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The aim of this study was to investigate the natural cycle of the new human pathogenic bacteria CandidatusNeoehrlichia mikurensis and Anaplasma phagocytophilumin Southern Hungary. We collected rodents with live-traps (2010–2013) and questing ticks with flagging in 2012. Small mammals were euthanized, tissue samples were collected and all the ectoparasites were removed and stored in 70% alcohol. We found relatively low overall prevalence of tick infestation (8%). Samples were analysed for A. phagocytophilumand CandidatusN. mikurensis with multiplex quantitative real-time PCR targeting a part of major surface protein 2 (msp2) and the heat shock protein groEL genes, respectively. The overall prevalence in tissue samples was 6.6% (skin) and 5.1% (spleen) for A. phagocytophilumand 1.7% (skin) and 3.4% (spleen) for CandidatusN. mikurensis. CandidatusN. mikurensis was only detected in Apodemus flavicollisand Apodemus agrarius, while A. phagocytophilumwas found in A. flavicollis, A. agrarius, Myodes glareolus, Microtus arvalisand Mus musculussamples. Prevalence of A. phagocytophilumin skin samples of A. flavicolliswas significantly higher than prevalence of N. mikurensis(p<0.05). Among questing Ixodes ricinusticks we found three (8.8%) individuals (female, male, nymph) infected with CandidatusN. mikurensis. Five (3.1%) questing ticks had A. phagocytophiluminfection (one I. ricinusmale, two Dermacentor reticulatusfemales and two Haemaphysalis concinnafemales). We found one I. ricinusnymph removed from a male A. flavicolliswith A. phagocytophiluminfection. Our study provides new data on the occurrence of these pathogens in rodent tissue samples, questing ticks and engorged ticks in Southern Hungary.

Three Palearctic members of the subgenus Pholeoixodes, i.e., Ixodes canisuga, Ixodes hexagonusand Ixodes kaiseriare frequently collected from dogs, cats, red foxes, badgers and other carnivorous/insectivorous hosts in Europe. While a pictorial identification key has been reported for female Pholeoixodesticks, a similar work has not been done on their male, nymphal and larval specimens. This study was initiated in order to clarify and re-examine those morphological characters of these three tick species, which can be used relatively easily to identify/distinguish them. In the case of larvae the aims included finding alternatives to chaetotaxy, which is difficult to observe and its usefulness is also affected by uncertainties in literature data.