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Hepatitis E virus (HEV) is prevalent worldwide and can cause persistent infection with severe morbidity. Antiviral treatment approaches can lead to the emergence of viral variants encoding escape mutations that may impede viral clearance. The frequency of these variants remains unknown in the human population as well as environment due to limited comprehensive data on HEV diversity. In this study, we investigated the HEV prevalence and diversity of circulating variants in environmental samples, that is, wastewater and rivers from North-Rhine Westphalia, Germany. HEV prevalence could be determined with 73% of samples tested positive for viral RNA via qRT-PCR. Using high-throughput sequencing, we were able to assess the overall genetic diversity in these samples and identified the presence of clinically relevant variants associated with drug resistance. In summary, monitoring variants from environmental samples could provide valuable insights into estimating HEV prevalence and identifying circulating variants that can impact treatment outcome.

Background: Carbapenem-resistant Enterobacterales and Acinetobacter baumannii are of major concern in terms of infection prevention and control. This study evaluated factors that may increase the frequency of Enterobacterales and A. baumannii with carbapenem resistance (CR) in patients admitted to a German University Hospital for implementation of optimized infection control management.Methods: A five-year-retrospective epidemiological cohort analysis was conducted on anamnestic risk factors for carrying Enterobacterales and/or A. baumannii with CR in patients who were first tested positive for these species at University Hospital Frankfurt (UHF) between January 2013 and June 2018. Results: 364 patients were tested positive for Enterobacterales and/or A. baumannii with CR, resulting in n=400 bacterial isolates in total, with Klebsiella pneumoniae being the most frequently detected species (n=146/400; 36.5%; 95% confidence interval: 31.8-41.4). In patients who were tested positive for Enterobacterales and/or A. baumannii with CR, any hospital stay within the previous 12 months was the most frequently reported common factor (n=275/364; 75.5%; 70.8-79.9).Conclusion: A hospital stay within the previous 12 months, including hospitals in Germany and abroad, is a frequent characteristic of patients who tested positive for Enterobacterales and/or A. baumannii with CR. Upon admission, any previous hospital stay of the given patient within the previous 12 months should be determined. Infection control strategies such as screening measures need to be adapted to these patient groups in hospital settings. In order to reflect the varying determinants in "nosocomial" cases in greater detail, the existing criteria used to characterize "nosocomial detection" of gram-negative bacteria with CR should be reviewed., Hintergrund: Enterobacterales und/oder Acinetobacter baumannii mit Carbapenem-Resistenz (CR) stellen in Kliniken eine enorme medizinische Herausforderung dar. Die vorliegende Studie untersucht anamnestische Charakteristika von Patienten des Universitätsklinikums Frankfurt am Main (UKF), die zum Risiko einer Trägerschaft von Enterobacterales und/oder A. baumannii mit CR beitragen. Ziel der Untersuchung ist die Etablierung einer zielgerichteten Strategie zur Infektionskontrolle.Methode: Retrospektive epidemiologischen Kohortenanalyse bei Patienten, die zwischen Januar 2013 und Juni 2018 am UKF positiv auf mindestens eine Spezies Enterobacterales und/oder A. baumannii mit CR getestet wurden. Ergebnisse: Innerhalb des Untersuchungszeitraums wurden bei 364 Patienten 400 Isolate von Enterobacterales und/oder A. baumannii mit CR nachgewiesen. Hiervon am häufigsten vertreten war Klebsiella pneumoniae (n=146/400; 36,5%; 95%-Konfidenzintervall: 31,8-41,4). Die anamnestische Angabe mindestens eines Krankenhausaufenthalts "innerhalb der letzten 12 Monate" wurde am häufigsten berichtet (n=275/364; 75,5%; 70,8-79,9).Zusammenfassung: Da sich ein Krankenhausaufenthalt innerhalb der letzten 12 Monate mit Nachweis von Enterobacterales und/oder A. baumannii mit CR als das am häufigsten berichtete Risikomerkmal herausstellte, sollte das bei Aufnahme eines Patienten sorgfältig abgefragt werden. Krankenhaushygienische Strategien zur Infektionsprävention, z.B. Implementierung von Screeningprotokollen für Patienten mit diesem Risikofaktor, können hilfreich sein und sollten situationsadaptiert eingesetzt werden. Um die derzeit als "nosokomiale Nachweise" zu bewertenden Fälle richtig darstellen zu können, ist eine Überarbeitung der derzeitigen Definitionskriterien notwendig.

Aim: Glutaraldehyde (GDA) is an active ingredient in many instrument disinfectants and is effective against a broad spectrum of microorganisms. In the past, the virus-inactivating properties of these products were mainly claimed based on quantitative suspension tests with different test viruses. Recently, however, a European Norm EN 17111:2018 has been published which allows examination of instrument disinfectants in a surface carrier test, simulating practical conditions. Therefore, it is of interest to evaluate GDA for the ability to inactivate the viruses used in this European Norm as test viruses. Methods: The virucidal efficacy of GDA as the active ingredient in instrument disinfectants was evaluated with 4 different test viruses in a method simulating practical conditions (EN 17111:2018).Results: With a fixed exposure time of five minutes at 20°C, 100 ppm GDA were necessary to inactivate vaccinia virus, classifying it as a limited spectrum virucidal activity for pre-cleaning products. For adenovirus, 125 ppm GDA were required, whereas for murine norovirus as a surrogate for human norovirus, 4,000 ppm GDA were required for a significant reduction of viral titres. Both non-enveloped viruses must be tested to prove virucidal activity in EN 17111:2018. But even 4,000 ppm were not enough to yield a 4 log10 reduction of the murine parvovirus at 20°C. This virus is only required as a test virus using this method if temperatures >=40°C are used. Conclusion: GDA, as the active ingredient of many instrument disinfectants, shows virucidal efficacy at 20°C. The necessary concentrations are strongly dependent on the stability of the test viruses. Due to the high stability of murine norovirus, GDA levels of 4,000 ppm were required to inactivate this virus within the 5-minute exposure time., Zielsetzung: Glutaraldehyd (GDA) besitzt ein breites Wirkspektrum gegen Mikroorganismen und wird daher in vielen Mitteln zur Instrumentendesinfektion verwendet. In der Vergangenheit wurden virusinaktivierende Eigenschaften dieser Produkte hauptsächlich auf Grundlage quantitativer Suspensionstests mit verschiedenen Testviren ermittelt. Inzwischen wurde die europäische Norm EN 17111:2018 veröffentlicht, welche die Untersuchung der Instrumentendesinfektionsmittel in einem Oberflächenträgertest unter praktischen Bedingungen ermöglicht. Daher ist es von Interesse, diesen wichtigen Wirkstoff vieler Instrumentendesinfektionsmittel auf die Fähigkeit zur Inaktivierung von Viren zu untersuchen, die in dieser europäischen Norm als Testviren ausgewählt wurden.Methode: Die viruzide Wirksamkeit von GDA als Wirkstoff in Instrumentendesinfektionsmitteln wurde mit 4 verschiedenen Testviren in einer Methode mit Simulation praktischer Bedingungen überprüft (EN 17111:2018).Ergebnisse: Die Ergebnisse der Studie mit einer festen Expositionszeit von fünf Minuten bei 20°C zeigten, dass 100 ppm GDA für die Inaktivierung des Vaccinia-Virus erforderlich waren. Daraus resultiert die Einstufung als "begrenzt viruzid" für Vorreinigungsprodukte. Für das Adenovirus wurden 125 ppm GDA benötigt, während für das murine Norovirus als Surrogat für das humane Norovirus 4.000 ppm GDA für eine signifikante Verringerung der Virustiter erforderlich waren. Beide nicht umhüllten Viren müssen gemäß EN 17111:2018 getestet werden, um den Nachweis einer viruziden Aktivität zu erbringen. Aber selbst 4.000 ppm reichten nicht aus, um eine Reduktion des murinen Parvovirus um 4 log10 bei 20°C nachzuweisen. Dieses Virus wird mit dieser Methode aber nur bei Temperaturen >=40°C eingesetzt.Fazit: Zusammenfassend lässt sich sagen, dass GDA als Wirkstoff vieler Instrumentendesinfektionsmittel eine viruzide Wirksamkeit bei 20°C besitzt. Die notwendigen Konzentrationen hängen stark von der Stabilität der Testviren ab. GDA- Ko

In situ hybridization (ISH) is a technique to determine potential correlations between viruses and lesions. The aim of the study was to compare ISH techniques for the detection of various viruses in different tissues. Tested RNA viruses include atypical porcine pestivirus (APPV) in the cerebellum of pigs, equine and bovine hepacivirus (EqHV, BovHepV) in the liver of horses and cattle, respectively, and Schmallenberg virus (SBV) in the cerebrum of goats. Examined DNA viruses comprise canine bocavirus 2 (CBoV-2) in the intestine of dogs, porcine bocavirus (PBoV) in the spinal cord of pigs and porcine circovirus 2 (PCV-2) in cerebrum, lymph node, and lung of pigs. ISH with self-designed digoxigenin-labelled RNA probes

Objectives: Aspergillus fumigatus is the most prevalent filamentous fungus in the respiratory tract of patients with cystic fibrosis (CF). The aim of this prospective multicentre study was to investigate the prevalence of azole-resistant A. fumigatus (ARAF) in respiratory secretions from CF patients across Germany and to characterize ARAF isolates by phenotypic and molecular methods. Methods: Twelve tertiary care centres from Germany participated in the study. In total, 2888 A. fumigatus isolates from 961 CF patients were screened for ARAF by using azole-containing agar plates. Antifungal susceptibility testing of isolates was performed by broth microdilution according to EUCAST guidelines. Analysis of mutations mediating resistance was performed using PCR and sequencing of the cyp51A gene. Furthermore, genotyping by microsatellite PCR was performed. Results: Of a total of 2888 A. fumigatus isolates, 101 isolates from 51 CF patients were found to be azole resistant (prevalence per patient 5.3%). The Essen centre had the highest prevalence (9.1%) followed by Munich (7.8%), Munster (6.0%) and Hannover (5.2%). Most ARAF isolates (n = 89) carried the TR34/L98H mutation followed by eight G54E/R, one TR46/Y121F/T289A and one F219S mutation. In two isolates no mutation was found. Genotyping results showed no major clustering. Forty-five percent of CF patients with ARAF had previously received azole therapy. Conclusions: This is the first multicentre study analysing the prevalence of ARAF isolates in German CF patients. Because of a resistance rate of up to 9%, susceptibility testing of A. fumigatus isolates from CF patients receiving antifungal treatment should be part of standard diagnostic work-up.

Objectives: Aspergillus fumigatus is the most prevalent filamentous fungus in the respiratory tract of patients with cystic fibrosis (CF). The aim of this prospective multicentre study was to investigate the prevalence of azole-resistant A. fumigatus (ARAF) in respiratory secretions from CF patients across Germany and to characterize ARAF isolates by phenotypic and molecular methods.Methods: Twelve tertiary care centres from Germany participated in the study. In total, 2888 A. fumigatus isolates from 961 CF patients were screened for ARAF by using azole-containing agar plates. Antifungal susceptibility testing of isolates was performed by broth microdilution according to EUCAST guidelines. Analysis of mutations mediating resistance was performed using PCR and sequencing of the cyp51A gene. Furthermore, genotyping by microsatellite PCR was performed.Results: Of a total of 2888 A. fumigatus isolates, 101 isolates from 51 CF patients were found to be azole resistant (prevalence per patient 5.3%). The Essen centre had the highest prevalence (9.1%) followed by Munich (7.8%), Münster (6.0%) and Hannover (5.2%). Most ARAF isolates (n = 89) carried the TR34/L98H mutation followed by eight G54E/R, one TR46/Y121F/T289A and one F219S mutation. In two isolates no mutation was found. Genotyping results showed no major clustering. Forty-five percent of CF patients with ARAF had previously received azole therapy.Conclusions: This is the first multicentre study analysing the prevalence of ARAF isolates in German CF patients. Because of a resistance rate of up to 9%, susceptibility testing of A. fumigatus isolates from CF patients receiving antifungal treatment should be part of standard diagnostic work-up.