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In this article, we describe the development and evaluation of a double antigen sandwich enzyme-linked immunosorbent assay (ELISA) able to detect serotype 4-specific antibodies from BTV-4 infected or vaccinated animals using a recombinant BTV-4 VP2 protein. The coding sequence of VP2 was inserted into a pVote plasmid by recombination in the Gateway® cloning system. Vaccinia virus (VacV) was used as a vector for the expression of the recombinant VP2. After production in BSR cells, recombinant VP2 was purified by immunoprecipitation using a FLAG tag and then used both as the coated ELISA antigen and as the HRP-tagged conjugate. The performance of the ELISA was evaluated with 1,186 samples collected from BTV negative, infected or vaccinated animals. The specificity and sensitivity of the BTV-4 ELISA were above the expected standards for the detection of anti-BTV-4 VP2 antibodies in animals reared in Europe or in the Mediterranean basin. Cross-reactions were observed with reference sera for serotypes 10 and 20, and to a lesser extent with serotypes 12, 17 and 24, due to their genetic proximity to serotype 4. Nevertheless, these serotypes have never been detected in Europe and the Mediterranean area. This ELISA, which requires only the production of a recombinant protein, can be used to detect BTV serotype 4-specific antibodies and is therefore an attractive alternative diagnostic method to serum neutralization.
Emmanuel Bréard; Mathilde Turpaud; Georges Beaud; Lydie Postic; Aurore Fablet; Martin Beer; Corinne Sailleau; Grégory Caignard; Cyril Viarouge; Bernd Hoffmann; Damien Vitour; Stéphan Zientara. Development and Validation of an ELISA for the Detection of Bluetongue Virus Serotype 4-Specific Antibodies. Viruses 2021, 13, 1741 .
AMA StyleEmmanuel Bréard, Mathilde Turpaud, Georges Beaud, Lydie Postic, Aurore Fablet, Martin Beer, Corinne Sailleau, Grégory Caignard, Cyril Viarouge, Bernd Hoffmann, Damien Vitour, Stéphan Zientara. Development and Validation of an ELISA for the Detection of Bluetongue Virus Serotype 4-Specific Antibodies. Viruses. 2021; 13 (9):1741.
Chicago/Turabian StyleEmmanuel Bréard; Mathilde Turpaud; Georges Beaud; Lydie Postic; Aurore Fablet; Martin Beer; Corinne Sailleau; Grégory Caignard; Cyril Viarouge; Bernd Hoffmann; Damien Vitour; Stéphan Zientara. 2021. "Development and Validation of an ELISA for the Detection of Bluetongue Virus Serotype 4-Specific Antibodies." Viruses 13, no. 9: 1741.
Due to changing distemper issues worldwide and to inadequate results of an inter-laboratory study in Germany, it seems sensible to adapt and optimize the diagnostic methods for the detection of the canine distemper virus (CDV) to the new genetic diversity of virus strains. The goal of the project was the development, establishment and validation of two independent one-step reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) methods for the safe detection of CDV in domestic and wild animals. For this purpose, an existing CDV-RT-qPCR was decisively adapted and, in addition, a completely new system was developed. Both CDV-RT-qPCR systems are characterized by a very high, comparable analytical and diagnostic sensitivity and specificity and can be mutually combined with inhibition or extraction controls. The reduction in the master mix used allows for the parallel implementation of both CDV-RT-qPCR systems without significant cost increases. For validation of the new CDV-RT-qPCR duplex assays, a panel comprising 378 samples derived from Germany, several European countries and one African country were tested. A sensitivity of 98.9% and a specificity of 100% were computed for the new assays, thus being a reliable molecular diagnostic tool for the detection of CDV in domestic and wild animals.
Sabrina Halecker; Sabine Bock; Martin Beer; Bernd Hoffmann. A New Molecular Detection System for Canine Distemper Virus Based on a Double-Check Strategy. Viruses 2021, 13, 1632 .
AMA StyleSabrina Halecker, Sabine Bock, Martin Beer, Bernd Hoffmann. A New Molecular Detection System for Canine Distemper Virus Based on a Double-Check Strategy. Viruses. 2021; 13 (8):1632.
Chicago/Turabian StyleSabrina Halecker; Sabine Bock; Martin Beer; Bernd Hoffmann. 2021. "A New Molecular Detection System for Canine Distemper Virus Based on a Double-Check Strategy." Viruses 13, no. 8: 1632.
Lyssaviruses are the causative agents for rabies, a zoonotic and fatal disease. Bats are the ancestral reservoir host for lyssaviruses, and at least three different lyssaviruses have been found in bats from Germany. Across Europe, novel lyssaviruses were identified in bats recently and occasional spillover infections in other mammals and human cases highlight their public health relevance. Here, we report the results from an enhanced passive bat rabies surveillance that encompasses samples without human contact that would not be tested under routine conditions. To this end, 1236 bat brain samples obtained between 2018 and 2020 were screened for lyssaviruses via several RT-qPCR assays. European bat lyssavirus type 1 (EBLV-1) was dominant, with 15 positives exclusively found in serotine bats (Eptesicus serotinus) from northern Germany. Additionally, when an archived set of bat samples that had tested negative for rabies by the FAT were screened in the process of assay validation, four samples tested EBLV-1 positive, including two detected in Pipistrellus pipistrellus. Subsequent phylogenetic analysis of 17 full genomes assigned all except one of these viruses to the A1 cluster of the EBLV-1a sub-lineage. Furthermore, we report here another Bokeloh bat lyssavirus (BBLV) infection in a Natterer’s bat (Myotis nattereri) found in Lower Saxony, the tenth reported case of this novel bat lyssavirus.
Antonia Klein; Sten Calvelage; Kore Schlottau; Bernd Hoffmann; Elisa Eggerbauer; Thomas Müller; Conrad Freuling. Retrospective Enhanced Bat Lyssavirus Surveillance in Germany between 2018–2020. Viruses 2021, 13, 1538 .
AMA StyleAntonia Klein, Sten Calvelage, Kore Schlottau, Bernd Hoffmann, Elisa Eggerbauer, Thomas Müller, Conrad Freuling. Retrospective Enhanced Bat Lyssavirus Surveillance in Germany between 2018–2020. Viruses. 2021; 13 (8):1538.
Chicago/Turabian StyleAntonia Klein; Sten Calvelage; Kore Schlottau; Bernd Hoffmann; Elisa Eggerbauer; Thomas Müller; Conrad Freuling. 2021. "Retrospective Enhanced Bat Lyssavirus Surveillance in Germany between 2018–2020." Viruses 13, no. 8: 1538.
The development of new diagnostic methods resulted in the discovery of novel hepaciviruses in wild populations of the bank vole (Myodes glareolus, syn. Clethrionomys glareolus). The naturally infected voles demonstrate signs of hepatitis similar to those induced by hepatitis C virus (HCV) in humans. The aim of the present research was to investigate the geographical distribution of bank vole-associated hepaciviruses (BvHVs) and their genetic diversity in Europe. Real-time reverse transcription polymerase chain reaction (RT-qPCR) screening revealed BvHV RNA in 442 out of 1838 (24.0%) bank voles from nine European countries and in one of seven northern red-backed voles (Myodes rutilus, syn. Clethrionomys rutilus). BvHV RNA was not found in any other small mammal species (n = 23) tested here. Phylogenetic and isolation-by-distance analyses confirmed the occurrence of both BvHV species (Hepacivirus F and Hepacivirus J) and their sympatric occurrence at several trapping sites in two countries. The broad geographical distribution of BvHVs across Europe was associated with their presence in bank voles of different evolutionary lineages. The extensive geographical distribution and high levels of genetic diversity of BvHVs, as well as the high population fluctuations of bank voles and occasional commensalism in some parts of Europe warrant future studies on the zoonotic potential of BvHVs.
Julia Schneider; Bernd Hoffmann; Cristina Fevola; Marie Schmidt; Christian Imholt; Stefan Fischer; Frauke Ecke; Birger Hörnfeldt; Magnus Magnusson; Gert Olsson; Annapaola Rizzoli; Valentina Tagliapietra; Mario Chiari; Chantal Reusken; Elena Bužan; Maria Kazimirova; Michal Stanko; Thomas White; Daniela Reil; Anna Obiegala; Anna Meredith; Jan Drexler; Sandra Essbauer; Heikki Henttonen; Jens Jacob; Heidi Hauffe; Martin Beer; Gerald Heckel; Rainer Ulrich. Geographical Distribution and Genetic Diversity of Bank Vole Hepaciviruses in Europe. Viruses 2021, 13, 1258 .
AMA StyleJulia Schneider, Bernd Hoffmann, Cristina Fevola, Marie Schmidt, Christian Imholt, Stefan Fischer, Frauke Ecke, Birger Hörnfeldt, Magnus Magnusson, Gert Olsson, Annapaola Rizzoli, Valentina Tagliapietra, Mario Chiari, Chantal Reusken, Elena Bužan, Maria Kazimirova, Michal Stanko, Thomas White, Daniela Reil, Anna Obiegala, Anna Meredith, Jan Drexler, Sandra Essbauer, Heikki Henttonen, Jens Jacob, Heidi Hauffe, Martin Beer, Gerald Heckel, Rainer Ulrich. Geographical Distribution and Genetic Diversity of Bank Vole Hepaciviruses in Europe. Viruses. 2021; 13 (7):1258.
Chicago/Turabian StyleJulia Schneider; Bernd Hoffmann; Cristina Fevola; Marie Schmidt; Christian Imholt; Stefan Fischer; Frauke Ecke; Birger Hörnfeldt; Magnus Magnusson; Gert Olsson; Annapaola Rizzoli; Valentina Tagliapietra; Mario Chiari; Chantal Reusken; Elena Bužan; Maria Kazimirova; Michal Stanko; Thomas White; Daniela Reil; Anna Obiegala; Anna Meredith; Jan Drexler; Sandra Essbauer; Heikki Henttonen; Jens Jacob; Heidi Hauffe; Martin Beer; Gerald Heckel; Rainer Ulrich. 2021. "Geographical Distribution and Genetic Diversity of Bank Vole Hepaciviruses in Europe." Viruses 13, no. 7: 1258.
Emerging variants of concern (VOCs) drive the SARS-CoV-2 pandemic. We assessed VOC B.1.1.7, now prevalent in several countries, and VOC B.1.351, representing the greatest threat to populations with immunity to the early SARS-CoV-2 progenitors. B.1.1.7 showed a clear fitness advantage over the progenitor variant (wt-S614G) in ferrets and two mouse models, where the substitutions in the spike glycoprotein were major drivers for fitness advantage. In the superspreader hamster model, B.1.1.7 and wt-S614G had comparable fitness, whereas B.1.351 was outcompeted. The VOCs had similar replication kinetics as compared to wt-S614G in human airway epithelial cultures. Our study highlights the importance of using multiple models for complete fitness characterization of VOCs and demonstrates adaptation of B.1.1.7 towards increased upper respiratory tract replication and enhanced transmission in vivo.
Lorenz Ulrich; Nico Joel Halwe; Adriano Taddeo; Nadine Ebert; Jacob Schön; Christelle Devisme; Bettina Salome Trüeb; Bernd Hoffmann; Manon Wider; Meriem Bekliz; Manel Essaidi-Laziosi; Marie Luisa Schmidt; Daniela Niemeyer; Victor Max Corman; Anna Kraft; Aurélie Godel; Laura Laloli; Jenna Nicole Kelly; ANGELE Breithaupt; Claudia Wylezich; Inês Margarida Berenguer Veiga; Mitra Gultom; Kenneth Adea; Benjamin Meyer; Christiane Eberhardt; Lisa Thomann; Monika Gsell-Albert; Fabien Labroussaa; Jörg Jores; Artur Summerfield; Christian Drosten; Isabella Anne Eckerle; Ronald Dijkman; Donata Hoffmann; Volker Thiel; Martin Beer; Charaf Benarafa. Enhanced fitness of SARS-CoV-2 variant of concern B.1.1.7, but not B.1.351, in animal models. 2021, 1 .
AMA StyleLorenz Ulrich, Nico Joel Halwe, Adriano Taddeo, Nadine Ebert, Jacob Schön, Christelle Devisme, Bettina Salome Trüeb, Bernd Hoffmann, Manon Wider, Meriem Bekliz, Manel Essaidi-Laziosi, Marie Luisa Schmidt, Daniela Niemeyer, Victor Max Corman, Anna Kraft, Aurélie Godel, Laura Laloli, Jenna Nicole Kelly, ANGELE Breithaupt, Claudia Wylezich, Inês Margarida Berenguer Veiga, Mitra Gultom, Kenneth Adea, Benjamin Meyer, Christiane Eberhardt, Lisa Thomann, Monika Gsell-Albert, Fabien Labroussaa, Jörg Jores, Artur Summerfield, Christian Drosten, Isabella Anne Eckerle, Ronald Dijkman, Donata Hoffmann, Volker Thiel, Martin Beer, Charaf Benarafa. Enhanced fitness of SARS-CoV-2 variant of concern B.1.1.7, but not B.1.351, in animal models. . 2021; ():1.
Chicago/Turabian StyleLorenz Ulrich; Nico Joel Halwe; Adriano Taddeo; Nadine Ebert; Jacob Schön; Christelle Devisme; Bettina Salome Trüeb; Bernd Hoffmann; Manon Wider; Meriem Bekliz; Manel Essaidi-Laziosi; Marie Luisa Schmidt; Daniela Niemeyer; Victor Max Corman; Anna Kraft; Aurélie Godel; Laura Laloli; Jenna Nicole Kelly; ANGELE Breithaupt; Claudia Wylezich; Inês Margarida Berenguer Veiga; Mitra Gultom; Kenneth Adea; Benjamin Meyer; Christiane Eberhardt; Lisa Thomann; Monika Gsell-Albert; Fabien Labroussaa; Jörg Jores; Artur Summerfield; Christian Drosten; Isabella Anne Eckerle; Ronald Dijkman; Donata Hoffmann; Volker Thiel; Martin Beer; Charaf Benarafa. 2021. "Enhanced fitness of SARS-CoV-2 variant of concern B.1.1.7, but not B.1.351, in animal models." , no. : 1.
We identified a putative novel atypical BTV serotype ‘36’ in Swiss goat flocks. In the initial flock clinical signs consisting of multifocal purulent dermatitis, facial oedema and fever were observed. Following BTV detection by RT-qPCR, serotyping identified BTV-25 and also a putative novel BTV serotype in several of the affected goats. We successfully propagated the so-called “BTV-36-CH2019” strain in cell culture, developed a specific RT-qPCR targeting Segment 2, and generated the full genome by high-throughput sequencing. Furthermore, we experimentally infected goats with BTV-36-CH2019. Regularly, EDTA blood, serum and diverse swab samples were collected. Throughout the experiment, neither fever nor clinical disease was observed in any of the inoculated goats. Four goats developed BTV viremia, whereas one inoculated goat and the two contact animals remained negative. No viral RNA was detected in the swab samples collected from nose, mouth, eye, and rectum, and thus the experimental infection of goats using this novel BTV serotype delivered no indications for any clinical symptoms or vector-free virus transmission pathways. The subclinical infection of the four goats is in accordance with the reports for other atypical BTVs. However, the clinical signs of the initial goat flock did most likely not result from infection with the novel BTV-36-CH0219.
Christina Ries; Andrea Vögtlin; Daniela Hüssy; Tabea Jandt; Hansjörg Gobet; Monika Hilbe; Carole Burgener; Luzia Schweizer; Stephanie Häfliger-Speiser; Martin Beer; Bernd Hoffmann. Putative Novel Atypical BTV Serotype ‘36’ Identified in Small Ruminants in Switzerland. Viruses 2021, 13, 721 .
AMA StyleChristina Ries, Andrea Vögtlin, Daniela Hüssy, Tabea Jandt, Hansjörg Gobet, Monika Hilbe, Carole Burgener, Luzia Schweizer, Stephanie Häfliger-Speiser, Martin Beer, Bernd Hoffmann. Putative Novel Atypical BTV Serotype ‘36’ Identified in Small Ruminants in Switzerland. Viruses. 2021; 13 (5):721.
Chicago/Turabian StyleChristina Ries; Andrea Vögtlin; Daniela Hüssy; Tabea Jandt; Hansjörg Gobet; Monika Hilbe; Carole Burgener; Luzia Schweizer; Stephanie Häfliger-Speiser; Martin Beer; Bernd Hoffmann. 2021. "Putative Novel Atypical BTV Serotype ‘36’ Identified in Small Ruminants in Switzerland." Viruses 13, no. 5: 721.
Influenza A viruses (IAV) of subtype H9N2, endemic in world-wide poultry holdings, are reported to cause spill-over infections to pigs and humans and have also contributed substantially to recent reassortment-derived pre-pandemic zoonotic viruses of concern, such as the Asian H7N9 viruses. Recently, a H9N2 bat influenza A virus was found in Egyptian fruit bats (Rousettus aegyptiacus), raising the question of whether this bat species is a suitable host for IAV. Here, we studied the susceptibility, pathogenesis and transmission of avian and bat-related H9N2 viruses in this new host. In a first experiment, we oronasally inoculated six Egyptian fruit bats with an avian-related H9N2 virus (A/layer chicken/Bangladesh/VP02-plaque/2016 (H9N2)). In a second experiment, six Egyptian fruit bats were inoculated with the newly discovered bat-related H9N2 virus (A/bat/Egypt/381OP/2017 (H9N2)). While R. aegyptiacus turned out to be refractory to an infection with H9N2 avian-type, inoculation with the bat H9N2 subtype established a productive infection in all inoculated animals with a detectable seroconversion at day 21 post-infection. In conclusion, Egyptian fruit bats are most likely not susceptible to the avian H9N2 subtype, but can be infected with fruit bat-derived H9N2. H9-specific sero-reactivities in fruit bats in the field are therefore more likely the result of contact with a bat-adapted H9N2 strain.
Nico Halwe; Marco Gorka; Bernd Hoffmann; Melanie Rissmann; ANGELE Breithaupt; Martin Schwemmle; Martin Beer; Ahmed Kandeil; Mohamed Ali; Ghazi Kayali; Donata Hoffmann; Anne Balkema-Buschmann. Egyptian Fruit Bats (Rousettus aegyptiacus) Were Resistant to Experimental Inoculation with Avian-Origin Influenza A Virus of Subtype H9N2, But Are Susceptible to Experimental Infection with Bat-Borne H9N2 Virus. Viruses 2021, 13, 672 .
AMA StyleNico Halwe, Marco Gorka, Bernd Hoffmann, Melanie Rissmann, ANGELE Breithaupt, Martin Schwemmle, Martin Beer, Ahmed Kandeil, Mohamed Ali, Ghazi Kayali, Donata Hoffmann, Anne Balkema-Buschmann. Egyptian Fruit Bats (Rousettus aegyptiacus) Were Resistant to Experimental Inoculation with Avian-Origin Influenza A Virus of Subtype H9N2, But Are Susceptible to Experimental Infection with Bat-Borne H9N2 Virus. Viruses. 2021; 13 (4):672.
Chicago/Turabian StyleNico Halwe; Marco Gorka; Bernd Hoffmann; Melanie Rissmann; ANGELE Breithaupt; Martin Schwemmle; Martin Beer; Ahmed Kandeil; Mohamed Ali; Ghazi Kayali; Donata Hoffmann; Anne Balkema-Buschmann. 2021. "Egyptian Fruit Bats (Rousettus aegyptiacus) Were Resistant to Experimental Inoculation with Avian-Origin Influenza A Virus of Subtype H9N2, But Are Susceptible to Experimental Infection with Bat-Borne H9N2 Virus." Viruses 13, no. 4: 672.
Outbreaks of the three capripox virus species, namely lumpy skin disease virus, sheeppox virus, and goatpox virus, severely affect animal health and both national and international economies. Therefore, the World Organization for Animal Health (OIE) classified them as notifiable diseases. Until now, discrimination of capripox virus species was possible by using different conventional PCR protocols. However, more sophisticated probe-based real-time qPCR systems addressing this issue are, to our knowledge, still missing. In the present study, we developed several duplex qPCR assays consisting of different types of fluorescence-labelled probes that are highly sensitive and show a high analytical specificity. Finally, our assays were combined with already published diagnostic methods to a diagnostic workflow that enables time-saving, reliable, and robust detection, differentiation, and characterization of capripox virus isolates.
Janika Wolff; Martin Beer; Bernd Hoffmann. Probe-Based Real-Time qPCR Assays for a Reliable Differentiation of Capripox Virus Species. Microorganisms 2021, 9, 765 .
AMA StyleJanika Wolff, Martin Beer, Bernd Hoffmann. Probe-Based Real-Time qPCR Assays for a Reliable Differentiation of Capripox Virus Species. Microorganisms. 2021; 9 (4):765.
Chicago/Turabian StyleJanika Wolff; Martin Beer; Bernd Hoffmann. 2021. "Probe-Based Real-Time qPCR Assays for a Reliable Differentiation of Capripox Virus Species." Microorganisms 9, no. 4: 765.
The genus capripoxvirus (CaPV), family Poxviridae, includes three virus species: goatpox virus (GPV), sheeppox virus (SPV) and lumpy skin disease virus (LSDV). CaPV causes disease outbreaks with consequent economic losses in Africa and the Middle East. LSDV has recently spread to Southeast Europe. As CaPVs share 96–97% genetic similarity along the length of the entire genome and are difficult to distinguish using serological assays, simple, reliable and fast methods for diagnosis and species differentiation are crucial in cases of disease outbreak. The present study aimed to develop a field-applicable CaPV differentiation method. Nanopore technology was used for whole genome sequencing. A local database of complete CaPV genomes and partial sequences of three genes (RPO30, P32 and GPCR) was established for offline Basic Local Alignment Search Tool (BLAST). Specificities of 98.04% in whole genome and 97.86% in RPO30 gene runs were obtained among the three virus species, while other databases were less specific. The total run time was shortened to approximately 2 h. Functionality of the developed procedure was proved by samples with high host background sequences. Reliable differentiation options for the quality and capacity of hardware, and sample quality of suspected cases, were derived from these findings. The whole workflow can be performed rapidly with a mobile suitcase laboratory and mini-computer, allowing application at the point-of-need with limited resource settings.
Kamal Eltom; Anna Althoff; Sören Hansen; Susanne Böhlken-Fascher; Ausama Yousif; Hussein El-Sheikh; Ahmed ElWakeel; Mahmoud Elgamal; Hadeer Mossa; Emad Aboul-Soud; Janika Wolff; Christian Korthase; Bernd Hoffmann; Nabawia Adam; Sanaa Abdelaziz; Mohamed Shalaby; Ahmed Abd El Wahed. Differentiation of Capripox Viruses by Nanopore Sequencing. Vaccines 2021, 9, 351 .
AMA StyleKamal Eltom, Anna Althoff, Sören Hansen, Susanne Böhlken-Fascher, Ausama Yousif, Hussein El-Sheikh, Ahmed ElWakeel, Mahmoud Elgamal, Hadeer Mossa, Emad Aboul-Soud, Janika Wolff, Christian Korthase, Bernd Hoffmann, Nabawia Adam, Sanaa Abdelaziz, Mohamed Shalaby, Ahmed Abd El Wahed. Differentiation of Capripox Viruses by Nanopore Sequencing. Vaccines. 2021; 9 (4):351.
Chicago/Turabian StyleKamal Eltom; Anna Althoff; Sören Hansen; Susanne Böhlken-Fascher; Ausama Yousif; Hussein El-Sheikh; Ahmed ElWakeel; Mahmoud Elgamal; Hadeer Mossa; Emad Aboul-Soud; Janika Wolff; Christian Korthase; Bernd Hoffmann; Nabawia Adam; Sanaa Abdelaziz; Mohamed Shalaby; Ahmed Abd El Wahed. 2021. "Differentiation of Capripox Viruses by Nanopore Sequencing." Vaccines 9, no. 4: 351.
African swine fever (ASF) is a contagious viral hemorrhagic disease of domestic pigs and wild boars. The disease is notifiable to the World Organisation for Animal Health (OIE) and is responsible for high mortality and serious economic losses. PCR and real-time PCR (qPCR) are the OIE-recommended standard methods for the direct detection of African swine fever virus (ASFV) DNA. The aim of our work was the simplification and standardization of the molecular diagnostic workflow in the lab. For validation of this “easy lab” workflow, different sample materials from animal trials were collected and analyzed (EDTA blood, serum, oral swabs, chewing ropes, and tissue samples) to identify the optimal sample material for diagnostics in live animals. Based on our data, the EDTA blood samples or bloody tissue samples represent the best specimens for ASFV detection in the early and late phases of infection. The application of prefilled ready-to-use reagents for nucleic acid extraction or the use of a Tissue Lysis Reagent (TLR) delivers simple and reliable alternatives for the release of the ASFV nucleic acids. For the qPCR detection of ASFV, different published and commercial kits were compared. Here, a lyophilized commercial kit shows the best results mainly based on the increased template input. The good results of the “easy lab” strategy could be confirmed by the ASFV detection in field samples from wild boars collected from the 2020 ASFV outbreak in Germany. Appropriate internal control systems for extraction and PCR are key features of the “easy lab” concept and reduce the risk of false-negative and false-positive results. In addition, the use of easy-to-handle machines and software reduces training efforts and the misinterpretation of results. The PCR diagnostics based on the “easy lab” strategy can realize a high sensitivity and specificity comparable to the standard PCR methods and should be especially usable for labs with limited experiences and resources.
Ahmed Elnagar; Jutta Pikalo; Martin Beer; Sandra Blome; Bernd Hoffmann. Swift and Reliable “Easy Lab” Methods for the Sensitive Molecular Detection of African Swine Fever Virus. International Journal of Molecular Sciences 2021, 22, 2307 .
AMA StyleAhmed Elnagar, Jutta Pikalo, Martin Beer, Sandra Blome, Bernd Hoffmann. Swift and Reliable “Easy Lab” Methods for the Sensitive Molecular Detection of African Swine Fever Virus. International Journal of Molecular Sciences. 2021; 22 (5):2307.
Chicago/Turabian StyleAhmed Elnagar; Jutta Pikalo; Martin Beer; Sandra Blome; Bernd Hoffmann. 2021. "Swift and Reliable “Easy Lab” Methods for the Sensitive Molecular Detection of African Swine Fever Virus." International Journal of Molecular Sciences 22, no. 5: 2307.
Schmallenberg virus (SBV) is the cause of severe fetal malformations when immunologically naïve pregnant ruminants are infected. In those malformed fetuses, a “hot-spot”-region of high genetic variability within the N-terminal region of the viral envelope protein Gc has been observed previously, and this region co-localizes with a known key immunogenic domain. We studied a series of M-segments of those SBV variants from malformed fetuses with point mutations, insertions or large in-frame deletions of up to 612 nucleotides. Furthermore, a unique cell-culture isolate from a malformed fetus with large in-frame deletions within the M-segment was analyzed. Each Gc-protein with amino acid deletions within the “hot spot” of mutations failed to react with any neutralizing anti-SBV monoclonal antibodies or a domain specific antiserum. In addition, in vitro virus replication of the natural deletion variant could not be markedly reduced by neutralizing monoclonal antibodies or antisera from the field. The large-deletion variant of SBV that could be isolated in cell culture was highly attenuated with an impaired in vivo replication following the inoculation of sheep. In conclusion, the observed amino acid sequence mutations within the N-terminal main immunogenic domain of glycoprotein Gc result in an efficient immune evasion from neutralizing antibodies in the special environment of a developing fetus. These SBV-variants were never detected as circulating viruses, and therefore should be considered to be dead-end virus variants, which are not able to spread further. The observations described here may be transferred to other orthobunyaviruses, particularly those of the Simbu serogroup that have been shown to infect fetuses. Importantly, such mutant strains should not be included in attempts to trace the spatial-temporal evolution of orthobunyaviruses in molecular-epidemiolocal approaches during outbreak investigations.
Kerstin Wernike; Ilona Reimann; Ashley C. Banyard; Franziska Kraatz; S. Anna La Rocca; Bernd Hoffmann; Sarah McGowan; Silke Hechinger; Bhudipa Choudhury; Andrea Aebischer; Falko Steinbach; Martin Beer. High genetic variability of Schmallenberg virus M-segment leads to efficient immune escape from neutralizing antibodies. PLOS Pathogens 2021, 17, e1009247 .
AMA StyleKerstin Wernike, Ilona Reimann, Ashley C. Banyard, Franziska Kraatz, S. Anna La Rocca, Bernd Hoffmann, Sarah McGowan, Silke Hechinger, Bhudipa Choudhury, Andrea Aebischer, Falko Steinbach, Martin Beer. High genetic variability of Schmallenberg virus M-segment leads to efficient immune escape from neutralizing antibodies. PLOS Pathogens. 2021; 17 (1):e1009247.
Chicago/Turabian StyleKerstin Wernike; Ilona Reimann; Ashley C. Banyard; Franziska Kraatz; S. Anna La Rocca; Bernd Hoffmann; Sarah McGowan; Silke Hechinger; Bhudipa Choudhury; Andrea Aebischer; Falko Steinbach; Martin Beer. 2021. "High genetic variability of Schmallenberg virus M-segment leads to efficient immune escape from neutralizing antibodies." PLOS Pathogens 17, no. 1: e1009247.
Between 2015 and 2018, we identified the presence of three so-far-unknown Bluetongue virus (BTV) strains (BTV-MNG1/2018, BTV-MNG2/2016, and BTV-MNG3/2016) circulating in clinical healthy sheep and goats in Mongolia. Virus isolation from EDTA blood samples of BTV-MNG1/2018 and BTV-MNG3/2016 was successful on the mammalian cell line BSR using blood collected from surveillance. After experimental inoculation of goats with BTV-MNG2/2016 positive blood as inoculum, we observed viraemia in one goat and with the EDTA blood of the experimental inoculation, the propagation of BTV-MNG2/2016 in cell culture was successful on mammalian cell line BSR as well. However, virus isolation experiments for BTV-MNG2/2016 on KC cells were unsuccessful. Furthermore, we generated the complete coding sequence of all three novel Mongolian strains. For atypical BTV, serotyping via the traditional serum neutralization assay is not trivial. We therefore sorted the ‘putative novel atypical serotypes’ according to their segment-2 sequence identities and their time point of sampling. Hence, the BTV-MNG1/2018 isolate forms the ‘putative novel atypical serotype’ 33, the BTV-MNG3/2016 the ‘putative novel atypical serotype’ 35, whereas the BTV-MNG2/2016 strain belongs to the same putative novel atypical serotype ‘30′ as BTV-XJ1407 from China.
Christina Ries; Tumenjargal Sharav; Erdene-Ochir Tseren-Ochir; Martin Beer; Bernd Hoffmann. Putative Novel Serotypes ‘33′ and ‘35′ in Clinically Healthy Small Ruminants in Mongolia Expand the Group of Atypical BTV. Viruses 2020, 13, 42 .
AMA StyleChristina Ries, Tumenjargal Sharav, Erdene-Ochir Tseren-Ochir, Martin Beer, Bernd Hoffmann. Putative Novel Serotypes ‘33′ and ‘35′ in Clinically Healthy Small Ruminants in Mongolia Expand the Group of Atypical BTV. Viruses. 2020; 13 (1):42.
Chicago/Turabian StyleChristina Ries; Tumenjargal Sharav; Erdene-Ochir Tseren-Ochir; Martin Beer; Bernd Hoffmann. 2020. "Putative Novel Serotypes ‘33′ and ‘35′ in Clinically Healthy Small Ruminants in Mongolia Expand the Group of Atypical BTV." Viruses 13, no. 1: 42.
Capripox virus (CaPV)-induced diseases (lumpy skin disease, sheeppox, goatpox) are described as the most serious pox diseases of livestock animals, and therefore are listed as notifiable diseases under guidelines of the World Organisation for Animal Health (OIE). Until now, only live-attenuated vaccines are commercially available for the control of CaPV. Due to numerous potential problems after vaccination (e.g., loss of the disease-free status of the respective country, the possibility of vaccine virus shedding and transmission as well as the risk of recombination with field strains during natural outbreaks), the use of these vaccines must be considered carefully and is not recommended in CaPV-free countries. Therefore, innocuous and efficacious inactivated vaccines against CaPV would provide a great tool for control of these diseases. Unfortunately, most inactivated Capripox vaccines were reported as insufficient and protection seemed to be only short-lived. Nevertheless, a few studies dealing with inactivated vaccines against CaPV are published, giving evidence for good clinical protection against CaPV-infections. In our studies, a low molecular weight copolymer-adjuvanted vaccine formulation was able to induce sterile immunity in the respective animals after severe challenge infection. Our findings strongly support the possibility of useful inactivated vaccines against CaPV-infections, and indicate a marked impact of the chosen adjuvant for the level of protection.
Janika Wolff; Tom Moritz; Kore Schlottau; Donata Hoffmann; Martin Beer; Bernd Hoffmann. Development of a Safe and Highly Efficient Inactivated Vaccine Candidate against Lumpy Skin Disease Virus. Vaccines 2020, 9, 4 .
AMA StyleJanika Wolff, Tom Moritz, Kore Schlottau, Donata Hoffmann, Martin Beer, Bernd Hoffmann. Development of a Safe and Highly Efficient Inactivated Vaccine Candidate against Lumpy Skin Disease Virus. Vaccines. 2020; 9 (1):4.
Chicago/Turabian StyleJanika Wolff; Tom Moritz; Kore Schlottau; Donata Hoffmann; Martin Beer; Bernd Hoffmann. 2020. "Development of a Safe and Highly Efficient Inactivated Vaccine Candidate against Lumpy Skin Disease Virus." Vaccines 9, no. 1: 4.
Capripox viruses (CaPVs) cause a highly contagious poxvirus disease of livestock animals. Working with CaPVs requires laboratories with a high biosecurity level (BSL 3), and reliable inactivation of these viruses is therefore necessary for working in areas or laboratories with a lower biosecurity status. Heat treatment provides a simple and well-established tool for the inactivation due to its substantial advantages (e.g., easy to perform, fast, cheap, and robust). In our study, we determined the time–temperature profiles needed for a fail-safe inactivation procedure using four different CaPV isolates in aqueous solution with and without the addition of protective serum. All four tested CaPV isolates were completely inactivated after 30 min at 56 °C or 10 min at 60 °C. Since different thermal stabilities of other CaPV isolates could not be fully excluded, we recommend an inactivation procedure of 1 h at 56 °C for safe shipment or working in laboratories with lower biosecurity levels than BSL 3.
Janika Wolff; Martin Beer; Bernd Hoffmann. Thermal Inactivation of Different Capripox Virus Isolates. Microorganisms 2020, 8, 2053 .
AMA StyleJanika Wolff, Martin Beer, Bernd Hoffmann. Thermal Inactivation of Different Capripox Virus Isolates. Microorganisms. 2020; 8 (12):2053.
Chicago/Turabian StyleJanika Wolff; Martin Beer; Bernd Hoffmann. 2020. "Thermal Inactivation of Different Capripox Virus Isolates." Microorganisms 8, no. 12: 2053.
Lumpy skin disease (LSD) is a viral disorder of cattle caused by the lumpy skin disease virus (LSDV) which can induce severe infections leading to high economic losses. Being of African origin, the first LSD outbreaks in Europe occurred in Greece and later in the Balkan region. Little is known about the mode of transmission, especially in relation to the potential role of arthropods vectors. The purpose of our study was to investigate the role of Stomoxys calcitrans in the transmission of LSDV and their presence at different farms in Switzerland. Laboratory-reared flies were exposed to LSDV spiked-blood and incubated under a realistic fluctuating temperature regime. Body parts, regurgitated blood, and faecal samples were analysed by qPCR for the presence of viral DNA and infectious virus at different time points post-feeding (p.f.). LSDV DNA was detected in heads, bodies, and regurgitated blood up to three days p.f. and up to two days p.f. in the faeces. Infectious virus was isolated from bodies and faeces up to two days and in the regurgitated blood up to 12 h p.f. There was no increase in viral load, consolidating the role of S. calcitrans as mechanical vectors for LSDV. Stomoxys flies were present at all eight farms investigated, including a farm located at 2128 m asl. The persistence of LSDV in S. calcitrans in combination with the long flight ranges of this abundant and widespread fly might have implications on LSD epidemiology and on implementing control measures during disease outbreaks.
Anca I. Paslaru; Niels O. Verhulst; Lena M. Maurer; Alexsandra Brendle; Nicole Pauli; Andrea Vögtlin; Sandra Renzullo; Yelena Ruedin; Bernd Hoffmann; Paul R. Torgerson; Alexander Mathis; Eva Veronesi. Potential mechanical transmission of Lumpy skin disease virus (LSDV) by the stable fly (Stomoxys calcitrans) through regurgitation and defecation. Current Research in Insect Science 2020, 1, 100007 .
AMA StyleAnca I. Paslaru, Niels O. Verhulst, Lena M. Maurer, Alexsandra Brendle, Nicole Pauli, Andrea Vögtlin, Sandra Renzullo, Yelena Ruedin, Bernd Hoffmann, Paul R. Torgerson, Alexander Mathis, Eva Veronesi. Potential mechanical transmission of Lumpy skin disease virus (LSDV) by the stable fly (Stomoxys calcitrans) through regurgitation and defecation. Current Research in Insect Science. 2020; 1 ():100007.
Chicago/Turabian StyleAnca I. Paslaru; Niels O. Verhulst; Lena M. Maurer; Alexsandra Brendle; Nicole Pauli; Andrea Vögtlin; Sandra Renzullo; Yelena Ruedin; Bernd Hoffmann; Paul R. Torgerson; Alexander Mathis; Eva Veronesi. 2020. "Potential mechanical transmission of Lumpy skin disease virus (LSDV) by the stable fly (Stomoxys calcitrans) through regurgitation and defecation." Current Research in Insect Science 1, no. : 100007.
Sheeppox virus (SPPV) together with goatpox virus and lumpy skin disease virus form the genus Capripoxvirus of the Poxviridae family. Due to their great economic importance and major impact on livelihood of small-scale farmers, OIE guidelines classify capripox viruses as notifiable diseases. In the present study, we examined pathogenesis of an Indian SPPV isolate and an Egyptian SPPV isolate in sheep. Three different infection routes were tested: (i) intravenous infection, (ii) intranasal infection and (iii) contact transmission between infected and naïve sheep. Clinical course, viremia and viral shedding as well as seroconversion were analyzed in order to establish a challenge model for SPPV infections that can be used in future vaccine studies. Next to in vivo characterization, both SPPV strains underwent next- and third-generation sequencing to obtain high quality full-length genomes for genetic characterization and comparison to already published SPPV sequences.
Janika Wolff; Sahar Abd El Rahman; Jacqueline King; Mohamed El-Beskawy; Anne Pohlmann; Martin Beer; Bernd Hoffmann. Establishment of a Challenge Model for Sheeppox Virus Infection. Microorganisms 2020, 8, 2001 .
AMA StyleJanika Wolff, Sahar Abd El Rahman, Jacqueline King, Mohamed El-Beskawy, Anne Pohlmann, Martin Beer, Bernd Hoffmann. Establishment of a Challenge Model for Sheeppox Virus Infection. Microorganisms. 2020; 8 (12):2001.
Chicago/Turabian StyleJanika Wolff; Sahar Abd El Rahman; Jacqueline King; Mohamed El-Beskawy; Anne Pohlmann; Martin Beer; Bernd Hoffmann. 2020. "Establishment of a Challenge Model for Sheeppox Virus Infection." Microorganisms 8, no. 12: 2001.
We present the complete genome sequence of bovine alphaherpesvirus 2 (BoHV-2), a member of the family Herpesviridae, subfamily Alphaherpesvirinae, genus Simplexvirus. BoHV-2 is the causative agent of bovine ulcerative mammillitis (bovine herpes mammillitis) and pseudo-lumpy skin disease. The genomic architecture of BoHV-2 is typical of most simplexvirus genomes and congruent with that of human alphaherpesvirus 1 (HHV-1). The genome comprises a total of 131,245 base pairs and has an overall G+C content of 64.9 mol%. A total of 75 open reading frames are predicted. The gene repertoire of BoHV-2 is analogous to that of HHV-1, although the coding region of US12 is missing. A phylogenetic analysis supported BoHV-2 as a member of the genus Simplexvirus.
Florian Pfaff; Antonie Neubauer-Juric; Stefan Krebs; Andreas Hauser; Stefanie Singer; Helmut Blum; Bernd Hoffmann. Full genome sequence of bovine alphaherpesvirus 2 (BoHV-2). Archives of Virology 2020, 166, 639 -643.
AMA StyleFlorian Pfaff, Antonie Neubauer-Juric, Stefan Krebs, Andreas Hauser, Stefanie Singer, Helmut Blum, Bernd Hoffmann. Full genome sequence of bovine alphaherpesvirus 2 (BoHV-2). Archives of Virology. 2020; 166 (2):639-643.
Chicago/Turabian StyleFlorian Pfaff; Antonie Neubauer-Juric; Stefan Krebs; Andreas Hauser; Stefanie Singer; Helmut Blum; Bernd Hoffmann. 2020. "Full genome sequence of bovine alphaherpesvirus 2 (BoHV-2)." Archives of Virology 166, no. 2: 639-643.
Ovine gammaherpesvirus-2 (OvHV-2) causes a lethal disease in cattle and some wild ruminants called malignant catarrhal fever (MCF), which affects the epithelial and lymphoid tissues of the respiratory and digestive tracts and has an important impact on the livestock industry. In this study, MCF was diagnosed in 18 of 427 cattle from different sites in Egypt by its typical clinical signs, found in all 18 animals: corneal opacity, fever, erosions in the buccal cavity, lymphadenitis, and purulent nasal discharge. All affected cattle had been reared in contact with clinically inconspicuous sheep. Of the 18 clinically ill cattle, 13 succumbed to the disease, resulting in estimated morbidity and case fatality rates of 4.2% and 72.2%, respectively. Five samples collected from the affected cattle were positive for OvHV-2 by real-time PCR and were used for sequencing of an 832-bp fragment of the ORF27/gp48 gene. The ORF27 nucleotide sequence of all Egyptian samples was identical, but distinct from viruses found in other parts of Africa and the Mediterranean.
Sahar Abd El Rahman; Ahmed Ateya; Mohamed El-Beskawy; Kerstin Wernike; Bernd Hoffmann; Michael Eschbaumer. Field Observations and Genetic Characterization of Sheep-Associated Malignant Catarrhal Fever in Egypt, 2018. Veterinary Sciences 2020, 7, 201 .
AMA StyleSahar Abd El Rahman, Ahmed Ateya, Mohamed El-Beskawy, Kerstin Wernike, Bernd Hoffmann, Michael Eschbaumer. Field Observations and Genetic Characterization of Sheep-Associated Malignant Catarrhal Fever in Egypt, 2018. Veterinary Sciences. 2020; 7 (4):201.
Chicago/Turabian StyleSahar Abd El Rahman; Ahmed Ateya; Mohamed El-Beskawy; Kerstin Wernike; Bernd Hoffmann; Michael Eschbaumer. 2020. "Field Observations and Genetic Characterization of Sheep-Associated Malignant Catarrhal Fever in Egypt, 2018." Veterinary Sciences 7, no. 4: 201.
To assist the global eradication of peste des petits ruminants virus (PPRV), a molecular test for the rapid and reliable detection of PPRV was developed which additionally enables the detection of pathogens relevant for differential diagnostics. For this purpose, the necessary time frame of a magnetic bead-based nucleic acid extraction protocol was markedly shortened to 7 min and 13 s. The optimized extraction was run on a BioSprint 15 platform. Furthermore, a high-speed multi-well RT-qPCR for the genome detection of PPRV and additional important pathogens such as Foot-and-mouth disease virus, Parapoxvirus ovis, Goatpox virus, and Mycoplasma capricolum subsp. capripneumoniae was established and combined with suitable internal control assays. The here-described qPCR is based on a lyophilized master mix and takes only around 30 to 40 min. Several qPCR cyclers were evaluated regarding their suitability for fast-cycling approaches and for their diagnostic performance in a high-speed RT-qPCR. The final evaluation was conducted on the BioRad CFX96 and also on a portable Liberty16 qPCR cycler. The new molecular test designated as “FastCheckFLI PPR-like”, which is based on rapid nucleic acid extraction and high-speed RT-qPCR, delivered reliable results in less than one hour, allowing its use also in a pen-side scenario.
Sabrina Halecker; Thomas C. Mettenleiter; Martin Beer; Bernd Hoffmann. “FastCheckFLI PPR-like”—A Molecular Tool for the Fast Genome Detection of PPRV and Differential Diagnostic Pathogens. Viruses 2020, 12, 1227 .
AMA StyleSabrina Halecker, Thomas C. Mettenleiter, Martin Beer, Bernd Hoffmann. “FastCheckFLI PPR-like”—A Molecular Tool for the Fast Genome Detection of PPRV and Differential Diagnostic Pathogens. Viruses. 2020; 12 (11):1227.
Chicago/Turabian StyleSabrina Halecker; Thomas C. Mettenleiter; Martin Beer; Bernd Hoffmann. 2020. "“FastCheckFLI PPR-like”—A Molecular Tool for the Fast Genome Detection of PPRV and Differential Diagnostic Pathogens." Viruses 12, no. 11: 1227.
Capripox viruses, with their members “lumpy skin disease virus (LSDV)”, “goatpox virus (GTPV)” and “sheeppox virus (SPPV)”, are described as the most serious pox diseases of production animals. A GTPV isolate and a SPPV isolate were sequenced in a combined approach using nanopore MinION sequencing to obtain long reads and Illumina high throughput sequencing for short precise reads to gain full-length high-quality genome sequences. Concomitantly, sheep and goats were inoculated with SPPV and GTPV strains, respectively. During the animal trial, varying infection routes were compared: a combined intravenous and subcutaneous infection, an only intranasal infection, and the contact infection between naïve and inoculated animals. Sheep inoculated with SPPV showed no clinical signs, only a very small number of genome-positive samples and a low-level antibody reaction. In contrast, all GTPV inoculated or in-contact goats developed severe clinical signs with high viral genome loads observed in all tested matrices. Furthermore, seroconversion was detected in nearly all goats and no differences concerning the severity of the disease depending on the inoculation route were observed. Conclusively, the employed SPPV strain has the properties of an attenuated vaccine strain, consistent with the genetic data, whereas the GTPV strain represents a highly virulent field strain.
Janika Wolff; Jacqueline King; Tom Moritz; Anne Pohlmann; Donata Hoffmann; Martin Beer; Bernd Hoffmann. Experimental Infection and Genetic Characterization of Two Different Capripox Virus Isolates in Small Ruminants. Viruses 2020, 12, 1098 .
AMA StyleJanika Wolff, Jacqueline King, Tom Moritz, Anne Pohlmann, Donata Hoffmann, Martin Beer, Bernd Hoffmann. Experimental Infection and Genetic Characterization of Two Different Capripox Virus Isolates in Small Ruminants. Viruses. 2020; 12 (10):1098.
Chicago/Turabian StyleJanika Wolff; Jacqueline King; Tom Moritz; Anne Pohlmann; Donata Hoffmann; Martin Beer; Bernd Hoffmann. 2020. "Experimental Infection and Genetic Characterization of Two Different Capripox Virus Isolates in Small Ruminants." Viruses 12, no. 10: 1098.