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Parasites of vectors--Ixodiphagus Hookeri and Its Wolbachia Symbionts in Ticks in The Netherlands

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Parasites of vectors--Ixodiphagus Hookeri and Its Wolbachia Symbionts in Ticks in The Netherlands

Ellen Tijsse-Klasen et al. Parasit Vectors.

Abstract

Background: Ixodiphagus hookeri is a parasitic wasp of ixodid ticks around the world. It has been studied as a potential bio-control agent for several tick species. We suspected that the presence of Wolbachia infected I. hookeri eggs in ticks is responsible for incidental detection of Wolbachia DNA in tick samples.

Methods: The 28S rRNA and 16S rRNA genes of a specimen of I. hookeri was amplified and sequenced. PCR on part of the 28S rRNA gene was used to detect parasitic wasp DNA in 349 questing Ixodes ricinus ticks from various sampling sites. Furthermore, the wsp gene of Wolbachia was sequenced from the I. hookeri specimen and a subset of ticks was tested using this marker.

Results: Several sequences from tick specimens were identical to the Wolbachia sequence of the I. hookeri specimen. Ixodiphagus hookeri was detected in 9.5% of all tested ticks, varying between 4% and 26% depending on geographic location. Ten out of eleven sampling sites throughout the Netherlands were positive for I. hookeri. Eighty-seven percent of I. hookeri-positive but only 1.6% of I. hookeri-negative ticks were Wolbachia positive. Detection of I. hookeri DNA was strongly associated with the detection of Wolbachia in ticks.

Conclusion: This is the first reported case of I. hookeri in the Netherlands. Furthermore I. hookeri harbours Wolbachia species and is broadly distributed in the Netherlands. While detection of Wolbachia DNA in ticks might often be due to parasitism with this wasp, other sources of Wolbachia DNA in ticks might exist as well.

Figures

Figure 1
Figure 1
Ixodiphagus hookeri (Howard) hatched from an Ixodes ricinus nymph (Photo: Oscar Vorst, Naturalis).
Figure 2
Figure 2
Phylogenetic analysis of Wolbachia from various invertebrate host species. Neighbor-joining trees with Kimura correction were based on wsp genes from this study and mined from GeneBank. Bootstrap proportions were calculated based on 500 replicates, only values < 80 are indicated.
Figure 3
Figure 3
Locations in the Netherlands with PCR verified presence of Ixodiphagus hookeri infested Ixodes ricinus ticks. Drents-Friese Wold; 2. Boswachterij Hardenberg; 3. Landgoed Singraven; 4. Duin en Kruidberg; 5. Pyramide van Austerlitz; 6. Koninklijke Houtvesterijen Hoog Soeren; 7. Hullenberg; 8. Rijk van Nijmegen; 9. Kop van Schouwen; 10. Vrouwenpolder; 11. Dintelse Gorzen; 12. Vijlenerbos; Two of these sites (7 and 10) were found positive in the pilot study; the remaining sites were investigated in the prevalence study.
Figure 4
Figure 4
Prevalence of the parasitic wasp Ixodiphagus hookeri in various locations spread over the Netherlands. Error bars indicate 95% confidence intervals calculated with Mid-P exact test.

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