Use of Molecular Methods for DNA Detection of Babesia canis vogeli in Blood Samples and Rhipicephalus sanguineus Ticks Collected in Dogs from Mexico

Abstract

Three different subspecies of Babesia canis have been detected on a global scale: B. canis canis in Europe, B. canis rossi in Africa, and B. canis vogeli, whose presence has been confirmed in America.  However, B. canis rossi has recently been reported in the United States, and B. canis vogeli has recently been identified in South Africa and Europe. The objective of this work was to optimize two methods for the molecular detection of the B. canis subspecies causing canine babesiosis in Mexico. Canine babesiosis is reported as a cosmopolitan tick-borne disease affecting domestic dogs, and its distribution is related to vector presence being more prevalent in regions with tropical and subtropical climates.  Blood samples and Rhipicephalus sanguineus ticks, 30 blood samples, and 18 tick specimens were taken from dogs with clinical manifestations compatible with canine babesiosis and a history of exposure to ticks.  The analysis of Restriction Fragment Length Polymorphisms in PCR-amplified and digested DNA with restriction enzyme TaqI and HinfI (PCR-RFLP) allowed the detection of the corresponding pattern for the B. canis vogeli subspecies (DNA fragments of 203 bp, 171 bp and 26 bp) in two blood samples and one of the tick specimens ticks subject to the PCR-RFLP analysis. Three blood samples and three of the ticks that were examined for the B. canis vogeli subspecies were found to have the 192 bp fragment of interest using a nested PCR assay. Although B. canis's presence in Mexico has earlier been shown microscopically, this is the first report of the subspecies B. canis vogeli's molecular detection in the country that has been verified by DNA sequencing.