Screening and Identification of Pathogen Box® Compounds with anti-Trypanosoma evansi Activity.

The development of new drugs targeting neglected animal diseases is imperative. In Asia and South America, Trypanosoma evansi is a pathogen that affects horses and other species, causing economic losses associated with reduced animal productivity and death. In order to accelerate the identification of drugs with activity against neglected diseases, Medicines for Malaria Venture has developed Pathogen Box®, a library of 400 different molecules. The present work aimed to identify compounds present in the Pathogen Box® library, measuring in vitro activity against T. evansi. Among the 400 compounds, 5 showed anti-T.evansi activity: pentamidine, MMV688410, MMV687273, MMV022478 and auranofin. Suramin, a trypanocidal activity molecule present on the Pathogen Box® reference compound list, demonstrated no anti-T. evansi activity in the in vitro assays. MMV688410 is the most promising candidate because it induces death and reduces the number of parasites in cell culture, and mainly because its mechanism of action is probably associated with inhibition of trypanosomal reductase enzyme, an exclusive target of trypanosomatides. Further in vitro and in vivo assays are needed to determine the efficacy of the compounds identified in this work, especially by associating tissue distribution and the ability of drugs to cross the blood brain barrier, as T. evansi is able to invade the central nervous system.

Prevalence of Anaplasma marginale, Babesia bovis, and Babesia bigemina in cattle in the Campos de Lages region, Santa Catarina state, Brazil, estimated by multiplex-PCR.

This study aimed to determine the prevalence of three common hemoparasites (Anaplasma marginale, Babesia bovis and Babesia bigemina) in cattle from 16 counties in the Campos de Lages region, Santa Catarina state, Brazil, and the factors affecting disease occurrence. The study population consisted of 257 clinically healthy animals from 21 rural farms. Bovine blood samples were collected by jugular venipuncture. DNA was extracted from whole blood by the phenol/ chloroform method. Genomic DNA extracted from blood samples was subjected to Multiplex PCR for screening of B. bovis, B. bigemina, and A. marginale using specific primers. Prevalences of A. marginale, B. bigemina, and B. bovis were 27%, 16%, and 29%, respectively. Mixed infection was observed in 17.5% of samples. The most frequent was Babesia bovis and Babesia bigemina in 6.62% of samples. A. marginale infection rates were statistically correlated with age groups of cattle. The infections detected in the study population were considered to be subclinical, based on the presence pathogen DNA and absence of clinical symptoms. Seasonality of the pathogens resulted in various degrees of infection, related to the age of the animals and the season. The Campos de Lages region is characterized by enzootic instability for these pathogens because of its climatic and geographic features.

First evaluation of an outbreak of bovine babesiosis and anaplasmosis in Southern Brazil using multiplex PCR.

Outbreaks of tick-borne disease cases in Santa Catarina, Brazil are known, but the presence of the pathogen DNA has never been determined. In this study, the first survey of Anaplasma marginale, Babesia bigemina, and Babesia bovis DNA on blood samples of 33 cattle from an outbreak in Ponte Alta Municipality, Santa Catarina, Brazil, has been carried out. A multiplex PCR detected 54.5% of animals were co-infected with 2 or 3 parasites, while 24.2% were infected with only 1 species. The most prevalent agent was B. bigemina (63.6%) followed by A. marginale (60.6%). This is the first report of tick-borne disease pathogens obtained by DNA analysis in Southern Brazil.