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Kampen, H.

Publications and source records attributed to Kampen, H..

5 recordsLinked to original sources

Experimental BTV-3 and BTV-8 infection of Culicoides sonorensis biting midges

BTV-3 emerged in the Netherlands in 2023 and spread rapidly to neighboring countries. Compared to the BTV-8 outbreak in 2006, the course of the BTV-3 epizootic is more severe. Experimental infection of laboratory-reared Culicoides sonorensis midges showed a slightly higher replication rate of BTV-3 than of BTV-8. One sentence summary lineWe experimentally infected laboratory-reared Culicoides sonorensis biting midges with bluetongue virus (BTV) serotypes 3 and 8 using virus-containing blood meal and found slightly higher replication rates of BTV-3.

microbiology↗

Schmallenberg virus non-structural proteins NSs and NSm are not essential for experimental infection of Culicoides sonorensis biting midges

The teratogenic orthobunyavirus Schmallenberg virus (SBV) is transmitted between its mammalian hosts by Culicoides biting midges. The genome of circulating SBV, i.e. variants present in viraemic ruminants or insect vectors, is very stable, while variants found in malformed ruminant foetuses display a high genetic variability. It was suggested that foetal infection provides an environment that favours viral mutations that enable immune escape in the unborn, but at the costs of limiting the ability of the virus to spread further. To investigate infection and dissemination rates of different SBV variants in the insect vectors, we fed laboratory-reared Culicoides sonorensis with blood containing the prototype strain BH80/11-4 from a viraemic cow or strain D281/12, which was isolated from the brain of a sheep foetus and harbours multiple mutations in all three genome segments. Further, virus variants lacking NSs, NSm or both non-structural proteins were included. Six days after feeding, virus replication was found in about 2% of the midges exposed to wild-type strain BH80/11-4. The absence of the non-structural proteins had no obvious effect on the oral susceptibility to virus infection, as 2.78% of the midges fed with the NSs-deletion mutant displayed after six days viral loads higher than the respective day-0-group, 1.92% of the midges exposed to the NSm-deletion mutant and 1.55% of midges exposed to the NSs/NSm-deletion mutant. In contrast, strain D281/12 did not replicate at all in the midges, supporting the assumption that SBV variants arising in infected foetuses are unable to enter the normal insect-mammalian host cycle. ImportanceBiting midges are responsible for the transmission of Schmallenberg virus (SBV), a pathogen of veterinary importance that primarily infects ruminants. Although SBV has been extensively studied in the mammalian host, the virus-intrinsic factors allowing infection of, and replication in, biting midges are largely unknown. Therefore, we infected laboratory-reared Culicoides sonorensis midges with SBV variants by feeding them with virus-containing blood. The SBV variants differed in their genome composition, as we used the prototype wild-type strain, a strain with multiple mutations that was isolated from the brain of a malformed foetus, and recombinants lacking either NSs or NSm or both of these non-structural proteins. While the non-structural proteins had no obvious effect, the variant from the malformed foetus did not replicate at all, indicating that virus variants with characteristic genomic mutations present in foetuses lose their ability to infect the insect vector and will be excluded from the natural transmission cycle.

microbiology↗

Emergence of bluetongue virus serotype 3 in western Germany, October 2023, and ad-hoc monitoring in Culicoides biting midges

Bluetongue virus serotype 3 emerged in October 2023 in Germany, where Schmallenberg virus is enzootic. BTV-3 was detected in a pool of Culicoides biting midges at the same time as infections were reported from ruminants. Schmallenberg virus was frequently found in vector pools, reflecting the epidemiological situation. One sentence summary lineCoinciding with the emergence and low-level circulation of bluetongue virus serotype 3 in ruminants in Germany in October 2023, the virus was detected in a pool of Culicoides biting midges of the Obsoletus Group, while enzootic Schmallenberg virus was found in numerous midge pools.

microbiology↗

Large-Scale COI Gene Data Analysis for the Development of a Gel-Based Multiplex PCR Test Capable of Identifying Biting Midge Vector Species and Haplotypes (Diptera: Ceratopogonidae) of the Culicoides subgenus Avaritia Fox, 1955

The emergence of culicoid-transmitted bluetongue and Schmallenberg viruses in several European countries demonstrated the ability of indigenous biting midge species to transmit pathogens. Entomologic research programmes identified members of the Obsoletus Group (Culicoides subgenus Avaritia) as keyplayers in disease epidemiology in Europe. However, morphological identification of potential vectors to species level is challenging due to the existence of isomorphic species. PCR tests developed to facilitate genetic species determination have been dismantled by the discovery of new genetic variants (haplotypes) of C. obsoletus sensu stricto (s.s.), forming distinct clades. In this study, 4,422 GenBank entries of the mitochondrial cytochrome c oxidase subunit I (COI) gene of subgenus Avaritia members of the genus Culicoides were analyzed to develop a conventional multiplex PCR, capable of detecting all vector species and recently described clades of the western Palaearctic in this subgenus. Numerous GenBank entries incorrectly assigned to a species were identified, analyzed and reassigned. The results suggest that the three clades of C. obsoletus represent independent species, whereas C. montanus should rather be regarded as a genetic variant of C. obsoletus s.s.. Based on these findings, specific primers were designed and validated with DNA material from field-caught biting midges which achieved very high diagnostic sensitivity (100%) when compared to an established reference PCR (82.6%). Hence, the newly developed multiplex PCR represents the first molecular tool which enables both the identification of accepted species and of the three clades of C. obsoletus s.s. and could provide new insights into the ecology of the latter.

genetics↗

African swine fever virus-like integrated elements in a soft tick genome - an ancient virus vector arms race?

BackgroundAfrican swine fever virus (ASFV) is the only known DNA-arbovirus and a most devastating suid pathogen that, originating from a sylvatic cycle in Africa, has spread to eastern Europe and recently reached western Europe and Asia, leading to a socio-economic crisis of global proportion. However, since neither closely related viruses nor integrated viral elements have yet been identified, ASFV evolution remains a mystery. ResultsHere, we show that soft ticks of the Ornithodoros moubata group, the natural arthropod vector of ASFV, harbour African swine fever virus-like integrated (ASFLI)-elements corresponding to up to 10% (over 20 kb) of the ASFV genome. Through orthologous dating and molecular clock analyses, we provide data suggesting that integration occurred over 1.47 million years ago. Furthermore, our data indicate that these elements, showing high sequence identities to modern ASFV, are maintained in the tick genome to protect the tick from infection with specific ASFV-strains through RNA interference. ConclusionWe suggest that this mechanism of protection, shaped through many years of co-evolution, is part of an evolutionary virus-vector "arms race", a finding that has not only high impact on our understanding of the co-evolution of viruses with their hosts but also provides a glimpse into the evolution of ASFV.

microbiology↗