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Zusammenfassung Hintergrund In den vergangenen Jahren hat sich die Gesamtgenomsequenzierung („whole genome sequencing“; WGS) in Kombination mit bioinformatischen Analysen zum Stand der Technik bei der Bewertung des Pathogenitäts- und Resistenzpotenzials sowie der Verwandtschaftsgrade zwischen Bakterien entwickelt. Die WGS-Analyse stellt somit ein zentrales Instrument bei der Typisierung von Erregern und der Untersuchung von Krankheits- und Ausbruchsclustern im Rahmen der molekularen Epidemiologie dar. Ziel der Studie war die Generierung eines Überblicks der in Deutschland auf Landes- und Bundesebene verfügbaren Erregertypisiermethoden von Salmonellen und Shiga-Toxin-bildenden bzw. enterohämorrhagischen Escherichia coli (STEC/EHEC) und den angewandten geno- und phänotypischen Methoden sowie über die Verfügbarkeit der genombasierten Typisierung und entsprechenden Analyseverfahren. Methoden Im Zeitraum vom Februar bis Juni 2020 wurde eine elektronische Umfrage bei Laboratorien durchgeführt, die für den öffentlichen Gesundheitsschutz und gesundheitlichen Verbraucherschutz tätig sind. Ergebnisse und Fazit Die Ergebnisse der Umfrage zeigten, dass viele der teilnehmenden Laboratorien über eine große Auswahl an phänotypischen und molekularbiologischen Methoden verfügen. Molekularbiologische Typisierungen werden am häufigsten für die Speziesidentifizierung von Salmonellen herangezogen. WGS-Verfahren sind vielfach schon bei Einrichtungen auf Bundes- und Landesebene etabliert oder befinden sich im Aufbau. Die Illumina-Sequenzierung ist dabei die am weitesten verbreitete Technologie. Die Umfrage bestätigt die Bedeutung von molekularbiologischen und genombasierten Typisierungstechnologien für die Laboratorien bei der Diagnostik von bakteriellen zoonotischen Erregern.

Zusammenfassung Einleitung Für eine Verbesserung der Patientenversorgung und die Erhöhung der Lebensmittelsicherheit im Sinne von One Health wurden im Rahmen des Projekts „Integrierte Genomische Surveillance von Zoonoseerregern (IGS-Zoo)“ Konzepte für eine sektorübergreifende genomische Surveillance von Shiga Toxin(Stx)-bildenden bzw. enterohämorrhagischen Escherichia coli (STEC/EHEC) in Deutschland entwickelt. Methoden Mittels Onlineumfrage erfolgte zunächst eine Bestandsaufnahme der aktuell in den Laboratorien im Bereich der staatlichen Untersuchungsämter im Veterinärwesen, der Lebensmittelüberwachung und des Öffentlichen Gesundheitsdienstes der Länder verfügbaren und der tatsächlich angewandten Typisierungsmethoden für diese Erreger. Ergebnisse Von 33 Teilnehmenden konnten 26 Fragebögen hinsichtlich STEC/EHEC ausgewertet werden. Pro Jahr werden in den Laboratorien zwischen 10 und 3500 Proben bearbeitet, daraus werden zwischen 3 und 1000 Erregerisolate gewonnen. Die aktuell am häufigsten verwendete Typisierungsmethode ist dabei die Bestimmung der Stx- und Intimin-kodierenden Gene mittels Polymerase-Kettenreaktion (PCR). Genomsequenzierung (Whole Genome Sequencing, WGS) ist in 8 staatlichen Laboratorien der Länder etabliert und zusätzlich in 9 Einrichtungen geplant. Als häufigstes Hindernis für eine weiterführende STEC/EHEC-Typisierung wurde angeführt, dass auch bei eindeutigem Nachweis des stx-Gens mittels PCR eine Isolierung aus Probenmaterial oftmals nicht erfolgreich ist. Diskussion Die Ergebnisse der Befragung sollen dazu beitragen, die im Projekt entwickelten Analyseverfahren bei der Zielgruppe zu integrieren, und zudem aufzeigen, wo Schwerpunkte bei der bedarfsgerechten Entwicklung von entsprechenden Schulungskonzepten gesetzt werden sollen.

The historical King Solomon has been discussed and debated by many scholars over the years. It is interesting, however, to see that the historicity of the city list of Hazor, Megiddo, and Gezer has been accepted by traditional and more radical scholars alike, who have suggested historical contexts in the 10th, 9th, or 8th century BCE for it. In this article we examine the list from a primarily literary point of view, placing it in the broader context of royal ideology in the ancient Near East and arguing that it may preserve memories of great cities from the Canaanite era.

In recent years, whole genome sequencing (WGS) in combination with bioinformatic analyses has become state of the art in evaluating the pathogenicity/resistance potential and relatedness of bacteria. WGS analysis thus represents a central tool in the investigation of the resistance and virulence potential of pathogens, as well as their dissemination via outbreak clusters and transmission chains within the framework of molecular epidemiology. In order to gain an overview of the available genotypic and phenotypic methods used for pathogen typing of Salmonella and Shiga toxin-producing and enterohemorrhagic Escherichia coli (STEC/EHEC) in Germany at state and federal level, along with the availability of WGS-based typing and corresponding analytical methods, a survey of laboratories was conducted.An electronic survey of laboratories working for public health protection and consumer health protection was conducted from February to June 2020.The results of the survey showed that many of the participating laboratories provide a wide range of phenotypic and molecular methods. Molecular typing is most commonly used for species identification of Salmonella. In many cases, WGS-based methods have already been established at federal and state institutions or are in the process of being established. The Illumina sequencing technology is the most widely used technology. The survey confirms the importance of molecular biology and whole genome typing technologies for laboratories in the diagnosis of bacterial zoonotic pathogens.HINTERGRUND: In den vergangenen Jahren hat sich die Gesamtgenomsequenzierung („whole genome sequencing“; WGS) in Kombination mit bioinformatischen Analysen zum Stand der Technik bei der Bewertung des Pathogenitäts- und Resistenzpotenzials sowie der Verwandtschaftsgrade zwischen Bakterien entwickelt. Die WGS-Analyse stellt somit ein zentrales Instrument bei der Typisierung von Erregern und der Untersuchung von Krankheits- und Ausbruchsclustern im Rahmen der molekularen Epidemiologie dar. Ziel der Studie war die Generierung eines Überblicks der in Deutschland auf Landes- und Bundesebene verfügbaren Erregertypisiermethoden von Salmonellen und Shiga-Toxin-bildenden bzw. enterohämorrhagischen Escherichia coli (STEC/EHEC) und den angewandten geno- und phänotypischen Methoden sowie über die Verfügbarkeit der genombasierten Typisierung und entsprechenden Analyseverfahren.Im Zeitraum vom Februar bis Juni 2020 wurde eine elektronische Umfrage bei Laboratorien durchgeführt, die für den öffentlichen Gesundheitsschutz und gesundheitlichen Verbraucherschutz tätig sind.Die Ergebnisse der Umfrage zeigten, dass viele der teilnehmenden Laboratorien über eine große Auswahl an phänotypischen und molekularbiologischen Methoden verfügen. Molekularbiologische Typisierungen werden am häufigsten für die Speziesidentifizierung von Salmonellen herangezogen. WGS-Verfahren sind vielfach schon bei Einrichtungen auf Bundes- und Landesebene etabliert oder befinden sich im Aufbau. Die Illumina-Sequenzierung ist dabei die am weitesten verbreitete Technologie. Die Umfrage bestätigt die Bedeutung von molekularbiologischen und genombasierten Typisierungstechnologien für die Laboratorien bei der Diagnostik von bakteriellen zoonotischen Erregern.

In order to improve patient care and to increase food safety within the framework of One Health, the project "Integrated Genomic Surveillance of Zoonotic Agents (IGS-Zoo)" aims to develop concepts for a genomic surveillance of Shiga toxin(Stx)-producing and enterohemorrhagic Escherichia coli (STEC/EHEC) in Germany.An online survey was conducted to assess the currently available and applied STEC/EHEC typing methods in the federal laboratories of veterinary regulation, food control, and public health service.Twenty-six questionnaires from 33 participants were evaluated with regard to STEC/EHEC. The number of STEC/EHEC-suspected samples that the laboratories process per year ranges between 10 and 3500, and out of these they obtain between 3 and 1000 pathogenic isolates. Currently the most frequently used typing method is the determination of Stx- and intimin-coding genes using polymerase chain reaction (PCR). Whole genome sequencing (WGS) is currently used by eight federal state laboratories, and nine are planning to implement it in the future. The most common obstacle for further typing of STEC/EHEC is that isolation from sample material is often unsuccessful despite apparent PCR detection of the stx genes.The results of the survey should facilitate the integration of the analysis methods developed in the project and emphasize the target groups' individual needs for corresponding training concepts.EINLEITUNG: Für eine Verbesserung der Patientenversorgung und die Erhöhung der Lebensmittelsicherheit im Sinne von One Health wurden im Rahmen des Projekts „Integrierte Genomische Surveillance von Zoonoseerregern (IGS-Zoo)“ Konzepte für eine sektorübergreifende genomische Surveillance von Shiga Toxin(Stx)-bildenden bzw. enterohämorrhagischen Escherichia coli (STEC/EHEC) in Deutschland entwickelt.Mittels Onlineumfrage erfolgte zunächst eine Bestandsaufnahme der aktuell in den Laboratorien im Bereich der staatlichen Untersuchungsämter im Veterinärwesen, der Lebensmittelüberwachung und des Öffentlichen Gesundheitsdienstes der Länder verfügbaren und der tatsächlich angewandten Typisierungsmethoden für diese Erreger.Von 33 Teilnehmenden konnten 26 Fragebögen hinsichtlich STEC/EHEC ausgewertet werden. Pro Jahr werden in den Laboratorien zwischen 10 und 3500 Proben bearbeitet, daraus werden zwischen 3 und 1000 Erregerisolate gewonnen. Die aktuell am häufigsten verwendete Typisierungsmethode ist dabei die Bestimmung der Stx- und Intimin-kodierenden Gene mittels Polymerase-Kettenreaktion (PCR). Genomsequenzierung (Whole Genome Sequencing, WGS) ist in 8 staatlichen Laboratorien der Länder etabliert und zusätzlich in 9 Einrichtungen geplant. Als häufigstes Hindernis für eine weiterführende STEC/EHEC-Typisierung wurde angeführt, dass auch bei eindeutigem Nachweis des stx-Gens mittels PCR eine Isolierung aus Probenmaterial oftmals nicht erfolgreich ist.Die Ergebnisse der Befragung sollen dazu beitragen, die im Projekt entwickelten Analyseverfahren bei der Zielgruppe zu integrieren, und zudem aufzeigen, wo Schwerpunkte bei der bedarfsgerechten Entwicklung von entsprechenden Schulungskonzepten gesetzt werden sollen.

Objectives Infections as a result of extended-spectrum β-lactamase-producing Enterobacterales (ESBL-E) are considered infections with a high public health burden. In this study, we aimed to identify incidences of and risk factors for healthcare-associated infections (HAIs) after rectal colonization with ESBL-producing Escherichia coli (ESBL-EC) or Klebsiella pneumoniae (ESBL-KP). Methods This prospective cohort study was performed in 2014 and 2015. Patients colonized with ESBL-EC or ESBL-KP were monitored for subsequent HAI with ESBL-E and other pathogens. In the case of an ESBL-E infection, rectal and clinical isolates were compared using pulsed-field gel electrophoresis (PFGE), and whole-genome sequencing (WGS) for ESBL-KP isolates. Proportional hazard models were applied to identify risk factors for HAIs, and to analyse competing risks. Results Among all patients admitted to the hospital during the study period, 13.6% were rectally screened for third-generation cephalosporin-resistant Enterobacterales (3GCREB). A total of 2386 rectal carriers of ESBL-EC and 585 of ESBL-KP were included in the study. Incidence density (ID) for HAI with ESBL-E was 2.74 per 1000 patient days at risk (95% confidence interval (CI) 2.16–3.43) among carriers of ESBL-EC, while it was 4.44 per 1000 patient days at risk (95% CI 3.17–6.04) among carriers of ESBL-KP. In contrast, ID for HAI with other pathogens was 4.36 per 1000 patient days at risk (95% CI 3.62–5.21) among carriers of ESBL-EC, and 5.00 per 1000 patient days at risk (95% CI 3.64–6.69) among carriers of ESBL-KP. Cox proportional hazard regression analyses identified colonization with ESBL-KP (HR = 1.58, 95% CI 1.068–2.325) compared with ESBL-EC as independent risk factor for HAI with ESBL-E. The results were consistent over all competing risk analyses. Conclusions Clinicians should be aware of the increased risk of ESBL-E infections among patients colonized with ESBL-KP compared with ESBL-EC that might be caused by underlying diseases, higher pathogenicity of ESBL-KP and other factors.

Non-typhoidal Salmonella enterica is an important gastrointestinal pathogen causing a considerable burden of disease. Resistance to third generation cephalosporins poses a serious threat for treatment of severe infections. In this study occurrence, phylogenetic relationship, and mechanisms of third generation cephalosporin resistance were investigated for clinical non-typhoidal S. enterica isolates in Germany. From 2017 to 2019, we detected 168 unique clinical S. enterica isolates with phenotypic resistance to third generation cephalosporins in a nation-wide surveillance. Compared to previous years, we observed a significant (P=0.0002) and consistent increase in resistant isolates from 0.41 % in 2005 to 1.71 % in 2019. In total, 34 different serovars were identified, most often S. Infantis (n=41; 24.4 %), S. Typhimurium (n=27; 16.1 %), S. Kentucky (n=21; 12.5 %), and S. Derby (n=17; 10.1 %). Whole genome analyses revealed extended-spectrum β-lactamase (ESBL) genes as main cause for third generation cephalosporin resistance, and most prevalent were bla CTX-M-1 (n=55), bla CTX-M-14 (n=25), and bla CTX-M-65 (n=23). There was no strict correlation between serovar, phylogenetic lineage, and ESBL type but some serovar/ESBL gene combinations were detected frequently, such as bla CTX-M-1 and bla CTX-M-65 in S. Infantis or bla CTX-M-14b in S. Kentucky. The ESBL genes were mainly located on plasmids, including IncI, IncA/C variants, emerging pESI variants, and a novel bla CTX-M-1harbouring plasmid. We conclude that third generation cephalosporin resistance is on the rise among clinical S. enterica isolates in Germany, and occurrence in various S. enterica serovars is most probably due to multiple acquisition events of plasmids.

The aim of this study was to gain an overview of the genetic diversity of Salmonella found in wildlife in Germany. We were particularly interested in exploring whether wildlife acts as a reservoir of certain serovars/subtypes or antimicrobial resistance (AMR) genes. Moreover, we wanted to explore the potential of Salmonella in spreading from wildlife to livestock and humans. To answer these questions, we sequenced 260 Salmonella enterica subsp. enterica isolates sampled between 2002 and 2020 from wildlife across Germany, using short-read whole genome sequencing. We found, consistent with previous findings, that some Salmonella sequence types are associated with certain animal species, such as S. Choleraesuis ST145 with wild boar and S. Enteritidis ST183 with hedgehogs. Antibiotic resistance was detected in 14.2% of all isolates, with resistance against important WATCH group antibiotics present in a small number of isolates. We further found that wildlife isolates do not form separate phylogenetic clusters distant to isolates from domestic animals and foodstuff, thus indicating frequent transmission events between these reservoirs. Overall, our study shows that Salmonella in German wildlife are diverse, with a low AMR burden and close links to Salmonella populations of farm and food-production environments.

Impfreaktionen bedingt durch Allergien gegen das Impfantigen oder enthaltene Hilfsstoffe wie Huhnereiweis oder Gelatine sind schon langer bekannt. Neuere Erkenntnisse bezuglich Reaktionen vermittelt uber das Kohlenhydratepitop Galaktose-alpha-1,3-Galaktose (Alpha-Gal) als Bestandteil tierischer Gelatine erweitern das Spektrum gelatinebedingter Allergien. In dieser Fallserie werden vier Patienten mit einer anaphylaktischen Reaktion nach einer Tollwutimfpung mit dem Impfstoff Rabipur® vorgestellt. Nach entsprechender allergologischer Testung mittels Pricktestung und Bestimmung des spezifischen IgE auf die im Impfstoff enthaltenen Allergene konnte eine Auslosung durch Alpha-Gal ausgeschlossen und eine Allergie gegenuber Gelatine nachgewiesen werden. Eine fehlende allergische Symptomatik nach dem Verzehr von Gelatine lasst sich eventuell durch die Hydrolyse im Darm und dem damit verbundenen Verlust der Allergenen Potenz erklaren. Weitere Implikationen beim Einsatz von gelatinehaltiger Infusionen in der Notfallmedizin werden diskutiert.

Side effects due to allergic reactions to vaccine antigen or to additives such as chicken protein or gelatin have been known for some time. Recent findings regarding reactions mediated via the carbohydrate epitope galactose-alpha-1,3-galactose (alpha-gal), a constituent of animal gelatin, broaden the spectrum of gelatin-related allergies. This case series presents four patients who developed anaphylactic reactions following rabies vaccination using the vaccine Rabipur®. After appropriate allergy testing by skin prick testing and the determination of specific IgE to allergens in the vaccine, triggering by alpha-gal could be excluded and an allergy to gelatin was detected. The absence of allergic symptoms following the consumption of gelatin could potentially be explained through intestinal hydrolysis resulting in a loss of allergenic potency. Further implications related to the use of gelatin-containing infusions in emergency medicine are discussed.

When looking at the literary history of the dynastic oracle in 2 Sam 7:1-17 and its narrative embedding in the historiographic concept of Samuel and Kings, one can argue that in the beginning, there was a royal oracle of salvation shaped by a common Ancient Near Eastern royal ideology. Even though the exact wording or the original ‘Sitz im Leben’ of the oracle cannot be identified with any certainty, some observations in the literary structure of the narrative indicate that the ‘deuteronomistic’ narrator in 2 Sam 7 used an earlier form of the dynastic oracle in order to combine it with the motive of building the temple and creating a coherent story that plays a key role in the historiographic structure of his narrative work. Later (dtr) editors enhanced the concept of building the temple over the dynastic promise and interpreted the time of David as the final stage of Israel’s conquest of the land. The dynastic oracle has not been revoked, but the political reign of the Davidides has been relativised, so that the text was opened up for new interpretations and applications, which can be studied in the wide range of its history of reception.

Despite extensive monitoring programs and preventative measures, Salmonella spp. continue to cause tens of thousands human infections per year, as well as many regional and international food-borne outbreaks, that are of great importance for public health and cause significant socio-economic costs. In Germany, salmonellosis is the second most common cause of bacterial diarrhea in humans and is associated with high hospitalization rates. Whole-genome sequencing (WGS) combined with data analysis is a high throughput technology with an unprecedented discriminatory power, which is particularly well suited for targeted pathogen monitoring, rapid cluster detection and assignment of possible infection sources. However, an effective implementation of WGS methods for large-scale microbial pathogen detection and surveillance has been hampered by the lack of standardized methods, uniform quality criteria and strategies for data sharing, all of which are essential for a successful interpretation of sequencing data from different sources. To overcome these challenges, the national GenoSalmSurv project aims to establish a working model for an integrated genome-based surveillance system of Salmonella spp. in Germany, based on a decentralized data analysis. Backbone of the model is the harmonization of laboratory procedures and sequencing protocols, the implementation of open-source bioinformatics tools for data analysis at each institution and the establishment of routine practices for cross-sectoral data sharing for a uniform result interpretation. With this model, we present a working solution for cross-sector interpretation of sequencing data from different sources (such as human, veterinarian, food, feed and environmental) and outline how a decentralized data analysis can contribute to a uniform cluster detection and facilitate outbreak investigations.

(1) Background: Resistance plasmids are under selective conditions beneficial for the bacterial host, but in the absence of selective pressure, this carriage may cause fitness costs. Compensation of this fitness burden is important to obtain competitive ability under antibiotic-free conditions. In this study, we investigated fitness effects after a conjugative transfer of plasmids containing various beta-lactamase genes transferred into Escherichia coli. (2) Methods: Fourteen beta-lactamase-encoding plasmids were transferred from clinical donor strains to E. coli J53. Growth rates were compared for all transconjugants and the recipient. Selected transconjugants were challenged in long-term growth experiments. Growth rates were assessed at different time points during growth for 500 generations. Whole-genome sequencing (WGS) of initial and evolved transconjugants was determined. Results: Most plasmid acquisitions resulted in growth differences, ranging from &minus, 4.5% to 7.2%. Transfer of a single blaCMY-16-carrying plasmid resulted in a growth burden and a growth benefit in independent mating. Long-term growth led to a compensation of fitness burdens and benefits. Analyzing WGS revealed genomic changes caused by Single Nucleotide Polymorphisms (SNPs) and insertion sequences over time. Conclusions: Fitness effects associated with plasmid acquisitions were variable. Potential compensatory mutations identified in transconjugants&rsquo, genomes after 500 generations give interesting insights into aspects of plasmid&ndash, host adaptations.

(1) Background: Resistance plasmids are under selective conditions beneficial for the bacterial host, but in the absence of selective pressure, this carriage may cause fitness costs. Compensation of this fitness burden is important to obtain competitive ability under antibiotic-free conditions. In this study, we investigated fitness effects after a conjugative transfer of plasmids containing various beta-lactamase genes transferred into Escherichia coli. (2) Methods: Fourteen beta-lactamase-encoding plasmids were transferred from clinical donor strains to E. coli J53. Growth rates were compared for all transconjugants and the recipient. Selected transconjugants were challenged in long-term growth experiments. Growth rates were assessed at different time points during growth for 500 generations. Whole-genome sequencing (WGS) of initial and evolved transconjugants was determined. Results: Most plasmid acquisitions resulted in growth differences, ranging from −4.5% to 7.2%. Transfer of a single blaCMY-16-carrying plasmid resulted in a growth burden and a growth benefit in independent mating. Long-term growth led to a compensation of fitness burdens and benefits. Analyzing WGS revealed genomic changes caused by Single Nucleotide Polymorphisms (SNPs) and insertion sequences over time. Conclusions: Fitness effects associated with plasmid acquisitions were variable. Potential compensatory mutations identified in transconjugants’ genomes after 500 generations give interesting insights into aspects of plasmid–host adaptations.

This Position Statement from the European Society of Gastrointestinal Endoscopy (ESGE) and the European Society of Gastroenterology Nurses and Associates (ESGENA) sets standards for the reprocessing of flexible endoscopes and endoscopic devices used in gastroenterology. An expert working group of gastroenterologists, endoscopy nurses, chemists, microbiologists, and industry representatives provides updated recommendations on all aspects of reprocessing in order to maintain hygiene and infection control.

Zusammenfassung Leitfaden In der operativen Medizin stellen infektiöse Patienten ein besonderes Risiko dar, ­insbesondere beim Auftreten multiresistenter Erreger. Einerseits kann es durch das invasive ­chirurgische und anästhesiologische Vorgehen zum Freisetzen von Erregern im Operationssaal und im Aufwachraum kommen, andererseits dürfen die dann erforderlichen Hygienemaßnahmen aber nicht zu einer relevanten Beeinträchtigung der internen Abläufe führen. Kalkulierte und mit den Betroffenen abgesprochene Hygienemaßnahmen sollten deshalb im Hygieneplan eines ­Klinikums festgelegt werden. Die Konsensbildung erleichtert dann deren Umsetzung.

The increase of Escherichia coli producing extended-spectrum β-lactamases (ESBL) in hospitals and their emergence as intestinal colonisers of healthy humans is of concern. Transmission ways and the extent of spread of distinct E. coli clones or ESBL genes among humans and animals via the food chain or the environment is a matter of debate. In this study we determined ESBL genotypes in E. coli isolates (n=233) resistant to 3rd generation cephalosporins from hospitals and medical practices using PCR and sequencing. Bacterial strain typing was performed by PCR-based phylogrouping, multilocus sequence typing (MLST) and a ST131-specific PCR. Results showed that CTX-M-15 (50.4%), CTX-M-1 (28.4%) and CTX-M-14 (5.6%) were the most common ESBL types. Especially, CTX-M-15 was associated with E. coli ST131 of phylogenetic group B2, which was the dominant sequence type among our isolates (35.8%). MLST typing revealed 40 different sequence types (STs), with ST131, ST410, ST10 and ST38 as the most prevalent ones. Our findings give an overview of the current distribution of ESBL-producing E. coli isolates from humans in Germany. E. coli O25b:H4-ST131 was confirmed to be the most common clone, which is known for its successful dissemination worldwide. Although heterogeneity among the isolates was found, several successful clones previously described in animals (ST410, ST10) also occurred in our isolate collection. Further detailed investigations of ESBL-producing isolates from different habitats are needed to evaluate possible transfer ways.

Main goal of this study was to determine the prevalence and molecular epidemiology of extended-spectrum β-lactamase (ESBL)-producing Enterobacteriaceae among 156 nursing home residents in Bavaria and to compare the results with healthy individuals from the Bavarian community. Intestinal colonisation by ESBL-producing Escherichia coli was detected in 23 nursing home residents (14.7%) using MacConkey agar supplemented with cefotaxime (1mg/L) for screening and the combined disc method for ESBL confirmation. Antimicrobial susceptibility testing revealed co-resistance to ciprofloxacin in 86.9% of the ESBL-producers. All isolates harboured CTX-M-ESBL with CTX-M-15 (65.2%) and CTX-M-27 (21.7%) as the most common types. Moreover, 16 isolates (69.6%) could be assigned by PCR-typing to the epidemic clonal lineage E. coli O25b-ST131. Further typing by rep-PCR and XbaI-macrorestriction with subsequent pulsed-field gel electrophoresis, respectively, revealed that two or more residents shared the same ESBL-producing E. coli clone in four nursing homes. In conclusion, we could show a high prevalence of ESBL-producing E. coli in Bavarian nursing homes (14.7%) compared to the healthy population (6.3%). Although the prevalence of ESBL-type CTX-M-15 in E. coli was similar in nursing home residents (65.2%) and healthy individuals (46%) the presence of E. coli O25b-ST131 clones differed substantially (69.6% and 14.2%, respectively). Furthermore, this study demonstrates that a person-to-person transmission or a common source of infection for ESBL-producing microorganisms may occur in these facilities. Therefore, basic hygiene measures should be assiduously implemented to prevent the further spread of these multidrug-resistant bacteria.

One of the most demanding challenges in infection control is the worldwide dissemination of multidrug-resistant (MDR) bacteria in clinical settings. Especially the increasing prevalence of carbapenemase producing Gram-negative pathogens poses an urgent threat to public health, as these enzymes confer resistance to almost all β-lactam antibiotics including carbapenems. In this study, we report a prolonged nosocomial outbreak of various NDM-1-producing Enterobacterales species due to clonal spread and cross-species exchange of plasmids and possibly transposons. Between July 2015 and September 2017, a total of 51 carbapenemase-positive isolates were collected from 38 patients and three environmental sources in a single German hospital. Combining molecular typing methods and whole genome sequencing, the metallo-β-lactamase gene blaNDM–1 was found to be present in 35 isolates of which seven additionally carried the carbapenemase gene blaKPC–2. Core genome MLST (cgMLST) revealed different clusters of closely related isolates of Escherichia coli, Klebsiella pneumoniae, Citrobacter freundii, Morganella morganii or Enterobacter cloacae indicating clonal spread. The detailed reconstruction of the plasmid sequences revealed that in all outbreak-associated isolates blaNDM–1 was located on similar composite transposons, which were also very similar to Tn125 previously described for Acinetobacter baumannii. In contrast to Tn125, these structures were flanked by IS26 elements, which could facilitate horizontal gene transfer. Moreover, the identical plasmid was found to be shared by E. coli and M. morganii isolates. Our results highlight the importance of detailed genome-based analyses for complex nosocomial outbreaks, allowing the identification of causal genetic determinants and providing insights into potential mechanisms involved in the dissemination of antibiotic resistances between different bacterial species.

One of the most demanding challenges in infection control is the worldwide dissemination of multidrug-resistant (MDR) bacteria in clinical settings. Especially the increasing prevalence of carbapenemase producing Gram-negative pathogens poses an urgent threat to public health, as these enzymes confer resistance to almost all β-lactam antibiotics including carbapenems. In this study, we report a prolonged nosocomial outbreak of various NDM-1-producing Enterobacterales species due to clonal spread and cross-species exchange of plasmids and possibly transposons. Between July 2015 and September 2017, a total of 51 carbapenemase-positive isolates were collected from 38 patients and three environmental sources in a single German hospital. Combining molecular typing methods and whole genome sequencing, the metallo-β-lactamase gene blaNDM–1 was found to be present in 35 isolates of which seven additionally carried the carbapenemase gene blaKPC–2. Core genome MLST (cgMLST) revealed different clusters of closely related isolates of Escherichia coli, Klebsiella pneumoniae, Citrobacter freundii, Morganella morganii or Enterobacter cloacae indicating clonal spread. The detailed reconstruction of the plasmid sequences revealed that in all outbreak-associated isolates blaNDM–1 was located on similar composite transposons, which were also very similar to Tn125 previously described for Acinetobacter baumannii. In contrast to Tn125, these structures were flanked by IS26 elements, which could facilitate horizontal gene transfer. Moreover, the identical plasmid was found to be shared by E. coli and M. morganii isolates. Our results highlight the importance of detailed genome-based analyses for complex nosocomial outbreaks, allowing the identification of causal genetic determinants and providing insights into potential mechanisms involved in the dissemination of antibiotic resistances between different bacterial species.