Abstract
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 Culex. pipiens pipiens biotypes pipiens and molestus. To facilitate detection of different Culex pipiens forms and their hybrids we established a multiplex real-time PCR. Culex pipiens 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 Culex pipiens pipiens biotype pipiens and Culex. torrentium, but we also detected Culex pipiens pipiens biotype molestus and hybrids of the two pipiens 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.
Highlights
In the past decades, West Nile virus (WNV) has spread from Africa and conquered various regions of temperate climate with substantial outbreaks in North America and Europe
In Europe and other temperate regions, members of the Culex pipiens complex are the most ubiquitous mosquito species [1,2] which serve as principal vectors for various arboviruses including WNV [3,4,5,6,7]
The results indicate an uneven distribution of Cpp. and Cx. torrentium throughout the country, sympatric occurrence of Cpp. biotype pipiens and biotype molestus and first detection of Cpp. biotype pipiens and biotype molestus hybrids at different locations in Germany
Summary
West Nile virus (WNV) has spread from Africa and conquered various regions of temperate climate with substantial outbreaks in North America and Europe. We report on a newly developed multiplex real-time PCR that allows discrimination of the various morphologically indistinguishable Culex species and biotypes. DNA extraction and multiplex real-time PCR assay Mosquitoes collected at the various study sites were frozen at
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