Identifying breeding hosts of Ixodes ricinus ticks using stable isotope analysis of their larvae - Proof of concept.

Ticks are important vectors of zoonotic pathogens. Ticks are parasites that are dependent on their hosts for blood meal to develop and reproduce. The abundance of ticks is dependent on the availability of suitable breeding hosts, often medium- and large-sized mammals. So far there has been a shortage of direct methods identifying the breeding hosts for the female ticks. In this study, we introduce a stable isotope analysis (SIA) method that enables us to identify the trophic group of the breeding host, i.e. the host on which the tick mother fed, by sampling larval ticks from the field. We established a reference database on the stable isotope (SI) values (δC and δN) of the blood of potential tick host species, and of larvae from Ixodes ricinus females, which have fed on known hosts. By comparing the SI values from field collected larval ticks to our reference data, we can determine their most likely host species group. Our results show that the isotopic signatures of I. ricinus tick larvae reflect the diet of the breeding host of the mother tick. SIA proved reliable in categorizing the breeding hosts of I. ricinus into two distinguishable trophic groups; herbivores and carni-omnivores. To our knowledge, this is the first time that stable isotope analyses have been applied to detect transovarial (i.e. over-generational) traces of a blood meal in ticks. The method provides an efficient, novel tool for directly identifying tick breeding hosts by sampling field collected larvae. Ixodes ricinus is the most important vector of TBPs (tick-borne pathogens) in Europe, and to predict and mitigate against the future risks that TBPs pose, it is crucial to have detailed knowledge on the hosts that support tick reproduction in nature.