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Patients with psychiatric comorbidity have been shown to experience high rates of burn injury. Burn epidemiology, etiology, and outcomes have been sparsely documented for patients with major psychiatric disorders. The aim of this study was to analyze the epidemiologic characteristics and outcomes in intensive care burn patients with pre-existing and acute major psychiatric disorders .A retrospective study was performed including intensive care burn patients admitted between March 2007 and December 2020. Demographic, clinical and epidemiological data were collected and analyzed. Major psychiatric co-morbidities were collected according to ICD-9 and ICD-10 classifications. Patients were stratified according to F-diagnoses.A total of 1325 patients were included. 16.6 % of all patients had one or more major psychiatric disorders- 9.3 % with anxiety, dissociative, stress-related, somatoform and other nonpsychotic mental disorders, 9.2 % with mood (affective) disorders, 3.5 % with schizophrenia and other non-mood psychotic disorders, and 1.8 % with disorders of adult personality and behavior. Patients with major psychiatric disorders presented with significantly higher burn severity, reflected by higher abbreviated burn severity index (ABSI) scores (5.9 vs. 5.3, p 0.001) and larger total body surface area (TBSA) affected (15.9 vs. 12.5 %, p = 0.002). Burned TBSA ≥ 30 and inhalation injuries were observed more frequently in patients with MDP, however without statistical significance. They also experienced prolonged hospital length-of-stay (LOS) (25.5 vs. 16.3 days, p 0.001), prolonged intensive care unit LOS (14.8 vs. 7.7 days, p 0.001), underwent surgical interventions (3.5 vs. 2.3, p 0.001) and mechanical ventilation more frequently (34.1 % vs. 16.5 %, p = 0.43) and had significantly longer ventilation durations (73.5 vs. 31.2 h, p = 0.002). Mortality rates were lower compared to patients without major psychiatric disorders (5.9 vs. 8.1, p 0.001).The prevalence of major psychiatric disorders in burn patients is considerably high. Patients with psychiatric comorbidities were found to have greater burn severity, prolonged total hospital and ICU LOS, underwent surgical interventions and mechanical ventilation more frequently and had prolonged ventilation duration. Our results highlight the importance of identifying burn patients with major psychiatric disorders who may necessitate additional resources and require extensive inpatient psychiatric care and counseling.

Older donors have the potential to close the gap between demand and supply in solid organs transplantation. Utilizing older organs, at the same time, has been associated with worse short- and long-term outcomes. Here, we introduce potential mechanisms on how treatments during machine perfusion (MP) may safely improve the utilization of older organs.Consequences of ischemia reperfusion injury (IRI), a process of acute, sterile inflammation leading to organ injury are more prominent in older organs. Of relevance, organ age and IRI seem to act synergistically, leading to an increase of damage associated molecular patterns that trigger innate and adaptive immune responses. While cold storage has traditionally been considered the standard of care in organ preservation, accumulating data support that both hypothermic and normothermic MP improve organ quality, particularly in older organs. Furthermore, MP provides the opportunity to assess the quality of organs while adding therapeutic agents. Experimental data have already demonstrated the potential of applying treatments during MP. New experimental show that the depletion of senescent cells that accumulate in old organs improves organ quality and transplant outcomes.As the importance of expanding the donor pool is increasing, MP and novel treatments bear the potential to assess and regenerate older organs, narrowing the gap between demand and supply.

The interactions between plants and soil microorganisms are fundamental for ecosystem functioning. However, it remains unclear if seasonality of plant growth impacts plant-microbial interactions, such as by inducing shifts in the microbial community composition, their biomass, or changes in the microbial uptake of plant-derived carbon. Here, we investigate the stability of microbial biomass of different functional groups and their net assimilation of plant-derived carbon over an entire growing season. Using a C3-C4 vegetation change experiment, and taking advantage of natural abundances of13C, we measured the plant-derived carbon in lipid biomarkers of soil microorganisms in rhizosphere and non-rhizosphere soil. We found that temporal and spatial stability was higher in bacterial than in fungal biomass, while the high temporal stability of all bacterial groups even increased in close proximity to roots. Moreover, differences in the association to plants, i.e., symbionts vs. free-living microorganisms, tend to determine the stability in the uptake of plant-derived carbon. Our results indicate, the inputs of plant-derived carbon over the growing season did not result in a shift in the microbial community composition, but instead, functional groups that are not in obligate symbiosis with plants showed a varying use of soil- and plant-derived carbon.

Recently, a new annotation tool “FungalTraits” was created based on the previous FUNGuild and FunFun databases, which has attracted high attention in the scientific community. These databases were widely used to gain more information from fungal sequencing datasets by assigning fungal functional traits. More than 1500 publications so far employed FUNGuild and the aim of this study is to compare this successful database with the recent FungalTraits database. Quality and quantity of the assignment by FUNGuild and FungalTraits to a fungal internal transcribed spacer (ITS)–based amplicon sequencing dataset on amplicon sequence variants (ASVs) were addressed. Sequencing dataset was derived from leaves and needles of 12 temperate broadleaved and coniferous tree species. We found that FungalTraits assigned more functional traits than FUNGuild, and especially the coverage of saprotrophs, plant pathogens, and endophytes was higher while lichenized fungi revealed similar findings. Moreover, ASVs derived from leaves and needles of each tree species were better assigned to all available fungal traits as well as to saprotrophs by FungalTraits compared to FUNGuild in particular for broadleaved tree species. Assigned ASV richness as well as fungal functional community composition was higher and more diverse after analyses with FungalTraits compared to FUNGuild. Moreover, datasets of both databases showed similar effect of environmental factors for saprotrophs but for endophytes, unidentical patterns of significant corresponding factors were obtained. As a conclusion, FungalTraits is superior to FUNGuild in assigning a higher quantity and quality of ASVs as well as a higher frequency of significant correlations with environmental factors.

Currently, lichen surveys are generally based on the examination of fruiting bodies. Lichens in the mycelial stage, in spores, or awaiting conditions for fruiting body formation are usually overlooked, even though they are important for maintaining biodiversity and ecosystem functions. This study aimed to explore the lichenized fungal community composition and richness associated with leaves and needles of 12 temperate tree species using Illumina MiSeq-based amplicon sequencing of the internal transcribed spacer (ITS) 2 region. Picea abies harbored the highest richness and number of lichenized fungal species. We found that the lichenized fungus Physcia adscendens dominated the leaves and needles of the most temperate tree species. Eleven lichenized fungal species detected in this study were recorded for the first time on leaves and needles. In addition, we identified Athallia cerinella, Fellhanera bouteillei, and Melanohalea exasperata that are on the German national red lists. Lichenized fungal richness was higher in conifer compared to broadleaf trees. Overall, tree species (within coniferous trees) and tree types (broadleaved vs. coniferous trees) harbored significantly different lichenized fungal community compositions pointing out the importance of host species. Diversity and community composition patterns of lichenized fungi were correlated mainly with tree species. Our study demonstrates that the diversity of foliicolous lichens associated with leaves and needles of 12 temperate tree species can be appropriately analyzed and functionally assigned using the ITS-based high-throughput sequencing. We highlighted the importance of conifers for maintaining the biodiversity of foliicolous lichens. Based on the discovery of many red list lichens, our methodological approach and results are important contributions to subsequent actions in the bio-conversation approaches.

Future climate scenarios indicate increasing drought intensity that threatens ecosystem functioning. However, the behavior of ecosystems during intense drought, such as the 2018 drought in Northern Europe, and their respective response following rewetting is not fully understood. We investigated the effect of drought on four different vegetation types in a temperate climate by analyzing dissolved organic matter (DOM) concentration and composition present in soil leachate, and compared it to two accompanying years. DOM is known to play an important role in ecosystem recovery and holds information on matter flows between plants, soil microorganisms and soil organic matter. Knowledge about DOM opens the possibility to better disentangle the role of plants and microorganisms in ecosystem recovery. We found that the average annual DOM concentration significantly decreased during the 2018 drought year compared to the normal year. This suggests a stimulation of DOM release under normal conditions, which include a summer drought followed by a rewetting period. The rewetting period, which holds high DOM concentrations, was suppressed under more intense drought. Our detailed molecular analysis of DOM using ultrahigh resolution mass spectrometry showed that DOM present at the beginning of the rewetting period resembles plant matter, whereas in later phases the DOM molecular composition was modified by microorganisms. We observed this pattern in all four vegetation types analyzed, although vegetation types differed in DOM concentration and composition. Our results suggest that plant matter drives ecosystem recovery and that increasing drought intensity may lower the potential for ecosystem recovery.

Despite the abundance of observations of foliar pathogens, our knowledge is severely lacking regarding how the potential fungal pathobiome is structured and which processes determine community assembly. In this study, we addressed these questions by analysing the potential fungal pathobiome associated with the senescing leaves and needles of 12 temperate tree species. We compared fungal plant pathogen load in the senescing leaves/needles and demonstrated that healthy-looking leaves/needles are inhabited by diverse and distinct fungal plant pathogens. We detected 400 fungal plant pathogenic ASVs belonging to 130 genera. The fungal plant pathogenic generalist, Mycosphaerella, was found to be the potential most significant contributor to foliar disease in seedlings. The analyses of assembly process and co-occurrence network showed that the fungal plant pathogenic communities in different tree types are mainly determined by stochastic processes. However, the homogenising dispersal highly contributes in broadleaf trees, whereas ecological drift plays an important role in coniferious trees. The deterministic assembly processes (dominated by variable selection) contributed more in broadleaf trees as compared to coniferous trees. We found that pH and P level significantly corresponded with fungal plant pathogenic community compositions in both tree types. Our study provides the first insight and mechanistic understanding into the community assembly, networks, and complete taxonomy of the foliar fungal pathobiome in senescing leaves and needles.

Globally, burn-related morbidity and mortality still remain high. In order to identify regional high-risk populations and to suggest appropriate prevention measure allocation, we aimed at analyzing epidemiological characteristics, etiology and outcomes of our 14-year experience with an intensive care unit (ICU) burn patient population.A retrospective observational study was conducted including patients treated between March 2007 and December 2020 in our intensive care burn unit. Demographic, clinical and epidemiological data were collected and analyzed.A total of 1359 patients were included. 68% of the subjects were males and the largest age group affected entailed 45-64-year-old adults (34%). Regarding etiology, flame and contact burns were the most common in all age groups. Mean affected total body surface area (TBSA) was 13 ± 14.5% in all subjects. Most of the burns occurred domestically or during recreational activities. Mean hospital stay was 17.77 ± 19.7 days. The average mortality was 7.7%. The mortality rate showed an overall decreasing trend whilst burn severity remained consistent from 2007 to 2020.Despite consistent burn severity presentations of annual ICU admissions, burn injury mortality showed a decreasing trend, which was in part attributed to substantial progress in burn care and treatment and improved burn prevention awareness. Statistically significant age and gender differences could be detected with regard to burn etiology and seasonality, as well as outcomes, which highlight the importance of individualized primary prevention programs.

Drought is an ever-increasing threat; its negative effects on ecosystems and their functioning directly impact our food security. It is therefore critical to understand the mechanisms that affect drought resilience of ecosystems. Many ecosystem functions depend on plant-soil interactions and are mediated by dissolved organic molecules, which are then recorded in the dissolved organic carbon (DOC) that leaches from plants and soils. In particular, DOC properties during and after rewetting can reveal if and how ecosystems are affected by drought. We therefore investigated the concentration of DOC in four different plant communities on sandy soils in Germany over three years that differed in drought intensity, including the extreme 2018 drought. We also analysed the molecular composition of DOC using ultrahigh-resolution mass spectrometry to identify the carbon sources during the rewetting period. A linear mixed effect model revealed that drought intensity significantly affected DOC concentration. DOC concentration in soil leachate was slightly increased following medium drought intensity, but was significantly reduced following high drought intensity. This suggests that medium intensity drought may stimulate DOC release, however, high intensity drought reduces DOC release. Molecular composition analysis of the DOC present during the rewetting period revealed an initial release of plant-derived carbon followed by an increase in soil organic matter-like compounds. Our findings indicate that the initial release of plant-derived carbon into soil leachate might be crucial for the ability of ecosystems to quickly recover from drought. High intensity drought may interrupt plant functioning to the point of preventing the accumulation and subsequent release of plant-derived carbon during drought, and therefore hamper ecosystem recovery. This suggests the presence of tipping points with respect to the ability of ecosystems to recover from drought. As such, monitoring DOC concentrations could lead to better assessements of the drought resilience of ecosystems.

Intrahepatic cholangiocarcinoma (ICC) is a rare disease with poor outcome, despite advances in surgical and non-surgical treatment. Recently, studies have reported a favorable long-term outcome of “very early” ICC (based on tumor size and absence of extrahepatic disease) after hepatic resection and liver transplantation, respectively. However, the prognostic value of tumor size and a reliable definition of early disease remain a matter of debate. Patients undergoing resection of histologically confirmed ICC between February 1996 and January 2021 at our institution were reviewed for postoperative morbidity, mortality, and long-term outcome after being retrospectively assigned to two groups: “very early” (single tumor ≤ 3 cm) and “advanced” ICC (size &gt, 3 cm, multifocality or extrahepatic disease). A total of 297 patients were included, with a median follow-up of 22.8 (0.1–301.7) months. Twenty-one (7.1%) patients underwent resection of “very early” ICC. Despite the small tumor size, major hepatectomies (defined as resection of ≥3 segments) were performed in 14 (66.7%) cases. Histopathological analyses revealed lymph node metastases in 5 (23.8%) patients. Patients displayed excellent postoperative outcome compared to patients with “advanced” disease: intrahospital mortality was not observed, and patients displayed superior long-term survival, with a 5-year survival rate of 58.2% (versus 24.3%) and a median postoperative survival of 62.1 months (versus 25.3 months, p = 0.013). In conclusion, although the concept of a “very early” ICC based solely on tumor size is vague as it does not necessarily reflect an aggressive tumor biology, our proposed definition could serve as a basis for further studies evaluating the efficiency of either surgical resection or liver transplantation for this malignant disease.

Older organs provide a substantial unrealized potential with the capacity to close the gap between demand and supply in organ transplantation. The potential of senolytics in improving age-related conditions has been shown in various experimental studies and early clinical trials. Those encouraging data may also be of relevance for transplantation. As age-differences between donor and recipients are not uncommon, aging may be accelerated in recipients when transplanting older organs; young organs may, at least in theory, have the potential to ‘rejuvenate’ old recipients. Here, we review the relevance of senescent cells and the effects of senolytics on organ quality, alloimmune responses and outcomes in solid organ transplantation. This article is part of the Special Issue - Senolytics - Edited by Joao Passos and Diana Jurk.

Objectives The characterization of CTX-M-₁₅ β-lactamase-producing Escherichia coli and Salmonella isolates originating mainly from German livestock and food. Methods E. coli (526, mainly commensals) and Salmonella (151) non-human isolates resistant to third-generation cephalosporins, originating from routine and monitoring submissions (2003-12) to the Federal Institute for Risk Assessment and different national targeted studies (2011-12), were examined for the presence of blaCTX-M-₁₅ genes by PCR amplification/sequencing. Additional resistance and virulence genes were screened by DNA microarray and PCR amplification. E. coli isolates with blaCTX-M-₁₅ were characterized by phylogenetic grouping, PFGE and multilocus sequence typing (MLST). The blaCTX-M-15 plasmids were analysed by replicon typing, plasmid MLST, S1 nuclease PFGE and Southern blot hybridization experiments. Results Twenty-one E. coli (livestock, food and a toy; 4.0%) and two Salmonella (horse and swine; 1.3%) isolates were CTX-M-₁₅ producers. E. coli isolates were mainly ascribed to three clonal lineages of sequence types ST678 (German outbreak with enteroaggregative Shiga-toxin-producing E. coli O104:H4; salmon, cucumber and a toy), ST410 (poultry, swine and cattle farms) and ST167/617 (swine farms and turkey meat). The blaCTX-M-₁₅ genes were located on IncI1 and multireplicon IncF plasmids or on the chromosome of E. coli ST410 isolates. Conclusions The prevalence of CTX-M-₁₅-producing isolates from non-human sources in Germany is still low. The blaCTX-M-₁₅ gene is, however, present in multidrug-resistant E. coli clones with pathogenic potential in livestock and food. The maintenance of the blaCTX-M-₁₅ gene due to chromosomal carriage is noteworthy. The possibility of an exchange of CTX-M-₁₅-producing isolates or plasmids between livestock and humans (in both directions) deserves continuous surveillance.

Livestock-associated methicillin-resistant Staphylococcus aureus (MRSA) have been found in various farm animal species throughout the world. It was the objective of this study to estimate the prevalence of MRSA in different cattle food chains (milk, beef, and veal) in Germany, to analyze the MRSA diversity along each food chain and to compare the characteristics of the different subtypes. Samples were collected between 2009 and 2012 from dairy herds (bulk tank milk), veal herds (dust from the stables), veal calves, and beef cattle at slaughter (nasal swabs) and carcasses of veal calves (surface cuts) and beef as well as veal at retail. Sampling was proportionally distributed over the country according to the cattle population (on-farm sampling), slaughterhouse capacity (abattoir samples), and the human population (meat at retail). Methicillin-resistant S. aureus were isolated using harmonized methods from all sample types and populations investigated. The highest proportion of positive samples was found in nasal swabs from veal calves at slaughter in 2012 (144/320; 45.0%) and the lowest rate in bulk tank milk in 2009 (14/388; 4.1%). Most isolates, irrespective of the origin, were from spa types t011 and t034. Both have been assigned to the clonal complex (CC) 398. Few isolates (15/632; 2.4%) were from spa types not associated with the CC398. Spa-type patterns were similar along individual food chains but differed between food chains. Antimicrobial resistance patterns differed between isolates from the different food chains and spa types. Isolates from the veal chain displayed the highest resistance rates. We conclude that there is substantial diversity in the MRSA prevalence across different cattle production sectors.

Salmonella enterica subsp. enterica serovar 4,[5],12:i:- is a monophasic variant of Salmonella Typhimurium. In this study, a total of 651 human and 107 food and environmental isolates of serovar 4,[5],12:i:- recovered from 2007 through 2011 in Switzerland were characterized by antibiotic resistance profiles and pulsed-field gel electrophoresis (PFGE). In addition, a selection of isolates belonging to the most frequent PFGE patterns was further subjected to multilocus variable-number tandem-repeat analysis (MLVA) and phage typing. Over the years 2007-2011, the reports of salmonellosis caused by Salmonella enterica serovar 4,[5],12:i:- significantly increased. A high prevalence of multidrug-resistant isolates, mainly showing an ampicillin-streptomycin-sulfonamide-tetracycline resistance pattern (ASSuT), was observed. In addition, four extended spectrum beta lactamase (ESBL) (CTX-M-55)-producing isolates were found. XbaI PFGE analysis of all isolates revealed over 150 different pulsotypes, and generally showed a considerable diversity within the monophasic isolates. Nevertheless, among these we identified seven dominant profiles, which encompassed 66% of all isolates tested. The PFGE type STYMXB.0131 dominated among human as well as food isolates. Multilocus variable-number tandem-repeat analysis profile 3-12-10-0-0211, which, in many cases, coincided with PFGE type STYMXB.0131 and phage type DT193 were the most prevalent types found for the isolates further characterized by these typing methods. Our data provide strong evidence for a spread of two specific Salmonella serovar 4,[5],12:i:- clones (PFGE pattern STYMXB.0131, resistance type ASSuT) and (PFGE pattern STYMXB.0131, resistance type SSuT). In contrast to the human isolates, the pork/poultry isolates expressed predominantly the SSuT resistance type.

Zur Bewertung der Verbreitung und Entwicklungstendenzen von Zoonosen und Zoonoseerregern werden Daten aus aktiven und passiven Monitoring- und Uberwachungssystemen in enger Zusammenarbeit mit den Landern gewonnen, zusammengefasst und der Risikobewertung zugrunde gelegt. In diesem Bericht werden die Rechtsgrundlagen und die Meldestrukturen fur ihre Durchfuhrung im foderalen System Deutschlands beschrieben. Zudem werden die Meldewege in der Europaischen Union aufgezeigt.

The objective of the study was to describe the Salmonella prevalence in turkey flocks before and after the implementation of the Salmonella control program in Germany and to identify factors that are potentially associated with the presence of Salmonella in the flocks. To achieve this, all breeding flocks and a representative sample of the fattening flocks were tested for Salmonella . None of the 98 turkey breeding flocks but 31 (10.3%) of 300 turkey fattening flocks were positive for Salmonella spp. in the baseline study during 2006/2007. In 11 (3.7%) fattening flocks S. Enteritidis (1 flock; 0.3%) or S. Typhimurium (8 flocks; 2.7%) or monophasic S. Typhimurium (2 flocks; 0.3%), which are of special public health relevance in Germany, were detected. Logistic regression analysis confirmed that production type and season were significant risk factors for the presence of Salmonella spp. in fattening turkey flocks in Germany. Data from mandatory official testing within the Salmonella control program in 2010 and 2011 revealed that Salmonella prevalence in turkey fattening flocks has decreased significantly to 3.3% and 2.6%. In line with this result, prevalence of S. Enteritidis or S. Typhimurium had decreased to 2.6% and 1.5%. Results indicate that the prevalence of Salmonella in turkey fattening flocks has decreased significantly.

In 2009, 1462 Escherichia coli isolates were collected in a systematic resistance monitoring approach from primary production, slaughterhouses and at retail and evaluated on the basis of epidemiological cut-off values. Besides resistance to antimicrobial classes that have been extensively used for a long time (e.g. sulphonamides and tetracyclines), resistance to (fluoro)quinolones and third-generation cephalosporins was observed. While in the poultry production chain the majority (60%) of isolates from laying hens was susceptible to all antimicrobials tested, most isolates from broilers, chicken meat and turkey meat showed resistance to at least one (85-93%) but frequently even to several antimicrobial classes (73-84%). In the cattle and pig production chain, the share of isolates showing resistance to at least one antimicrobial was lowest (16%) in dairy cows, whereas resistance to at least one antimicrobial ranged between 43% and 73% in veal calves, veal and pork. Resistance rates to ciprofloxacin and nalidixic acid in isolates from broilers were 41.1% and 43.1%, respectively. Likewise, high resistance rates to (fluoro)quinolones were observed in isolates from chicken meat and turkey meat. In contrast, ciprofloxacin resistance was less frequent in E. coli isolates from the cattle and pig production chain with highest rate in veal calves (13.3%). Highest resistance rates to cephalosporins were observed in broilers and chicken meat, with 5.9% and 6.2% of the isolates showing resistance. In dairy cattle and veal, no isolates with cephalosporin resistance were detected, whereas 3.3% of the isolates from veal calves showed resistance to ceftazidime. Resistance to (fluoro)quinolones and cephalosporins in E. coli isolates is of special concern because they are critically important antimicrobials in human antimicrobial therapy. The emergence of this resistance warrants increased monitoring. Together with continuous monitoring of antimicrobial usage, management strategies should be regularly assessed and adapted.

Salmonella enterica serovar Infantis (Salmonella Infantis) is consistently isolated from broiler chickens, pigs, and humans worldwide. This study investigated 93 epidemiologically unrelated Salmonella Infantis strains isolated in Germany between 2005 and 2008 in respect to their transmission along the food chain. Various phenotypic and genotypic methods were applied, and the pathogenicity and resistance gene repertoire was determined. Phenotypically, 66% of the strains were susceptible to all 17 antimicrobials tested, while the others were almost all multidrug-resistant (two or more antimicrobial resistances), with different resistance profiles and preferentially isolated from broiler chickens. A number of phage types (PTs) were shared by strains from pigs, broiler chickens, and humans (predominated by PT 29). One, PT 1, was only detected in strains from pigs/pork and humans. Pulsed-field gel electrophoresis (PFGE) subdivided strains in seven different clusters, named A-G, consisting of 35 various XbaI profiles with coefficient of similarity values of 0.73-0.97. The majority of XbaI profiles were assigned to clusters A and C, and two predominant XbaI profiles were common in strains isolated from all sources investigated. Multi-locus sequence typing (MLST) analysis of selected strains representing the seven PFGE clusters revealed that they all belonged to ST32. The pathogenicity gene repertoire of 37 representative Salmonella Infantis strains analyzed by microarray was also identical. The resistance gene repertoire correlated perfectly with the phenotypic antimicrobial resistance profiles, and multidrug-resistant strains were associated with class 1 integrons. Overall, this study showed that two major closely related genotypes of Salmonella Infantis can transmit in Germany to humans through contaminated broiler meat or pork, and consequently presents a hazard for human health.

Salmonella enterica serovar Derby (S. Derby) is one of the most prevalent serovars in pigs in Europe and in the U.S. and ranks among the 10 most frequently isolated serovars in humans. Therefore, a set of 82 epidemiologically unrelated S. Derby strains isolated between 2006 and 2008 from pigs, pork and humans in Germany was selected and investigated in respect to the transmission of clonal groups of the serovar along the food chain. Various phenotypic and genotypic methods were applied and the pathogenicity and resistance gene repertoire was determined. Phenotypically 72% of the strains were susceptible to all 17 antimicrobials tested while the others were monoresistant to tetracycline or multi-resistant with different resistance profiles. Four major clonal groups were identified based on PFGE, sequence data of the virulence genes sopA, sopB and sopD, VNTR-locus STTR5 and MLST revealing also the new sequence type ST774. Thirty different PFGE profiles were detected resulting in four clusters representing the four groups. The pathogenicity gene repertoire of 32 representative S. Derby strains analyzed by microarray showed six types with differences in the Salmonella pathogenicity islands, pathogenicity genes on smaller islets or prophages and fimbriae coding genes. The pathogenicity gene repertoire of the predominant types PAT DE1 and DE2 were most similar to the ones of S. Paratyphi B (dT+, O5-) and to a minor degree to S. Infantis and S. Virchow PATs. Overall this study showed that in Germany currently one major S. Derby clone is frequently isolated from pigs and humans. Contaminated pork was identified as one vehicle and consequently is a risk for human health. To prevent this serovar from entering the food chain, control measurements should be applied at the farm level.

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

Objectives This study was initiated to collect retrospective information on the occurrence of plasmid-mediated quinolone resistance (PMQR) in Salmonella enterica and Escherichia coli isolates in Europe and to identify the responsible genes. Methods Databases of national reference laboratories containing MIC values for Salmonella and E. coli isolated between 1994 and 2009 in animals, humans, food and the environment from 13 European countries were screened for isolates exhibiting a defined quinolone resistance phenotype, i.e. reduced susceptibility to fluoroquinolones and nalidixic acid. PCR and sequence analysis were performed to identify the responsible PMQR genes. Results Screening of databases of 13 European countries resulted in a selection of 1215 Salmonella and 333 E. coli isolates. PMQR genes were identified in 59% of the Salmonella isolates and 15% of the E. coli isolates selected. In Salmonella, qnrS1 (n¿=¿125) and variants of qnrB (n¿=¿138) were frequently identified, whereas qnrA1 (n¿=¿3) and aac(6')-1b-cr (n¿=¿3) were rarely found. qnrD was detected in 22 Salmonella isolates obtained from humans and animals. In E. coli, qnrS1 was identified in 19 isolates and qnrB19 was found in one isolate. No qnrC or qepA genes were detected in either Salmonella or E. coli. Conclusions This study shows the occurrence and dissemination of PMQR genes in Salmonella and E. coli in Europe with a defined quinolone resistance phenotype. We also report the first detection of qnrD in Salmonella collected in Europe.

This paper investigates the relationship between different types of personal experience and start-up success. We propose four extensions to earlier studies on the subject. First, we distinguish between management, technical, industry, and entrepreneurial experience of founders. Second, we consider decreasing marginal effects of different types of experience on start-up success. Third, our measures for start-up success correspond to the different stages of the entrepreneurial process. Fourth, we measure the "breadth" of experience in different fields in addition to the "depth" of experience per field. In the empirical part of the paper, we test all hypotheses using a sample of 137 German start-ups.

This paper presents a resource-based theoretical model for an extended version of the network success hypothesis. It derives four main hypotheses for the relationships between resources obtained from personal networks of the founders and a new venture's success. The model takes into account a broad range of control variables. In the empirical study, based on a sample of 123 German entrepreneurs, we do not find much support for our hypotheses. We conclude that, quite in contrast to most existing theories, network links have close to no impact on getting cheap or exclusive resources.

To study the molecular characteristics of the quinolone and associated ampicillin resistance mechanisms present in Salmonella enterica serovar Virchow isolated from Turkish foods.Nine epidemiologically unrelated Salmonella Virchow strains isolated from foods (chicken and minced meat) sold in different markets in Ankara were analysed for their susceptibility to 17 antimicrobials. The strains were typed by PFGE and plasmid profiling and investigated by molecular methods (PCR/sequencing) for the presence of several resistance genes, class 1 integrons and mutations in the quinolone resistance-determining regions. Plasmids conferring quinolone resistance were analysed by restriction fragment length polymorphism (RFLP) analysis, DNA hybridization, sequencing, replicon-typing PCR and mating experiments.All strains showed nalidixic acid resistance (MICor= 128 mg/L) together with a decreased susceptibility to ciprofloxacin (three strains with an MIC of 1 mg/L and six with an MIC of 0.25 mg/L), associated with mutations within the gyrA gene (Asp-87 --Tyr-87). In three strains, qnrS1 genes were detected. Ampicillin resistance encoded by a bla(CTX-M3) gene and/or bla(TEM-1-like) gene was found in four strains. Three of these strains carried an approximately 45 kb conjugative plasmid, designated pRQ2006, harbouring qnrS1 and a Tn3-like transposon. Partial sequencing and RFLP of pRQ2006 indicated its similarity to the qnrS1 plasmid pAH03786 found in a Japanese Shigella flexneri 2b isolate.This is the first study describing the presence of qnrS1 genes in bacterial isolates from Turkey. The pRQ2006 plasmid seems to be more related to the S. flexneri 2b qnrS1 plasmid pAH0376 than to the Salmonella qnrS1-carrying plasmids pINF5 and TPqnrS-2.

The present study was undertaken to identify and characterize integrons and integrated resistance gene cassettes among multidrug resistant (MDR) Salmonella isolates from slaughter animals and food products of animal origin in Ethiopia. A total of 98 epidemiologically unrelated Salmonella isolates comprising 13 serovars were characterized using serotyping, phage typing, antimicrobial resistance testing and the pulsed-field gel electrophoresis (PFGE) method. Integron-PCR was used to detect the presence of class 1 and class 2 integrons in the MDR strains. The associated individual resistance gene cassettes were identified using specific PCRs and DNA sequencing. The location of the integrons was determined by Southern blot hybridization analysis. Among the Salmonella serovars, a high level of antimicrobial resistance was found to streptomycin (82.6%), tetracycline (75.5%), sulfamethoxazole (60.2%), spectinomycin (53.1%), ampicillin (42.8%), nalidixic acid (34.7%), nitrofurantoin (30.6%), trimethoprim (27.5%), gentamicin (20.4%) and ciprofloxacin (19.4%). Class 1 integrons were detected in 53.1% of the MDR isolates comprising serovars Anatum, Braenderup, Kentucky, Saintpaul and Typhimurium. Of the class 1 integron positive isolates 61.5% harboured the integron-associated gene cassettes: aadA2, aadA2 + bla PSE−1 , dfrA1- aadA1 and dfrA12-orf-aadA2 (amplicon sizes 1000 bp, 1000 + 1200 bp, 1600 bp and 1900 bp, respectively). The chromosomally located aadA2 and aadA2 + bla PSE−1 resistance gene cassettes occurred exclusively in S . Typhimurium DT104 isolates, the other cassettes were found on large plasmids in several serovars. An aacCA5-aadA7 gene cassette array (amplicon size 1600 bp) was exclusively found in all MDR S . Kentucky strains of R type Str/SpeSmxGenNalAmpTetCipCef and this integron was shown to be chromosomally located. Results of the present study indicate that class 1 integrons carrying gene cassettes, which confer resistance to different classes of antimicrobials such as aminoglycosides, s-lactams and trimethoprim are widespread among the MDR Salmonella serovars isolated from slaughter animals and food products of animal origin in Ethiopia indicating the important role of these genetic elements in the dissemination of multidrug resistance.

Context: Mutations in PPARG are associated with insulin resistance and familial partial lipodystrophy, a disease characterized by altered distribution of sc fat and symptoms of the metabolic syndrome. The encoded protein, peroxisome proliferator-activated receptor (PPAR)-γ, plays a pivotal role in regulating lipid and glucose metabolism, the differentiation of adipocytes, and other cellular regulatory processes.Objectives: The objective of the study was to detect a novel PPARG mutation in a kindred with partial lipodystrophy and analyze the functional characteristics of the mutant protein.Patients and Methods: In three subjects with partial lipodystrophy, one unaffected family member, and 124 unaffected subjects, PPARG was screened for mutations by direct sequencing. Body composition, laboratory abnormalities, and hepatic steatosis were assessed in each affected subject. Transcriptional activity was determined, and EMSA was performed to investigate DNA binding capacity of the mutant protein.Results: We identified a PPARG mutation, C190S, causing partial lipodystrophy with metabolic alterations in three affected family members. The mutation was absent in the unaffected family member and unaffected controls. The mutation is located within zinc-finger 2 of the DNA binding domain. C190S PPARγ has a significantly lower ability to activate a reporter gene than wild-type PPARγ in absence and presence of rosiglitazone. A dominant-negative effect was not observed. Compared with wild-type PPARγ, C190S PPARγ shows a reduced capacity to bind DNA.Conclusion: Mutation of a zinc-binding amino acid of PPARγ leads to an altered protein-DNA binding pattern, resulting in a partial loss of function, which in turn is associated with partial lipodystrophy.

Animal derived food is a relevant source of human infections with Salmonella enterica. In this paper we analyse the presence of Salmonella in meat with respect to the observed serovars and their resistance to the fluoroquinolone ciprofloxacin and 3rd generation cephalosporins in the years 2003 to 2012. Data originated from 8176 isolates that were isolated from meat, characterized in the National Reference Laboratory for Salmonella and tested for antimicrobial resistance in the National Reference Laboratory for antimicrobial resistance in this time period. The analysis reveals substantial differences in resistance patterns between isolates from different types of meat and different serovars. Frequent serovars were mostly associated with one type of meat, suggesting an additional influence of specific characteristics of the serovars besides the effect of selection pressure excerted by antimicrobial treatments. Results show a clear increase in resistance to fluoroquinolones and 3rd generation cephalosporins that was most prominent in isolates from poultry meat. Although the number of human infections with Salmonella in Germany decreased sharply in recent years, results indicate a substantial exposure of consumers to Salmonella that are resistant to important antimicrobials via meat.

The objectives of this study were to determine antimicrobial susceptibility and to characterize the molecular mechanisms of multidrug resistance among German food-borne Salmonella isolates of different serovars.A total of 319 epidemiologically independent multidrug-resistant isolates from German foodstuffs comprising 25 different serovars were tested for their antimicrobial susceptibility by broth microdilution. The presence of antimicrobial resistance genes, integrons of classes 1 and 2 and their integrated resistance gene cassettes as well as the Salmonella genomic island 1 (SGI1) was investigated by PCR and DNA sequencing. Localization of integrons and relevant resistance genes was done by Southern hybridization. Sequence analysis revealed mutations in the quinolone resistance-determining region of the gyrA gene.The most prevalent resistances found in the multidrug-resistant serovars of Salmonella enterica from foods were to streptomycin (94%), sulfamethoxazole (92%), tetracycline (81%), ampicillin (73%), spectinomycin (72%), chloramphenicol (48%) and trimethoprim (27%). Twenty-four resistance genes covering six antimicrobial families (beta-lactams, aminoglycosides, phenicols, sulphonamides, tetracycline, and trimethoprim) were identified in the food isolates, many of them integrated as gene cassettes in class 1 and class 2 integrons. Class 1 integrons were detected in 65% of the multidrug-resistant Salmonella isolates comprising 16 different serovars, while class 2 integrons were found in 10% of the isolates belonging to two serovars only. The results demonstrate a clear predominance of both SGI1-borne resistance genes and class 1 integrons in Salmonella serovar Typhimurium DT104 and of class 2 integrons in Salmonella serovar Paratyphi B (d-tartrate positive). Nalidixic acid resistance found in 15% of the isolates was associated with single mutations in the gyrA gene.This study confirms the role of foods of animal and other origin as a reservoir of multidrug-resistant Salmonella and underlines the need for continuing surveillance of food-borne zoonotic bacterial pathogens along the food chain.

This paper reports the susceptibility to the quinolone nalidixic acid and the fluoroquinolone ciprofloxacin of 14,514 strains of Salmonella isolated in Germany from poultry, cattle and pigs between 1998 and 2001. Quinolone-resistant salmonellae were most frequently isolated from poultry, with a prevalence of 10.2 to 16-8 per cent. Poultry-associated serotypes, such as Salmonella Paratyphi B (d-tartrate positive), Salmonella Hadar and Salmonella Virchow, had the highest prevalence of quinolone resistance, ranging between 35 and 74 per cent. All the nalidixic acid-resistant strains also had a reduced susceptibility to ciprofloxacin, with minimum inhibitory concentrations (MICS) of 0.125 to 2 μg/ml. A comparison of the Mics for ciprofloxacin of the strains of these poultry-associated serotypes and Salmonella Enteritidis phage type 4 isolated in 1998/99 and 2000/01 indicated that there had been a shift towards higher Mic values of up to 2 μg/mI. The quinolone resistance-determining region (QRDR) of the gyrA gene and the homologue region of the parC gene of 31 selected strains were sequenced. Several different amino acid changes were observed in gyrA of the quinolone-resistant isolates at positions 83 and 87, but no substitutions were observed in parC .

The presence of integrons in 85 multiresistant German isolates of the predominating Salmonella enterica subsp. enterica serovar Paratyphi B dT + clone was investigated. All isolates possessed a chromosomally located Tn 7 -like class 2 integron carrying the same dfrA1 - sat1 - aadA1 array of gene cassettes. Only four isolates (4.7%) revealed an additional class 1 integron with two strains each containing the aadA1 or dfrA1-aadA1 gene cassettes.

A 1.6kb DNA fragment identified by random amplifiable polymorphic DNA differentiation (RAPD) from a Salmonella enterica serotype Typhimurium phage type DT104 isolate was used to investigate the prevalence of the region in 160 DT104 isolates, 83 other epidemiological important S. Typhimurium phage types and 20 strains selected from 17 other Salmonella serotypes. PCR screening tests using two different primer-sets derived from the RAPD fragment's nucleotide sequence showed that 76% of the 160 DT104 isolates investigated, including subtypes DT104A, DT104B, DT104B low, DT104H and DT104L, reacted positively. High sensitivity was shown for DT104 strains expressing at least the penta-resistance pattern ACSSuT (97% of 104 strains tested). DT104 susceptible strains showed only a sensitivity of 35% (17 strains tested). In contrast, 83% of the 83 strains from the other S. Typhimurium phage types reacted negatively. Strains from five out of the 17 other serotypes showed a positive signal with one primer-set. The other primer-set exhibited only a positive reaction with one S. Dublin isolate. The analysis of a 2415bp extended sequence revealed homologies to genes encoded by Escherichia coli O157:H7 prophages, suggesting that the described region contains genes of a prophage specific for DT104 and related phage types.

A total of 24,591 nonhuman salmonella strains isolated in Germany between 1986 and 1998 were examined for their resistance to nalidixic acid by an agar diffusion method. The rate of resistance (inhibition zone, ≤13 mm) ranged from 0.2% in 1986 to a peak of 14.8% in 1990. Between 1991 and 1998 the MICs for nalidixic acid-resistant strains ranged from more than 256 μg/ml for nalidixic acid to between 0.25 and 128 μg/ml for enrofloxacin. In the early 1990s a particularly high incidence of fluoroquinolone resistance (49.5%) was seen among isolates of Salmonella enterica serotype Typhimurium ( Salmonella Typhimurium) definitive phage type 204c that mainly originated from cattle. Among isolates from poultry an increase in the incidence of nalidixic acid resistance to a peak of 14.4% was observed in 1994. This peak was due to the presence of specific resistant serotypes, mainly serotypes Hadar, Saintpaul, Paratyphi B ( d -tartrate positive; formerly serotype Java) and Newport. Such strains exhibited a decreased susceptibility to enrofloxacin (MIC, 1 μg/ml). Among isolates from pigs the peak incidence of resistance was reached in 1993, with 7.5% of isolates resistant to nalidixic acid and enrofloxacin. The study demonstrates an increase in the incidence of strains that are resistant to nalidixic acid and that have decreased susceptibility to enrofloxacin after the licensing of enrofloxacin. In addition, the number of other serotypes that exhibited nalidixic acid resistance or reduced enrofloxacin susceptibility increased among the total number of isolates investigated between 1992 and 1998.

In recent years in France, England, Wales, Denmark and the USA about 500 human infections occurred, which were caused by multidrug-resistant Salmonella enterica Serovar (S.) Kentucky isolates displaying high-level resistance to fluoroquinolones (ciprofloxacin, MICor = 4 mg/l). The responsible clone was referred to as ST198-X1.To determine whether this clone is also present in German S. Kentucky isolates, the National Reference Laboratory for Salmonella (NRL-Salm) at the BfR analyzed the trend of S. Kentucky isolates received over the past years. Since 2010 the first entries of highly ciprofloxacin resistant S. Kentucky isolates, especially from turkey meat products, were recorded. 15 isolates originating from animal or food as well as one human isolate displayed MIC values ofor = 8 mg/l to ciprofloxacin. Molecular biological typing methods showed the in Germany isolated S. Kentucky isolates to be identical to the clone described by Le Hello et al. (2011) and to carry a multidrug resistance conferring region (SGI1). Since fluoroquinolones are considered by the WHO in human and veterinary medicine as drugs of critical importance, this trend demands attention. The implementation of mitigation strategies for this highly resistant clone seems to be required.

Campylobacter is a poorly recognized foodborne pathogen, leading the statistics of bacterially caused human diarrhoea in Europe during the last years. In this review, we present qualitative and quantitative German data obtained in the framework of specific monitoring programs and from routine surveillance. These also comprise recent data on antimicrobial resistances of food isolates. Due to the considerable reduction of in vitro growth capabilities of stressed bacteria, there is a clear discrepancy between the detection limit of Campylobacter by cultivation and its infection potential. Moreover, antimicrobial resistances of Campylobacter isolates established during fattening of livestock are alarming, since they constitute an additional threat to human health. The European Food Safety Authority (EFSA) discusses the establishment of a quantitative limit for Campylobacter contamination of broiler carcasses in order to achieve an appropriate level of protection for consumers. Currently, a considerable amount of German broiler carcasses would not comply with this future criterion. We recommend Campylobacter reduction strategies to be focussed on the prevention of fecal contamination during slaughter. Decontamination is only a sparse option, since the reduction efficiency is low and its success depends on the initial contamination concentration.

The Lao People’s Democratic Republic (Lao PDR) features in the group of least developed countries (LDCs), with a UNDP Global Human Development Index (HDI) rank of 131 (out of 177 countries) (UN Lao PDR 2009). Significant improvements are primarily impeded by geographic and demographic conditions: 70% of the country is mountainous and thickly forested (Mongabay 2009), with highly dispersed and ethnically diverse population (82.9% of approximately 5.6 million) (Messerli et al. 2008) living in rural and remote areas lacking access to basic social infrastructure, communication, transport links, and professional opportunities, and where poverty tends to be more acute (Government of Lao PDR/UN Country Team 2006).

Background The purpose of this study was to investigate the prevalence of MRSA in herds of fattening pigs in different regions of Germany, and to determine factors associated with the occurrence of this pathogen. For this purpose pooled dust samples were collected, and a questionnaire covered information regarding herd characteristics and management practices. Samples were pre-enriched in high-salt medium followed by selective enrichment containing cefoxitin/aztreonam, and culturing. Presumptive colonies were confirmed by multiplex-PCR targeting nuc-, mecA- and 16S rRNA-genes. Isolates were spa- and SCCmec-, and in selected cases, multilocus sequence-typed. Susceptibilities to 13 antimicrobials were determined by broth microdilution. Statistical analysis was carried out using backward stepwise logistic regression to calculate odds ratios with the MRSA test result as the outcome and herd characteristics as categorical covariates. Results Overall, 152 of 290 (52%) fattening pig farms tested positive for MRSA. The prevalence in the east, north- and south-west of Germany ranged from 39 to 59%. t011 (66%) and t034 (23%) were the most commonly identified spa-types, and 85% of isolates carried SCCmec Type V. Identified spa-types were all associated with clonal complex CC398. Susceptibility testing revealed that all isolates were resistant to tetracycline. High resistance rates were also found for sulfamethoxazole/trimethoprim (40%), and quinupristin/dalfopristin (32%). In addition, 83% of strains displayed multidrug resistant (> 3 substance classes) phenotypes. Logistic regression revealed herd size (large farms OR: 5.4; CI: 2.7-11.2; p < 0.05), and production type (wean-to-finish OR: 4.0; CI: 1.6-10.4; p < 0.05) as risk factors associated with a positive MRSA finding in fattening pig operations. Conclusions MRSA CC398 is widely distributed among herds of fattening pigs in Germany. Farm management plays a crucial role in the dissemination of MRSA with herd size, and production type representing potential major indicators.