A survey on ticks and tick-borne zoonotic pathogens was carried out in 2009 and 2010 in Colli Euganei Regional Park (Veneto Region, Italy). Fiftyfive sites were sampled once or monthly to assess the presence and seasonality of ticks. In 67 out of 191 dragging 359 ticks were collected: Ixodes ricinus was the most abundant species with 341 specimens (54 adults, 95 nymphs, 192 larvae), followed by Rhipicephalus sanguineus (11 larvae), Dermacentor marginatus (6 adults) and I. acuminatus (1 adult). The overall average density for I. ricinus was 2.05 ticks/100m2. PCR was applied to identify the zoonotic agents. B. burgdorferi s.l. was found in 16/61 adults (26.2%; 95%CI 15.2% - 37.2%), in 2/32 pools of nymphs (5.0%; 95%CI 1.6% - 11.3%) and in 5/35 pools of larvae (1.0%; 95%CI 0.1% - 3.6%). Rickettsia spp. was found in all the tick species excepted for I. acuminatus. The species of Rickettsia identified in I. ricinus were R. monacensis (26/118 – 22.0%) and R. helvetica (4/118 – 3.4%). R. monacensis was identified in one pool of Rh. sanguineus larvae. R. slovaca was identified in one adult of D. marginatus. All the examined ticks were negative for TBE virus, A. phagocitophilum, Bartonella spp. and piroplasms.