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The Pirbright Institute publication directory contains details of selected publications written by our researchers.

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Tuppurainen E S, Lubinga J C, Stoltsz W H, Troskie M, Carpenter S T, Coetzer J A W, Venter E H, Oura C A L (2013)

Mechanical transmission of lumpy skin disease virus by Rhipicephalus appendiculatus male ticks

Epidemiology and Infection 141 (2), 425-430
Publisher’s version: http://dx.doi.org/10.1017/S0950268812000805

Abstract

Lumpy skin disease (LSD) is an economically important, acute or sub-acute, viral disease of cattle that occurs across Africa and in the Middle East. The aim of this study was to investigate if lumpy skin disease virus (LSDV) can be transmitted mechanically by African brown ear ticks (Rhipicephalus appendiculatus Neum.). Laboratory-bred R. appendiculatus males were fed on experimentally infected viraemic ‘donor’ cattle. Partially fed male ticks were then transferred to feed on an uninfected ‘recipient’ cow. The recipient animal became viraemic, showed mild clinical signs of LSD and seroconverted. Additionally, R. appendiculatus males were found to transmit LSDV through feeding on skin lacking visible lesions, demonstrating that viraemic animals without lesions at the feeding site of ticks may be a source of infection. This is the first time that transmission of poxviruses by a tick species has been demonstrated and the importance of this mode of transmission in the spread of LSDV in endemic settings is discussed.
Tuppurainen E S M, Lubinga J C, Stoltsz W H, Troskie M, Carpenter S T, Coetzer J A W, Venter E H, Oura C A L (2013)

Evidence of vertical transmission of lumpy skin disease virus in Rhipicephalus decoloratus ticks

Ticks and Tick-borne Diseases 4 (4), 329-333
Publisher’s version: http://dx.doi.org/10.1016/j.ttbdis.2013.01.006

Abstract

Lumpy skin disease (LSD) is an economically important acute or sub-acute disease of cattle that occurs across Africa and in the Middle East. The aim of this study was to assess whether Rhipicephalus decoloratus ticks were able to transmit lumpy skin disease virus (LSDV) transovarially. Uninfected, laboratory-bred R. decoloratus larvae were placed to feed on experimentally infected “donor” cattle. After completion of the life cycle on donor animals, fully engorged adult female ticks were harvested and allowed to lay eggs. Larvae that hatched from these eggs were then transferred to feed on uninfected “recipient” cattle. The latter became viraemic and showed mild clinical disease with characteristic skin lesions and markedly enlarged precrural and subscapular lymph nodes. This is the first report of transovarial transmission of poxviruses by R. decoloratus ticks, and the importance of this mode of transmission in the spread of LSDV in endemic settings requires further investigation.
Valdazo-Gonzalez B, Timina A, Scherbakov A, Abdul-Hamid N F, Knowles N J, King D P (2013)

Multiple introductions of serotype O foot-and-mouth disease viruses into East Asia in 2010-2011

Veterinary Research 44, e76
Publisher’s version: http://dx.doi.org/10.1186/1297-9716-44-76

Abstract

Foot-and-mouth disease virus (FMDV) is a highly contagious and genetically variable virus. Sporadic introductions of this virus into FMD-free countries may cause outbreaks with devastating consequences. In 2010 and 2011, incursions of the FMDV O/SEA/Mya-98 strain, normally restricted to countries in mainland Southeast Asia, caused extensive outbreaks across East Asia. In this study, 12 full genome FMDV sequences for representative samples collected from the People's Republic of China (PR China) including the Hong Kong Special Administrative Region (SAR), the Republic of Korea, the Democratic People's Republic of Korea, Japan, Mongolia and The Russian Federation were generated and compared with additional contemporary sequences from viruses within this lineage. These complete genomes were 8119 to 8193 nucleotides in length and differed at 1181 sites, sharing a nucleotide identity >= 91.0% and an amino acid identity >= 96.6%. An unexpected deletion of 70 nucleotides within the 5'-untranslated region which resulted in a shorter predicted RNA stem-loop for the S-fragment was revealed in two sequences from PR China and Hong Kong SAR and five additional related samples from the region. Statistical parsimony and Bayesian phylogenetic analysis provide evidence that these outbreaks in East Asia were generated by two independent introductions of the O/SEA/Mya-98 lineage sometime between August 2008 and March 2010. The rapid emergence of these viruses from Southeast Asia highlights the importance of adopting approaches to closely monitor the spread of this lineage that now poses a threat to livestock industries in other regions.
van Knippenberg I, Fragkoudis R, Elliott R M (2013)

The transient nature of Bunyamwera orthobunyavirus NSs protein expression: effects of increased stability of NSs protein on virus replication

PLoS ONE 8 (5), e64137
Publisher’s version: http://dx.doi.org/10.1371/journal.pone.0064137

Abstract

The NSs proteins of bunyaviruses are the viral interferon antagonists, counteracting the host's antiviral response to infection. During high-multiplicity infection of cultured mammalian cells with Bunyamwera orthobunyavirus (BUNV), NSs is rapidly degraded after reaching peak levels of expression at 12hpi. Through the use of inhibitors this was shown to be the result of proteasomal degradation. A recombinant virus (rBUN4KR), in which all four lysine residues in NSs were replaced by arginine residues, expresses an NSs protein (NSs4KR) that is resistant to degradation, confirming that degradation is lysine-dependent. However, despite repeated attempts, no direct ubiquitylation of NSs in infected cells could be demonstrated. This suggests that degradation of NSs, although lysine-dependent, may be achieved through an indirect mechanism. Infection of cultured mammalian cells or mice indicated no disadvantage for the virus in having a non-degradable NSs protein: in fact rBUN4KR had a slight growth advantage over wtBUNV in interferon-competent cells, presumably due to the increased and prolonged presence of NSs. In cultured mosquito cells there was no difference in growth between wild-type BUNV and rBUN4KR, but surprisingly NSs4KR was not stabilised compared to the wild-type NSs protein.
Veronesi E, Antony F, Gubbins S, Golding N, Blackwell A, Mertens P P C, Brownlie J, Darpel K E, Mellor P S, Carpenter S (2013)

Measurement of the infection and dissemination of bluetongue virus in Culicoides biting midges using a semi-quantitative RT-PCR assay and isolation of infectious virus

PLoS ONE 8 (8), e70800
Publisher’s version: http://dx.doi.org/10.1371/journal.pone.0070800

Abstract

Background: Culicoides biting midges (Diptera: Ceratopogonidae) are the biological vectors of globally significant arboviruses of livestock including bluetongue virus (BTV), African horse sickness virus (AHSV) and the recently emerging Schmallenberg virus (SBV). From 2006-2009 outbreaks of BTV in northern Europe inflicted major disruption and economic losses to farmers and several attempts were made to implicate Palaearctic Culicoides species as vectors. Results from these studies were difficult to interpret as they used semi-quantitative RT-PCR (sqPCR) assays as the major diagnostic tool, a technique that had not been validated for use in this role. In this study we validate the use of these assays by carrying out time-series detection of BTV RNA in two colony species of Culicoides and compare the results with the more traditional isolation of infectious BTV on cell culture. Methodology/Principal Findings: A BTV serotype 1 strain mixed with horse blood was fed to several hundred individuals of Culicoides sonorensis (Wirth & Jones) and C. nubeculosus (Mg.) using a membrane-based assay and replete individuals were then incubated at 25 degrees C. At daily intervals 25 Culicoides of each species were removed from incubation, homogenised and BTV quantified in each individual using sqPCR (C-q values) and virus isolation on a KC-C. sonorensis embryonic cell line, followed by antigen enzyme-linked immunosorbent assay (ELISA). In addition, comparisons were also drawn between the results obtained with whole C. sonorensis and with individually dissected individuals to determine the level of BTV dissemination. Conclusions/Significance: C-q values generated from time-series infection experiments in both C. sonorensis and C. nubeculosus confirmed previous studies that relied upon the isolation and detection of infectious BTV. Implications on the testing of field-collected Culicoides as potential virus vectors by PCR assays and the use of such assays as front-line tools for use in diagnostic laboratories in this role are discussed.
Veronesi E, Henstock M, Gubbins S, Batten C, Manley R, Barber J, Hoffmann B, Beer M, Attoui H, Mertens P P C, Carpenter S (2013)

Implicating Culicoides biting midges as vectors of Schmallenberg virus using semi-quantitative RT-PCR

PLoS ONE 8 (3), e57747
Publisher’s version: http://dx.doi.org/10.1371/journal.pone.0057747

Abstract

The recent unprecedented emergence of arboviruses transmitted by Culicoides biting midges in northern Europe has necessitated the development of techniques to differentiate competent vector species. At present these techniques are entirely reliant upon interpretation of semi-quantitative RT-PCR (sqPCR) data in the form of Cq values used to infer the presence of viral RNA in samples. This study investigates the advantages and limitations of sqPCR in this role by comparing infection and dissemination rates of Schmallenberg virus (SBV) in two colony lines of Culicoides. Through the use of these behaviorally malleable lines we provide tools for demarcating arbovirus infection and dissemination rates in Culicoides which to date have prevented clear implication of primary vector species in northern Europe. The study demonstrates biological transmission of SBV in an arthropod vector, supporting the conclusions from field-caught Culicoides and provides a general framework for future assessment of vector competence of Culicoides for arboviruses using sqPCR. When adopting novel diagnostic technologies, correctly implicating vectors of arboviral pathogens requires a coherent laboratory framework to fully understand the implications of results produced in the field. This study illustrates these difficulties and provides a full examination of sqPCR in this role for the Culicoides-arbovirus system.
Volpato G, Di Nardo A, Rossi D, Saleh S M L, Broglia A (2013)

'Everybody knows', but the rest of the world: the case of a caterpillar-borne reproductive loss syndrome in dromedary camels observed by Sahrawi pastoralists of Western Sahara

Journal of Ethnobiology and Ethnomedicine 9, e5
Publisher’s version: http://dx.doi.org/10.1186/1746-4269-9-5

Abstract

Background: The traditional knowledge of local communities throughout the world is a valuable source of novel ideas and information to science. In this study, the ethnoveterinary knowledge of Sahrawi pastoralists of Western Sahara has been used in order to put forward a scientific hypothesis regarding the competitive interactions between camels and caterpillars in the Sahara ecosystem. Methods: Between 2005 and 2009, 44 semi-structured interviews were conducted with Sahrawi pastoralists in the territories administered by the Sahrawi Arab Democratic Republic, Western Sahara, using a snow-ball sampling design. Results: Sahrawi pastoralists reported the existence of a caterpillar-borne reproductive loss syndrome, known locally as duda, affecting their camels. On the basis of Sahrawi knowledge about duda and of a thorough literature review, we built the hypothesis that: 1) caterpillars of the family Lasiocampidae (genera Lasiocampa, Psilogaster, or Streblote) have sudden and rare outbreaks on Acacia treetops in the Western Sahara ecosystem after heavy rainfall; 2) during these outbreaks, camels ingest the caterpillars while browsing; 3) as a consequence of this ingestion, pregnant camels have sudden abortions or give birth to weaklings. This hypothesis was supported by inductive reasoning built on circumstantiated evidence and analogical reasoning with similar syndromes reported in mares in the United States and Australia. Conclusions: The possible existence of a caterpillar-borne reproductive loss syndrome among camels has been reported for the first time, suggesting that such syndromes might be more widespread than what is currently known. Further research is warranted to validate the reported hypothesis. Finally, the importance of studying folk livestock diseases is reinforced in light of its usefulness in revealing as yet unknown biological phenomena that would deserve further investigation.
Weesendorp E, Morgan S, Stockhofe-Zurwieden N, Popma-De Graaf D J, Graham S P, Rebel J M J (2013)

Comparative analysis of immune responses following experimental infection of pigs with European porcine reproductive and respiratory syndrome virus strains of differing virulence

Veterinary Microbiology 163 (1-2), 1-12
Publisher’s version: http://dx.doi.org/10.1016/j.vetmic.2012.09.013

Abstract

Porcine reproductive and respiratory syndrome virus (PRRSV) is difficult to control due to a high mutation rate and the emergence of virulent strains. The objective of this study was to analyze the immunological and pathological responses after infection with the European subtype 3 strain Lena in comparison to subtype 1 strains Belgium A and Lelystad-Ter Huurne (LV). Sixteen pigs were inoculated per strain, and sixteen pigs with PBS. At days 7 and 21 post-inoculation (p.i.), four pigs per group were immunized with an Aujeszky disease vaccine (ADV) to study the immune competence after PRRSV infection. Infection with the Lena strain resulted in fever and clinical signs. This was not observed in the Belgium A or LV-infected pigs. Infection with the Lena strain resulted in high virus titers in serum, low numbers of IFN-gamma secreting cells, a change in leukocyte populations and a delayed antibody response to immunization with ADV. Levels of IL-1 beta, IFN-alpha, IL-10, IL-12, TNE-alpha and IFN-gamma mRNA of the Lena-infected pigs were increased during the first week of infection. For pigs infected with the Belgium A or LV strain, the effects of infection on these parameters were less pronounced, although for the Belgium A-infected pigs, the level of the analyzed cytokines, except for TNF-alpha, and leukocyte populations were comparable to the Lena-infected pigs. These results suggest that while the outcome of infection for the three strains was comparable, with mostly clearance of viremia at day 33 p.i, differences in immune responses were observed, perhaps contributing to their virulence.
Wensman J J, Munir M, Thaduri S, Hornaeus K, Rizwan M, Blomstrom A-L, Briese T, Lipkin W I, Berg M (2013)

The X proteins of bornaviruses interfere with type I interferon signalling

Journal of General Virology 94, 263-269
Publisher’s version: http://dx.doi.org/10.1099/vir.0.047175-0

Abstract

Borna disease virus (BDV) is a neurotropic, negative-stranded RNA virus causing persistent infection and progressive neurological disorders in a wide range of warm-blooded animals. The role of the small non-structural X protein in viral pathogenesis is not completely understood. Here we investigated whether the X protein of BDV and avian bornavirus (ABV) interferes with the type I interferon (IFN) system, similar to other non-structural proteins of negative-stranded RNA viruses. In luciferase reporter assays, we found that the X protein of various bornaviruses interfered with the type I IFN system at all checkpoints investigated, in contrast to previously reported findings, resulting in reduced type I IFN secretion.
Willcocks S, Offord V, Seyfert H-M, Coffey T J, Werling D (2013)

Species-specific PAMP recognition by TLR2 and evidence for species-restricted interaction with Dectin-1

Journal of Leukocyte Biology 94 (3), 449-458
Publisher’s version: http://dx.doi.org/10.1189/jlb.0812390

Abstract

TLRs mediate recognition of a wide range of microbial products, including LPS, lipoproteins, flagellin, and bacterial DNA, and signaling through TLRs leads to the production of inflammatory mediators. In addition to TLRs, many other surface receptors have been proposed to participate in innate immunity and microbial recognition, and signaling through some of these, for example, C-type lectins, is likely to cooperate with TLR signaling in defining inflammatory responses. In the present study, we examined the importance of the ECD and intracellular TIR domain of boTLR2 and huTLR2 to induce a species-specific response by creating a chimeric TLR2 protein. Our results indicate that the strength of the response to any TLR2 ligand tested was dependent on the extracellular, solenoid structure, but not the intracellular TIR domain. Furthermore, we examined whether the recognition of two PAMPs by Dectin-1, a CLR, depends on the interaction with TLR2 from the same species. TLR2 expression seemed to affect the Dectin-1-dependent production of CXCL8 to ?-glucan containing zymosan as well as Listeria monocytogenes. Furthermore, the interaction of Dectin-1 with TLR2 seemed to require that both receptors are from the same species. Our data demonstrate that the differences in the TLR2 response seen between the bovine and human system depend on the ECD of TLR2 and that collaborative recognition of distinct microbial components by different classes of innate-immune receptors is crucial in orchestrating inflammatory responses.

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