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Ticks are one of the principal arthropod vectors of human and animal infectious diseases. Whereas the prevalence of tick-borne encephalitis virus in ticks in Europe is well studied, there is less information available on the prevalence of the other tick-borne viruses (TBVs) existing worldwide. The aim of this study was to improve the epidemiological survey tools of TBVs by the development of an efficient high-throughput test to screen a wide range of viruses in ticks.In this study, we developed a new high-throughput virus-detection assay based on parallel real-time PCRs on a microfluidic system, and used it to perform a large scale epidemiological survey screening for the presence of 21 TBVs in 18,135 nymphs of I. ricinus collected from five European countries. This extensive investigation has (i) evaluated the prevalence of four viruses present in the collected ticks, (ii) allowed the identification of viruses in regions where they were previously undetected.In conclusion, we have demonstrated the capabilities of this new screening method that allows the detection of numerous TBVs in a large number of ticks. This tool represents a powerful and rapid system for TBVs surveillance in Europe and could be easily customized to assess viral emergence.

Background: The epidemiology of the zoonotic tick-transmitted parasite Babesia spp. and its occurrence in wild reservoir hosts in Sweden is unclear. In European deer, several parasite species, including Babesia capreoli and the zoonotic B. venatorum and B. divergens has been reported previously. The European roe deer, Capreolus capreolus, is an important and common part of the indigenous fauna in Europe, as well as an important host for Ixodes ricinus ticks, the vector of several Babesia spp. in Europe. Here, we aimed to investigate the occurrence of Babesia spp. in roe deer in Sweden. Findings: Roe deer (n = 77) were caught and sampled for blood. Babesia spp. was detected with a PCR assay targeting the 18S rRNA gene. The prevalence of Babesia spp. was 52 %, and two species were detected; B. capreoli and B. venatorum in 44 and 7.8 % of the individuals, respectively. Infection occurred both in summer and winter. Conclusions: We showed that roe deer in Sweden, close to the edge of their northern inland distributional range, are infected with Babesia spp. The occurrence of B. venatorum in roe deer imply that it is established in Sweden and the zoonotic implication of this finding should be regarded to a greater extent in future., Funding Agencies|Lawski foundation (Sven och Lilly Lawskis fond for naturvetenskaplig forskning); Royal Physiographic Society in Lund; Royal Swedish Academy of Agriculture and Forestry [H14-0069-ALF]; Swedish Environmental Protection Agency project; Swedish hunters organization; foundation Marie-Clair Cronstedts stiftelse: RaFast; EU Interreg - ScandTick Innovation

Due to increased travel, climatic, and environmental changes, the incidence of tick-borne disease in both humans and animals is increasing throughout Europe. Therefore, extended surveillance tools are desirable. To accurately screen tick-borne pathogens, a large scale epidemiological study was conducted on 7050 Ixodes ricinus nymphs collected from France, Denmark, and the Netherlands using a powerful new high-throughput approach. This advanced methodology permitted the simultaneous detection of 25 bacterial, and 12 parasitic species (including; Borrelia, Anaplasma, Ehrlichia, Rickettsia, Bartonella, Candidatus Neoehrlichia, Coxiella, Francisella, Babesia, and Theileria genus) across 94 samples. We successfully determined the prevalence of expected (Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum, Rickettsia helvetica, Candidatus Neoehrlichia mikurensis, Babesia divergens, Babesia venatorum), unexpected (Borrelia miyamotoi) and rare (Bartonella henselae) pathogens in the three European countries. Moreover we detected Borrelia spielmanii, Borrelia miyamotoi, Babesia divergens, and Babesia venatorum for the first time in Danish ticks. This surveillance method represents a major improvement in epidemiological studies, able to facilitate comprehensive testing of tick-borne pathogens, and which can also be customized to monitor emerging diseases.

The following interpretive summary refers to an oral presentation given at the 2002 Annual Meeting of the Livestock Insect's Workers Conference. The New World screwworm, Cochliomyia hominivorax (Coquerel) has been successfully eradicated from the United States, Mexico and Central America. The program involves monitoring infestations and the mass rearing and release of sterile screwworms. In order for the program to be successful, research must be conducted to make the identification and rearing of the insect more efficient. Methods have been developed to distinguish the screwworm from other fly larvae that may be found in animal wounds using Polymerase Chain Reaction (PCR) and Enzyme Linked Immunoassay (ELISA) techniques. The development of a genetic sexing strain would help to reduce the number of insects reared at the mass rearing facility. In order to conduct such research, numerous strains with identifiable genetic markers must be maintained in the laboratory. A cryopreservation technique would allow our laboratory to reduce the number of strains that are actively being maintained. This would allow more space and labor to be devoted to research. The procedure could also aid in reducing the frequency in which new production strains need to be established. It may be an important tool in investigating strain deterioration in the mass rearing process. A method for cryopreserving screwworm embryos has been developed. This procedure allowed an average of 52.5% of the eggs to hatch. About 22% of the larvae survived to pupation. The developmental time was greater for individuals from frozen embryos and the resulting pupae were smaller. A colony with normal developmental characteristics was re-established within two generations. Additional research is required to determine the effects of cryopreservation on other strains of New World screwworm.