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Assessment of vancomycin-resistant Enterococcus faecium (VREfm) prevalence upon hospital admission and analysis of risk factors for colonization.From 2014 to 2018, patients were recruited within 72 hours of admission to seven participating German university hospitals, screened for VREfm and questioned for potential risk factors (prior multidrug-resistant organism detection, current/prior antibiotic consumption, prior hospital, rehabilitation or long-term care facility stay, international travel, animal contact and proton pump inhibitor [PPI]/antacid therapy). Genotype analysis was done using cgMLST typing. Multivariable analysis was performed.In 5 years, 265 of 17,349 included patients were colonized with VREfm (a prevalence of 1.5%). Risk factors for VREfm colonization were age (adjusted OR [aOR], 1.02; 95% CI, 1.01-1.03), previous (aOR, 2.71; 95% CI, 1.87-3.92) or current (aOR, 2.91; 95% CI, 2.60-3.24) antibiotic treatment, prior multidrug-resistant organism detection (aOR, 2.83; 95% CI, 2.21-3.63), prior stay in a long-term care facility (aOR, 2.19; 95% CI, 1.62-2.97), prior stay in a hospital (aOR, 2.91; 95% CI, 2.05-4.13) and prior consumption of PPI/antacids (aOR, 1.29; 95% CI, 1.18-1.41). Overall, the VREfm admission prevalence increased by 33% each year and 2% each year of life. 250 of 265 isolates were genotyped and 141 (53.2%) of the VREfm were the emerging ST117. Multivariable analysis showed that ST117 and non-ST117 VREfm colonized patients differed with respect to admission year and prior multidrug-resistant organism detection.Age, healthcare contacts and antibiotic and PPI/antacid consumption increase the individual risk of VREfm colonization. The VREfm admission prevalence increase in Germany is mainly driven by the emergence of ST117.

Species within the Enterobacter cloacae complex (ECC) include globally important nosocomial pathogens. A three-year study of ECC in Germany identified Enterobacter xiangfangensis as the most common species (65.5%) detected, a result replicated by examining a global pool of 3246 isolates. Antibiotic resistance profiling revealed widespread resistance and heteroresistance to the antibiotic colistin and detected the mobile colistin resistance (mcr)−9 gene in 19.2% of all isolates. We show that resistance and heteroresistance properties depend on the chromosomal arnBCADTEF gene cassette whose products catalyze transfer of L-Ara4N to lipid A. Using comparative genomics, mutational analysis, and quantitative lipid A profiling we demonstrate that intrinsic lipid A modification levels are genospecies-dependent and governed by allelic variations in phoPQ and mgrB, that encode a two-component sensor-activator system and specific inhibitor peptide. By generating phoPQ chimeras and combining them with mgrB alleles, we show that interactions at the pH-sensing interface of the sensory histidine kinase phoQ dictate arnBCADTEF expression levels. To minimize therapeutic failures, we developed an assay that accurately detects colistin resistance levels for any ECC isolate.

Enterobacter cloacae complex (ECC) represent globally important nosocomial pathogens. A three-year study of ECC in Germany identified Enterobacter xiangfangensis as the most common species (65.5%) detected, a result replicated by examining a global pool of 3246 isolates. Antibiotic resistance profiling revealed widespread resistance and heteroresistance to the antibiotic colistin and detected the mobile colistin resistance (mcr)-9 gene in 19.2% of all isolates. We show that resistance and heteroresistance properties depend on the chromosomal arnBCADTEF gene cassette whose products catalyze transfer of L-Ara4N to lipid A. Using comparative genomics, mutational analysis, and quantitative lipid A profiling we demonstrate that intrinsic lipid A modification levels are genospecies-dependent and governed by allelic variations in phoPQ and mgrB, that encode a two-component sensor-activator system and specific inhibitor peptide. By generating phoPQ chimeras and combining them with mgrB alleles, we show that interactions at the pH-sensing interface of the sensory histidine kinase phoQ dictate arnBCADTEF expression levels. To minimize therapeutic failures, we developed an assay that accurately detects colistin resistance levels for any ECC isolate.

With newly rising coronavirus disease 2019 (COVID-19) cases, important data gaps remain on (i) long-term dynamics of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection rates in fixed cohorts (ii) identification of risk factors, and (iii) establishment of effective surveillance strategies. By polymerase chain reaction and antibody testing of 1% of the local population and >90,000 app-based datasets, the present study surveilled a catchment area of 300,000 inhabitants from March 2020 to February 2021. Cohort (56% female; mean age, 45.6 years) retention was 75 to 98%. Increased risk for seropositivity was detected in several high-exposure groups, especially nurses. Unreported infections dropped from 92 to 29% during the study. “Contact to COVID-19–affected” was the strongest risk factor, whereas public transportation, having children in school, or tourism did not affect infection rates. With the first SARS-CoV-2 cohort study, we provide a transferable model for effective surveillance, enabling monitoring of reinfection rates and increased preparedness for future pandemics.

Purpose: This in vitro study was designed to determine if standard antiseptics used for skin and environmental surface cleansing can disrupt the metabolic activity (as a measure of viability) of multidrug-resistant gram-negative bacteria, methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococcus isolates within their native biofilms. Methods: Sixty clinical isolates of multidrug-resistant bacteria were selected for testing in different chlorhexidine gluconate, octenidine, polyhexanide and chloroxylenol concentrations. Metabolic inhibition of biofilm for each clinical isolate was analysed using a biofilm viability assay. Results: Chlorhexidine gluconate (mean = 83.8% ± 9.8%) and octenidine (mean = 84.5% ± 6.8%) showed the greatest efficacy against biofilms of the tested microorganisms, with the greatest efficacies against MRSA. The antiseptics demonstrated the least efficacy against biofilms of Pseudomonas aeruginosa. Conclusion: Chlorhexidine gluconate and octenidine showed the greatest level of bacterial metabolic inhibition and were statistically equivalent. Polyhexanide was more effective than chloroxylenol, but both were inferior to chlorhexidine gluconate and octenidine against the tested organisms.

ObjectivesTo analyse the rectal carriage rate and the molecular epidemiology of vancomycin-resistant Enterococcus faecium (VREfm) recovered from patients upon hospital admission.MethodsAdult patients were screened at six German university hospitals from five different federal states upon hospital admission for rectal colonization with VREfm between 2014 and 2018. Molecular characterization of VREfm was performed by WGS followed by MLST and core-genome MLST analysis.ResultsOf 16350 patients recruited, 263 were colonized with VREfm, with increasing prevalence rates during the 5 year study period (from 0.8% to 2.6%). In total, 78.5% of the VREfm were vanB positive and 20.2% vanA positive, while 1.2% harboured both vanA and vanB. The predominant ST was ST117 (56.7%) followed by ST80 (15%), ST203 (10.9%), ST78 (5.7%) and ST17 (3.2%). ST117/vanB VREfm isolates formed a large cluster of 96 closely related isolates extending across all six study centres and four smaller clusters comprising 13, 5, 4 and 3 isolates each. In contrast, among the other STs inter-regional clonal relatedness was rarely observed.ConclusionsTo our knowledge, this is the largest admission prevalence and molecular epidemiology study of VREfm. These data provide insight into the epidemiology of VREfm at six German university hospitals and demonstrate the remarkable inter-regional clonal expansion of the ST117/vanB VREfm clone.

In the present study, we were interested in the decline over time of anti SARS-CoV-2 antibodies and SARS-CoV-2 specific T-cell responses after two doses of mRNA vaccines in total and by age group and comorbidity. The second goal was to suggest an immunological correlate for protection on an individual basis and to describe the probability of protection over time after second vaccination.We analysed blood samples from 228 residents (median age 83.8 years) and from 273 Health Care Workers (HCW; median age 49.7 years) of five nursing homes and one home for the elderly with assisted living support. Participants had received two vaccinations. The blood samples were analysed for SARS-CoV-2 specific antibody and T-cell responses. We compared outcomes in the HCW and residents in the respective institutions. No breakthrough infections occurred during the study period. The initial immune responses in the younger participants were about 30 % higher than in the older ones. Over time, all parameters dropped continuously in all groups within the maximum observation period of 232 days. Comorbidities such as coronary heart disease or diabetes mellitus reduced the initial immune responses, regardless of age. In contrast to an almost linear decline in antibody levels, we observed that the interferon-gamma response remained at a constant level between about day 120 and 180, only to decline further thereafter.Based on our data, we propose on an individual level a correlate of protection: Persons who have a neutralizing capacity of 75 % (which would correspond to approx. 200 BAU/ml) and an interferon-gamma response above 200 mIU/ml should be considered to be protected resp. sufficiently immunized.

Enterococci with special resistance patterns (mainly vancomycin-resistant enterococci) play an important role in everyday clinical practice. Rising resistance rates to linezolid, daptomycin or tigecycline are also increasingly reported. Therapeutically, linezolid and daptomycin are the most important substances mainly in infections due to vancomycin-resistant enterococci. Several systematic meta-analyses of bloodstream infections showed discrepant results in the comparison of mortality of linezolid and daptomycin-treated bacteraemias. The containment of enterococci with special resistance patterns is currently receiving great attention. The key hygienic issue in all recommendations for dealing with multidrug-resistant enterococci can be summarized very simply: current scientific evidence is often inconsistent and studies that have clearly tested a single intervention for efficacy are lacking. The present work gives an insight into the current epidemiology and therapeutic strategies. Furthermore, the recently published German KRINKO recommendations are presented.

BackgroundMore than one year into the COVID-19 pandemic, important data gaps remain on longitudinal prevalence of SARS-CoV-2 infection at the population level and in defined risk groups, efficacy of specific lockdown measures, and on (cost-)effective surveillance.MethodsThe ELISA (LübeckLongitudinalInvestigation ofSARS-CoV-2 Infection) study invited adult inhabitants (n=∼300,000) from the Lübeck area (Northern Germany) and enrolled 3051 participants (∼1%); 1929 population-matched and 1645 with high-exposure based on profession. The one-year study period (03/2020-02/2021) spanned massive influx of tourism in the summer, rise of infection rates in the fall/winter 2020/2021, and two lockdowns. Participants were screened seven times for SARS-CoV-2 infection using PCR and antibody testing and monitored with an app-based questionnaire (n=∼91,000).ResultsCohort (56% female; mean age: 45.6 years) retention was 75%-98%; 89 persons (3.5%) were antibody- and/or PCR-positive. Seropositivity was almost 2-fold higher in men and increased risk detected in several high-exposure groups (highest for nurses, followed by police, army, firemen, and students). In May 2020, 92% of the infections were missed by PCR testing; by February 2021, only 29% remained undiagnosed. “Contact to COVID-19-affected” was the most relevant risk factor. Other factors, such as frequent use of public transportation, shopping, close contacts at work, and extensive tourism in the summer did not impact infection rates.ConclusionsWe i) provide a model for effective, regional surveillance; ii) identify infection risk factors informing public health measures; iii) demonstrate that easing of lockdown measures appears safe at times of low prevalence in the presence of continuous monitoring.

Background: Housing and access to healthcare pose particular challenges to asylum seekers and refugees. The study aim was to assess their infection risk during the COVID-19 pandemic. Methods: We provide the first event-free, prospective study on SARS-CoV-2 cases among adult asylum seekers/refugees in Europe. SARS-CoV-2 genome and antibody titers were determined in adult asylum seekers/refugees living in shared accommodation in Lubeck, Germany at two time-points and compared to the results from a local population-based cohort. Findings: In November/December 2020, we detected 2/97 PCR- (2·1%; 95% confidence interval [CI]: 0·4-6·3%) and 4/97 (4·1%; CI: 1·4-9·2%) seropositive asylum seekers/refugees compared to 3/2547 (0·1%; CI: 0·0-0·3%) PCR-positive and 12/2547 (0·5%; CI: 0·3-0·8%) seropositive probands in the control sample. In February 2021, 2/67 (3·0%; CI: 0·5-9·1%) PCR-, and 25/67 (37·3%; CI: 27·4-48·1) antibody-positive individuals were found in the study group in comparison to 2/2371 (0·1%; CI: 0·0-0·3% and 38/2371 (1·6%; CI: 1·2-2·1%) in the control group. Age, sex, or facility equipment did not impact the results. "Living-with-own-children-in-the-shelter” was significantly positively correlated with infection risk. Importantly, none of the PCR-positive refugees were aware of their infection. Only 32·9% of the asylum seekers were willing to be vaccinated compared to 85·5% in the control population. Interpretation: Refugees residing in shared accommodations represent a high-risk group for SARS-CoV-2 infection and transmission. The present study suggests a need for (i) tailored testing strategies, (ii) improved information of this subgroup, and (iii) high-priority vaccination. Funding Information: The study was funded by the Federal Ministry of Education and Research. Declaration of Interests: C.K. serves as medical advisor to Centogene for genetic testing reports in the fields of movement disorders and dementia, excluding Parkinson's disease and serves on the Scientific Advisory Board of Retromer Therapeutics. Neither activity represents a conflict of interest. Likewise, none of the other authors declares any financial conflict of interest. Ethics Approval Statement: The ethics committee of the University of Lubeck approved the study (Az. 20-150). All participants gave written informed consent. All study materials were translated into the refugees' native languages, and interpreters were available on-site at the study center.

Die rasche Zunahme multiresistenter Gram-negativer Erreger (MRGN) ist ein drangendes und weitgehend ungelostes globales Problem. Die Behandlungsoptionen bei diesen Erregern sind sehr stark eingeschrankt. Nur wenige neue Substanzen sind zugelassen worden oder befinden sich aktuell in klinischen Phase-II/III-Studien. Ubersichtliche Vorstellung der bislang vorliegenden Daten zu den neuen β‑Laktam-Antibiotika und β‑Laktamase-Inhibitor-Kombinationen. Die neuen Makrolide, Ketolide und Aminoglykoside werden nicht adressiert. Selektive Literaturrecherche zu den Substanzen Ceftazidim/Avibactam, Ceftolozan/Tazobactam, Imipenem/Cilastatin + Relebactam, Meropenem/Vaborbactam, Aztreonam/Avibactam und Cefiderocol unter Einbeziehung aktuell registrierter Studien mit klinischer Auswertung und Datenanalyse. Die Entwicklung neuer Substanzen zur Therapie von Infektionen durch MRGN eroffnet neue Optionen bei Infektionen durch besonders schwierig zu behandelnde Erreger, insbesondere Erreger, die Carbapenemasen vom Klebsiella-pneumoniae-Carbapenemase (KPC) und OXA-48-Typ bilden. β‑Laktamase-Bildner werden durch die neuen Substanzen oder Kombinationen unterschiedlich stark gehemmt; allerdings fehlen noch immer ausreichende Therapieoptionen fur Metallo-β-Laktamase-Bildner sowie Infektionen durch multiresistente Pseudomonas-aeruginosa- und Acinetobacter-spp.-Stamme. Vielfach sind die klinischen Daten noch indifferent und stammen aus uneinheitlich definierten Patientenkollektiven. Direkte Vergleiche mit etablierten Behandlungsstrategien wie dem „Last-resort-Einsatz“ von Polymyxinen sind kaum moglich. Leider sind auch bereits Falle einer raschen Resistenzentwicklung beschrieben. Der Stellenwert der Toxizitat und optimalen Dosierung – auch bei Organversagen oder Organersatzverfahren wie Dialyse – ist vielfach noch unklar.

ZusammenfassungDie Ausbreitung von resistenten Gram-negativen Erregern weltweit ist besorgniserregend. Von besonderem Interesse ist die Verbreitung von Resistenzen gegen Beta-Laktam-Antibiotika, die in vielen Fällen Mittel der Wahl zur Therapie sind. Im European Antimicrobial Resistance Surveillance Network (EARS-Net) wird die Resistenzentwicklung bei schweren Infektionskrankheiten in Europa regelmäßig überwacht. In Deutschland können Isolate zur Abklärung und/oder Bestätigung von Carbapenemasen an das Nationale Referenzzentrum (NRZ) für Gram-negative Krankenhauserreger an der Uniklinik Bochum eingeschickt werden.Im ersten Teil der Arbeit werden die aktuell publizierten Daten für Europa ausgewertet und dargestellt sowie mit den aktuell verfügbaren Daten aus Deutschland verglichen. Im zweiten Teil werden Therapieoptionen von Infektionen durch resistente Erreger dargestellt.

Liver abscess is a rare but potentially fatal disease with mortality rates of 2 – 12 %. The spectrum of causative microorganisms varies across the world and is changing due to increasing worldwide travel activity. As a result, previously less frequent pathogens must increasingly be considered in the differential diagnosis. The most common pathogens of liver abscess are so-called abscess streptococci, anaerobes and Gram-negative bacteria such as Escherichia coli and Klebsiella pneumoniae. Certain clones among the Gram-negatives, typically among Klebsiella spp., harbour specific virulence factors and are therefore referred to as hypervirulent clones which spread worldwide. In any case, the causative agent should be identified before anti-infective treatment is initiated. Several pathogens such as Candida spp., Pseudomonas aeruginosa or Staphylococcus aureus are uncommon and only detected in a specific context. Various therapeutic regimens are available for anti-infective therapy. Usually an empirical therapy is started, combining mostly β-lactam antibiotics or quinolones with anaerobically active substances.

We report a case of severe infection with liver abscess and endophthalmitis caused by a hypervirulent Klebsiella pneumoniae strain in an immunocompetent German male patient without travel history to Asia. Phenotypic and molecular characterization showed high similarity to the reference genome NTUH-K2044 isolated in Asia. The isolate was assigned as ST2398 (clonal complex 66). The findings underline global spread of hypervirulent Klebsiella pneumoniae strains to Europe.

Objectives: This study aimed at determining the prevalence of rifaximin resistance in a large collection of Enterobacterales resistant to third-generation cephalosporins. A simple agar screen was d...

Objectives To assess the admission prevalence of third-generation cephalosporin-resistant Enterobacterales (3GCREB) and to assess whether risk factors vary by β-lactamase genotype. Methods Adult patients were recruited within 72 h of admission to general wards of six university hospitals in 2014 and 2015. Rectal swabs were screened for 3GCREB and isolates were analysed phenotypically and genotypically. Patients were questioned on potential risk factors. Multivariable analyses were performed to identify risk factors for 3GCREB colonization and for specific β-lactamases. Results Of 8753 patients screened, 828 were 3GCREB positive (9.5%). Eight hundred and thirteen isolates were available for genotyping. CTX-M-15 was the most common ESBL (38.0%), followed by CTX-M-1 (22.5%), CTX-M-14 (8.7%), CTX-M-27 (7.5%) and SHV-ESBL (4.4%). AmpC was found in 11.9%. Interestingly, 18 Escherichia coli isolates were AmpC positive, 12 of which (67%) contained AmpC on a gene of plasmid origin [CMY (n = 10), DHA (n = 2)]. Risk factors for 3GCREB colonization varied by genotype. Recent antibiotic exposure and prior colonization by antibiotic-resistant bacteria were risk factors for all β-lactamases except CTX-M-14 and CTX-M-27. Travel outside Europe was a risk factor for CTX-M-15 and CTX-M-27 [adjusted OR (aOR) 3.49, 95% CI 2.88–4.24 and aOR 2.73, 95% CI 1.68–4.43]. A previous stay in a long-term care facility was associated with CTX-M-14 (aOR 3.01, 95% CI 1.98–4.59). A preceding hospital stay in Germany increased the risk of CTX-M-15 (aOR 1.27, 95% CI 1.14–1.41), while a prior hospital stay in other European countries increased the risk of SHV-ESBL colonization (aOR 3.85, 95% CI 1.67–8.92). Conclusions The detection of different ESBL types is associated with specific risk factor sets that might represent distinct sources of colonization and ESBL-specific dissemination routes.

Enterococci with special resistance patterns (mainly vancomycin-resistant enterococci) play an important role in everyday clinical practice. Rising resistance rates to linezolid, daptomycin or tigecycline are also increasingly reported. Therapeutically, linezolid and daptomycin are the most important substances mainly in infections due to vancomycin-resistant enterococci. Several systematic meta-analyses of bloodstream infections showed discrepant results in the comparison of mortality of linezolid and daptomycin-treated bacteraemias. The containment of enterococci with special resistance patterns is currently receiving great attention. The key hygienic issue in all recommendations for dealing with multidrug-resistant enterococci can be summarized very simply: current scientific evidence is often inconsistent and studies that have clearly tested a single intervention for efficacy are lacking. The present work gives an insight into the current epidemiology and therapeutic strategies. Furthermore, the recently published German KRINKO recommendations are presented.

Purpose International guidelines for antibiotic treatment of spontaneous bacterial peritonitis (SBP) are based on studies conducted decades ago and do not reflect regional differences of bacterial epidemiology. Methods We retrospectively analyzed epidemiology of agents, antibiotic resistance patterns, and survival in liver cirrhosis patients with their first episode of SBP during the years 2007-2013. Results Of the 311 patients included, 114 patients had a positive ascites culture, and 197 had an ascitic neutrophil count >250 μL. Gram-positive bacteria (47.8%) were more frequently found than Gram-negatives (44.9%), fungi in 7.2%. Enterobacter spp. (40.6%), Enterococcus spp. (26.1%), and Staphylcoccus spp. (13.8%) were the most frequently isolated agents. Third-generation cephalosporins covered 70.2% of non-nosocomial and 56.3% of nosocomial-acquired SBP cases.When SBP was diagnosed by a positive ascitic culture, survival was highly significantly reduced (mean: 13.9 ± 2.9 months; 95% confidence interval [CI]: 8.1-19.8) compared with culture-negative SBP patients (mean: 44.1 ± 5.4 months; 95% CI: 33.4-54.9; P = 0.000). Along with model of end-stage liver disease score and intensive care unit contact, a positive ascites culture remained an independent risk factor associated with poor survival (odds ratio: 1.49; 95% CI: 1.09-2.03) in multivariate analysis; piperacillin/tazobactam proved to be an adequate antibiotic for nosocomial and non-nosocomial SBP in 85.1% and 92.5%, respectively. SBP infection with Enterococcus spp. was associated with poor patient survival (P = 0.048). Conclusions Third-generation cephalosporins have poor microbial coverage for treatment of SBP. Current guidelines need to adapt for the emerging number of Gram-positive infectious agents in SBP patients.

Infektionen durch Carbapenem-resistente Enterobakterien (CRE) stellen aufgrund ihrer hohen Morbiditat und Mortalitat weltweit eine wachsende Bedrohung fur Lebertransplantatempfanger (LTR) dar, v. a. bei Nachweis von Klebsiella pneumoniae Carbapenemase (KPC)-bildenden Erregern. Nach der aktuellen Literatur entwickeln 3 bis 13 % der Organtransplantierten in Endemiegebieten Infektionen durch CRE, wobei der primare Infektionsort eng mit dem transplantierten Organ im Zusammenhang steht. Die kumulative 30-Tage-Sterblichkeit von LTR mit Infektionen durch Carbapenem-resistente K. pneumoniae liegt bei 36 %, und die 180-Tage-Mortalitatsrate betragt 58 %. Die hohe Anfalligkeit von LTR fur potenziell todlich verlaufende bakterielle Infektionen mit multiresistenten Erregern (MRE) fuhrt in der Praxis zu einer haufigeren empirischen Anwendung von erweiterten Breitspektrum-Antibiotikatherapien, die wiederum die Selektion extremer Resistenzen fordert. Dennoch besteht eine relevante Gefahr des Scheiterns der herkommlichen empirischen Behandlung, da kulturbasierte Techniken zur mikrobiologischen Identifikation von CRE eine 48- bis 72-stundige Verzogerung von der Abnahme der Blutkulturen bis zur mikrobiologischen Befunderstellung mit Resistenztestung beinhalten, sodass zielgerichtete Therapien nur verzogert begonnen werden konnen. Dieser Teufelskreis ist schwer zu durchbrechen und fuhrt zu einer fortwahrenden Einengung der antibiotischen Behandlungsmoglichkeiten. Da die derzeit verfugbaren therapeutischen Optionen auserst begrenzt sind, kommt der Infektionspravention eine herausragende Bedeutung zu, v. a. in der Fruhphase nach Lebertransplantation mit intensiver Immunsuppression. Die Verbesserung klinischer Behandlungsergebnisse ist eine grose Herausforderung und erfordert einen mehrgliedrigen Ansatz, bei dem stringent praktizierte Hygienemasnahmen, Screening-Tests auf MRE, zeitsparende molekularbiologische Untersuchungstechniken und Antibiotic Stewardship (ABS) kombiniert werden. Dieser Artikel stellt die aktuelle Literatur in Bezug auf die Haufigkeit und das klinische Outcome von CRE-Infektionen bei LTR vor und fasst aktuelle praventive und therapeutische Empfehlungen zusammen, um die Bedrohung durch CRE im Real-Life-Setting der klinischen Transplantationsmedizin zu minimieren.

To enable an efficient and enduring decontamination of the rectal mucosa during transanal endosocopic procedures, we developed a device for reversible endolumenal colon occlusion (ColoShield). The aim of this study was to assess the value of ColoShield in reducing peritoneal contamination during a transrectal procedure. Sixteen pigs underwent transrectal hybrid NOTES cholecystectomy after standardized disinfective rectal washout either with endolumenal colon occlusion using ColoShield (N = 8) or without colon occlusion (N = 8). Rectal swab samples were taken before and after rectal washout and at the end of the procedure. Peritoneal biopsies for microbiological evaluation were obtained at the end of the procedure and at necropsy 7 days after surgery. Peritoneal contamination at the end of surgery was significantly lower using ColoShield compared to not using colon occlusion [13 (1/8) vs. 75 % (6/8); P = 0.012]. No significant differences were found regarding contamination of rectal swabs and peritoneal contamination at necropsy. The application of ColoShield may increase the safety of transrectal NOTES and transanal endoscopic procedures by reducing peritoneal contamination and consecutive infectious complications.

Komplizierte Infektionen durch Vancomycin-resistente Enterokokken (VRE) sind ein groses therapeutischs Problem. Zur Therapie kommen da nur wenige Reservesubstanzen in Frage. Resistenzen gegenuber den Reserveantibiotika wurden allerdings mittlerweile auch beschrieben. Von den zugelassenen Substanzen ist Linezolid das am haufigsten eingesetzte Antibiotikum. Fur die Therapiealternative Tigecyclin liegen insbesondere bei lebensbedrohlichen Verlaufen wie Sepsis oder Endokarditis wenige klinische Daten vor. Daptomycin ist eine Alternative, die jedoch fur Enterokokken-Infektionen in Deutschland nicht zugelassen ist. In der vorliegenden Arbeit wird der Stellenwert der verschiedenen Therapieoptionen im klinischen Alltag auf Basis vorhandener Daten kritisch dargestellt.

Background This study was designed as a quasi-experiment to evaluate automatic inoculation of fecal specimens, using the automated specimen inoculator Previ Isola (bioMérieux, France). Methods We evaluated the quality of cultures, recovery rates of enteropathogenic bacteria (Salmonella, Shigella, Campylobacter, and Yersinia species), and cost-effectiveness in terms of technical time. The Previ Isola recovery rates for the two-year period from August 2009 to July 2011 were compared with historical manual inoculation data of the previous two years (August 2007 to July 2009). The regional (Baden-Württemberg) and nationwide (Germany) trends of recovery rates for this four-year period were referred. Results A total of 5,884 fecal specimens were collected over the study period. Most positive cultures were for Salmonella, followed by Campylobacter. Compared with the historical data, the numbers of Campylobacter-positive specimens for a year between August and July were increased significantly, from 19 in 2007-2008 and 10 in 2008-2009 to 32 in 2009-2010 (P=0.002) and 32 in 2010-2011 (P=0.003), respectively. During the study period, the official data for our region and nationwide did not show this increase in the recovery rate of Campylobacter. For Salmonella, Shigella, and Yersinia, no significant changes were observed. Compared with manual inoculation, the mean hands-on time with Previ Isola inoculation was significantly shortened, from 37:30 min to 8:42 min per 15 fecal specimens. Conclusions Inoculation by Previ Isola improves the quality of routine culture of fecal specimens, with better sensitivity for Campylobacter and less hands-on time.

Liver abscess is a rare but potentially fatal disease with mortality rates of 2 - 12 %. The spectrum of causative microorganisms varies across the world and is changing due to increasing worldwide travel activity. As a result, previously less frequent pathogens must increasingly be considered in the differential diagnosis. The most common pathogens of liver abscess are so-called abscess streptococci, anaerobes and Gram-negative bacteria such as Escherichia coli and Klebsiella pneumoniae. Certain clones among the Gram-negatives, typically among Klebsiella spp., harbour specific virulence factors and are therefore referred to as hypervirulent clones which spread worldwide. In any case, the causative agent should be identified before anti-infective treatment is initiated. Several pathogens such as Candida spp., Pseudomonas aeruginosa or Staphylococcus aureus are uncommon and only detected in a specific context. Various therapeutic regimens are available for anti-infective therapy. Usually an empirical therapy is started, combining mostly β-lactam antibiotics or quinolones with anaerobically active substances.

In humans, Cryptococcus mainly infects individuals with HIV infection or other types of immunosuppression. Here, we report the first case of disseminated cryptococcosis in a simian immunodeficiency virus-negative 27-year-old female Gorilla gorilla presenting with lethargy, progressive weight loss and productive cough. The diagnosis was confirmed by positive lung biopsy culture, serum cryptococcal antigen, and cerebral histopathology demonstrating encapsulated yeasts. Molecular characterisation of lung culture isolate yielded Cryptococcus neoformans var. grubii. An immune-deficiency could not be demonstrated.

Die Kryptokokkose ist eine Pilzinfektion, die meist durch Cryptococcus neoformans verursacht wird. Neben den seltener werdenden Erkrankungen bei HIV-Infizierten werden zunehmend Falle bei anderen Patientenkollektiven wie Organtransplantierten, Patienten mit chronischen Organerkrankungen oder auch bei Patienten ohne Immundefekt diagnostiziert. Das Krankheitsbild reicht von lokalisierten Infektionen der Atemwege oder der Haut bis hin zur typischen Meningoenzephalitis nach hamatogener Dissemination. Da aber auch alle anderen Organe beteiligt sein konnen, resultiert eine vielgestaltige Prasentation der Erkrankung. Daher muss die Kryptokokkose oft in die differenzialdiagnostischen Uberlegungen einbezogen werden. Aufgrund sensitiver Labortests kann die Erkrankung bei disseminierter Infektion rasch gesichert werden. Fur therapeutische Entscheidungen sind Patientenfaktoren und die jeweilige Organbeteiligung ausschlaggebend. Wahrend in der ersten Therapiephase bei disseminierten oder schweren lokalen Erkrankungen in der Regel Amphotericin B-Derivate und 5-Flucytosin in Kombination eingesetzt werden, ist nach dieser Induktionstherapie oder bei leichten lokalisierten Infektionen eine Therapie mit Fluconazol indiziert. Echinocandine sind bei der Kryptokokkose nicht wirksam.

As part of the multicentre Antibiotic Therapy Optimisation Study (ATHOS), minimum inhibitory concentrations (MICs) were determined for cephalosporins alone and in combination with the β-lactamase inhibitors tazobactam, clavulanic acid and avibactam against third-generation cephalosporin-resistant Escherichia coli, Klebsiella spp. and Enterobacter spp. isolates collected in German hospitals. MIC50/90 values were 0.25–4 mg/L for cefepime/tazobactam, 0.25–2 mg/L for ceftazidime/avibactam, 0.125–0.5 mg/L for ceftaroline/avibactam, 0.5–4 mg/L for cefpodoxime/clavulanic acid and 0.25–1 mg/L for aztreonam/avibactam, depending on the underlying resistance mechanism and organism. Based on in vitro testing, β-lactam antibiotics play an important role in the treatment of infections due to β-lactamase-producing organisms.

The National Reference Centre for Staphylococci and Enterococci in Germany has received an increasing number of clinical linezolid-resistant E. faecium isolates in recent years. Five isolates harbored a cfr(B) variant gene locus the product of which is capable of conferring linezolid resistance. The cfr(B)-like methyltransferase gene was also detected in Clostridium difficile. Antimicrobial susceptibility was determined for cfr(B)-positive and linezolid-resistant E. faecium isolates and two isogenic C. difficile strains. All strains were subjected to whole genome sequencing and analyzed with respect to mutations in the 23S rDNA, rplC, rplD and rplV genes and integration sites of the cfr(B) variant locus. To evaluate methyltransferase function, the cfr(B) variant of Enterococcus and Clostridium was expressed in both E. coli and Enterococcus spp. Ribosomal target site mutations were detected in E. faecium strains but absent in clostridia. Sequencing revealed 99.9% identity between cfr(B) of Enterococcus and cfr of Clostridium. The methyltransferase gene is encoded by transposon Tn6218 which was present in C. difficile Ox3196, truncated in some E. faecium and absent in C. difficile Ox3206. The latter finding explains the lack of linezolid and chloramphenicol resistance in C. difficile Ox3206 and demonstrates for the first time a direct correlation of elevated linezolid MICs in C. difficile upon cfr acquisition. Tn6218 insertion sites revealed novel target loci for integration, both within the bacterial chromosome and as an integral part of plasmids. Importantly, the very first plasmid-association of a cfr(B) variant was observed. Although we failed to measure cfr(B)-mediated resistance in transformed laboratory strains the occurrence of the multidrug resistance gene cfr on putatively highly mobile and/or extrachromosomal DNA in clinical isolates is worrisome with respect to dissemination of antibiotic resistances.

Automation of plate streaking is ongoing in clinical microbiological laboratories, but evaluation for routine use is mostly open. In the present study, the recovery of microorganisms from the Previ Isola system plated polyurethane (PU) swab samples is compared to manually plated control viscose swab samples from wounds according to the CLSI procedure M40-A (quality control of microbiological transport systems). One hundred twelve paired samples (224 swabs) were analyzed. In 80/112 samples (71%), concordant culture results were obtained with the two methods. In 32/112 samples (29%), CFU recovery of microorganisms from the two methods was discordant. In 24 (75%) of the 32 paired samples with a discordant result, Previ Isola plated PU swabs were superior. In 8 (25%) of the 32 paired samples with a discordant result, control viscose swabs were superior. The quality of colony growth on culture media for further investigations was superior with Previ Isola inoculated plates compared to manual plating techniques. Gram stain results were concordant between the two methods in 62/112 samples (55%). In 50/112 samples (45%), the results of Gram staining were discordant between the two methods. In 34 (68%) of the 50 paired samples with discordant results, Gram staining of PU swabs was superior to that of control viscose swabs. In 16 (32%) of the 50 paired samples, Gram staining of control viscose swabs was superior to that of PU swabs. We report the first clinical evaluation of Previ Isola automated specimen inoculation for wound swab samples. This study suggests that use of an automated specimen inoculation system has good results with regard to CFU recovery, quality of Gram staining, and accuracy of diagnosis.

In October 2009, a 55-year-old male with diabetes mellitus, probably since 2007, peripheral artery occlusive disease (PAOD) without any notion of peripheral neuropathy, and a history of alcoholism in 2003 presented with right lower leg pain accompanied by local hyperthermia, swelling, and reddening without fever. The medical history revealed that, in August 2000, the patient had suffered from a grade III open comminuted tibia fracture associated with a compartment syndrome. The fracture had been fixed using intramedullary nailing and fasciotomy at the lower leg had been performed. An intraoperative swab had yielded aerobic spore-forming bacteria after enrichment culture without further differentiation, but no anaerobic bacteria. In August 2001, the patient presented with a pus-filled fistula at the distal third of the lower leg. At this time point, the intramedullary nail was removed and the tibia was reamed for the treatment of osteomyelitis. Staphylococcus aureus (S. aureus) was grown from intraoperative specimens with resistance to penicillin and ampicillin only. Intravenous cefuroxime was given for 34 days. After removal of the implant, including reaming of the long bone and aggressive debridement, the fistula at the lower leg healed slowly. In the following 8 years, no event of trauma, manipulation, or surgery to the right lower leg was recorded. Upon readmission, plain radiographs of the right lower leg showed the radiological sequelae of comminuted lower leg fracture with full consolidation of the fracture lines. Typical signal alterations were found at magnetic resonance imaging (MRI) scanning within the medullary canal, including enhancement after the application of contrast agent, suggesting osteomyelitis within the distal third of the tibia. The tibia was opened proximally and a phlegmon of the medullary cavity was drained. Aggressive wound debridement, including reaming and jet lavage, was done. A rod was formed from polymethylmethacrylate (PMMA) bone cement and introduced as a local drug delivery system for gentamicin. Intraoperative swab of the right tibia and tissue specimens taken from the medullary cavity tissue revealed a Gram-positive anaerobic rod. The bacterium was identified as Clostridium clostridioforme (C. clostridioforme) at 99.8% using API rapid 32 A strips (bioMerieux, France). In contrast, matrix-assisted laser desorption/ionization timeof-flight (MALDI-TOF) identification detected C. sphenoides with an identification score of 1.6. For exact species determination, 16S rRNA gene sequence comparisons were performed (GATC, Germany), and the isolated bacterium was identified as C. celerecrescens (99% identity from a 512-nt fragment). Antibiotic susceptibilities were determined by Etest (bioMerieux, France). The bacillus was susceptible to amoxicillin/clavulanic acid (minimum inhibitory concentration [MIC] 2 mg/L), clindamycin (MIC 2 mg/L), imipenem (MIC 2 mg/L), and metronidazole (MIC 0.06 mg/L). To provide broad coverage, a treatment including cefuroxime (1,500 mg three times/day) plus metronidazole (400 mg three times/day) with intravenous administration for 15 days was begun. Twelve days later, the tibia was revised. The bone cement rod was removed and repetitive surgical debridement, including A. Mischnik (&) S. Zimmermann I. Bekeredjian-Ding Department of Infectious Diseases, University of Heidelberg, Medical Microbiology and Hygiene, Im Neuenheimer Feld 324, 69120 Heidelberg, Germany e-mail: alexander.mischnik@med.uni-heidelberg.de

International guidelines for antibiotic treatment of spontaneous bacterial peritonitis (SBP) are based on studies conducted decades ago and do not reflect regional differences of bacterial epidemiology.We retrospectively analyzed epidemiology of agents, antibiotic resistance patterns, and survival in liver cirrhosis patients with their first episode of SBP during the years 2007-2013.Of the 311 patients included, 114 patients had a positive ascites culture, and 197 had an ascitic neutrophil count250 μL. Gram-positive bacteria (47.8%) were more frequently found than Gram-negatives (44.9%), fungi in 7.2%. Enterobacter spp. (40.6%), Enterococcus spp. (26.1%), and Staphylcoccus spp. (13.8%) were the most frequently isolated agents. Third-generation cephalosporins covered 70.2% of non-nosocomial and 56.3% of nosocomial-acquired SBP cases.When SBP was diagnosed by a positive ascitic culture, survival was highly significantly reduced (mean: 13.9 ± 2.9 months; 95% confidence interval [CI]: 8.1-19.8) compared with culture-negative SBP patients (mean: 44.1 ± 5.4 months; 95% CI: 33.4-54.9; P = 0.000). Along with model of end-stage liver disease score and intensive care unit contact, a positive ascites culture remained an independent risk factor associated with poor survival (odds ratio: 1.49; 95% CI: 1.09-2.03) in multivariate analysis; piperacillin/tazobactam proved to be an adequate antibiotic for nosocomial and non-nosocomial SBP in 85.1% and 92.5%, respectively. SBP infection with Enterococcus spp. was associated with poor patient survival (P = 0.048).Third-generation cephalosporins have poor microbial coverage for treatment of SBP. Current guidelines need to adapt for the emerging number of Gram-positive infectious agents in SBP patients.

Complicated infections caused by vancomycin-resistant enterococci (VRE) represent a therapeutic challenge, since adequate treatment options are limited and antibiotic resistance to the available agents has already been described. The most frequently used antibiotic in VRE treatment is linezolid. Tigecycline is an alternative to linezolid, however, clinical data for severe infections such as sepsis or endocarditis are scarce. Daptomycin on the one hand is an option but has not yet been approved for the treatment of enterococcal infections in Germany on the other hand. The present review critically evaluates the clinical significance of the antibiotics in question for VRE therapy based on existing data.

Linezolid is an antibiotic of last resort for the treatment of infections with vancomycin-resistant enterococci (VRE). Here we report the increasing prevalence of linezolid resistance among clinical Enterococcus faecium strains from German hospital patients. Linezolid minimum inhibitory concentrations (MICs) were determined for 4461 clinical E. faecium strains isolated between 2008 and 2014. Isolates originated from the network of diagnostic laboratories collaborating with the National Reference Centre (NRC) for Staphylococci and Enterococci covering all German federal states. All linezolid-resistant isolates were determined by broth microdilution and confirmed by Etest as well as by analysing the 23S rDNA for putative mutations. Marker genes were determined by PCR. Genotyping was performed by SmaI macrorestriction analysis in pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) for selected isolates. An increase in linezolid resistance was observed, from1% in 2008 to9% in 2014. Occasionally, outbreaks with linezolid-resistant VRE (ST117) were observed. In total, 232 (92.4%) of 251 linezolid-resistant E. faecium isolates (including 61 vanA and 29 vanB) contained the G2576T 23S rDNA mutation and showed a varying mixture of wild-type and mutated alleles per genome sufficient to confer linezolid resistance. In vitro growth experiments revealed a stable linezolid MIC. Of the 251 linezolid-resistant isolates, 5 were cfr-positive. In conclusion, these NRC data identified a country-wide ongoing trend of increasing linezolid resistance among clinical E. faecium isolates within the last 5 years.

A case of a patient suffering from tuberculosis of the pubic symphysis and pubic bone is presented. There were no symptoms other than pain in the right groin area for 12 months. An X-ray of the pelvis showed an osteodestructive lesion of the pubic bone, and an MRI revealed an abscess formation of the pubic symphysis. Tissue samples were collected via CT-guided needle biopsy. Histological evaluation of tissue and analysis by PCR prompted the diagnosis of musculoskeletal tuberculosis. Despite antituberculous chemotherapy according to the current guidelines, the osteodestructive lesion progressed. This case highlights the difficulty of treating bone infections in general. Moreover, Mycobacterium tuberculosis as a rare causative agent of bone infections is discussed.

Tuberculosis (TB) infection is a major concern in patients with chronic autoimmune conditions under immunosuppressive therapy. Gastrointestinal tuberculosis can be misdiagnosed as Crohn’s disease with detrimental consequences for the patient. We report on a 40-year old ethnic Turkish patient with HLA-B27 positive spondyloarthritis who developed gastrointestinal symptoms under immunosuppressive treatment with infliximab. Crohn’s disease was diagnosed at a primary care hospital and immunosuppressive treatment was escalated. Initial diagnostic tests for tuberculosis were negative. When the clinical condition deteriorated, the patient was transferred to our intensive care unit for further diagnosis and treatment. Tuberculosis was suspected due to clinical presentation and radiological signs and anti-tuberculous treatment was initiated. After the onset of treatment, first microbiological results confirmed the diagnosis of miliary TB with Mycobacterium bovis. As an infection route we assume primary gastrointestinal infection with M. bovis during the patient’s annual holidays in Turkey with a rapid development of miliary TB under infliximab and escalated immunosuppressive therapy. This case report demonstrates the difficulties in differentiating intestinal TB from other granulomatous conditions such as Crohn’s disease. The diagnostic tools for gastrointestinal tuberculosis are discussed in detail regarding their sensitivity, specificity as well as positive and negative predictive values.

Salmonella enterica serovar Minnesota is a rarely isolated organism in clinical samples mainly grown from stool cultures. Sepsis due to Salmonella is known in severely immunocompromised patients, but so far urosepsis due to S. enterica serovar Minnesota has not been described. We report a case of a 31-year-old patient suffering from Crohn’s disease treated with infliximab and azathioprine, in whom was implanted a double-J ureteric catheter for urolithiasis. The patient presented with urinary tract infection and severe sepsis. S. enterica serovar Minnesota was grown from urine and blood cultures. After empiric antimicrobial treatment with meropenem and vancomycin, treatment was changed to ceftriaxone. Antimicrobial treatment was continued for a total of 3 weeks without evidence of Salmonella recurrence on follow-up visits. Salmonella spp. rarely cause urinary tract infection and sepsis. However, in immunocompromised patients, non-typhoidal salmonellosis merits a thorough clinical and microbiological evaluation.

Septicaemia and septic arthritis due to Yersinia pseudotuberculosis are rare diseases with high mortality rates. Reactive arthritis caused by Yersinia infection is a well-known complication but septic arthritis is found at a much lower frequency. It has already been established that there is a relationship between yersiniosis and iron but there are currently no data about yersiniosis and haematological disorders such as haemophilia. We report a case of septic arthritis due to Y. pseudotuberculosis as an early manifestation of Yersinia septicaemia in a patient with severe haemophilia A. The patient had no history of immunosuppression and presented with a repeat case of haemarthrosis with a fever of unknown origin. Furthermore, he suffered from acute-on-chronic renal failure and non-ST segment elevation myocardial infarction. Arthrocentesis and blood culture tested positive for Y. pseudotuberculosis. Iron deposits at localized sites in the synovium in patients with haemophilia have been described, and as Yersinia infections are common in patients with secondary iron overload, we felt that a review of the literature was in order. Severe Yersinia infection is often associated with iron overload, a condition that might occur as a side effect in the treatment of haemophilia. Iron overload, which plays an important role in the pathogenesis of haemophilic arthropathy, may have increased the virulence of Y. pseudotuberculosis in the present case.

Sepsis is a major cause of mortality in hospitalized patients worldwide, with lethality rates ranging from 30 to 70 %. Sepsis is caused by a variety of different pathogens, and rapid diagnosis is of outstanding importance, as early and adequate antimicrobial therapy correlates with positive clinical outcome. In recent years, matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) fingerprinting has become a powerful tool in microbiological diagnostics. The direct identification of micro-organisms in a positive blood culture by MALDI-TOF MS can shorten the diagnostic procedure significantly. Therefore, the aim of the present study was to evaluate whether identification rates could be improved by using the new Sepsityper kit from Bruker Daltonics for direct isolation and identification of bacteria from positive blood cultures by MALDI-TOF MS compared with the use of conventional separator gel columns, and to integrate the MALDI-TOF MS-based identification method into the routine course of blood culture diagnostics in the setting of a microbiological laboratory at a university hospital in Germany. The identification of Gram-negative bacteria by MALDI-TOF MS was significantly better using the Sepsityper kit compared with a separator gel tube-based method (99 and 68 % correct identification, respectively). For Gram-positive bacteria, only 73 % were correctly identified by MALDI-TOF with the Sepsityper kit and 59 % with the separator gel tube assay. A major problem of both methods was the poor identification of Gram-positive grape-like clustered cocci. As differentiation of Staphylococcus aureus from coagulase-negative staphylococci is of clinical importance, a PCR was additionally established that was capable of identifying S. aureus directly from positive blood cultures, thus closing this diagnostic gap. Another benefit of the PCR approach is the possibility of directly detecting the genes responsible for meticillin resistance in staphylococci and for vancomycin resistance in enterococci, which is of high importance for early adequate treatment. Both of the described methods were finally integrated into a protocol for fast and effective identification of bacteria from positive blood cultures.