Characteristics of two zoonotic swine influenza A(H1N1) viruses isolated in Germany from diseased patients.

Interspecies transmission of influenza A viruses (IAV) from pigs to humans is a concerning event as porcine IAV represent a reservoir of potentially pandemic IAV. We conducted a comprehensive analysis of two porcine A(H1N1)v viruses isolated from human cases by evaluating their genetic, antigenic and virological characteristics. The HA genes of those human isolates belonged to clades 1C.2.1 and 1C.2.2, respectively, of the A(H1N1) Eurasian avian-like swine influenza lineage. Antigenic profiling revealed substantial cross-reactivity between the two zoonotic H1N1 viruses and human A(H1N1)pdm09 virus and some swine viruses, but did not reveal cross-reactivity to H1N2 and earlier human seasonal A(H1N1) viruses. The solid-phase direct receptor binding assay analysis of both A(H1N1)v showed a predominant binding to α2-6-sialylated glycans similar to human-adapted IAV. Investigation of the replicative potential revealed that both A(H1N1)v viruses grow in human bronchial epithelial cells to similar high titers as the human A(H1N1)pdm09 virus. Cytokine induction was studied in human alveolar epithelial cells A549 and showed that both swine viruses isolated from human cases induced higher amounts of type I and type III IFN, as well as IL6 compared to a seasonal A(H1N1) or a A(H1N1)pdm09 virus. In summary, we demonstrate a remarkable adaptation of both zoonotic viruses to propagate in human cells. Our data emphasize the needs for continuous monitoring of people and regions at increased risk of such trans-species transmissions, as well as systematic studies to quantify the frequency of these events and to identify viral molecular determinants enhancing the zoonotic potential of porcine IAV.

Evaluation of 17 years of MERIN (Meningitis and Encephalitis register in Lower Saxony, Germany) surveillance system: participants acceptability survey, completeness and timeliness of data.

BACKGROUND: A Meningitis and Encephalitis Surveillance (MERIN) was implemented in 2003 in Lower Saxony, Germany as an alternative to acute flaccid paralyses surveillance, as the latter did not reach WHO sensitivity criteria. The system provides information on circulating enterovirus (EV) serotypes by focussing on patients with suspected aseptic meningitis, encephalitis or acute flaccid paralysis and contributes to the national surveillance in documenting polio free status. MERIN is based on voluntary participation of hospitals. Therefore, our evaluation focusses on acceptability of the system's objectives and performance, and identifying areas for improvement. METHODS: To assess acceptability, 32 contributing hospitals were invited to an online-based survey (11/2021 to 01/2022) to rate the MERIN objectives, laboratory's performance, their workload, modes of processes and communication. Ideas for improvement were collected in open fields. In addition, data completeness and timeliness of laboratory diagnostics were assessed. RESULTS: Of 32 hospitals, 21 responded (66% response rate), sending 30 questionnaires, 25 from pediatric and 5 from neurological departments. High levels of satisfaction with the communication (≥ 96%), timeliness (≥ 81%), and distribution of the results (≥ 85%) were reported, 97% of participants judged the required workload as adequate. The median proportion of eligible patients included in MERIN was 75%. Participants gave rapid and reliable diagnostic testing the highest priority (96%), while monitoring of Germany's polio-free status was rated the lowest (61%). Providing medical reports digitally as well as regular updates about circulating EV serotypes were identified as areas for improvement. Data completeness of selected variables ranged from 78.3 to 99.9%. Median time between sample collection and arrival at laboratory was 2 days [IQR 1-3], EV diagnostics via PCR took one day [IQR 0-6] and EV isolation on cell culture 11 days [IQR 10-13]. CONCLUSION: MERIN is a highly accepted surveillance system. Its quality was enhanced further by addressing the suggested improvements such as regular reports on circulating EV serotypes and facilitating digital access to laboratory results. Our results emphasise the importance of recognizing and considering participants' motivations and expectations, and addressing their priorities, even if this is not the surveillance system's main focus. This approach can be applied to surveillance systems of other non-mandatory notifiable diseases.

Risk factors for Leptospira seropositivity in rural Northern Germany, 2019.

We investigated seroprevalence and factors associated with Leptospira spp. infections in humans in rural Northern Germany. Sera of 450 participants were tested for leptospira-reactive IgG antibodies by two enzyme-linked immunosorbent assays (ELISA). A narrow (specific) and a broad (sensitive) case definition were applied and results compared in the analysis. Personal data were collected via questionnaire and associations with the serostatus were investigated by multivariable logistic regression. The seroprevalence estimates were 1.6% (95%-confidence interval (CI) = 0.63-3.2) under the narrow and 4.2% (95%-CI = 2.6-6.5%) under the broad case definition. Few (14%) participants knew about the pathogen. No seropositive participant recalled a prior leptospirosis diagnosis. Spending more than two hours a week in the forest was significantly associated with anti-leptospira IgG in both models (broad case definition: adjusted odds ratio (aOR) = 2.8, 95%-CI = 1.2-9.1; narrow case definition: aOR = 11.1, 95%-CI = 1.3-97.1). Regular cleaning of storage rooms was negatively associated in the broad (aOR = 0.17, 95%-CI = 0.03-0.98) and touching a dead rodent in the past 10 years in the narrow case definition model (aOR = 0.23, 95%-CI = 0.05-1.04). Our findings support risk factors identified in previous investigations. To counter the low awareness for the pathogen, we recommend that health authorities communicate risks and preventive measures to the public by using target-group specific channels.

New and Confirmed Foci of Tick-Borne Encephalitis Virus (TBEV) in Northern Germany Determined by TBEV Detection in Ticks.

Tick-borne encephalitis (TBE) is a tick-transmitted, virus-induced neurological disease with potentially fatal outcomes in humans and animals. Virus transmission takes places in so-called tick-borne encephalitis virus (TBEV) microfoci, which constitute small areas of sustained virus circulation. In southern Germany, TBEV has been endemic for decades; however, a northward expansion of risk areas, based on disease incidence in the human population, has been observed in recent years. The present study investigated TBEV occurrence in questing ticks at eight locations in the federal state of Lower Saxony, northwestern Germany, chosen due to reported associations with human TBE cases (N = 4) or previous virus detection (N = 4). A total of 20,056 ticks were collected in 2020 and 2021 and tested for TBEV RNA in pools of ten nymphs or five adults by quantitative reverse transcription-PCR (RT-qPCR). Positive results were confirmed by RT amplification of the viral E gene. In total, 18 pools from five different sampling locations were positive for TBEV RNA. One previously unknown transmission focus was detected, while ongoing virus circulation was confirmed at the four further locations. Phylogenetic analysis showed that two different virus strains with different origins circulate in the locations identified as natural foci.

[Four years after the Ebola crisis : Challenges, experiences, and implications in the German public health context]. / Vier Jahre nach der Ebolakrise : Herausforderungen, Erfahrungen und Schlussfolgerungen für den Öffentlichen Gesundheitsdienst in Deutschland.

The Ebola virus disease outbreak in West Africa in 2014/2015 was by far the biggest, most prolonged, and geographically most widespread outbreak of this disease since the discovery of the Ebola virus in 1976. Although no cases of Ebola virus disease were confirmed in Germany, a number of crisis management activities were initiated.Based on a combination of local, national, and international lessons learned, literature research, and a large number of discussions among German colleagues as well as German and foreign colleagues, the experiences of selected German public health actors as well as implications for health protection activities in Germany are presented.On the one hand, preparedness for managing unusual high consequence health events-caused by rare, highly pathogenic biological agents-including the provision of adequate material and personnel resources remains important in Germany. On the other hand, more German engagement in global health is necessary, because the dividing line between global health and local health is increasingly disappearing.

First Isolation and Phylogenetic Analyses of Tick-Borne Encephalitis Virus in Lower Saxony, Germany.

Tick-borne encephalitis (TBE) is the most important tick-borne arboviral disease in Europe. Presently, the main endemic regions in Germany are located in the southern half of the country. Although recently, sporadic human TBE cases were reported outside of these known endemic regions. The detection and characterization of invading TBE virus (TBEV) strains will considerably facilitate the surveillance and assessment of this important disease. In 2018, ticks were collected by flagging in several locations of the German federal state of Lower Saxony where TBEV-infections in humans (diagnosed clinical TBE disease or detection of TBEV antibodies) were reported previously. Ticks were pooled according to their developmental stage and tested for TBEV-RNA by RT-qPCR. Five of 730 (0.68%) pools from Ixodes spp. ticks collected in the areas of "Rauher Busch" and "Barsinghausen/Mooshuette" were found positive for TBEV-RNA. Phylogenetic analysis of the whole genomes and E gene sequences revealed a close relationship between the two TBEV isolates, which cluster with a TBEV strain from Poland isolated in 1971. This study provides first data on the phylogeny of TBEV in the German federal state of Lower Saxony, outside of the known TBE endemic areas of Germany. Our results support the hypothesis of an east-west invasion of TBEV strains in Western Europe.

Feasibility of a birth cohort study dedicated to assessing acute infections using symptom diaries and parental collection of biomaterials.

BACKGROUND: A birth cohort dedicated to studying infections in early childhood may be assisted by parental recording of symptoms on a daily basis and a collection of biomaterials. We aimed at testing the feasibility of this approach for use in a long-term study focusing on infections in children in Germany. METHODS: Parents of 1- to 3-year-old children (n = 75) were recruited in nursery schools. They were asked to complete a symptom diary on a daily basis and to take monthly and symptom-triggered nasal swabs and stool samples from their child over the study period of three months. Feasibility was measured by means of the return proportions of symptom diaries and bio samples; acceptance was assessed by a questionnaire delivered to participants at the end of the study. RESULTS: The majority of the participants filled in the symptom diary during the three months study for 75 or more days (77.3%), and provided the monthly nasal swabs (62.7%) and stool samples (65.3%). The time needed for the tasks was acceptable for most participants (symptom diary: 92.3%, nasal swabs: 98.5%, stool samples: 100.0%). In 64.3% of the symptom-triggered nasal swabs, respiratory viruses were found compared to 55.5% in throat swabs taken by health-care professionals within the "ARE surveillance Lower Saxony", a special project by the Governmental Institute of Public Health of Lower Saxony to investigate causal pathogens for acute respiratory infections in children. CONCLUSIONS: The parental assessment of symptoms and collection of biomaterials in a birth cohort dedicated to studying infections appears feasible in a middle class German population. The success of the study will depend on the ability to maintain these activities over a long time period.

Hepatitis B outbreak in a nursing home associated with reusable lancet devices for blood glucose monitoring, Northern Germany 2010.

In September 2010, an outbreak of acute hepatitis B virus (HBV) infections in a nursing home was notified to public health authorities in Northern Germany. To identify the route of transmission and prevent further cases a retrospective cohort study was conducted. Blood samples of residents were tested for serologic markers of HBV infection and HBV subgenotypes and sequences were analyzed. Outbreak-related cases were defined as residents of the nursing home with detection of hepatitis B surface antigen (HBsAg) and the HBV DNA sequence of the outbreak strain in 2010. Information on possible risk factors as patient care, invasive diagnostic, and therapeutical procedures was collected using a standardized questionnaire. Risk ratios (RR) and 95% confidence intervals (CI) were estimated with exact Poisson regression and binomial regression. Sixty-four residents were included in the study, 5 of them were outbreak-related cases, 12 had a past HBV infection. The outbreak strain belonged to HBV genotype D2 (HBsAg subtype ayw3, Ala118) which is not prevalent in Germany but in Eastern Europe. All cases (median age 81) were female, had diabetes, blood glucose monitoring, and chiropody. In multivariable analysis only blood glucose monitoring was associated with HBV infection (RR = 22, 95%CI 3.0-∞). Blood glucose monitoring was reported to be done by nursing home staff with patient-based reusable lancet devices. In nursing home settings the use of single use lancets for blood glucose monitoring is recommended strongly to prevent transmission. National guidelines on the handling of point-of-care devices and reusable equipment in long-term care facilities should be developed.

The first German map of georeferenced ixodid tick locations.

BACKGROUND: Georeferenced locations of ixodid ticks are required to depict the observed distribution of species. Further, they are used as input data for species distribution models also known as niche models. The latter were applied to describe current and future (projected) tick distributions. Beside model assumptions and selected climate parameters, the number of georeferenced tick locations available as a digital dataset is of fundamental importance for the reliability of such models. For Germany, however, no comprehensive dataset of ixodid tick species exists. The goal of this study was to put together all the available information on ixodid tick locations in Germany to produce such a digital dataset and to visualize it in a map. FINDINGS: A total of 2,044 georeferenced locations of ixodid ticks in Germany were compiled from two existing datasets (altogether 993 locations) and an extensive literature study (1,051 locations). The resulting digital dataset comprises the following tick species: Ixodes ricinus (1,855 locations), Ixodes apronophorus (1), Ixodes frontalis (1), Ixodes hexagonus (1), Ixodes trianguliceps (4), Dermacentor marginatus (77), Dermacentor reticulatus (96), Haemaphysalis concinna (8) and Hyalomma marginatum (1). The data were used to draw a tick map for Germany, showing I. ricinus occurring in the whole federal territory, while D. marginatus has been restricted to the climatically favoured region of the Rhine valley. Clustered locations of D. reticulatus were also documented in the Rhine valley as well as in Berlin and its vicinity. CONCLUSIONS: The introduced map depicts for the first time the available geographical coordinates of ixodid tick locations in Germany. The digital dataset used to draw the map is provided to the scientific community as a basis for further investigations such as species distribution modelling.

High rate of spontaneous clearance of acute hepatitis C virus genotype 3 infection.

Treating acute hepatitis C with interferon alpha prevents chronicity in nearly all cases when therapy is initiated within 3 months after infection. However, 15-50% of untreated patients may clear the hepatitis C virus (HCV) spontaneously. Therefore, factors are needed to identify patients prior to therapy who have a higher or lower risk for developing a chronic course to avoid unnecessary treatment. The role of the HCV genotype for spontaneous recovery from acute hepatitis C has been discussed controversially. In the year 2002, all 1,176 new incoming prisoners in a Northern German prison for young men (age 16-24) were screened for anti-HCV antibodies and 92 tested positive. Ninety eight percent of these reported i.v.-drug abuse for a median of 32 months prior to imprisonment. HCV-RNA negative individuals (21%) were serotyped and HCV-RNA positive patients were genotyped. The prevalence of HCV genotype 3 was significantly higher among individuals who had cleared HCV spontaneously as compared to chronically infected patients (86% vs. 38%; P = 0.002). Ninety three percent of individuals exposed to HCV genotype 1 but only 63% of individuals exposed to genotype 3 experienced a chronic course of the infection (P = 0.006). Thus, acute infection in young Caucasian men with HCV genotype 3 leads more often to spontaneous clearance than infection with HCV genotype-1. Considering also the high chance of successful treatment of chronic HCV genotype 3 infection with pegylated-interferon in combination with ribavirin, we suggest not to treat acute hepatitis C genotype 3 infection early but rather to wait at least 3 months after the onset of symptoms when chronicity becomes likely.

Comparison of a shiga toxin enzyme-linked immunosorbent assay and two types of PCR for detection of shiga toxin-producing Escherichia coli in human stool specimens.

Shiga toxin (Stx)-producing Escherichia coli (STEC) is a major cause of sporadic cases of disease as well as serious outbreaks worldwide. The spectrum of illnesses includes mild nonbloody diarrhea, hemorrhagic colitis, and hemolytic-uremic syndrome. STEC produces one or more Stxs, which are subdivided into two major classes, Stx1 and Stx2. The ingestion of contaminated food or water, person-to-person spread, and contact with animals are the major transmission modes. The infective dose of STEC may be less than 100 organisms. Effective prevention of infection is dependent on rapid detection of the causative bacterial pathogen. In the present study, we examined 295 stool specimens for the presence of Stx-producing E. coli by three different methods: an Stx enzyme-linked immunosorbent assay, a conventional PCR assay, and a LightCycler PCR (LC-PCR) assay protocol recently developed by our laboratory at the Institute of Medical Microbiology at Hannover Medical School. Our intent was to compare these three methods and to examine the utility of the STEC LC-PCR protocol in a clinical laboratory. The addition of a control DNA to each sample to clearly discriminate inhibited specimens from negative ones enhanced the accuracy of the LC-PCR protocol. From our results, it can be concluded that LC-PCR is a very useful tool for the rapid and safe detection of STEC in clinical samples.