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Background  Bluetongue disease of ruminants is a typical insect-borne disease caused by bluetongue virus (BTV) of the genus  Orbivirus  (family  Reoviridae ) and transmitted by some species of  Culicoides  (Diptera: Ceratopogonidae). Recently, the detection of BTV in yaks in high altitude meadows of the Shangri-La district of Yunnan Province, China, prompted an investigation of the  Culicoides  fauna as potential vectors of BTV.    Methods  A total of 806  Culicoides  midges were collected by light trapping at three sites at altitudes ranging from 1800 to 3300 m. The species were identified based on morphology and the DNA sequences of cytochrome  c  oxidase subunit 1 ( cox 1). PCR and quantitative PCR following reverse transcription were used to test for the presence of BTV RNA in  Culicoides  spp. A phylogenetic analysis was used to analyze the  cox 1 sequences of some specimens.    Results  Four species dominated these collections and  cox 1 barcoding revealed that at least two of these appear to belong to species new to science.  Culicoides tainanus  and a cryptic species morphologically similar to  C. tainanus  dominated low altitude valley collections while  C. nielamensis  was the most abundant species in the high-altitude meadow. A species related to  C. obsoletus  occurred at all altitudes but did not dominate any of the collections. BTV RT-qPCR analysis detected BTV RNA in two specimens of  C. tainanus , in one specimen closely related to  C. tainanus  and in one specimen closely related to  C. obsoletus  by barcode sequencing.    Conclusions  This study suggests that BTV in high altitude areas of Yunnan is being transmitted by three species of  Culicoides , two of which appear to be new to science. This research may be useful in improving understanding of the effects of global warming on arboviral disease epidemiology and further study is important in research into disease control and prevention.

Potential vectors of bluetongue virus in high altitude areas of Yunnan Province, China