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Background    Haemaphysalis erinacei  is one of the few ixodid tick species for which valid names of subspecies exist. Despite their disputed taxonomic status in the literature, these subspecies have not yet been compared with molecular methods. The aim of the present study was to investigate the phylogenetic relationships of  H. erinacei  subspecies, in the context of the first finding of this tick species in Romania.    Results  After morphological identification, DNA was extracted from five adults of  H. e. taurica  (from Romania and Turkey), four adults of  H. e. erinacei  (from Italy) and 17 adults of  H. e. turanica  (from China). From these samples fragments of the cytochrome  c  oxidase subunit 1 ( cox 1) and 16S rRNA genes were amplified via PCR and sequenced. Results showed that  cox 1 and 16S rRNA gene sequence divergences between  H. e. taurica  from Romania and  H. e. erinacei  from Italy were below 2%. However, the sequence divergences between  H. e. taurica  from Romania and  H. e. turanica  from China were high (up to 7.3% difference for the 16S rRNA gene), exceeding the reported level of sequence divergence between closely related tick species. At the same time, two adults of  H. e. taurica  from Turkey had higher 16S rRNA gene similarity to  H. e. turanica  from China (up to 97.5%) than to  H. e. taurica  from Romania (96.3%), but phylogenetically clustered more closely to  H. e. taurica  than to  H. e. turanica .    Conclusions  This is the first finding of  H. erinacei  in Romania, and the first (although preliminary) phylogenetic comparison of  H. erinacei  subspecies. Phylogenetic analyses did not support that the three  H. erinacei  subspecies evaluated here are of equal taxonomic rank, because the genetic divergence between  H. e. turanica  from China and  H. e. taurica  from Romania exceeded the usual level of sequence divergence between closely related tick species, suggesting that they might represent different species. Therefore, the taxonomic status of the subspecies of  H. erinacei  needs to be revised based on a larger number of specimens collected throughout its geographical range.

Phylogenetic analysis of Haemaphysalis erinacei Pavesi, 1884 (Acari: Ixodidae) from China, Turkey, Italy and Romania