Philip S. Mellor

Bluetongue: Epidemiology in the European Union

Central and northern Europe are now at risk from bluetongue virus.

Bluetongue in Europe: past, present and future

The recent arrival in Northern and Western (NW) Europe of bluetongue virus (BTV), which causes the ruminant disease ‘bluetongue’, has raised the profile of this vector-borne ruminant disease and sparked discussions on the reasons for its sudden emergence so far north. This expansion has not happened in isolation and the disease has been expanding into Southern and Eastern Europe for the last decade. This shifting disease distribution is being facilitated by a number of different introduction mechanisms including the movement of infected livestock, the passive movement of infected Culicoides on the wind and, in NW Europe, an unknown route of introduction. The expansion of BTV in Europe has forced a re-evaluation of the importance of Palaearctic Culicoides species in transmission, as well as the importance of secondary transmission routes, such as transplacental transmission, in facilitating the persistence of the virus. The current European outbreak of BTV-8 is believed to have caused greater economic damage than any previous single-serotype outbreak. Although attempts are being made to improve the capacity of European countries to cope with future BTV incursions, the options available are limited by a lack of basic entomological data and limited virological surveillance.

Bluetongue virus: virology, pathogenesis and immunity

Bluetongue (BT) virus, an orbivirus of the Reoviridae family encompassing 24 known serotypes, is transmitted to ruminants via certain species of biting midges (Culicoides spp.) and causes thrombo-hemorrhagic fevers mainly in sheep. During the 20th century, BTV was endemic in sub-tropical regions but in the last ten years, new strains of BTV (serotypes 1, 2, 4, 8, 9, 16) have appeared in Europe leading to a devastating disease in naive sheep and bovine herds (serotype 8). BTV enters into insect cells via the viral inner core VP7 protein and in mammalian cells via the external capsid VP2 haemagglutinin, which is the major determinant of BTV serotype and neutralization. BTV replicates in mononuclear phagocytes and endothelial cells where it induces expression of inflammatory cytokines as well as apoptosis. BTV can remain as nonreplicating entities concealed in erythrocytes for up to five months. Homologous protection against one BTV serotype involves neutralizing antibodies and T cell responses directed to the external VP2 and VP5 proteins, whereas heterologous protection is supported by T cells directed to the NS1 non structural protein and inner core proteins. Classical inactivated vaccines directed to a specific serotype generate protective immunity and may help control current epidemic situations. New recombinant vaccine strategies that allow differentiating infected from vaccinated animals and that generate cross protective immunity are urgently needed to efficiently combat this worldwide threatening disease.

Culicoides and the emergence of bluetongue virus in northern Europe

In June 2006, bluetongue virus, an arboviral pathogen of ruminants, appeared in northern Europe for the first time, successfully overwintered and subsequently caused substantial losses to the farming sector in 2007 and 2008. This emergence served as a test of how the probability of arboviral incursion into new regions is assessed and has highlighted the reliance of decision making on paradigms that are not always underpinned by basic biological data. In this review, we highlight those areas of the epidemiology of bluetongue that are poorly understood, reflect upon why certain vital areas of research have received little attention and, finally, examine strategies that could aid future risk assessment and intervention.

Identification of a novel bluetongue virus vector species of Culicoides in Sicily

The vectors of bluetongue virus are certain species of Culicoides biting midges, and in the Mediterranean area Culicoides imicola has long been considered to be the only field vector. In Sicily an entomological and serological surveillance programme has been in operation since the autumn of 2000, which has shown that the prevalence and abundance of C imicola is lower than in many other Italian regions. Moreover, in 2002, there were outbreaks of bluetongue in the absence of C imicola, and in these regions bluetongue viral RNA was detected by means of a nested reverse-transcriptase PCR in wild-caught, non-blood-engorged, parous Culicoides pulicaris. Furthermore, bluetongue virus serotype 2 was isolated on five occasions from extracts of non-blood-engorged parous C pulicaris by using embryonated hens eggs and BHK-21 cells as assay systems. These findings suggest that in parts of Italy and possibly in other areas of Europe, where C imicola is absent or rare, C pulicaris may act as a fully competent vector of bluetongue virus.

Clinical signs and pathology shown by British sheep and cattle infected with bluetongue virus serotype 8 derived from the 2006 outbreak in northern Europe

Four poll Dorset sheep and four Holstein-Friesian cattle were infected with the northern European strain of bluetongue virus (btv), btv-8, to assess its pathogenicity in uk breeds. The time course of infection was monitored in both species by using real-time reverse transcriptase-pcr (rt-pcr), conventional rt-pcr and serology. Two of the sheep developed severe clinical signs that would have been fatal in the field; the other two were moderately and mildly ill, respectively. The cattle were clinically unaffected, but had high levels of viral rna in their bloodstream. Real-time rt-pcr detected viral rna as early as one day after infection in the cattle and three days after infection in the sheep. Antibodies against btv were detected by six days after infection in the sheep and eight days after infection in the cattle. Postmortem examinations revealed pathology in the cattle that was more severe than suggested by the mild clinical signs, but the pathological and clinical findings in the sheep were more consistent.

Assessing the risk of bluetongue to UK livestock: uncertainty and sensitivity analyses of a temperature-dependent model for the basic reproduction number

Since 1998 bluetongue virus (BTV), which causes bluetongue, a non-contagious, insect-borne infectious disease of ruminants, has expanded northwards in Europe in an unprecedented series of incursions, suggesting that there is a risk to the large and valuable British livestock industry. The basic reproduction number, R0, provides a powerful tool with which to assess the level of risk posed by a disease. In this paper, we compute R0 for BTV in a population comprising two host species, cattle and sheep. Estimates for each parameter which influences R0 were obtained from the published literature, using those applicable to the UK situation wherever possible. Moreover, explicit temperature dependence was included for those parameters for which it had been quantified. Uncertainty and sensitivity analyses based on Latin hypercube sampling and partial rank correlation coefficients identified temperature, the probability of transmission from host to vector and the vector to host ratio as being most important in determining the magnitude of R0. The importance of temperature reflects the fact that it influences many processes involved in the transmission of BTV and, in particular, the biting rate, the extrinsic incubation period and the vector mortality rate.

Observations on breeding sites and light-trap collections of Culicoides during an outbreak of bluetongue in Cyprus.

During an epizootic of bluetongue virus disease of sheep in Cyprus in the autumn of 1977, 16 species of Culicoides were collected in light-traps and 4 species were collected from breeding sites. Two of the species found breeding in close association with sheep and goats are potential vectors of bluetongue virus. The possibility of a third species being a vector in Cyprus is discussed.

The Replication of Bluetongue Virus in Culicoides Vectors

BTV is maintained in nature by an endless series of alternating cycles of replication in Culicoides midges and various mammalian ruminant species. Experimentation has shown that the ability of the virus to infect Culicoides persistently and be transmitted by them is restricted to a relatively small number of species. In essence, therefore, the world distribution map of BTV is little more than a distribution map of competent insect vectors. Once ingested by a competent vector, BTV attaches to the luminal surface of the mid-gut cells, infects these cells and replicates in them. Progeny virus is then released through the basement lamina into the haemocoel from where the secondary target organs including the salivary glands are infected. Subsequent to virus replication in the salivary glands transmission can taken place. The whole cycle from infection to transmission takes between 10-15 days at 25 degrees C and individual vectors once infected usually remain so for life. Not all female midges within a vector species are susceptible to infection with BTV, or if infected, are competent to transmit the virus. A series of barriers or constraints exists within certain individuals of a vector species which either prevents virus infection or else restricts it in such a way as to stop transmission. Each population of a vector species of Culicoides has a variable proportion of these so-called refractory midges. The refractory and susceptible traits for BTV within a vector species are under genetic control, and by selective breeding, highly susceptible or completely insusceptible populations can be obtained. However, the mechanisms by which these traits are expressed are poorly understood. Further studies are therefore urgently required to determine the precise biochemical nature of these mechanisms and their mode of operation.

Oral susceptibility to bluetongue virus of Culicoides (Diptera: Ceratopogonidae) from the United Kingdom.

Abstract Oral susceptibility to infection with bluetongue virus (family Resviridae, genus Orbivirus, BTV) serotype 9 was characterized in three Palaearctic species of Culicoides (Diptera: Ceratopogonidae). Variation in susceptibility to infection by using a recently described feeding technique was shown to occur between populations of Culicoides obsoletus Meigen complex midges from different geographic regions of the United Kingdom with virus infection rates varying from 0.4 to 7.4% of those tested. Susceptibility to infection was consistent on an annual basis at selected sites. Prevalence of infection in the most susceptible populations of both the C. obsoletus and Culicoides pulicaris L. complexes was comparable with that of Culicoides imicola Kieffer, the major vector of BTV in southern Europe and throughout Africa, when using the same feeding method and virus. These results are discussed with reference to the potential threat of the virus to susceptible livestock in northern Europe.