Larval habitats characterization and species composition of Anopheles mosquitoes in Tunisia, with particular attention to Anopheles maculipennis complex.

In Tunisia, malaria transmission has been interrupted since 1980. However, the growing number of imported cases and the persistence of putative vectors stress the need for additional studies to assess the risk of malaria resurgence in the country. In this context, our aim was to update entomological data concerning Anopheles mosquitoes in Tunisia. From May to October of 2012, mosquito larval specimens were captured in 60 breeding sites throughout the country and identified at the species level using morphological keys. Environmental parameters of the larval habitats were recorded. Specimens belonging to the An. maculipennis complex were further identified to sibling species by the ribosomal deoxyri- bonucleic acid (rDNA)-internal transcribed spacer 2 (ITS2) polymerase chain reaction (PCR) technique. In total, 647 Anopheles larvae were collected from 25 habitats. Four species, including An. labranchiae, An. multicolor, An. sergentii ,a nd An. algeriensis, were morphologically identified. rDNA-ITS2 PCR confirmed that An. labranchiae is the sole member of the An. maculipennis complex in Tunisia. An. labranchiae was collected throughout northern and central Tunisia, and it was highly associated with rural habitat, clear water, and sunlight areas. Larvae of An. multicolor and An. sergentii existed separately or together and were collected in southern Tunisia in similar types of breeding places. The first map of Anopheles distribution in Tunisia was based on data collected between 1968 and 1974 during the malaria eradication campaign. A literature review by Brunhes and others 7 compiled 12 species in 1999: An. algeriensis, An. cinereus, An. claviger, An. dthali, An. labranchiae, An. marteri, An. multicolor, An. petragnani, An. plumbeus, An. sergentii, An. superpictus ,a ndAn. ziemanni. The most recent investiga- tion dating back to the 1990s detected only six species, includ- ing those suspected as malaria vectors in Tunisia. 8-10