Detection of Anaplasma phagocytophilum, Candidatus Neoehrlichia sp., Coxiella burnetii and Rickettsia spp. in questing ticks from a recreational park, Portugal

Tick-borne agents with medical relevance have been recorded in Portugal but little is known about their occurrence in urban outdoor leisure areas. This study aimed to investigate ticks and tick-borne agents in three public parks of Lisbon's metropolitan area. A total of 234 questing ticks belonging to eight species were found in Parque Florestal de Monsanto (PFM). Ixodes ventalloi represented 40% of collections. Mitochondrial genes confirmed Ixodes morphological identification, evidencing the intraspecific variability of I. ricinus and particularly I. frontalis populations. Regarding tick-borne agents, Rickettsia massiliae DNA were found in 21 (9.0%) ticks, Coxiella burnetii in 15 (6.4%), Anaplasma phagocytophilum in five (2.1%), an agent closely related to Candidatus Neoehrlichia mikurensis in two (0.9%), Rickettsia sibirica mongolitimonae and Rickettsia monacensis each in one (0.4%). Active enzootic cycles were suggested for these agents by the detection of positives in different time periods. Five tick species were founded with C. burnetii, including I. ventalloi which seems to be a new association record. This tick was also the only species found positive for A. phagocytophilum and the Candidatus Neoehrlichia mikurensis-like agent. Two A. phagocytophilum variants were detected in PFM, one of them representing a potentially new ecotype already found in I. ventalloi from another Portuguese area. To the authors´ knowledge, this is also the first report of such a Candidatus Neoehrlichia mikurensis-like microorganism. These data show an interesting diversity of ticks and tick-borne agents with potential public health relevance in PFM, an urban recreational area commonly frequented by humans and their pets.