The present study examines the mating and breeding behavior as well as the genetic differentiation of Culex pipiens biotype pipiens and Cx. pipiens biotype molestus. Firstly, the mating behavior of Cx. pipiens s.l. originating from larval populations of various epigeous and hypogeous breeding sites in Germany was examined. Autogeny was prevailing in underground populations, occasionally found in semi-open water reservoirs like drains, rarely in containers, but never in ponds and ditches. Secondly, in a multilocus enzyme electrophoretic study the gene flow among seven geographic populations of Cx. pipiens biotype pipiens and the biotype molestus from several European countries was quantified. For comparison, five populations of Cx. quinquefasciatus from Asia, Africa and North America, three populations of Cx. torrentium (Germany) and other outgroup species were also examined. Thirdly, the mitochondrial cytochrome c oxidase submit I gene of both biotypes from Germany was analysed by a polymerase chain reaction - restriction fragment length polymorphism assay and the ascertained DNA-sequences were aligned with genebank data of Russian populations. The population genetic analyses revealed much higher genetic distances between local populations of Cx. pipiens biotype pipiens and Cx. pipiens biotype molestus compared to the low differentiation between geographically remote populations within each taxon. The UPGMA (unweighted pair group method with arithmetic mean) analysis and F-statistics positioned the biotypes pipiens on one side and molestus on the other side in discrete monophyletic clusters. Gene flow between local populations of the biotypes pipiens and molestus could be shown to be lower than gene flow between geographically distant populations within each of the two groups, leading to the conclusion that Cx. pipiens biotype molestus could be a distinct taxon. Culex quinquefasciatus was genetically well-separated, in particular by the diagnostic enzyme marker malate dehydrogenase (nicotinamide adenine dinucleotide phosphate). The genetic markers adenylatekinase and hydrobutyrate dehydrogenase allowed to screen thousands of morphologically similar samples of either Cx. pipiens s.l. and Cx torrentium and it could be shown that Cx. torrentium is a very frequent species in central Europe.

The efficacy and safety of artemether-lumefantrine for the treatment of malaria in nonimmune populations are not well defined. In this study, 165 nonimmune patients from Europe and non-malarious areas of Colombia with acute, uncomplicated falciparum malaria or mixed infection including P. falciparum were treated with the six-dose regimen of artemether-lumefantrine. The parasitologic cure rate at 28 days was 96.0% for the per protocol population (119/124 patients). Median times to parasite clearance and fever clearance were 41.5 and 36.8 hours, respectively. No patient had gametocytes after Day 7. Treatment was well tolerated; most adverse events were mild to moderate and seemed to be related to malaria. There were few serious adverse events, none of which were considered to be drug-related. No significant effects on ECG or laboratory parameters were observed. In conclusion, the six-dose regimen of artemether-lumefantrine was effective and well tolerated in the treatment of acute uncomplicated falciparum malaria in nonimmune patients.

The Culex pipiens complex consists of several species, subspecies, forms, races, physiological variants, or biotypes according to different authors and includes the 2 holarctic variants Cx. pipiens biotype pipiens and Cx. pipiens biotype molestus. Differences in morphological characters are overlapping and thus are delimited in their taxonomic value, even when behavioral and reproductive specializations are apparent. Our enzyme electrophoretic study included 7 geographic populations of Cx. pipiens biotype pipiens and 7 of the biotype molestus from several European countries. For comparison, 5 populations of Culex quinquefasciatus from Asia, Africa, and North America were examined. The aim was an assessment of the extent of genetic differences between local populations of the biotypes pipiens and molestus versus the degree of differentiation between geographic populations of both groups. Culex torrentium, Cx. modestus, Culex stigmatosoma, and Culex territans were studied for comparison as taxonomical well-defined species. The population genetic analyses revealed much higher genetic distances between local populations of Cx. pipiens biotype pipiens and Cx. pipiens biotype molestus compared to the low differentiation between geographic populations within each taxon. The UPGMA analysis and F-statistics position the geographic populations in discrete monophyletic clusters. Gene flow between local populations of the biotypes pipiens and molestus could be shown to be lower than gene flow between geographically distant populations within each of the 2 groups, leading to the conclusion that Cx. pipiens biotype molestus could be a distinct taxon. Culex quinquefasciatus could be diagnosed as genetically well separated, in particular by the diagnostic enzyme marker MDH (NADP). Two genetic enzyme markers were identified to differentiate Cx. torrentium from Cx. pipiens s.l. Culex modestus, Cx. stigmatosoma, and Cx. territans showed considerable genetic distances to the species of the Culex pipiens complex and between each other, and several genetic markers could be identified.

Malaria is one of the most severe health problems facing the world today. Until the mid-twentieth century, Europe was an endemic area of malaria, with the Balkan countries being heavily infested. Sibling species belonging to the Anopheles maculipennis complex are well-known as effective vectors of Plasmodium in Europe. A vast number of human malaria cases in the past in the former Yugoslavia territory have stressed the significance of An. maculipennis complex species as primary and secondary vectors. Therefore, the present study evaluates the species composition, geographic distribution and abundance of these malaria vector species. Mosquitoes were collected in the northern Serbian province of Vojvodina and analysed by PCR-RFLP, multiplex PCR and sequencing of the ITS2 intron of genomic rDNA. Four sibling species of the An. maculipennis complex were identified. Both larvae and adults of the recently described species An. daciae were identified for the first time in Serbia. In 250 larval samples, 109 (44%) An. messeae, 90 (36%) An. maculipennis s.s., 33 (13%) An. daciae and 18 (7%) An. atroparvus were identified. In adult collections, 81 (47%) An. messeae, 55 (32%) An. daciae, 33 (19%) An. maculipennis s.s., and 3 (2%) An. atroparvus were recorded. The most abundant species in Vojvodina was An. messeae, whereas An. atroparvus was confirmed a rare species in all parts. Since this species is a potentially, highly competent malarial vector, low population density could be crucial to prevent a new establishment of endemic malaria transmission in Serbia.

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.