation on fattening pig farms conducted in 2011 and 2012, 48 fattening farms in different agricultural regions of Germany were sampled. Primary cultures of boot swabs and collective faecal samples were stored at -80 °C and screened for the presence of the mcr-1 colistin-resistance gene. The laboratory results were linked to farm-related data collected via questionnaire. Logistic regression models were used to investigate the association between occurrence of mcr-1 and farm-related data. Escherichia coli carrying the mcr-1 gene were isolated from 26 of 216 (12.0%) mixed bacterial cultures originating from 12 of 48 (25.0%) farms. Results of the logistic regression analyses indicate that the transmission between pigs or their direct environment is crucial for the occurrence of these resistant bacteria. However, there was no statistically significant association between antimicrobial use and the occurrence of the mcr-1 gene.

The zoonotic potential of Clostridium difficile from small companion animals and their owners.

BACKGROUND: Clostridium difficile infections (CDI) in humans range from asymptomatic carriage to life-threatening intestinal disease. Findings on C. difficile in various animal species and an overlap in ribotypes (RTs) suggest potential zoonotic transmission. However, the impact of animals for human CDI remains unclear. METHODS: In a large-scale survey we collected 1,447 fecal samples to determine the occurrence of C. difficile in small companion animals (dogs and cats) and their owners and to assess potential epidemiological links within the community. The Germany-wide survey was conducted from July 2012-August 2013. PCR ribotyping, Multilocus VNTR Analysis (MLVA) and PCR detection of toxin genes were used to characterize isolated C. difficile strains. A database was defined and logistic regression used to identify putative factors associated with fecal shedding of C. difficile. RESULTS: In total, 1,418 samples met the inclusion criteria. The isolation rates for small companion animals and their owners within the community were similarly low with 3.0% (25/840) and 2.9% (17/578), respectively. PCR ribotyping revealed eight and twelve different RTs in animals and humans, respectively, whereas three RTs were isolated in both, humans and animals. RT 014/0, a well-known human hospital-associated lineage, was predominantly detected in animal samples. Moreover, the potentially highly pathogenic RTs 027 and 078 were isolated from dogs. Even though, C. difficile did not occur simultaneously in animals and humans sharing the same household. The results of the epidemiological analysis of factors associated with fecal shedding of C. difficile support the hypothesis of a zoonotic potential. CONCLUSIONS: Molecular characterization and epidemiological analysis revealed that the zoonotic risk for C. difficile associated with dogs and cats within the community is low but cannot be excluded.

Association of farm-related factors with characteristics profiles of extended-spectrum ß-lactamase- / plasmid-mediated AmpC ß-lactamase-producing Escherichia coli isolates from German livestock farms.

Resistance to ß-lactam antibiotics, including third-generation cephalosporins, is of major concern for animal and human health. In this study, extended-spectrum ß-lactamase (ESBL) / plasmid-mediated AmpC (pAmpC) ß-lactamase -producing Escherichia coli isolates from German livestock farms were characterised and associations of these isolate characteristics with farm-related factors were investigated across different types of livestock. A total of 469 isolates originating from 150 farms (34 broiler farms, 38 fattening pig farms, 43 dairy cattle farms, 35 beef cattle farms) was included in the analyses. ESBL-gene family, phylogroup and phenotypic antimicrobial susceptibility for several antimicrobial agents were determined. This data was used to define different profiles characterising the isolates. Multivariate analyses using a distance-based non-parametric approach were performed to investigate associations between the profiles of the isolates and farm-related factors (e.g. management, husbandry, and environment of the farms). Co-occurrence of ESBL-gene families were not found in any of the isolates analysed. Sixty-eight percent of the isolates carried blaCTX-M variant genes. The frequency of phylogroups was as follows: A (55%), B1 (35%), D (17%) and B2 (3%). The most frequent phenotypic non-wildtype profile was non-wildtype status of solely cefepime (27%). Profiles of isolates from broilers differed substantially from those of other isolates. Associations between farm-related factors and characteristics profiles differed, depending on the isolate characteristics included in the analyses. Some factors describing the farm environment, like waterfowl in the surrounding of the farm, were associated with all tested profiles. The epidemiological method applied defines distances between isolates on basis of isolate characteristics data and is capable of analysing associations between isolate characteristics and epidemiological factors. As additional data, such as plasmid characteristics, gene type, or sequence information could be included in future studies, the method is suitable to identify points of action to reduce the occurrence of antimicrobial resistant bacteria.

Simultaneous occurrence of MRSA and ESBL-producing Enterobacteriaceae on pig farms and in nasal and stool samples from farmers.

Methicillin-resistant Staphylococcus aureus (MRSA) and extended-spectrum ß-lactamase (ESBL) producing enterobacteria (ESBL-E) have emerged in livestock. This study prospectively investigates the prevalence of MRSA and ESBL-E on pig farms and in nasal and stool samples from farmers and compares molecular characteristics of these ESBL-E isolates. In 2014, samples were derived at 51 pig farms in Germany. Per farm, five dust and five fecal samples were collected; one nasal and one stool sample were retrieved from farmers. ESBL-E isolates from humans and environmental isolates from the respective farms were characterized using whole genome sequencing for classical multilocus sequence typing (MLST), determination of ESBL-encoding genes and an ad hoc core genome MLST (cgMLST) analysis. MRSA and ESBL-E were detected on 49 (96%) and 31 (61%) of the farms, respectively; in most cases (59%) simultaneously. Nasal MRSA carriage was detected in 72 of 85 (84.7%) farmers and five of 84 (6.0%) farmers carried ESBL-E. ESBL-Escherichia coli isolates from farmers belonged to MLST STs/ESBL-genes ST10/CTX-M-1, ST196/TEM-52, ST278/TEM-52, ST410/CTX-M-15 and ST453/CTX-M-1. In one case, the human ESBL-E isolate was clonally identical to isolates from the farm environment; in the other four cases typing results indicated potential exchange of resistance determinants between human and environmental isolates, but, comparing the isolates within a minimum spanning tree indicated differences in cgMLST-patterns between the farms (p=0.076). This study demonstrated rectal ESBL-E carriage rates among farmers, which were similar to those in the general population. Molecular typing suggested that cross-transmission between the farmers and the farm environment is possible.

Antibiotic Resistances in Livestock: A Comparative Approach to Identify an Appropriate Regression Model for Count Data.

Antimicrobial resistance in livestock is a matter of general concern. To develop hygiene measures and methods for resistance prevention and control, epidemiological studies on a population level are needed to detect factors associated with antimicrobial resistance in livestock holdings. In general, regression models are used to describe these relationships between environmental factors and resistance outcome. Besides the study design, the correlation structures of the different outcomes of antibiotic resistance and structural zero measurements on the resistance outcome as well as on the exposure side are challenges for the epidemiological model building process. The use of appropriate regression models that acknowledge these complexities is essential to assure valid epidemiological interpretations. The aims of this paper are (i) to explain the model building process comparing several competing models for count data (negative binomial model, quasi-Poisson model, zero-inflated model, and hurdle model) and (ii) to compare these models using data from a cross-sectional study on antibiotic resistance in animal husbandry. These goals are essential to evaluate which model is most suitable to identify potential prevention measures. The dataset used as an example in our analyses was generated initially to study the prevalence and associated factors for the appearance of cefotaxime-resistant Escherichia coli in 48 German fattening pig farms. For each farm, the outcome was the count of samples with resistant bacteria. There was almost no overdispersion and only moderate evidence of excess zeros in the data. Our analyses show that it is essential to evaluate regression models in studies analyzing the relationship between environmental factors and antibiotic resistances in livestock. After model comparison based on evaluation of model predictions, Akaike information criterion, and Pearson residuals, here the hurdle model was judged to be the most appropriate model.

Cefotaxime-resistant E. coli in dairy and beef cattle farms-Joint analyses of two cross-sectional investigations in Germany.

Resistance to third-generation cephalosporins and other beta-lactam antibiotics is of major concern for animal and human health. Knowledge of the prevalence of resistant bacteria in primary production is an important element to estimate transmission along the stages in the food production chain and the exposure of the human population. The primary objective of this study was to determine the prevalence of cefotaxime-resistant commensal E. coli in dairy and beef cattle production units throughout Germany. Secondarily, the association between management factors and the presence of cefotaxime resistance was investigated. In total, 60 beef cattle and 52 dairy cattle production units all over Germany were included. Cefotaxime-resistant E. coli were isolated from at least one sample in 70% (95% CI: 58-83%) of the farms keeping beef cattle and 85% (95% CI: 75-94%) of the farms keeping dairy cattle. The sample prevalence was 35% (161/455; 95% CI: 31-40%) and 48% (156/323; 95% CI: 43-54%), respectively. Most factors associated with resistance to cefotaxime indicate that less intensive production results in a lower number of positive samples. For beef cattle, antimicrobial treatment of the whole animal group was significantly associated with an increased proportion of samples containing cefotaxime resistant E. coli. In addition, our results indicate that better hygiene management could improve the resistance situation on cattle farms.

Cefotaxime-resistant Escherichia coli in broiler farms-A cross-sectional investigation in Germany.

In this investigation the farm prevalence of cefotaxime-resistant Escherichia coli (CREC) in German broiler farms was evaluated. In total, 59 flocks on 34 broiler farms were sampled in four agricultural regions of Germany. Per broiler flock, three faecal samples, a pair of boot swabs and one dust sample were taken and examined for the presence of CREC. After pre-enrichment of sample material in Luria-Bertani-broth, the broth was streaked onto MacConkey agar containing 1mg/l cefotaxime (CTX). CREC isolates were detected in at least one sample from each flock resulting in a farm prevalence of 100%. The proportion of positive samples was high in all three sample types. Of 177 collective faecal samples 81.9% were positive, of 59 boot swabs and 59 dust samples 79.7% and 62.7% were positive. In conclusion, the prevalence of broiler farms with cefotaxime-resistant E. coli in Germany is very high. We suggest that the analysis of collective faecal samples is sufficient to determine the CREC farm status. In addition to other studies our study supports the finding that cefotaxime resistance is a good proxy for the presence of ESBL- or plasmidic AmpC-beta-lactamases.

Circulation of clonal populations of fluoroquinolone-resistant CTX-M-15-producing Escherichia coli ST410 in humans and animals in Germany.

Multidrug-resistant Escherichia coli encoding CTX-M-type extended-spectrum ß-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.

Prevalence and potential risk factors for the occurrence of cefotaxime resistant Escherichia coli in German fattening pig farms--a cross-sectional study.

A cross-sectional study concerning farm prevalence and risk factors for the count of cefotaxime resistant Escherichia coli (E. coli) (CREC) positive samples per sampling group on German fattening pig farms was performed in 2011 and 2012. Altogether 48 farms in four agricultural regions in the whole of Germany were investigated. Faecal samples, boot swabs and dust samples from two sampling groups per farm were taken and supplemental data were collected using a questionnaire. On 85% of the farms, at least one sample contained cefotaxime resistant E. coli colonies. Positive samples were more frequent in faeces (61%) and boot swabs (54%) than in dust samples (11%). Relevant variables from the questionnaire were analysed in a univariable mixed effect Poisson regression model. Variables that were related to the number (risk) of positive samples per sampling group with a p-value <0.2 were entered in a multivariable model. This model was reduced to statistically significant variables via backward selection. Factors that increased the risk for positive samples involved farm management and hygienic aspects. Farms that had a separate pen for diseased pigs had a 2.8 higher mean count of positive samples (95%-CI [1.71; 4.58], p=0.001) than farms without an extra pen. The mean count was increased on farms with under-floor exhaust ventilation compared to farms with over floor ventilation (2.22 [1.43; 3.46], p=0.001) and more positive samples were observed on farms that controlled flies with toxin compared to farms that did not (1.86 [1.24; 2.78], p=0.003). It can be concluded, that CREC are wide spread on German fattening pig farms. In addition the explorative approach of the present study suggests an influence of management strategies on the occurrence of cefotaxime resistant E. coli.

[On the occurence of extended-spectrum- and AmpC-beta-lactamase-producing Escherichia coli in livestock: results of selected European studies]. / Zum Vorkommen von Extended-Spektrum- und AmpC-Beta-Laktamase-produzierenden Escherichia coli in Nutztierbeständen: Ergebnisse ausgewählter europäischer Studien.

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.