DETECTION AND IDENTIFICATION OF Rickettsia AGENTS IN TICKS COLLECTED FROM WILD BIRDS IN BRAZIL BY POLYMERASE CHAIN REACTION-RESTRICTION FRAGMENT LENGTH POLYMORPHISM (PCR-RFLP) ANALYSIS
ABSTRACT. Santolin I.D.A.C., Famadas K.M. & McIntosh D. Detection and identification of Rickettsia agents in ticks collected from wild birds in Brazil by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis. [Detecção e identificação de espécies de Rickettsia em carrapatos coletados de aves silvestres no Brasil pela PCR-RFLP.] Revista Brasileira de Medicina Veterinária, 35(Supl.2):68-73, 2013. Programa de Pós-Graduação em Ciências Veterinárias, Instituto de Veterinária, Anexo 1, Universidade Federal Rural do Rio de Janeiro, Campus Seropédica, BR 465 Km 7, Seropédica, RJ 23897-970, Brasil. E-mail: email@example.com The detection and identification of Rickettsia bacteria in tick populations have become easier and more precise during the last 25 years as result of the developmental of robust molecular biology based methods. Currently, seven species of Ricketssia have been described in Brazilian ticks, using PCR for detection and amplicon sequencing for identification. In silico analysis of sequences encoding the gltA, htrA and ompB genes of those species was performed and revealed the basis for a novel PCR-RFLP method that allows their differential identification. The method was evaluated using larvae and nymphs of Amblyomma longirostre, A. ovale and A. varum collected from birds in the Tinguá Biological Reserve, in Rio de Janeiro State. The species “Candidatus Rickettsia amblyommii” was identified in 100% of the A. longirostre examined, while the other two tick species were PCR negative. The basic method employs the restriction endonucleases MspI and RsaI and can be performed in the course of a single working day. It offers a convenient and cost effective means to perform large scale analysis of tick populations, and should be of benefit to researchers who lack the financial or technical resources necessary for sequence based identification.