Molecular detection of six high importance endosymbiotic bacteria in Belgian wild-caught mosquitoes

Introduction
Several disease vectors presented a resistance to various pesticides currently used. One of an alternative solution was to use endosymbiotic bacteria because their probably interactive effects with their host. According to the introduction risks of mosquito born disease and their dispersion, we propose to investigate the prevalence of six endosymbiontic bacteria in wild-caught Culicidae in Belgium.

Methods
Eleven species of Belgian fields mosquitoes (Culex pipiens s.l., Cx. torrentium, Cx. hortensis, Anopheles claviger, An. maculipennis s.l., An. plumbeus, Culiseta annulata, Ochlerotatus geniculatus, Oc. dorsalis, Aedes albopictus and Coquillettidia richiardii) were used for the screening of six genera endosymbiotic bacteria (Wolbachia sp., Commamonas sp., Delftia sp., Pseudomonas sp., Acinetobacter sp. and Asaia sp.) according to their possible impact in mosquito biology. PCR was done for the screening and positives bands were sequenced and deposited in GenBank.

Results
Total of 144 larvae and 32 adults were used. Wolbachia, Pseudomonas, Acinetobacter and Asaia were found in mosquitoes with different proportions, according to stages (adults, larvae) with a predominance of Pseudomonas in all species, as far as Acinetobacter and Asaia also have a high prevalence. Commamonas and Delftia were absent from all species tested, either in larvae and in adults.

Discussion
Choice of endosymbiotic bacteria studied here was allowing of their importance in literature. For Pseudomonas, it showed that this bacteria could produced ovipositon attractants for mosquito. Acinetobacter was suggested efficient in transmission and maintenance within host populations. Asaia was capable of efficiently crossing body barriers and colonizing different organs. Wolbachia was currently the most studied bacteria which plays an important role in the genetic manipulation of the host. Present advances in understanding the mosquito–microbiota relationships may have a great impact in a better understanding of some traits of mosquito biology and in the development of innovative mosquito-borne disease-control strategies.