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Article ; Online: Next-generation diagnostics

Claudia Wylezich / Sten Calvelage / Kore Schlottau / Ute Ziegler / Anne Pohlmann / Dirk Höper / Martin Beer

Microbiome, Vol 9, Iss 1, Pp 1-

virus capture facilitates a sensitive viral diagnosis for epizootic and zoonotic pathogens including SARS-CoV-2

2021 Volume 13

Abstract: Abstract Background The detection of pathogens in clinical and environmental samples using high-throughput sequencing (HTS) is often hampered by large amounts of background information, which is especially true for viruses with small genomes. Enormous ... ...

Abstract	Abstract Background The detection of pathogens in clinical and environmental samples using high-throughput sequencing (HTS) is often hampered by large amounts of background information, which is especially true for viruses with small genomes. Enormous sequencing depth can be necessary to compile sufficient information for identification of a certain pathogen. Generic HTS combining with in-solution capture enrichment can markedly increase the sensitivity for virus detection in complex diagnostic samples. Methods A virus panel based on the principle of biotinylated RNA baits was developed for specific capture enrichment of epizootic and zoonotic viruses (VirBaits). The VirBaits set was supplemented by a SARS-CoV-2 predesigned bait set for testing recent SARS-CoV-2-positive samples. Libraries generated from complex samples were sequenced via generic HTS (without enrichment) and afterwards enriched with the VirBaits set. For validation, an internal proficiency test for emerging epizootic and zoonotic viruses (African swine fever virus, Ebolavirus, Marburgvirus, Nipah henipavirus, Rift Valley fever virus) was conducted. Results The VirBaits set consists of 177,471 RNA baits (80-mer) based on about 18,800 complete viral genomes targeting 35 epizootic and zoonotic viruses. In all tested samples, viruses with both DNA and RNA genomes were clearly enriched ranging from about 10-fold to 10,000-fold for viruses including distantly related viruses with at least 72% overall identity to viruses represented in the bait set. Viruses showing a lower overall identity (38% and 46%) to them were not enriched but could nonetheless be detected based on capturing conserved genome regions. The internal proficiency test supports the improved virus detection using the combination of HTS plus targeted enrichment but also points to the risk of cross-contamination between samples. Conclusions The VirBaits approach showed a high diagnostic performance, also for distantly related viruses. The bait set is modular and expandable according to ...
Keywords	Metagenomics virus detection ; Capture enrichment ; RNA baits ; VirBaits 1.0 ; Diagnostic tool ; Proficiency test ; Microbial ecology ; QR100-130
Subject code	500
Language	English
Publishing date	2021-02-01T00:00:00Z
Publisher	BMC
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Neurological disease suspected to be caused by tick‐borne encephalitis virus infection in 6 horses in Switzerland

Ioannis Magouras / Angelika Schoster / Nathalie Fouché / Vinzenz Gerber / Martin H. Groschup / Ute Ziegler / Raffael Fricker / Christian Griot / Andrea Vögtlin

Journal of Veterinary Internal Medicine, Vol 36, Iss 6, Pp 2254-

2022 Volume 2262

Abstract: Abstract Background Reports on acute tick‐borne encephalitis virus (TBEV) infections with signs of neurologic disease in horses are limited. Objectives To describe the epidemiological, clinical, and laboratory findings of suspected acute TBEV infections ... ...

Abstract	Abstract Background Reports on acute tick‐borne encephalitis virus (TBEV) infections with signs of neurologic disease in horses are limited. Objectives To describe the epidemiological, clinical, and laboratory findings of suspected acute TBEV infections in 6 horses. Animals Six horses originating from TBEV endemic regions of Switzerland were presented to equine hospitals with acute onset of neurologic disease between 2011 and 2019. Methods Retrospective case series. Horses with acute onset of signs of neurologic disease that were subjected to clinical and microbiological examinations to rule out infectious diseases affecting the central nervous system. Results All horses exhibited acute signs of neurologic disease including ataxia and proprioceptive deficits. Horses tested positive for TBEV using virus neutralization test and samples were further tested for TBEV‐specific IgM. The presence of TBEV‐specific IgM antibodies was confirmed in 5 horses (cases 1‐5, Laboratory Unit [LU] values ranging from 30 to 56). One horse (case no. 6) with an LU value just below the test threshold (LU = 22.3) was also included under the hypothesis that the horse was transitioning from acute to chronic infection. All horses originated from areas where humans with confirmed tick‐borne encephalitis reported to have been bitten by ticks. Conclusions and Clinical Importance Acute TBEV infection should be a differential diagnosis in horses with signs of neurologic disease and originating from TBEV endemic areas. The establishment of harmonized diagnostic criteria would help to overcome the diagnostic challenges associated with TBEV and other Flavivirus infections in horses.
Keywords	ataxia ; flavivirus ; meningoencephalitis ; ticks ; vector‐borne ; Veterinary medicine ; SF600-1100
Subject code	630
Language	English
Publishing date	2022-11-01T00:00:00Z
Publisher	Wiley
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Vaccination of Zoo Birds against West Nile Virus—A Field Study

Felicitas Bergmann / Dominik Fischer / Luisa Fischer / Heike Maisch / Tina Risch / Saskia Dreyer / Balal Sadeghi / Dietmar Geelhaar / Lisa Grund / Sabine Merz / Martin H. Groschup / Ute Ziegler

Vaccines, Vol 11, Iss 652, p

2023 Volume 652

Abstract: West Nile virus (WNV) is known to cause disease and death in humans and various animals worldwide. WNV has circulated in Germany since 2018. In 2020, four birds tested positive for the WNV genome at Zoopark Erfurt (Thuringia). Moreover, virus ... ...

Abstract	West Nile virus (WNV) is known to cause disease and death in humans and various animals worldwide. WNV has circulated in Germany since 2018. In 2020, four birds tested positive for the WNV genome at Zoopark Erfurt (Thuringia). Moreover, virus neutralization assays detected neutralizing antibodies (nAb) against WNV in 28 birds. In addition, nAb against WNV and Usutu virus (USUV) were found in 14 birds. To protect valuable animals and to reduce the risk of viral transmission from birds to humans, we performed a field study on WNV vaccination at the zoo. To conduct the study, 61 birds from the zoo were categorized into three groups and subjected to a vaccination regimen, where each bird received either 1.0 mL, 0.5 mL, or 0.3 mL of a commercial inactivated WNV vaccine three times. The vaccinations were administered at three-week intervals, or as per modified vaccination schedules. Furthermore, 52 birds served as non-vaccinated controls. Adverse vaccination reactions were absent. The greatest increase in nAb titres was observed in birds that received 1.0 mL of vaccine. However, pre-existing antibodies to WNV and USUV appeared to have a major effect on antibody development in all groups and in all bird species, whereas sex and age had no effect. After vaccination, no death was detected in vaccinated birds for more than 1 year.
Keywords	West Nile virus (WNV) ; inactivated vaccine ; zoological garden ; zoo bird ; Germany ; Medicine ; R
Subject code	590
Language	English
Publishing date	2023-03-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Longitudinal Study of the Occurrence of Usutu Virus and West Nile Virus Infections in Birds in a Zoological Garden in Northern Germany

Felicitas Bergmann / Martina Schmoock-Wellhausen / Christine Fast / Cora M. Holicki / Friederike Michel / Patrick Wysocki / Balal Sadeghi / Martin H. Groschup / Ute Ziegler

Pathogens, Vol 12, Iss 753, p

2023 Volume 753

Abstract: Usutu virus (USUV) and West Nile virus (WNV) are known to cause diseases and mortalities in bird populations. Since 2010/2011, USUV has circulated in Germany and spread nationwide, while WNV was only introduced into East Germany in 2018. The zoological ... ...

Abstract	Usutu virus (USUV) and West Nile virus (WNV) are known to cause diseases and mortalities in bird populations. Since 2010/2011, USUV has circulated in Germany and spread nationwide, while WNV was only introduced into East Germany in 2018. The zoological garden investigated is located in Northern Germany, where USUV infections in wild birds have been detected for several years. In this longitudinal study conducted over a four-year period, zoo birds were sampled biannually and screened for molecular and serological evidence of USUV and WNV. USUV genomes were detected in eight of the sampled birds and whole-genome sequences revealed the circulation of USUV lineages Europe 3 and Africa 3. Of the eight birds infected with USUV during the study period, four died after the infection, while four survived without displaying clinical signs. Furthermore, in a few of the birds, a USUV (re-)infection was confirmed on a serological level with three birds producing USUV-neutralizing antibodies (nAbs) over a period of four years. Nonetheless, in two birds sampled throughout this longitudinal study, neither a USUV nor a WNV infection was evident. In 2022, WNV nAbs were detected for the first time in a juvenile zoo bird, indicating the introduction of the virus into this region.
Keywords	USUV ; WNV ; bird ; zoo ; monitoring ; surveillance ; Medicine ; R
Subject code	590
Language	English
Publishing date	2023-05-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Tick-borne encephalitis virus (TBEV) antibodies in animal sera – occurrence in goat flocks in Germany, longevity and ability to recall immunological information after more than six years

Christine Klaus / Ute Ziegler / Donata Hoffmann / Franziska Press / Christine Fast / Martin Beer

BMC Veterinary Research, Vol 15, Iss 1, Pp 1-

2019 Volume 6

Abstract: Abstract Background TBE is an important tick-borne viral zoonosis in Europe and some parts of Asia. Humans can become infected by tick bite and in some cases also by consumption of nonpasteurized raw milk and raw milk products from ruminants. Serological ...

Abstract	Abstract Background TBE is an important tick-borne viral zoonosis in Europe and some parts of Asia. Humans can become infected by tick bite and in some cases also by consumption of nonpasteurized raw milk and raw milk products from ruminants. Serological investigations of milking flocks can help to assess the risk of TBEV infection for humans. 735 blood samples from 50 goat flocks from four federal states of Germany were tested by TBEV-VNT to assess a potential risk for TBEV infection. There are some gaps in the knowledge about immunity in animals, for example with regard to the longevity of TBEV immunity. Two goats and two sheep were immunized and TBEV antibody titers could be detected for up to 7 years. Furthermore, nothing is known about a possible long-lasting immunological memory that could quickly be reactivated by an additional contact to TBEV. Seven years after the first immunization two goats and two sheep as well as two naïve goats and two sheep were boostered and TBEV antibody titers followed. Results Only one sample in each of the three states was TBEV-antibody positive (VNT), albeit with low titers. However, in Baden-Württemberg seven samples were positive, among them four goats of the same flock. TBEV-antibody positive titers were detected in goats for up to 6 years and 10 months, in sheep for up to 4 years and 7 months. Seven years after immunization a clear immunological recall occurred in response to administration of one dose of vaccine in two goats and two sheep. Conclusion It can be concluded that in the tested flocks the risk of an alimentary TBEV infection was low. However, in one single flock a considerably higher risk must be assumed. Antibody titers in goats and sheep can last very long after contact to TBEV, albeit at a low level. This should be taken into consideration in cases where the risk of an alimentary infection is assessed in a flock by serological investigations. The immunological recall gives rise to the suspicion that the immunological memory after a first contact to TBEV lasts for many years, probably lifelong.
Keywords	Tick borne encephalitis ; Animal sera ; Virus neutralization test ; Flaviviridae ; Veterinary medicine ; SF600-1100
Subject code	630
Language	English
Publishing date	2019-11-01T00:00:00Z
Publisher	BMC
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Reconstruction of the molecular evolution of Usutu virus in Germany

Felicitas Bergmann / Cora M Holicki / Friederike Michel / Sabine Bock / Nelly Scuda / Grit Priemer / Susanne Kenklies / Timo Siempelkamp / Jasmin Skuballa / Claudia Sauerwald / Louise Herms / Aemero Muluneh / Martin Peters / Andreas Hlinak / Martin H Groschup / Balal Sadeghi / Ute Ziegler

PLoS Neglected Tropical Diseases, Vol 17, Iss 10, p e

Insights into virus emersion and circulation.

2023 Volume 0011203

Abstract: Usutu virus (USUV) is a mosquito-borne flavivirus that is widely distributed in southern and central Europe. The zoonotic virus circulates primarily between birds and mosquitoes, can, however, in rare cases infect other mammals including humans. In the ... ...

Abstract	Usutu virus (USUV) is a mosquito-borne flavivirus that is widely distributed in southern and central Europe. The zoonotic virus circulates primarily between birds and mosquitoes, can, however, in rare cases infect other mammals including humans. In the past, USUV has been repeatedly associated with mass mortalities in birds, primarily blackbirds and owls. Birds commonly succumb either due to the peracute nature of the infection or due to severe encephalitis. In Germany, USUV has spread rapidly since its first detection in 2010 in mosquitoes under the presence of susceptible host and vector species. Nonetheless, there is to date limited access to whole genome sequences resulting in the absence of in-depth phylogenetic and phylodynamic analyses. In this study, 118 wild and captive birds were sequenced using a nanopore sequencing platform with prior target enrichment via amplicons. Due to the high abundancy of Europe 3 and Africa 3 in Germany an ample quantity of associated whole genome sequences was generated and the most recent common ancestor could be determined for each lineage. The corresponding clock phylogeny revealed an introduction of USUV Europe 3 and Africa 3 into Germany three years prior to their first isolation in the avifauna in 2011 and 2014, respectively. Based on the clustering and temporal history of the lineages, evidence exists for the genetic evolution of USUV within Germany as well as new introductions thereof into the country.
Keywords	Arctic medicine. Tropical medicine ; RC955-962 ; Public aspects of medicine ; RA1-1270
Subject code	590
Language	English
Publishing date	2023-10-01T00:00:00Z
Publisher	Public Library of Science (PLoS)
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Serological Hendra Virus Diagnostics Using an Indirect ELISA-Based DIVA Approach with Recombinant Hendra G and N Proteins

Anne Balkema-Buschmann / Kerstin Fischer / Leanne McNabb / Sandra Diederich / Nagendrakumar Balasubramanian Singanallur / Ute Ziegler / Günther M. Keil / Peter D. Kirkland / Maren Penning / Balal Sadeghi / Glenn Marsh / Jennifer Barr / Axel Colling

Microorganisms, Vol 10, Iss 1095, p

2022 Volume 1095

Abstract: Since the identification of Hendra virus (HeV) infections in horses in Australia in 1994, more than 80 outbreaks in horses have been reported, and four out of seven spillover infections in humans had a fatal outcome. With the availability of a subunit ... ...

Abstract	Since the identification of Hendra virus (HeV) infections in horses in Australia in 1994, more than 80 outbreaks in horses have been reported, and four out of seven spillover infections in humans had a fatal outcome. With the availability of a subunit vaccine based on the HeV-Glycoprotein (HeV-G), there is a need to serologically D ifferentiate the I nfected from the V accinated A nimals (DIVA). We developed an indirect ELISA using HeV-G expressed in Leishmania tarentolae and HeV-Nucleoprotein (HeV-N) expressed in recombinant baculovirus-infected insect cells as antigens. During evaluation, we tested panels of sera from naïve, vaccinated and infected horses that either originated from a Hendra-virus free region, or had been pre-tested in validated diagnostic tests. Our data confirm the reliability of this approach, as HeV-N-specific antibodies were only detected in sera from infected horses, while HeV-G-specific antibodies were detected in infected and vaccinated horses with a high level of specificity and sensitivity. Given the excellent correlation of data obtained for German and Australian HeV-negative horses, we assume that this test can be applied for the testing of horse serum samples from a variety of geographical regions.
Keywords	Hendra virus ; horse ; ELISA ; vaccination ; infection ; DIVA ; Biology (General) ; QH301-705.5
Subject code	630
Language	English
Publishing date	2022-05-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Low levels of classical BSE infectivity in rendered fat tissue

Christine Fast / Markus Keller / Martin Kaatz / Ute Ziegler / Martin H. Groschup

Veterinary Research, Vol 49, Iss 1, Pp 1-

2018 Volume 5

Abstract: Abstract BSE infectivity in mesentery fat is most likely associated with embedded nervous tissue. To prove this mesentery containing celiac ganglion was taken from oral BSE infected cattle in different stages of the disease and from one control animal. ... ...

Abstract	Abstract BSE infectivity in mesentery fat is most likely associated with embedded nervous tissue. To prove this mesentery containing celiac ganglion was taken from oral BSE infected cattle in different stages of the disease and from one control animal. Fat was rendered according to standard tallow production methods and the prion infectivity therein analysed in transgenic mouse bioassay. Rendered fat of the clinical animal revealed low infectivity levels, whereas preclinical and control animals remained negative. This study, although not representative, provides a proof of principle, indicating the potential contamination of melted mesenteric fat by embedded nervous structures during standard tallow production.
Keywords	Veterinary medicine ; SF600-1100
Language	English
Publishing date	2018-12-01T00:00:00Z
Publisher	BMC
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Pathogenicity of West Nile Virus Lineage 1 to German Poultry

Cora M. Holicki / Friederike Michel / Ana Vasić / Christine Fast / Martin Eiden / Cristian Răileanu / Helge Kampen / Doreen Werner / Martin H. Groschup / Ute Ziegler

Vaccines, Vol 8, Iss 507, p

2020 Volume 507

Abstract: West Nile virus (WNV) is a mosquito-borne virus that originates from Africa and at present causes neurological disease in birds, horses, and humans all around the globe. As West Nile fever is an important zoonosis, the role of free-ranging domestic ... ...

Abstract	West Nile virus (WNV) is a mosquito-borne virus that originates from Africa and at present causes neurological disease in birds, horses, and humans all around the globe. As West Nile fever is an important zoonosis, the role of free-ranging domestic poultry as a source of infection for humans should be evaluated. This study examined the pathogenicity of an Italian WNV lineage 1 strain for domestic poultry (chickens, ducks, and geese) held in Germany. All three species were subcutaneously injected with WNV, and the most susceptible species was also inoculated via mosquito bite. All species developed various degrees of viremia, viral shedding (oropharyngeal and cloacal), virus accumulation, and pathomorphological lesions. Geese were most susceptible, displaying the highest viremia levels. The tested waterfowl, geese, and especially ducks proved to be ideal sentinel species for WNV due to their high antibody levels and relatively low blood viral loads. None of the three poultry species can function as a reservoir/amplifying host for WNV, as their viremia levels most likely do not suffice to infect feeding mosquitoes. Due to the recent appearance of WNV in Germany, future pathogenicity studies should also include local virus strains.
Keywords	West Nile virus ; arbovirus ; experimental infection ; pathogenesis ; poultry ; chickens ; Medicine ; R
Subject code	630
Language	English
Publishing date	2020-09-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Spread of West Nile Virus and Usutu Virus in the German Bird Population, 2019–2020

Ute Ziegler / Felicitas Bergmann / Dominik Fischer / Kerstin Müller / Cora M. Holicki / Balal Sadeghi / Michael Sieg / Markus Keller / Rebekka Schwehn / Maximilian Reuschel / Luisa Fischer / Oliver Krone / Monika Rinder / Karolin Schütte / Volker Schmidt / Martin Eiden / Christine Fast / Anne Günther / Anja Globig /

Franz J. Conraths / Christoph Staubach / Florian Brandes / Michael Lierz / Rüdiger Korbel / Thomas W. Vahlenkamp / Martin H. Groschup

Microorganisms, Vol 10, Iss 4, p

2022 Volume 807

Abstract: West Nile virus (WNV) and Usutu virus (USUV) are important flaviviruses circulating in Germany. While USUV was first reported more than 10 years ago, WNV has only reached the country in 2018. Wild birds are important amplifying hosts for both viruses. ... ...

Abstract	West Nile virus (WNV) and Usutu virus (USUV) are important flaviviruses circulating in Germany. While USUV was first reported more than 10 years ago, WNV has only reached the country in 2018. Wild birds are important amplifying hosts for both viruses. Therefore, we have been monitoring the bird population in different regions of Germany by a previously established network for many years. This report summarizes the results of molecular and/or serological methods of 2345 blood samples from birds of 22 different orders and over 2900 bird carcasses from 2019 and 2020. USUV RNA circulation was found in different regions of Germany, with emphasis on USUV lineages Europe 3 and Africa 3. Increased evidence of USUV lineage Europe 2 was detected in eastern Germany. WNV RNA was found only in birds from the eastern part of the country. The seroprevalence for USUV was between 3.11% and 7.20% in all three regions investigated, whereas the WNV seroprevalence spanned from 14.77% to 16.15% in eastern Germany, with a noticeable tendency for a westward and southward expansion in both years. Thus, wild bird monitoring for WNV and USUV can serve as an early warning system for a human exposure risk.
Keywords	West Nile virus ; Usutu virus ; bird ; monitoring ; flavivirus ; Germany ; Biology (General) ; QH301-705.5
Subject code	590
Language	English
Publishing date	2022-04-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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