Molecular Detection of a Borrelia sp. in Nymphs of Amblyomma brasiliense Ticks (Acari: Ixodidae) from Iguaçu National Park, Brazil, Genetically Related to Borrelia from Ethiopia and Côte D’Ivoire

ABSTRACT Gram-negative spirochetes of the genus Borrelia are transmitted to vertebrate hosts through the tick bite during blood intake. Pathogenic Borrelia species may cause relapsing fever or Lyme borreliosis in humans. Our study aimed to molecularly detect and characterize bacteria of this genus in ticks collected in the Iguacu National Park, located in southern Brazil. Ticks were collected from the environment (free living) and from hosts from May 2015 to July 2017. In total, 829 ticks were tested, being 741 from the environment and 88 from hosts; 128 ticks were larvae, 523 were nymphs, and 178 were adults (80 males and 98 females). The species identified were: Amblyomma brasiliense (42.9%), Amblyomma coelebs (16.8%), Amblyomma sp. (15.0%), Amblyomma incisum (10.3%), Amblyomma ovale (8.7%), Haemaphysalis juxtakochi (5.5%), Rhipicephalus microplus (0.5%), Amblyomma longirostre (0.2%), and Rhipicephalus sanguineus sensu lato (0.1%). DNA extraction was performed with pools of larvae or nymphs, or individually with adults, for a total of 394 samples. The PCR technique used to detect Borrelia DNA was performed with two rounds of amplification reactions targeting a segment of the flagellin B gene (flaB). Amplification occurred in only one DNA sample, which was obtained from nymphs of A. brasiliense collected from the vegetation of a trail. The nucleotide sequence analysis revealed 90.8% identity to a sequence of Borrelia sp. from Cote d’Ivoire, and 89.1% identity to a sequence from Ethiopia. Phylogenetic analysis revealed that the Borrelia sequence from A. brasiliense was distinct from the Borrelia species of the groups belonging to Lyme borreliosis, relapsing fever and the one associated with reptile and echidna as hosts. The sequence is likely from a putative new species of Borrelia and was detected for the first time in A. brasiliense ticks.