Phylogenetic status and matrilineal structure of the biting midge, Culicoides imicola, in Portugal, Rhodes and Israel

Abstract

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.

First overview of the Culicoides Latreille (Diptera: Ceratopogonidae) livestock associated species of Reunion Island, Indian Ocean

“…Cytochrome Oxidase subunit I (COI) gene amplification was carried out using primers C1J1718 (forward) and C1N2191 (reverse) to obtain an approximately 600 bp product as described previously (Dallas et al, 2003). Amplification reactions by Polymerase Chain Reaction (PCR) were performed in 25 l of reaction volume with 5 l of buffer 5×, 0.5 l of dNTP (10 mM), 2 l of MgCl 2 (25 mM), 0.5 ml of each primer; 0.5 l of Taq polymerase (5 U/l), 15 l of sterile water and 1 l of template DNA.…”

A survey of biting midges of the genus Culicoides Latreille, 1809 (Diptera: Ceratopogonidae) in NE Bulgaria, with respect to transmission of avian haemosporidians

“…The initial PCR activation step for the used C1-J-1718-mod (GGWGGRTTTGGWAAYTGAYTAG) and C1-N-2191-mod (AGHWCCAAAAGTTTCYTTTTTCC) primer pair, modified from previously described primers (Dallas et al 2003), was performed at 94°C for 3 min followed by 35 cycles of denaturation at 94°C for 30 sec, annealing at 52°C for 30 sec and extension at 72°C for 30 sec. The final cycle was completed with 10 min of extension at 72°C.…”

“…Numerous studies (Dallas et al 2003;Li et al 2003;Cêtre-Sossah et al 2004;Wörheide et al 2004;Perrin et al 2006) dealing with a similar problem found the noncoding genetic region of the internal transcribed spacer 1 (ITS-1) of the mitochondrial ribosomal DNA and mitochondrial cytochrome c oxidase subunit 1 (COI) to be highly promising in order to establish a reliable polymerase chain reaction (PCR)-based method for species identification.…”

Morphological and molecular identification of species of the Obsoletus group (Diptera: Ceratopogonidae) in Scandinavia

“…4 Alignment of the ITS-1 sequences of C. obsoletus, C. scoticus, C. chiopterus, and C. dewulfi. Nucleotides that differ are indicated, and dots correspond to conserved nucleotides (Y=C or T, R=A or G, M=A or C, S=G or C, W=A or T) marker systems are very useful depending on the topic of the study (Dallas et al 2003;Cetre-Sossah et al 2004;Gomulski et al 2005). Mathieu et al (2007) established a very useful molecular identification method based on ITS-1 of the Western European Obsoletus group by looking at three specimens of different geographic origin from each of the species, and in an extension, this establish a quantitative method for identification of C. imicola ).…”

“…The molecular barcoding method based on mitochondrial cytochrome oxidase subunit I, COI, was reliable for inter-and intraspecies differentiation of biting midges (Dallas et al 2003;Linton et al 2002;Nolan et al 2007;Pages and Monteys 2005). Additionally, the development and implementation of molecular techniques with the two internal transcribed spacers of the nuclear ribosomal DNA ITS-1 and ITS-2 (Cetre- Sossah et al 2004;Gomulski et al 2005) have been valuable in elucidating the phylogenetic relationship within the genus Culicoides (Gomulski et al 2006;Mathieu et al 2007;Perrin et al 2006).…”