Bluetongue in Europe and the Mediterranean Basin: history of occurrence prior to 2006

Bluetongue virus (BTV) exists around the world in a broad band covering much of the Americas, Africa, southern Asia and northern Australia. Historically, it also occasionally occurred in the southern fringes of Europe. It is considered to be one of the most important diseases of domestic livestock. Recently BTV has extended its range northwards into areas of Europe never before affected and has persisted in many of these locations causing the greatest epizootic of bluetongue (BT), the disease caused by BTV, on record. Indeed, the most recent outbreaks of BT in Europe are further north than this virus has ever previously occurred anywhere in the world. The reasons for this dramatic change in BT epidemiology are complex but are linked to recent extensions in the distribution of its major vector, Culicoides imicola, to the involvement of novel Culicoides vector(s) and to on-going climate-change. This paper investigates these recent outbreaks in the European theatre, up to the beginning of 2006, highlights prospects for the future and sets the scene for the following papers in this special issue.     

Prediction of bluetongue vector distribution in Europe and north Africa using satellite imagery

Bluetongue is an infectious, non-contagious arboviral disease thought to infect all known ruminant species. Since 1998, an unprecedented epizootic of the disease has occurred in the Mediterranean region, resulting in the deaths of over 800,000 sheep to date. Bluetongue virus (BTV) is transmitted by biting midges of which one species, Culicoides imicola, is the major vector in the old world. C. imicola was trapped for 2 years at 87 sites across Portugal and models were developed for predicting the presence and abundance of the midge at these sites. Discriminant analysis was used to identify the best models from 40 temporally Fourier-processed 1 km spatial resolution remotely-sensed variables. The best models correctly predicted presence and absence at 83 of the 87 sites, and abundance at 76 sites. The models were then used to predict C. imicola presence and abundance elsewhere across Europe and north Africa. C. imicola was predicted to be present and in high abundance at the majority of areas affected in the recent bluetongue epizootic, including the Balearics, Sardinia, Corsica, Sicily, areas of mainland Italy, large areas of Greece, western Turkey and northern Algeria and Tunisia.     

Evidence for a new field Culicoides vector of African horse sickness in South Africa

Between February and May 1998, approximately 100 horses died of African horse sickness (AHS) in the cooler, mountainous, central region of South Africa. On 14 affected farms, 156,875 Culicoides of 27 species were captured. C. imicola Kieffer, hitherto considered the only field vector for AHS virus (AHSV), constituted <1% of the total Culicoides captured, and was not found on 29% of the farms. In contrast, 65% of the Culicoides were C. bolitinos Meiswinkel, and was found on all farms. Five isolations of AHSV were made from C. bolitinos, and none from 18 other species of Culicoides (including C. imicola).     

Prediction of areas around the Mediterranean at risk of bluetongue by modelling the distribution of its vector using satellite imaging.

Bluetongue is an infectious disease of ruminants caused by a virus transmitted by biting midges, one species of which, Culicoides imicola, is the major vector in the Old World. Following an epizootic of African horse sickness,a related disease, in Iberia and Morocco between 1987 and 1991, C imicola was trapped for two years at 44 sites in the affected region and models were developed for predicting the abundance of C imicola at these sites. Discriminant analysis was applied to identify the best model of three levels of abundance from 40 Fourier-processed remotely sensed variables and a digital elevation model. The best model correctly predicted the abundance level at 41 of the 44 sites. The single most important variable was the phase of the annual cycle of the normalised difference vegetation index. The model was used to predict the abundances of C imicola elsewhere around the Mediterranean and predicted high levels of abundance in many areas recently affected by bluetongue, including the Balearics, Sardinia, Sicily, eastern Greece, western Turkey, Tunisia and northern Algeria. The model suggests that eastern Spain, the island of Ibiza, the provinces of Lazio and Puglia in Italy, the Peloponnese and parts of northern Algeria and Libya may be at risk of bluetongue in 2001.     

Molecular detection of Culicoides spp. and Culicoides imicola, the principal vector of bluetongue (BT) and African horse sickness (AHS) in Africa and Europe

Bluetongue (BT) and African Horse Sickness (AHS) are infectious arthropod-borne viral diseases affecting ruminants and horses, respectively. Culicoides imicola Kieffer, 1913, a biting midge, is the principal vector of these livestock diseases in Africa and Europe. Recently bluetongue disease has re-emerged in the Mediterranean Basin and has had a devastating effect on the sheep industry in Italy and on the islands of Sicily, Sardinia, Corsica and the Balearics, but fortunately, has not penetrated onto mainland France and Spain. To survey for the presence of C. imicola, an extensive light-trap network for the collection of Culicoides, was implemented in 2002 in southern mainland France. The morphological identification of Culicoides can be both tedious and time-consuming because its size ranges from 1.5 to 3 mm. Therefore, an ITS1 rDNA polymerase chain reaction (PCR)-based diagnostic assay was developed to rapidly and reliably identify Culicoides spp. and C. imicola. The aim of this work was to set up a rapid test for the detection of C. imicola amongst a pool of insects collected in areas at risk for BT. The sequence similarity of the rDNA (nuclear ribosomal DNA), which is greater within species than between species, is the foundation of its utilisation in species-diagnostic assays. The alignment of the 11 ITS1 sequences of Culicoides obtained from Genbank and EMBL databases helped us to identify one region in the 5' end and one in the 3' end that appear highly conserved. PCR primers were designed within these regions to amplify genus-specific fragments. In order to set up a C. imicola-specific PCR, another forward primer was designed and used in combination with the previously designed reverse primer. These primers proved to be highly specific and sensitive and permitted a rapid diagnostic separation of C. imicola from Culicoides spp.     

Bluetongue virus isolations from midges belonging to the Obsoletus complex (Culicoides, Diptera: Ceratopogonidae) in Italy

Between July and September 2002 there were outbreaks of bluetongue on three sheep holdings in the communities of San Gregorio Magno (Salerno, Campania), Laviano (Salerno, Campania) and Carpino (Foggia, Puglia), and the involvement of bluetongue virus (btv) was confirmed serologically and virologically. The mortality rate was at least 11 per cent and involved btv serotype 2 (btv-2) and serotype 9 (btv-9). These holdings were also surveyed for the Culicoides (Diptera: Ceratopogonidae) vectors; approximately 10,000 midges belonging to 15 species were captured, but they did not include a single specimen of the classical Afro-Asiatic bluetongue vector, Culicoides imicola. Species belonging to the Obsoletus complex dominated the light-trap collections, and Culicoides obsoletus Meigen, Culicoides scoticus Downes and Kettle and Culicoides dewulfi Goetghebuer constituted 90 per cent of all the Culicoides species captured. Fifty-six pools of the Obsoletus complex (excluding C dewulfi), each containing 100 individual midges and containing only parous and gravid females, were assayed for virus. btv-2 was isolated from three pools from San Gregorio Magno and Carpino, and btv-9 was isolated from one pool from Laviano. These results indicate that a species other than C imicola is involved in the current re-emergence of bluetongue in the Mediterranean Basin, but whether it is C obsoletus sensu stricto or C scoticus, or both, is uncertain.     

Seasonal abundance and parity of Culicoides biting midges associated with livestock at Roma, Lesotho (Diptera: Ceratopogonidae)

A light-trap survey was undertaken of the species composition, seasonal abundance and parity of Culicoides at Roma, Lesotho, to establish whether the likely vectors for bluetongue and African horse sickness occur in this area as well as the chance of transmission. A total of 34 catches was made between 21 September 1985 and 24 September 1986; 32,819 Culicoides were caught belonging to 19 species. Culicoides numbers rapidly built up from December to a peak in February which implies that this may also be the optimum time for virus transmission. The number of Culicoides dropped sharply in April with the onset of cooler conditions. C. zuluensis was the dominant species forming 69.6% of the totalled catches, followed by C. pycnostictus with 11.7%. C. imicola, the only proven vector of bluetongue, was never abundant representing only 4.4% of the midges caught. The parous rate for each of the 2 commonest species was low, implying a low vector capacity.     

Identification of cryptic species of Culicoides (Diptera: Ceratopogonidae) in the subgenus Culicoides and development of species-specific PCR assays based on barcode regions

Culicoides biting midges (Diptera: Ceratopogonidae) are vectors of important diseases affecting wild and domestic animals. During the last decade they have played a major role in the epidemiology of the largest bluetongue epizootic ever recorded in Europe, the disease is transmitted between hosts almost exclusively by bites of Culicoides midges and affects both domestic and wild ruminants however severe disease usually occurs in certain breeds of sheep and some species of deer. An accurate vector identification is of major importance in arthropod borne diseases surveillance, as great differences in vectorial capacity are found even between close species. Unfortunately, specialized taxonomic knowledge of Culicoides identification is rarely available in routine surveillance, mainly based on wing morphology. Recently, some European species of Culicoides belonging to the subgenus Avaritia Fox, 1955 and Culicoides Latreille, 1809 have been described as new bluetongue virus vectors.In the present study, by using a fragment of the barcode region (COI gene) we report the presence of up to 11 species within the subgenus Culicoides in Catalonia (NE Spain), a region recently affected by a bluetongue epizootic. The molecular analysis revealed new non-described cryptic species which were grouped in three complexes of morphologically similar species, two in the Pulicaris complex resembling Culicoides pulicaris, two in the Fagineus complex resembling Culicoides fagineus and three in the Newsteadi complex resembling Culicoides newsteadi. The phylogenetic relationships among them showed that cryptic species detected in both Pulicaris and Fagineus complexes were closely related, whereas those in the Newsteadi complex were more distant. Accurate analysis of all species using morphological and molecular approaches resulted in the detection of diagnostic metric traits for cryptic species and the design of several new species-specific single and multiplex PCR assays to identify unambiguously all the species, most of them still lacking a specific molecular diagnosis.     

Identifying equine premises at high risk of introduction of vector-borne diseases using geo-statistical and space-time analyses

The identification of premises that may play an important role in the introduction or spread of animal diseases is fundamental to the development of risk-based surveillance and control programs. A combination of geo-statistical and cluster analysis methods was used to identify geographical areas and periods of time at highest risk for introduction of the African horse sickness virus (AHSV) into the Castile and Leon (CyL) region of Spain. Risk was estimated based on the predicted premises-specific abundance of Culicoides spp., a vector for AHSV, and on the frequency of equine introductions from outside regions. The largest abundance of Culicoides spp. was observed between May and September in the northern region of CyL. Six significant (P-value <0.01) space-time clusters of equine premises were found, at which presence of Culicoides spp. was predicted and live equidae were introduced from outside CyL. The clusters included 37 equine premises and took place between April and December. These results will contribute to updating plans for prevention of AHSV introduction and spread in Spain. The methodological approach developed here may be adapted and applied to design and establish risk-based surveillance and control programs for Spain and other European countries.     

The distribution of Culicoides imicola in Italy: application and evaluation of current Mediterranean models based on climate

In August 2000 bluetongue (BT) disease appeared amongst sheep on the island of Sardinia spreading later to Sicily and to mainland Italy. The majority of areas affected by BT were surveyed for Culicoides imicola, the only proven vector of the disease known to occur in the Mediterranean region. The data from 1456 light-trap collections, made in months with a mean temperature of 12.5 degrees C, were used to test the accuracy of current models predicting the prevalence and abundance of C. imicola across the region. For Italy, the distribution of C. imicola was found to be very irregular and did not fit the modelled predictions. The possible reasons for this are discussed, and suggestions made as to which variables may improve this fit in the development of future risk models. In Italy, past surveys failed to reveal the presence of C. imicola, and so could be construed as evidence of its recent invasion, and thus rampant spread northwards. Although equivocal, historical records indicate that C. imicola was overlooked in the past. Six recommendations are made as to the possible future course of Culicoides research in southern Europe.     

Bluetongue virus in the Mediterranean Basin 1998-2001

Bluetongue (BT) exists around the world in a broad band covering much of the Americas, Africa, southern Asia, northern Australia and, occasionally, the southern fringe of Europe. It is considered to be one of the most important diseases of domestic livestock. Recently the virus causing this disease has extended its range northwards into areas of Europe never before affected and has persisted in many of these locations causing the greatest epizootic of the disease on record. The reasons for this dramatic change in BT epidemiology are complex but are linked to recent extensions in the distribution of its major vector, Culicoidesimicola, to the involvement of novel Culicoides vector(s) and to an apparent ability of the virus to overwinter in the absence of adult vectors. In addition, the effects of these changes have been exacerbated by problems in control, particularly in relation to vaccination. This paper explores these areas and highlights prospects for the future.     

Climate change: effects on culicoides--transmitted viruses and implications for the UK

Changes in the distribution and abundance of insects are likely to be amongst the most important and immediate effects of climate change. We review here the risk that climate change poses to the UK's livestock industry via effects on Culicoides biting midges, the vectors of several arboviruses, including those that cause bluetongue (BT) and African horse sickness (AHS). The major old-world vector of BT and AHS viruses, C. imicola, occurs in southern Europe and will spread further north as global temperatures increase. It is unlikely, however, that in the foreseeable future it will reach and become established in the UK. As the distribution of C. imicola moves north, however, it may bring BT and AHS viruses into the range of other Culicoides species that are known to be competent vectors and which occur much further north. Once infected via this 'baton effect', these species may be able to spread the viruses over much of Europe, including the UK. Climate change may increase their vector competence further and will also increase the likelihood of viruses surviving from one year to the next. An additional risk is that the predicted increase in the frequency of short periods of hot temperatures may lead to the creation of novel vector species, by removing the barriers that in colder conditions make them refractory to viral infection.     

Prevalence of bluetongue virus antibodies in sheep in central Ethiopia

Competitive ELISA was applied to detect antibodies against bluetongue virus in sheep sera collected from different agro-climatic areas in Ethiopia. A total of 90 serum samples were tested and 42 (46.67%) were positive for bluetongue virus antibodies. A prevalence rate ranging from 9.67% for sheep sampled in the highland to 92.85% for sheep sampled in the lowland was recorded. The prevalence correlated with the probable distribution of the Culicoides vector. This is the first report indicating the presence of bluetongue virus infection in animals from Ethiopia.     

The phenology and population dynamics of Culicoides spp. in different ecosystems in The Netherlands

The Netherlands has enjoyed a relatively free state of vector-borne diseases of economic importance for more than one century. Emerging infectious diseases may change this situation, threatening the health of humans, domestic livestock and wildlife. In order to be prepared for the potential outbreak of vector-borne diseases, a study was undertaken to investigate the distribution and seasonal dynamics of candidate vectors of infectious diseases with emphasis on bluetongue vectors (Culicoides spp.). The study focused primarily on the relationship between characteristic ecosystems suitable for bluetongue vectors and climate, as well as on the phenology and population dynamics of these vectors. Twelve locations were selected, distributed over four distinct habitats: a wetland area, three riverine systems, four peat land areas and four livestock farms. Culicoides populations were sampled continuously using CO(2)-baited counterflow traps from July 2005 until August 2006, with an interruption from November 2005 to March 2006. All vectors were identified to species level. Meteorological and environmental data were collected at each location. Culicoides species were found in all four different habitat types studied. Wetland areas and peat bogs were rich in Culicoides spp. The taxonomic groups Culicoides obsoletus (Meigen) and Culicoides pulicaris (Linnaeus) were strongly associated with farms. Eighty-eight percent of all Culicoides consisted of the taxon C. obsoletus/Culicoides scoticus. On the livestock farms, 3% of Culicoides existed of the alleged bluetongue vector Culicoides dewulfi Goetghebuer. Culicoides impunctatus Goetghebuer was strongly associated with wetland and peat bog. Many Culicoides species were found until late in the phenological season and their activity was strongly associated with climate throughout the year. High annual variations in population dynamics were observed within the same study areas, which were probably caused by annual variations in environmental conditions. The study demonstrates that candidate vectors of bluetongue virus are present in natural and livestock-farm habitats in the Netherlands, distributed widely across the country. Under favourable climatic conditions, following virus introduction, bluetongue can spread among livestock (cattle, sheep and goats), depending on the nature of the viral serotype. The question now arises whether the virus can survive the winter conditions in north-western Europe and whether measures can be taken that effectively halt further spread of the disease.     

环状病毒的流行与传播

环状病毒是牲畜常见的重要病原体,主要包括有蓝舌病病毒、非洲马瘟病毒、马器质性脑病病毒和流行性出血热病毒等。这些病毒能够通过吸血性的库蠓传播。本文主要介绍了这几种病毒在世界各地的流行与传播情况。     

The use of a membrane feeding technique to determine the infection rate of Culicoides imicola (Diptera, Ceratopogonidae) for 2 bluetongue virus serotypes in South Africa.

Culicoides spp. in the Lowveld of the northern Transvaal, Republic of South Africa, were fed bluetongue virus serotypes 3 and 6 and African horsesickness virus serotype 1 through latex and chicken skin membranes. After an incubation period of 10 days at 25-27 degrees C, the infection rate of C. imicola for bluetongue virus serotypes 3 and 6 was established at 31% and 24% respectively. No African horsesickness virus could be recovered. The membrane feeding technique and handling procedures proved to be suitable for field studies.