Schmallenberg virus challenge models in cattle: infectious serum or culture-grown virus?

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Abstract

Schmallenberg virus (SBV), discovered in Europe in 2011, causes mild transient disease in adult ruminants, but fetal infection can lead to severe malformation in cattle, sheep and goats.

To elucidate the pathogenesis of this novel orthobunyavirus, considerable efforts are required. A reliable and standardized infection model is essential for in vivo studies. In the present study, two groups of four cattle were inoculated with either serum passaged in cattle only or cell culture-grown virus. The replication of culture-grown SBV in cattle was reduced compared to virus inoculated via infectious serum. In a second experiment, the infectious serum was titrated in calves; the tested batch contained 10 ^2.83 infectious doses per mL. Hence, serum-borne virus that was only passaged in the natural host is a suitable option for a standardized SBV infection model.

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Novel Orthobunyavirus in Cattle, Europe, 2011

Bernd Hoffmann, Matthias Scheuch, Dirk Höper … (2012)

In 2011, an unidentified disease in cattle was reported in Germany and the Netherlands. Clinical signs included fever, decreased milk production, and diarrhea. Metagenomic analysis identified a novel orthobunyavirus, which subsequently was isolated from blood of affected animals. Surveillance was initiated to test malformed newborn animals in the affected region.

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Rapid detection and quantification of RNA of Ebola and Marburg viruses, Lassa virus, Crimean-Congo hemorrhagic fever virus, Rift Valley fever virus, dengue virus, and yellow fever virus by real-time reverse transcription-PCR.

Christian Drosten, Stephan Göttig, Stefan Schilling … (2002)

Viral hemorrhagic fevers (VHFs) are acute infections with high case fatality rates. Important VHF agents are Ebola and Marburg viruses (MBGV/EBOV), Lassa virus (LASV), Crimean-Congo hemorrhagic fever virus (CCHFV), Rift Valley fever virus (RVFV), dengue virus (DENV), and yellow fever virus (YFV). VHFs are clinically difficult to diagnose and to distinguish; a rapid and reliable laboratory diagnosis is required in suspected cases. We have established six one-step, real-time reverse transcription-PCR assays for these pathogens based on the Superscript reverse transcriptase-Platinum Taq polymerase enzyme mixture. Novel primers and/or 5'-nuclease detection probes were designed for RVFV, DENV, YFV, and CCHFV by using the latest DNA database entries. PCR products were detected in real time on a LightCycler instrument by using 5'-nuclease technology (RVFV, DENV, and YFV) or SybrGreen dye intercalation (MBGV/EBOV, LASV, and CCHFV). The inhibitory effect of SybrGreen on reverse transcription was overcome by initial immobilization of the dye in the reaction capillaries. Universal cycling conditions for SybrGreen and 5'-nuclease probe detection were established. Thus, up to three assays could be performed in parallel, facilitating rapid testing for several pathogens. All assays were thoroughly optimized and validated in terms of analytical sensitivity by using in vitro-transcribed RNA. The >or=95% detection limits as determined by probit regression analysis ranged from 1,545 to 2,835 viral genome equivalents/ml of serum (8.6 to 16 RNA copies per assay). The suitability of the assays was exemplified by detection and quantification of viral RNA in serum samples of VHF patients.

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Organ distribution of Schmallenberg virus RNA in malformed newborns.

S Bilk, C Schulze, M. Fischer … (2012)

A novel orthobunyavirus was first detected in German dairy cows in autumn 2011 and was subsequently found in the brains of malformed lambs, kids and calves in the Netherlands, Germany, Belgium, France, Italy, Great Britain, Luxembourg and Spain. For rapid detection of this novel virus, named Schmallenberg virus, a real time quantitative reverse transcription PCR (RT-qPCR) was developed at the Friedrich-Loeffler-Institut and provided to the federal veterinary state laboratories in Germany. For diagnostic purposes, the organ distribution of this new virus was analyzed in several organs and body fluids of 15 lambs and two calves showing typical malformations. Spleen, cerebrum, meconium, spinal cord, rib cartilage, umbilical cord, placental fluid out of the stomach as well as external placental fluid scraped from the coat of the foetuses were collected during necropsy. All animals were tested RT-qPCR positive in the external placental fluid, and all but one were also RT-qPCR positive in the cerebrum, the umbilical and the spinal cord. Our results suggest that both the external placental fluid and the umbilical cord could be suitable sample materials for the confirmation of an infection with Schmallenberg virus in malformed newborns, at least in lambs. This is of special interest since those samples can be collected very easily on the farm without the need of a necropsy. Copyright © 2012 Elsevier B.V. All rights reserved.

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Author and article information

Contributors

Kerstin Wernike

Michael Eschbaumer

Angele Breithaupt

Bernd Hoffmann

Martin Beer

Journal

Journal ID (nlm-ta): Vet Res

Journal ID (iso-abbrev): Vet. Res

Title: Veterinary Research

Publisher: BioMed Central

ISSN (Print): 0928-4249

ISSN (Electronic): 1297-9716

Publication date (Print): 2012

Publication date (Electronic): 11 December 2012

Volume: 43

Issue: 1

Page: 84

Affiliations

[1 ]Institute of Diagnostic Virology, Friedrich-Loeffler-Institut (FLI), Suedufer 10, Greifswald, Insel Riems, 17493, Germany

[2 ]Department of Experimental Animal Facilities and Biorisk Management, Friedrich-Loeffler-Institut (FLI), Suedufer 10, Greifswald, Insel Riems, 17493, Germany

[3 ]Faculty of Veterinary Medicine, University of Calgary, Calgary, Canada

[4 ]Present address: Department of Veterinary Pathology, Faculty of Veterinary Medicine, Freie Universität Berlin, Berlin, Germany

Article

Publisher ID: 1297-9716-43-84

DOI: 10.1186/1297-9716-43-84

PMC ID: 3538505

PubMed ID: 23231006

SO-VID: f7d85618-6a1c-438e-a236-8cbdeeeddd3f

License:

This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Schmallenberg virus challenge models in cattle: infectious serum or culture-grown virus?

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Horses and Humans Research Foundation

Most cited references 11

Novel Orthobunyavirus in Cattle, Europe, 2011

Rapid detection and quantification of RNA of Ebola and Marburg viruses, Lassa virus, Crimean-Congo hemorrhagic fever virus, Rift Valley fever virus, dengue virus, and yellow fever virus by real-time reverse transcription-PCR.

Organ distribution of Schmallenberg virus RNA in malformed newborns.

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