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Introduction: Ticks (Acari: Ixodida) are vectors of many pathogens i. a: Babesia microti [Bm] and Toxoplasma gondii [Tg]. In Poland, Ixodes ricinus [Ir] is the main vector of both pathogens. This tick species and pathogens transmitted by them are a significant threat to human and animal health., Objectives of the Study: Detection of the protozoans Bm and Tg in ticks collected in the Tarnogórski district area., Material and Methods: The ticks were collected from vegetation and pets in the spring period of their activity on the territory of the Tarnogórski district. The parasites were preserved in 70% ethanol. DNA was isolated by ammonia method. Bm was detected by nested-PCR using specific primers for the 18S rRNA sequence. To detect the Tg a commercial kit was used. The PCR products were separated on 2% ethidium bromide stained agarose gels and visualised under UV light., Results: It was showed that all collected ticks belonged to the species Ir. Bm was detected in 50.87% and Tg in 64.91% of all examined ticks. Co-existence of these both protozoans in 36.84% of total examined ticks was noted., Conclusions: The study showed a high risk of exposure to Bm and Tg in the studied area. Ticks Ir play an important role in the transmission of Bm in this region. Demonstrating a high percentage of ticks collected from animals infected with Tg may indicate their important role in the transmission of this pathogen, but it requires a further studies.

Niepołomice Forest is located about 20 kilometers east of Cracow (Malopolska province, southern Poland). Its natural and touristic values, as well as wide range of hosts occurring within indicate this to be an area of high risk of exposure to Ixodes ricinus and tick-borne diseases it transfers. I. ricinus is a common species in Poland and Europe. Its seasonal activity begins in Poland in the early spring, and ends with late autumn. A total number of 129 specimens of I. ricinus was collected by flagging in Niepołomice Forest. DNA was isolated by ammonia method from 30 randomly-selected individuals. PCR was used to detect tick-borne pathogens with primers specific for Anaplasma phagocytophilum, Borrelia burgdorferi sensu lato and Babesia sp. Molecular studies confirmed the presence of all three pathogens in I. ricinus. A. phagocytophilum was found in 76.7%, Babesia sp., 60%, B. burgdorferi s. l., in 3.3% of studied ticks. A. phagocytophilum co-infection with Babesia sp., was found in 46.7% of the specimens. A co-infection of all three tested pathogens was recorded in one case (3.3%). In Poland the problem of tick-borne diseases is a growing issue, therefore people residing in southern Polish touristic areas should be informed about the prevention and protection against ticks.