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Background   Culicoides imicola  (Diptera: Ceratopogonidae) is an important Afrotropical and Palearctic vector of disease, transmitting viruses of animal health and economic significance including African horse sickness and bluetongue viruses. Maternally inherited symbiotic bacteria (endosymbionts) of arthropods can alter the frequency of  COI  (cytochrome c oxidase subunit I) mitochondrial haplotypes (mitotypes) in a population, masking the true patterns of host movement and gene flow. Thus, this study aimed to assess the mtDNA structure of  C. imicola  in relation to infection with  Candidatus  Cardinum hertigii (Bacteroides), a common endosymbiont of  Culicoides  spp.    Methods  Using haplotype network analysis,  COI  Sanger sequences from  Cardinium -infected and -uninfected  C. imicola  individuals were first compared in a population from South Africa. The network was then extended to include mitotypes from a geographic range where  Cardinium  infection has previously been investigated.    Results  The mitotype network of the South African population demonstrated the presence of two broad mitotype groups. All  Cardinium -infected specimens fell into one group (Fisher’s exact test,  P  = 0.00071) demonstrating a linkage disequilibrium between endosymbiont and mitochondria. Furthermore, by extending this haplotype network to include other  C. imicola  populations from the Mediterranean basin, we revealed mitotype variation between the Eastern and Western Mediterranean basins (EMB and WMB) mirrored  Cardinium -infection heterogeneity.    Conclusions  These observations suggest that the linkage disequilibrium of  Cardinium  and mitochondria reflects endosymbiont gene flow within the Mediterranean basin but may not assist in elucidating host gene flow. Subsequently, we urge caution on the single usage of the  COI  marker to determine population structure and movement in  C. imicola  and instead suggest the complementary utilisation of additional molecular markers.    Graphical Abstract

parasites and vectors

Cardinium symbiosis as a potential confounder of mtDNA based phylogeographic inference in Culicoides imicola (Diptera: Ceratopogonidae), a vector of veterinary viruses