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Anaplasma phagocytophilumand spotted fever group Rickettsiaare obligate intracellular Gram-negative tick-borne bacteria, among which several may cause clinical infections in humans. Several Rickettsiaspp. and A. phagocytophilumare transmitted in Europe by Ixodes ricinus, the most common tick species feeding on humans in this area. The aim of this study was to evaluate the annual prevalence of Rickettsiaspp. and A. phagocytophilumin I. ricinuscollected from humans during three consecutive years.

The aims of our study were to determine (i) which tick species bite humans in Romania and (ii) the prevalence of Borrelia (B.) burgdorferigenospecies in these ticks. All ticks collected from patients who presented to the Clinic of Infectious Diseases Cluj Napoca in spring/summer 2010 were morphologically identified by an entomologist and tested for B. burgdorferigenospecies prevalence by a real-time PCR assay targeting the hbbgene and melting curve analysis. Out of 532 ticks, 518 were Ixodes ricinus, 10 Dermacentor marginatus, and 3 Haemaphysalisspp. ticks, and one unidentified tick due to destruction. Since evaluation of the hbbPCR revealed that it was not possible to differentiate between B. spielmanii/B. valaisianaand B. garinii/B. bavariensis, sequencing of an 800-bp fragment of the ospAgene was performed in these cases. Out of 389 investigated ticks, 43 were positive by hbbPCR for B. burgdorferisensu lato. The positive samples were 42 Ixodes ricinus(11.1% B. burgdorferisensu lato prevalence) and the one unidentified tick. Species identification revealed the presence of mainly B. afzelii, but also of B. garinii, B. burgdorferisensu stricto, B. valaisiana, and B. lusitaniae. In 4 samples, differentiation between B. spielmanii/B. valaisianawas impossible. Our study shows that the most relevant human pathogenic B. burgdorferigenospecies – predominantly B. afzelii– are present in ticks collected from Romanian patients.