Abstract
Background: Phlebotomine sand flies (Diptera: Psychodiae) in the Republic of Moldova have been understudied for decades. Our study provides a first update on their occurrence, species composition and bloodmeal sources after fifty years.Methods: During 5 seasons (2013-2017), 58 localities from 20 regions were surveyed for presence of sand flies using CDC light traps and manual aspirators. Species identification was done by a combination of morphological and molecular approaches (DNA barcoding, MALDI-TOF MS protein profiling). In engorged females, host blood was identified by three molecular techniques (RFLP, cytb sequencing and MALDI-TOF peptide mass mapping). Population structure of most abundant species was studied by cox1 haplotyping, phylogenetic analyses of ITS2 and cox1 genetic markers were used to resolve relationships of other detected species.Results: In total, 780 sand flies were collected at 30 (51.7%) localities from 12 regions of Moldova. Three species were identified by an integrative morphological and molecular approach: Phlebotomus papatasi, P. perfiliewi and Adlerius sp., the first being the most abundant and widespread, markedly anthropophilic based of bloodmeal analyses and occurring also indoor, showing low structuring of population with only 5 haplotypes of cox1 detected. Distinct morphological and molecular characters of Adlerius sp. specimens suggest presence of yet undescribed species.Conclusions: Our study revealed a presence of stable sand fly population of three species in Moldova that pose a nuisance by biting as well as potential thread of pathogen transmission and shall be further studied.