Identification of Anopheles daciae in Germany through ITS2 sequencing

Abstract

Until the middle of the twentieth century, malaria was frequently endemic in parts of Germany; Anopheles maculipennis complex species were considered the primary vectors. Three species of this complex have been identified in Germany: A. maculipennis s.s., Anopheles messeae and Anopheles atroparvus; the last predominantly from the coastal regions of Northern Germany. Anopheles daciae is a recently described member of the A . maculipennis complex and resembles the well-characterised species A. messeae, although the two species can be distinguished through their egg morphology and sequencing of the internal transcribed spacer 2 (ITS2) region of their nuclear rDNA. In this study, we harvested larval and adult mosquito samples from five breeding sites and ten CO(2) trap collection sites in the Upper Rhine Valley of Southwestern Germany to analyse the complement of anopheline species present. Mosquito ITS2 DNA was extracted and polymerase chain reaction (PCR)-amplified using established protocols. Genomic analysis was performed by a species-diagnostic restriction fragment length polymorphism assay as well as by sequencing of PCR products; the data obtained were aligned against nucleic acid sequences from English mosquitoes retrieved from GenBank. Additionally, the larval breeding sites of A. messeae were characterised through water quality measurement. Forty-seven samples were successfully processed, of which 6 were identified as A. daciae and 41 as A. messeae. All samples of A. daciae, which has not previously been found in Central Europe, originated from one CO(2) trap collection site in Dettenheim, close to Karlsruhe, Southwestern Germany. The identification of this malarial vector in a novel area may have implications for the re-emergence of disease subsequent to climatic changes.

Molecular Characterization of Mosquitoes (Diptera: Culicidae) in Northwestern Iran by Using rDNA-ITS2

“…The molecular marker rDNA-ITS2 is used in molecular taxonomy for separating sibling species (2), in phylogeny for establishing evolutionary relationships among mosquito species (3), and in phylogeography and population genetics for assessing the geographic distribution of mosquitoes (4).…”

Distribution of individual members of the mosquito Anopheles maculipennis complex in Germany identified by newly developed real‐time PCR assays

“…maculipennis complex members produced in this study together with respective data from two other recent studies (Weitzel et al. , ; Kronefeld et al. , ) were used to model the distribution of the four species in Germany.…”

“…, ; Weitzel et al. , ); black = presence, white = absence. Prediction of species distribution: the probability of occurrence increases from red over orange to yellow, green indicates presence according to the selected threshold.…”

Anopheles (Anopheles) petragnani Del Vecchio 1939—a new mosquito species for Germany

“…albopictus (Becker et al 2013; Pluskota et al 2008), Ae. j. japonicus (Becker et al 2011; Schaffner et al 2009), Culiseta ( Allotheobaldia ) longiareolata (Macquart, 1838) (Becker and Hoffmann 2011), Anopheles ( Anopheles ) daciae Linton, Nicolescu & Harbach 2004 (Weitzel et al 2012) and Aedes ( Finlaya ) koreicus (Edwards, 1917) (Werner et al 2015). …”