Additional file 2: Table S2. of Host-feeding patterns of mosquito species in Germany

Kruskal-Wallis tests on the differences of the percentages of detected birds, non-human mammals and humans between the three land use classes (natural, rural and urban) for the three most frequent mosquito species (Fig. 3). (DOCX 14 kb

Additional file 3: Table S3. of Host-feeding patterns of mosquito species in Germany

Chi-square tests on the differences in the frequencies of detected mammalian or avian hosts among all pairs of used trapping methods with adjusted P-values for multiple comparisons. (DOCX 13 kb

Additional file 1 of Interaction of Mesonivirus and Negevirus with arboviruses and the RNAi response in Culex tarsalis-derived cells

Additional file 1: Table S1. List of primers used 5′ to 3′

Basta!: Für Saxophonquartett

<div><p>Mosquitoes and other arthropods may transmit medically important pathogens, in particular viruses such as West Nile virus. The presence of suitable hosts and competent vectors for those zoonotic viruses is essential for an enzootic transmission, which is a prerequisite for epidemics. To establish reliable risk projections, it is an urgent need for an exact identification of mosquito species, which is especially challenging in the case of sibling species, such as <i>Culex. pipiens pipiens</i> biotypes <i>pipiens</i> and <i>molestus</i>. To facilitate detection of different <i>Culex pipiens</i> forms and their hybrids we established a multiplex real-time PCR. <i>Culex pipiens</i> samples were obtained by egg raft collection and rearing until imago stage or adult sampling using CO₂ baited traps and gravid traps. In total, we tested more than 16,500 samples collected all over Germany in the years 2011 and 2012. The predominant species in Germany are <i>Culex pipiens pipiens</i> biotype <i>pipiens</i> and <i>Culex. torrentium</i>, but we also detected <i>Culex pipiens pipiens</i> biotype <i>molestus</i> and hybrids of the two <i>pipiens</i> biotypes at sites where both species occur sympatrically. This report of a potentially important bridge vector for West Nile virus might have major impact in the risk projections for West Nile virus in Germany.</p></div

Trapping-sites of the nationwide surveillance program selected for the study.

<p>Trap types: BT: Biogents Sentinel; GT: Gravid trap; EVS: Enceph. Vector Surveillance Trap (Bioquip); ex-La.: reared from larvae ; ex-Pu.: reared from pupae.</p

Classification of <i>Culex</i> samples from the German mosquito surveillance program.

<p>Graphical representation of the Culex species composition in Germany. 48 different trapping sites in Germany were combined according to their geographical relatedness to form 10 cluster areas shown representing Lower Rhine Valley and further sites in Palatine, Upper Rhine Valley and further sites in Baden-Württemberg, Lake Constance, Lake Chiemsee and other sites in Bavaria, Hesse, Upper Elbe Valley in Saxonia, Oder Valley in Brandenburg, Baltic Sea in Mecklenburg-West Pomerania, Metropolitan Region Hamburg and various sites in Schleswig-Holstein (for more details see also <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0071832#pone-0071832-t001" target="_blank">Table 1</a>)). White (<i>Cpp. pipiens</i>), black (<i>Cx. torrentium</i>) and red (<i>Cpp. molestus</i>) quarters indicate pools that were composed of a single species. Grey (<i>Cpp. pipiens</i>+<i>Cx. torrentium</i>) and dark-red (<i>Cpp. molestus</i>+<i>Cx. torrentium</i>) quarters indicate pools composed of two species. With the current set-up (i.e.using pooled samples) the composition of pink quarters could be either two biotypes <i>Cpp. pipiens</i> and <i>Cpp. molestus</i> or hybrids of both biotypes. The n-numbers given in the graphs notify total numbers of individuals analysed in each cluster.</p

Identification of <i>Cpp.</i> biotype hybrids in two German metropolitan areas.

<p>Detailed species composition at the 4 sampling sites of the Rhine-Main metropolitan area (right graph) and 5 sampling sites at Hamburg metropolitan area (left graph). DNA samples from single individuals collected at these sampling sites were subjected to the multiplex real-time PCR and analysed for the presence of biotype hybrids. Bars represent species distribution in percent at each site. White indicates <i>Cpp.</i> biotype <i>pipiens</i>; red <i>Cpp.</i> biotype <i>molestus</i>; yellow hybrids of biotypes <i>pipiens</i> and <i>molestus</i>. Biotype hybrids were found at Dirmstein and Bobenheim-Roxheim trapping sites in Southwest Germany and at the Wulksfelde trapping site in northern Germany. Additionally, the presence of <i>Cx. torrentium</i> was assed and is indicated by the grey bars in both graphs.</p