The biting midge Culicoides imicola Kieffer (Diptera: Ceratopogonidae) is the most important Old World vector of African horse sickness (AHS) and bluetongue (BT). Recent increases of BT incidence in the Mediterranean basin are attributed to its increased abundance and distribution. The phylogenetic status and genetic structure of C. imicola in this region are unknown, despite the importance of these aspects for BT epidemiology in the North American BT vector. In this study, analyses of partial mitochondrial cytochrome oxidase subunit I gene (COI) sequences were used to infer phylogenetic relationships among 50 C. imicola from Portugal, Rhodes, Israel, and South Africa and four other species of the Imicola Complex from southern Africa, and to estimate levels of matrilineal subdivision in C. imicola between Portugal and Israel. Eleven haplotypes were detected in C. imicola, and these formed one well-supported clade in maximum likelihood and Bayesian trees implying that the C. imicola samples comprise one phylogenetic species. Molecular variance was distributed mainly between Portugal and Israel, with no haplotypes shared between these countries, suggesting that female-mediated gene flow at this scale has been either limited or non-existent. Our results provide phylogenetic evidence that C. imicola in the study areas are potentially competent AHS and BT vectors. The geographical structure of the C. imicola COI haplotypes was concordant with that of BT virus serotypes in recent BT outbreaks in the Mediterranean basin, suggesting that population subdivision in its vector can impose spatial constraints on BT virus transmission.

A faunistic study of phlebotomine sandflies was carried out on the mainland and on four islands in Greece between 1999 and 2004. Sandflies were collected in 18 areas, and the population structure was observed and recorded. A total of 10 species were identified; their distribution is presented here. Of these, the species Phlebotomus neglectus Tonnoir and P. perfiliewi Parrot (Diptera: Psychodidae), epidemiologically the most important vectors of leishmaniases and sandfly fever in Greece, were shown to be present in the main endemic foci of the country.

Summary 1.The spread of vector-borne diseases into new areas, commonly attributed to environmental change or increased trade and travel, could be exacerbated if novel vector species in newly invaded areas spread infection beyond the range of traditional vectors. 2. By analysing the differential degree of overlap between the environmental envelopes for bluetongue, a devastating livestock disease, and its traditional (Afro-Asian) and potential new (Palearctic) midge vectors, we have implicated the latter in the recent dramatic northward spread of this disease into Europe. 3. The traditional vector of bluetongue virus, the Afro-Asian midge Culicoides imicola , was found to occur in warm (annual mean 12-20 ° C), thermally stable locations that were dry in summer (< 400 mm precipitation). The Palearctic C. obsoletus and C. pulicaris complexes were both found to occur in cooler (down to 7 ° C annual mean), thermally more variable and wetter (up to 700 mm summer precipitation) locations. 4. Of 501 recorded outbreaks from the 1998-2004 bluetongue epidemic in southern Europe, 40% fall outside the climate envelope of C. imicola , but within the species' envelopes of the C. obsoletus and C. pulicaris complexes. 5. The distribution in multivariate environmental space of bluetongue virus is closer to that of the Palaearctic vectors than it is to that of C. imicola . This suggests that Palearctic vectors now play a substantial role in transmission and have facilitated the spread of bluetongue into cooler, wetter regions of Europe. 6. Synthesis and applications . The risk to Northern Europe now depends on how much of the distributions of the widespread, abundant Palearctic midge vectors (the C. obsoletus and C. pulicaris complexes) bluetongue can occupy, perhaps determined by thermal constraints on viral replication. This was highlighted by the sudden appearance in summer 2006 of bluetongue virus at latitudes of more than 50 ° North -approximately 6 ° further North than previous outbreaks in southern Europe. Future surveillance for bluetongue and for related Culicoides -borne pathogens should include studies to record and explain the distributional patterns of all potential Palearctic vector species.

Culicoides biting midges (Diptera: Ceratopogonidae) were trapped between 1999 and 2004 at 122 locations in mainland Greece and on most of the larger Aegean and Ionian islands, using OVI light traps, in order to determine the distribution and seasonal activity of bluetongue virus vectors and other Culicoides species. Thirty-nine Culicoides species were identified, six of which (C. furcillatus, C. impunctatus, C. paolae, C. pictipennis, C. riethi, and C. scoticus) were identified for the first time in Greece. Two of these (C. impunctatus and C. scoticus) may be of veterinary importance due to their role as vectors of bluetongue virus and related orbiviruses. In addition, C. imicola was detected for the first time in mainland Greece.