Programa de Pós-Graduação em Ciências Farmacêuticas
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Doctoral Thesis Infecção natural e análise molecular de tripanosomatídeos em diferentes espécies animais no estado do Rio Grande do Norte(Universidade Federal do Rio Grande do Norte, 2023-03-17) Araújo Neto, Vicente Toscano de; Câmara, Antônia Cláudia Jácome da; Galvão, Lucia Maria da Cunha; https://orcid.org/0000-0002-6207-4502; http://lattes.cnpq.br/6437957961133013; https://orcid.org/0000-0001-7337-9320; http://lattes.cnpq.br/5445255186647578; https://orcid.org/0000-0002-0155-5230; http://lattes.cnpq.br/6780469943240040; Alves, Carlos Roberto; Salha, Daniella Regina Arantes Martins; Barbosa, David Soeiro; Mendonça, Vagner JoséThe study analyzed the occurrence of natural infection by Trypanosoma cruzi and Leishmania spp. using molecular markers and phylogenetic analysis in blood clots from domesticated animals in the state of Rio Grande do Norte. Blood clot samples were collected from 163 blood samples from sheep, goats, horses, cattle, and dogs, as well as 64 popliteal lymph node aspirates from dogs in nine municipalities in the state of Rio Grande do Norte (RN). Initially, natural infection by T. cruzi using 18S and kDNA was observed in blood clot samples from sheep, goats, and horses, with the parasite identified in 28.3% (17/60), 22.7% (5/22), and 15.4% (2/14) of the samples, respectively. These animals were distributed across the three mesoregions studied throughout the state of RN. At this point, infection in horses was described for the first time in Brazil. Subsequently, the report of a dog in the acute phase of T. cruzi infection was related to clinical, hematological, biochemical, parasitological, and characterization of the isolate using biological, molecular, and phylogenetic methods. A seemingly asymptomatic young dog exhibited a prolonged period of patent parasitemia, normocytic and hypochromic anemia. In addition, cardiac damage in this animal was observed due to the high concentration of biomarkers such as troponin I, NT-ProBN, and qualitative segmental hypokinesis. Other dog blood clot samples were also subjected to T. cruzi kDNA amplification and showed 10.9% (7/64) positivity. Phylogenetic analysis of a sheep blood clot sample revealed DNA sequences closely correlated with DTU II samples. Meanwhile, the parasite isolated from the dog in the acute phase was genetically characterized by three markers and phylogenetic analysis as TcIII. DTU III, initially described in the wild environment, is increasingly present in the domestic cycle in the Brazilian semiarid region. Data from dog samples revealed that popliteal cultures in biphasic medium were positive in 12.5% (8/64) of the samples, with flagellated forms resembling promastigotes. Sequencing and phylogenetic analysis of eight culture samples showed similarity above 99% with Leishmania infantum sequences. The widespread distribution of T. cruzi and Leishmania infantum infection in domesticated animals contributes to the expansion of the epidemiological landscape and the need to better understand the role of these animals in the transmission cycle as reservoirs and/or sentinels of infections in humans.Master Thesis Infecção pelo Trypanosoma cruzi em Canis familiares e Triatomíneos (Hemiptera, Reduviidae, Triatominae) na zona rural do Estado do Rio Grande do Norte(2018-03-16) Araújo Neto, Vicente Toscano de; ; ; Guedes, Paulo Marcos da Matta; ; Souza, Almir Pereira de;The infection by Trypanosoma cruzi in Canis familiaris and Triatomines was evaluated in the home and peridomiciliary environment of rural communities in the State of Rio Grande do Norte. Blood samples from 43 dogs from rural areas in the municipalities of Acari, Caicó, Caraúbas and Marcelino Vieira were evaluated for T. cruzi infection by serological techniques and by Polymerase Chain Reaction (PCR) to identify T. cruzi kDNA. Serology was initially performed to identify anti-T.cruzi antibodies by ELISA (Enzyme-Linked Immuno Sorbent Assay) for IgM and IgG in house and the Indirect Immunofluorescence (IFR) reaction. The presence of anti-Leishmania antibodies with DPP® (Dual Path Platform, Bio-Manguinhos / FIOCRUZ) and ELISA were also used. The catches of triatomines were carried out in the same locations and in the same period in which the dog samples were collected and the infection evaluated by direct examination and/or PCR. The criterion for positivity in T. cruzi serology was the agreement between two different techniques, which resulted in 16.3% (7/43) of the samples suggestive of acute phase and 11.6% (5/43) of chronic phase. The reactive serology for Leishmania was 9.3% of the animals in the four municipalities studied, being more reactive in the municipalities of Acari. These results of reactivity for Leishmania and T. cruzi may be related to the overlap of endemic areas. In the identification of kDNA by PCR in blood samples from dogs, 41.9% (18/43) of the samples present the band 330pb, of these 6.8% (4/43) are in the municipality of Acari, 2.3% (1/43) in Caicó, 20.9% (9/43) in Caraúbas and 9.3% (4/43) in Marcelino Vieira. The catches of triatomines identified adults and nymphs of T. brasiliensis, T. pseudomaculata, R. nasutus and adults of P. lutzi in peridomiciliary and/or intradomiciliary environments of all the studied localities. The analysis of the intestinal contents evaluated by PCR found positivity of 78.5% (95/121) of the triatomines that cohabitated these environments with the dogs. The natural T. cruzi infection was identified in dogs and triatomines in all municipalities studied, probably helping to maintain active T. cruzi infection in the domiciliary area and increasing the risk of infection in the human population.