First report and molecular characterization of Trypanosoma vivax in cattle from state of Pernambuco, Brazil

Abstract

This paper describes an outbreak of Trypanosoma vivax for the first time in the state of Pernambuco, Brazil, affecting dairy cattle in the municipality of Itambé in the northern coastal zone of the state. Clinical signs compatible with infection by blood protozoa and epidemic miscarriages were observed. The diagnosis of T. vivax was confirmed through biometric microscopy and molecular analysis with PCR and DNA sequencing. The T. vivax isolate detected in the present study proved to be genetically very close to other Brazilian isolates of the protozoan despite being geographically distant.

A New PCR-RFLP for Species-Specific Diagnosis of South American Animal Trypanosomiasis

“…The very low sensitivity of the parasitological method based on parasite visualization by light microscopy (MHCT, BCT and PBS) is widely recognized. Moreover, for Trypanosomiasis, the diagnostic tools based on amplification of the parasite's DNA have been extensively recognized for their high detection capability (below one parasite per mL), therefore their sensitivity is Science Publications AJAVS close to 100% (De Souza Pimentel et al, 2012). One of the most widely used methods for molecular characterization of the etiological species involved in Animal Trypanosomiasis is the assay previously described by .…”

“…Mekata et al (2009) designed a test to detect T. evansi from Bolivian and Peru bovines by PCR using Expression-Site-Associated Genes 6 (ESAG-6) as DNA template. In other work, the identification of T. vivax isolated from Pernambuco's cattle samples (Brazil) was performed by a previously described PCR coupled with amplicon sequencing (De Souza Pimentel et al, 2012). For T. vivax diagnosis in South American animals, PCR assays have also been carried out based on cathepsin L-like (CatL-like) cysteine proteases genes or on spliced-leader gene intergenic sequences (Cortez et al, 2009).…”

“…The principal disadvantage of this proach is the nonspecific symptoms of Trypanosomiasis, which are similar to other relevant hemoparasitosis present in the same regions, such as Anaplasmosis and Babesiosis (De Souza Pimentel et al, 2012;Desquesnes et al, 2013). Furthermore, the use of Science Publications AJAVS traditional parasitological methods, such as the Buffy Coat Technique-BCT (Murray et al, 1977), the MicroHematocrit Centrifugation Technique or MHCT (Woo, 1970) and the Peripheral Blood Smear (PBS) stained with Giemsa, are very limited because of their very low sensitivity, especially in the sub-acute and chronic phases of the infection, where the parasites are frequently hosted within tissues.…”

“…In the last two decades, molecular tools such as Polymerase Chain Reaction (PCR) have been shown to achieve higher rates of sensitivity (close to 100%) for Trypanosome DNA amplification De Souza Pimentel et al, 2012). However, most of the available methods are specific to the determination of species involved in African Animal Trypanosomiasis, with poor resolution for the circulating species in the South American variant Geysen et al, 2003).…”

“…However, most of the available methods are specific to the determination of species involved in African Animal Trypanosomiasis, with poor resolution for the circulating species in the South American variant Geysen et al, 2003). In other cases, the molecular tools described are specific to only one of the species (Mekata et al, 2009;De Souza Pimentel et al, 2012). In order to contribute to the generation of new tools for species-specific diagnosis of Trypanosomiasis from field samples of bovines and buffaloes, in this study we design a new PCR-RFLP adjusted to etiological species described in South America (T. vivax, T. evansi and T. theileri).…”