Determinação de patogenicidade e resistência à sulfadiazina de três isolados de Toxoplasma gondii do Estado do Rio Grande do Norte e atividade anti-toxoplasma do timol e estragol

Abstract

Toxoplasma gondii is an obligate intracellular protozoan widely distributed, it is the cause of toxoplasmosis, a usually benign disease that can have serious repercussions in immunosuppressed and congenitally infected fetuses. This disease is one of the main drivers of hospitalization related to food and, considering the atypical pattern by which this can be seen in Brazil, the adverse effects on the standard treatment and resistance emerging phenotypes present this study aimed to standardize the maintenance and determine Profile of pathogenicity and resistance to sulfadiazine of local isolates de T. gondii; check the immunogenic potential and behavioral change; Besides evaluating the antiparasitic potential of thymol and estragol versus clonal and local isolated of the parasite. The parasite-induced cytopathic effect was evaluated in RAW 264.7 cell culture and pathogenicity was determined in mice following infection with monitoring for 30 days and confirmed by PRC-RFLP using CS3 marker. The phenotypic resistance to sulfadiazine was measured using a curve doses in animals infected (100, 200 or 300 mg / kg) and anti-Toxoplasma activity of thymol and estragole against local isolates and standard strains were determined in infected Swiss mice and treated during 6 days. Changes in memory/learning and impulsiveness were assessed in plus-maze discriminative avoidance task. Polymorphisms were determined by sequencing of DHPS gene. Blood samples were collected from C57BL/6 mice for the ELISA and measurement of cytokines in C57BL/6 mice. The cytopathic effect and PCR-RFLP analysis showed that the chicken populations represent different isolates of the parasite. Isolated CK3 was the most pathogenic in vivo, moderately pathogenic isolated CK2 induced a humoral response and different behavior of ME49 strain changes. Treatment failures were observed in animals infected with CK3 and Pg 1 and treated with sulfadiazine but were not observed polymorphisms in the DHPS gene. It was possible to establish a model for determination of anti-Toxoplasma activity of new compounds and, through this, it was observable that thymol and estragole were effective against the strain ME49, but only estragole was active against the isolated CK2 with a strong Th1 response. Thymol, estragole, and sulfadiazine had no action against CK3 and Pg 1. These atypical pathogenicity and resistance profiles of local parasites were first observed in the literature and may explain the differential toxoplasmosis profiles observed in this region.