Biting midges (Culicoides species) are vectors of arboviruses and were responsible for the emergence and spread of Schmallenberg virus (SBV) in Europe in 2011 and are likely to be involved in the emergence of other arboviruses in Europe. Improved surveillance and better understanding of risks require a better understanding of the circulating viral diversity in these biting insects. In this study, we expand the sequence space of RNA viruses by identifying a number of novel RNA viruses from Culicoides impunctatus (biting midge) using a meta-transcriptomic approach. A novel metaviromic pipeline called MetaViC was developed specifically to identify novel virus sequence signatures from high throughput sequencing (HTS) datasets in the absence of a known host genome. MetaViC is a protein centric pipeline that looks for specific protein signatures in the reads and contigs generated as part of the pipeline. Several novel viruses, including an alphanodavirus with both segments, a novel relative of the Hubei sobemo-like virus 49, two rhabdo-like viruses and a chuvirus, were identified in the Scottish midge samples. The newly identified viruses were found to be phylogenetically distinct to those previous known. These findings expand our current knowledge of viral diversity in arthropods and especially in these understudied disease vectors.
Artificial infection of mosquitoes with the endosymbiont bacteria Wolbachia can interfere with malaria parasite development. Therefore, the release of Wolbachia-infected mosquitoes has been proposed as a malaria control strategy. However, Wolbachia effects on vector competence are only partly understood, as indicated by inconsistent effects on malaria infection reported under laboratory conditions. Studies of naturally-occurring Wolbachia infections in wild vector populations could be useful to identify the ecological and evolutionary conditions under which these endosymbionts can block malaria transmission. Here we demonstrate the occurrence of natural Wolbachia infections in three species of black fly (genus Simulium), which is a main vector of the avian malaria parasite Leucocytozoon. Prevalence of Leucocytozoon was high (25%), but the nature and magnitude of its association with Wolbachia differed between black fly species. Wolbachia infection was positively associated with avian malaria infection in S. cryophilum, negatively associated in S. aureum, and unrelated in S. vernum. These differences suggest that Wolbachia interacts with the parasite in a vector host species-specific manner. This provides a useful model system for further study of how Wolbachia influences vector competence. Such knowledge, including the possibility of undesirable positive association, is required to guide endosymbiont based control methods.
The species Lemonia taraxaci (Denis & Schiffermüller, 1755) is divided into three species based on distribution areas and cytochrome c oxidase subunit 1 (COI) distances above 2%: Central European Lemonia taraxaci (type locality: Austria, Vienna area), Eastern European to South Siberian Lemonia sibirica Wnukowsky, 1934 stat. rev., bona sp. (type locality: Russia, Novosibirsk Oblast, Ozero-Karachi and Russia, Altai Krai, Kornilovo), and Italian Lemonia italiana sp. n. (type locality: Italy, Calabria centr., Sila grd., Casali del Manco, San Nicola Silano (CS)). Authorship of Lemonia strigata Antoshin & Zolotuhin, 2011 is reconsidered. The close relation between the typical L. strigata and taraxaci-like adults from the Balkans is proven genetically. According to this, L. strigata is suggested to be a junior synonym of taraxaci-looking Lemonia taraxaci var. montana Buresch, 1915 (type locality: Bulgaria, Rhodope Mts, Musala), although further DNA investigation is needed to prove this hypothesis.
The Algerian-Tunisian Lemonia philopalus (Donzel, 1842) is reviewed and redescribed. The status of the Moroccan population named Lemonia philopalus rungsi Rougeot 1971 (type locality: Morocco, Merchouch) is revised, it is taken from the synonymy to the Iberian population and left as a subspecies of L. philopalus until the genetic relationship between the Moroccan and the Algerian-Tunisien populations is studied. The Iberian population is raised to a specific level as Lemonia vazquezi Oberthür, 1916 bona sp. (type locality: Spain, Madrid, Rivas-Vaciamadrid), it has a 2.13% genetic distance from the Moroccan L. philopalus rungsi. A lectotype for L. vazquezi is designated from the Natural History Museum (London, UK). The new Middle Eastern Lemonia levantina sp. n. is described (type locality: Jordan, Rift Valley, 40 km N of Aqaba, sands NW Rahma). The new species is externally similar to L. philopalus, L. philopalus rungsi, and L. vazquezi but has a 4.86% genetic distance from the Moroccan population and 5.55% from the Iberian one. The Levantine species is compared with Lemonia syriensis Daniel, 1953 because the two are sympatric and show external similarities. The ecology, phenology, and distribution of the species are discussed.
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