Detection and genotyping of Trypanosoma cruzi from açai products commercialized in Rio de Janeiro and Pará, Brazil

Abstract

BackgroundSeveral cases of food-borne acute Chagas disease (ACD) have been reported in the Brazilian Amazon so far. Up to 2004, the occurrence of ACD by oral transmission, associated with food consumption, was rare. Recent cases of ACD in Brazil have been attributed to the consumption of juice from the açai palm containing reservoir animals or insect vectors waste, infected with Trypanosoma cruzi. This study aimed to determine the T. cruzi contamination rate and to genotype the parasite in food samples prepared from açai, which are commercialized in Rio de Janeiro and the Pará States in Brazil.MethodsThe amplificability of DNA extracted from açai samples, and T. cruzi and Triatominae detection were performed by conventional PCR. Molecular characterization was done by multilocus PCR analysis, to determine the parasite discrete type units (DTUs) based on the size of PCR products in agarose gels, using the intergenic region of the spliced leader (SL), 24 Sα rDNA and nuclear fragment A10 as targets.ResultsFrom the 140 samples of açai-based products analyzed, T. cruzi DNA was detected in 14 samples (10%); triatomine DNA was detected in one of these 14 samples. The parasite genotyping demonstrated that food samples containing açai showed a mixture of T. cruzi DTUs with TcIII, TcV and TcI prevailing.ConclusionsIn this study, the molecular detection and identification of T. cruzi from açai-based manufactured food samples, was performed for the first time. Although parasite DNA is a marker of possible contamination during food manufacturing, our findings do not indicate that açai is a source of Chagas disease via oral transmission per se, as live parasites were not investigated. Nevertheless, a molecular approach could be a powerful tool in the epidemiological investigation of outbreaks, supporting previous evidence that açai-based food can be contaminated with T. cruzi. Furthermore, both food quality control and assessment of good manufacturing practices involving açai-based products can be improved, assuring the safety of açai products.

WHF IASC Roadmap on Chagas Disease

“…Because oral transmission occurs primarily through food, especially fruit contaminated with triatomine feces, implementation of safe food-handling practices combined with health promotion activities on the prevention of oral transmission are needed to prevent continued oral CD outbreaks. Promotion of safe food-handling practices among households and food vendors in areas at risk is essential, especially for high-risk foods such as sugar cane and acai [58,59]. Preventing transmission via blood transfusion, organ transplant, and laboratory accidents There has been growing acceptance of the need for screening of blood donations, in both endemic and nonendemic countries over the past 30 years, resulting in a significant decrease in known transfusion-related diseases.…”

Validation of a novel multiplex real-time PCR assay forTrypanosoma cruzidetection and quantification in açai pulp

“…Namely, the characteristic dark color of the fruit, which is associated with high anthocyanin concentrations and a large amount of organic matter [34,35], greatly limit the possibility of parasites detection during microscopic visualization. In this context, molecular methods can overcome these limitations regarding analysis directly from food, and provide specific diagnosis [26,41,50,52,63] and T. cruzi genotyping [51]. The qPCR developed in the present study showed improved sensitivity, in which was possible to detect the DNA corresponding to 0.01 T. cruzi equivalents/mL in the sample.…”

“…The qPCR screening of these GEA samples was shown to be more sensitive than microscopic examination, since the molecular method revealed a positivity of 20% (9/45), whereas all samples were negative for T. cruzi by direct observation at a light microscope. Before that, Ferreira et al [51] determined, in food samples commercialized in Rio de Janeiro and Pará states, T. cruzi contamination rates and molecular characterization through conventional PCR and multilocus PCR analysis, respectively. For T. cruzi DNA detection, a PCR amplifying the telomeric region of gp85/sialidase superfamily was performed.…”

“…Moreover, the use of PCR for the detection of T. cruzi DNA has already been described in literature, since several studies demonstrate that this methodology presents greater sensitivity and specificity in the face of conventional parasitological methods [53]. Nevertheless, it is still demanded the development of methodologies capable to detect T. cruzi in food samples and, thereby, plan strategies to get a better understanding of oral transmission and to assure the quality of the products thar are being commercialized [34,51].…”

Parasite, Compartments, and Molecules: Trick versus Treatment on Chagas Disease

“…The oral route, which is probably the most frequent mechanism among vectors and wild mammals, has recently become relevant, due to environmental changes caused by deforestation [12]. T. cruzi DNA was recently shown in 10% among 140 samples of açai-based products marketed in Rio de Janeiro and Pará States in Brazil [13].…”