Experimental bluetongue virus infection of Culicoides austropalpalis, collected from a farm environment in Victoria, Australia


Bluetongue virus
oral susceptibility
quantitative PCR
vector competence

How to Cite

Duchemin, J.-B., White, J. R., Di Rubbo, A., Shi, S., Venter, G. J., Holmes, I., & Walker, P. J. (2021). Experimental bluetongue virus infection of Culicoides austropalpalis, collected from a farm environment in Victoria, Australia. Veterinaria Italiana, 57(4), 341–345. https://doi.org/10.12834/VetIt.2114.12867.1


Following the emerging bluetongue virus transmission in European temperate regions, we question the vector competence of the abundant Culicoides austropalpalis Lee and Reye in South-East temperate Australia. Field collected Culicoides midges were membrane fed with a bluetongue virus serotype 1 (BTV-1). The average feeding rate was 50%. After 13 days, survival rate was 25% and virus RNA presence was checked by quantitative PCR targeting viral genome segment 10. Virus RNA was found in 7.4% of individually tested females with relative viral RNA load values lower than freshly fed females, indicating that viral replication was low or null. A second qPCR targeting viral genome segment 1 confirmed the presence of virus RNA in only four out of 29 previously positive specimens. After 10 days culture on Culicoides cells, none of these four confimed positive samples did show subsequent cytopathogenic effect on Vero cells or BTV antigen detection by ELISA. As control for this virus activity detection, 12 days after microinjection of BTV-1, Culex annulirostris mosquitoes showed, after culture on Kc cells, cytopathogenic effect on Vero cells, with ELISA-confirmed infection. Despite its abundance in farm environment of the temperate Australian regions, the results of this study make C. austropalpalis of unlikely epidemiological importance in the transmission of BTV in Australia.


Bellis G.A., Gibson D.S., Polkinghorne I.G., Johnson S.J. & Flanagan M. 1994. Infection of Culicoides brevitarsis and C. wadai (Diptera: Ceratopogonidae) with four Australian serotypes of bluetongue virus. J Med Entomol, 31(3), 382-387.

Carpenter S., McArthur C., Selby R., Ward R., Nolan D.V., Luntz A.J.M., Dallas J.F., Tripet F. & Mellor P.S. 2008. Experimental infection studies of UK Culicoides species midges with bluetongue virus serotypes 8 and 9. Vet Rec, 163(20), 589-592.

Carpenter S., Wilson A. & Mellor P.S. 2009. Culicoides and the emergence of bluetongue virus in northern Europe. Trends Microbiol, 17(4), 172-178.

Dyce A.L., Bellis G.A. & Muller M.J. 2007.Pictorial atlas of Australasian culicoides wings (diptera: ceratopogonidae). Australian Biological Resources Study, Canberra.

Firth C., Blasdell K.R., Amos-Ritchie R., Sendow I., Agnihotri K., Boyle D.B., Daniels P., Kirkland P.D. & Walker P.J. 2017. Genomic analysis of bluetongue virus episystems in Australia and Indonesia. Vet Res, 48(1), 82.

Hawkes R.A., Kirkland P.D., Sanders D.A., Zhang F., Li Z., Davis R.J., Zhang N. 2000. Laboratory and field studies of an antigen capture ELISA for bluetongue virus. J Virol Methods, 85(1-2), 137-149.

Hofmann M., Griot C., Chaignat V., Perler L. & Thür B. 2008. Blauzungenkrankheit erreicht die Schweiz. Schweizer Archiv Für Tierheilkunde, 150(2), 49-56.

Hooper P.T., Lunt R.A., Gould A.R., Hyatt A.D., Russell G.M., Kattenbelt J.A., Blacksell S.D., Reddacliff L.A., Kirkland P.D., Davis R.J., Durham P.J., Bishop A.L. & Waddington J. 1999. Epidemic of blindness in kangaroos--evidence of a viral aetiology. Austr Vet J, 77(8), 529-536.

Kay B.H., Boreham P.F.L., Dyce A.L. & Standfast H.A. 1978.- Blood Feeding of Biting Midges (Diptera: Ceratopogonidae) ast Kowanyama, Cape York Peninsula, North Queensland. Austr J Entomol, 17(2), 145-149.

Kettle D.S. & Elson M.M. 1975. Variation in larvae and adults of Culicoides austropalpalis Lee and Reye in S.E. Queensland. J Natural History, 9(3), 321-336.

Nevill E., Erasmus B.J., Venter G.J., Walton T. & Osburn B. 1992. A six-year survey of viruses associated with Culicoides biting midges throughout South Africa (Diptera: Ceratopogonidae) In Bluetongue, African horse sickness and related orbiviruses. Proc. 2nd International Symposium (T.E. Walton & B.I. Osburn, eds). CRC Press, Boca Raton, Florida, 314 319

Paweska J.T., Venter G.J. & Mellor P.S. 2002. Vector competence of South African Culicoides species for bluetongue virus serotype 1 (BTV-1) with special reference to the effect of temperature on the rate of virus replication in C. imicola and C. bolitinos. Med Vet Entomol, 16(1), 10-21.

R Core Team. 2017. R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. URL https://www.R-project.org/.

Shaw A.E., Monaghan P., Alpar H.O., Anthony S., Darpel K.E., Batten C.A., Guercio A., Alimena G., Vitale M., Bankowska K., Carpenter S., Jones H., Oura C.A.L., King D.P., Elliott H., Mellor P.S. & Mertens P.P.C. 2007. Development and initial evaluation of a real-time RT-PCR assay to detect bluetongue virus genome segment 1. J Virol Methods, 145(2), 115-126.

Shaw A.E., Veronesi E., Maurin G., Ftaich N., Guiguen F., Rixon F., Ratinier M., Mertens P., Carpenter S., Palmarini M., Terzian C. & Arnaud F. 2012. Drosophila melanogaster as a Model Organism for Bluetongue Virus Replication and Tropism. J Virol, 86(17), 9015-9024.

St George T.D., Cybinski D.H., Della-Porta A.J., McPhee D.A., Wark M.C. & Bainbridge M.H. 1980. The isolation of two bluetongue viruses from healthy cattle in Australia. Austr Vet J, 56(11), 562-563.

Standfast H.A., Dyce A.L., St George T.D., Muller M.J., Doherty R.L., Carley J.G. & Filippich C. 1984. Isolation of arboviruses from insects collected at Beatrice Hill, Northern Territory of Australia, 1974-1976. Austr J Biol Sci, 37(5-6), 351-366.

Stanislawek W.L., Lunt R.A,. Blacksell S.D., Newberry K.M., Hooper P.T., White J.R. 1996. Detection by ELISA of bluetongue antigen directly in the blood of experimentally infected sheep. Vet Microbiol, 52(1-2), 1-12.

Van Der Saag M.R., Gu X., Ward M.P. & Kirkland P.D. 2016. Development and evaluation of real-time PCR assays for bloodmeal identification in Culicoides midges. Med Vet Entomol, 30(2), 155-165.

Venter G.J., Paweska J.T., Van Dijk A.A., Mellor P.S. & Tabachnick W.J. 1998. Vector competence of Culicoides bolitinos and C. imicola for South African bluetongue virus serotypes 1, 3 and 4. Med Vet Entomol, 12(4), 378-385.

Veronesi E., Antony F., Gubbins S., Golding N., Blackwell A., Mertens P.P., 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.

Wechsler S.T., McHolland L.E. & Tabachnick W.J. 1989. Cell Lines from Culicoides variipennis (Diptera: Ceratopogonidae) Support Replication of Bluetongue Virus. J Invertebrate Pathol, 54(3), 385-393

Wickham H. 2016. ggplot2: Elegant Graphics for Data Analysis. Springer-Verlag, New York. https://ggplot2.tidyverse.org

Wild C. 1984. The influence of meteorological factors on capture of Culicoides in flight (Diptera: Ceratopogonidae). The University of Queensland, School of Biological Sciences.

Copyright (c) 2022 Veterinaria Italiana