Molecular detection of Trypanosoma sp. and Blastocrithidia sp. (Trypanosomatidae) in phlebotomine sand flies (Psychodidae) in the Federal District of Brazil

Abstract: INTRODUCTION : This study describes the occurrence of trypanosomatids in phlebotomines in Brasília, Brazil. METHODS : Two hundred and ten females of 13 sand fly species were analyzed by polymerase chain reaction (PCR) using different molecular markers (D7 24Sα rRNA, kDNA, and ITS1) and sequencing. RESULTS : PCR revealed trypanosomatid-positive samples from Nyssomyia whitmani and Evandromyia evandroi, which were negative by kDNA and ITS1 Leishmania-specific PCRs. DNA sequence analysis of D7 24Sα rRNA amplicons indicated the occurrence of Blastocrithidia sp. and Trypanosoma sp. in Nyssomyia whitmani and Evandromyia evandroi, respectively. CONCLUSIONS : Two trypanosomatid species other than Leishmania sp. were found to circulate in sand flies in Central Brazil.

Report of new cases of Trypanosoma caninum in Brazil

INTRODUCTION: Trypanosoma caninum is a protozoan species recently described in dogs, whose occurrence has been reported in areas of overlap with visceral leishmaniasis. METHODS: Trypanosoma sp. were isolated from nine dogs and characterized by molecular methods. RESULTS: PCR and sequencing confirmed the presence of T. caninum in all dogs, revealing two new areas of transmission: Barra Mansa and São João do Piauí. CONCLUSIONS: The nine new cases described, when added to those already published, account for 62 cases of natural infection by T. caninum and show the geographical spread of this species to new areas, where other trypanosomatids circulate.

Characterization of plasma menbrane polypeptides of trypanosoma from bats

Cell surface proteins of Trypanosoma dionisii, Trypanosoma vespertilionis and Trypanosoma sp. (M238) were radiodinated and their distribution both in the detergent-poor (DPP) and dertergent-enriched phase (DRP) was studied using a phase separation technique in Triton X-114 as well as polyacrylamide gel electrophoresis in sodium dodecyl sulphate (SDS-PAGE). Significant differences were observed in the proteins present in the DRP when the three species of trypanosoma were compared. Two major bands with 88 and 70 KDa were observed in T. sp. (M238) but were not detectable in T. dionisii and T. vespertilionis. Three polypeptides whith 96, 77 and 60 KDa were identified in the DRP of T. vespertilionis. Three major bands with 84, 72 and 60 KDa were observed in the DRP of T. dionisii. Two polypeptides with 34-36 KDa present in the DPP, were observed in the three Trypanosome species analyzed. Our observations show that T. sp. (M238) has characteristic surface polypeptides not found in T. vespertilionis.

Review on the Molecular Tools for the Understanding of the Epidemiology of Animal Trypanosomosis in West Africa

The epidemiology of animal trypanosomosis around Bobo-Dioulasso (Burkina Faso, West Africa) benefited a lot in the last years from the progress of molecular tools. The two most used molecular techniques were the polymerase chain reaction for the diagnosis of the disease in cattle and the characterization of the trypanosomes in the host and the vector on one hand, and the microsatellite DNA polymorphism in tsetse flies to study the intraspecific genetic variability of the vector on the other hand. The results obtained in the Sideradougou area during a recent two year survey with these techniques, associated with many other georeferenced informations concerning vector and cattle distribution, natural environment, landuse, ground occupation, livestock management, were combined in a Geographical Information System. This new approach of a complex pathogenic system led to a better evaluation of the risk of trypanosome transmission.

The rising importance of Triatoma rubrofasciata

The migration of invasive vector species has contributed to the worldwide extension of infectious diseases such as dengue (Aedes aegypti) and chikungunya (Aedes albopictus). It is probably a similar behaviour for certain vectors of Chagas disease which allowed it to become a continental burden in Latin America. One of them, Triatoma rubrofasciata has also been spreading throughout the tropical and subtropical world. Here, the recent and massive peridomestic presence of T. rubrofasciata in Vietnam cities is reported, and tentatively explained, highlighting the need for improved entomological surveillance.

Infecção experimental de macacos Cebus apella sp pelo Trypanosoma cruzi (cepa "y") I - curva da parasitemia na fase aguda da doença de Chagas

Sete macacos (Cebus apella sp) foram inoculados por via subcutânea peto Trypanosoma cruzi com um número de formas tripomastigotas que variou de 25 x 103 a 5 x 1O6. Um outro primata, também da mesma espécie, foi inoculado por via palpebral com fezes de Triatoma infestans contendo formas de T. cruzi provenientes da mesma cepa "Y". A presença do T. cruzi foi assinalada na maioria dos animais, pela primeira vez, no 8º dia após a infecção. Os picos máximos da parasitemia foram observados entre o 9º e o 12º dia da infecção. O número de tripanosomas caiu acentuadamente do 19º ao 25º dia. Todos os animais infectados sobreviveram. Estudos estão sendo conduzidos com o intuito de se observar o comportamento destes animais na fase crônica da doença.

Infecção experimental de macacos cebus apella sp pelo Trypanosoma cruzi: avaliação clínica, eletrocardiqgráfica e anatomopatológica

Dos trinta e dois macacos capturados no interior do Estado de São Paulo e mantidos em laboratório em gaiolas individuais (24 a 25 C° e 70% de umidificação) após vários xenodiagnósticos negativos, 12 foram infectados por via intraperitoneal com diferentes cepas de Trypanosoma cruzi, cujas formas tripomastigotas injetadas variaram de 1.10(5) a 5.10(6) Os 20 macacos restantes foram mantidos como controle. No período de 1 a 6 anos tanto os animais inoculados como os não inoculados, foram submetidos axenodiagnóstico e teste soro lógico de aglutinação direta, exame clínico e o eletrocardiograma. Posteriormente os macacos foram necropsiados e todos os órgãos submetidos a exame macro e microscópico. O exame clínico e o eletrocardiograma não revelaram alterações. Dos 12 macacos infectados, 4 apresentaram evidências de infecção ao exame histopatológico: utn com formas amastigotas nos tecidos e 3 com miocardite crônica de grau leve. A parasitemiafoi comprovada em 66,66% dos animais na fase aguda e a sorologia em 91,66% na fase crônica. Concluiu-se que os macacos Cebus não expressaram susceptibilidade ao desenvolvimento das lesões que caracterizam a fase . crônica da doença de Chagas mas poderiam ser usados para manter as cepas de T. cruzi e estudos de pesquisa sorológico a longo termo.

Caracterização biológica, histopatológica e análise de ácido nucléico de uma cepa Trypanosoma cruzi da região de Marília, SP

Estudou-se o comportamento biológico e histopatológico de uma cepa genuínamente mariliense de Trypanosoma cruzi, isolada em 1997 através de xenodiagnóstico artificial. Vinte e cinco camundongos swiss foram infectados intraperitonealmente, sendo 11 utilizados para a realização da curva parasitêmica e observação da morfologia dos tripomastigotas e 14 foram sacrificados após o 17, 23, 30, 60 e 180 dias pós-infecção e coletados coração, esôfago, fígado, cólon, e músculo esquelético (fragmento da coxa direita) para análise histopatológica. Cultura em meio LIT foi realizada para análise de DNA. Os resultados mostraram predomínio de formas largas, baixa parasitemia com picos médios de 860 tripomastigotas/5mil de sangue ao redor do 20º dia de infecção. Nenhum camundongo morreu na fase aguda da infecção. Exame histopatológico mostrou poucos ninhos de amastigotas em coração, raros em músculo esquelético e cólon com discreto processo inflamatório. Comparada com a cepa Y, que foi isolada de uma paciente da mesma região, notamos diferentes características biológicas e comportamentais, porém a análise de DNA as coloca no mesmo grupo, demonstrando a proximidade dessas cepas.

A new triatomine host of trypanosoma from the Central Amazon of Brazil: Cavernicola lenti n.sp. (Hemiptera, Reduviidae, Triatominae)

Adults and nymphs of Cavernicola lenti, new species, from Amazonas state, Brazil, are described and illustrated. Observations on the biology of the new species are presented. Preliminaty findings indicate that C. lenti is a probable vector of bat trypanosomes.

Trypanosoma guairaensis sp. n. (Protozoa, Kinetoplastida) parasita de megaloancistrus aculeatus (Perugia, 1891) (Pisces, Loricariidae)

Descreve-se a espécie Trypanosoma guairaensis sp. n. parasita de Megaloancistrus aculeatus do rio Paraná (reservatório de Itaipu). Características morfométricas como comprimento e largura do núcleo e comprimento da parte livre do flagelo, bem como os valores do índice nuclear, diferenciam T. guairaensis de todas as espécies de tripanosomas descritas para peixes de água doce do Brasil.

Trypanosoma nupelianus sp. n. (Protozoa, Kinetoplastida) parasitizing rhinelepis aspera (Osteichthyes, Loricariidae) from Paraná river, Brazil

It is described Trypanosoma nupelianus sp. n. parasitizing Rhinelepis aspera taken from Paraná river (Itaipu reservoir, Brazil). Morphometric characters such as body length, width of the body and nucleus dimensions show that T. nupelianus sp. n. is different from other trypanosomes described for fishes from tropical regions.

Chronic myocardial damage in experimental T. cruzi infection of a new world primate, Cebus sp. monkey

Eighteen Cebus apella monkeys, (juvenile and adult of both sexes) were inoculated five years ago, with three Trypanosoma cruzi strains (CA1, n = 10; Colombian, n=4 and Tulahuen, n=4), either by conjunctival or intraperitoneal route, once or repeatedly. Parasitological, hematological, serological, enzymatic, radiographic, electro and echocardiographic findings have been peviously published15 and they are similar to those observed in human pathology. The most frequent electrocardiographic alteration was right branch bundle block. Six animals, chosen at random, were sacrificed. Those sacrificed 20 to 25 months post-first inoculation showed focal accumuli of leukocytes with myocytolysis. Foci of diffuse interstitial fibrosis with mild infiltrate of leukocytes among fibers were observed in the animals sacrificed 36 to 47 months post-inoculation. No parasites were seen. The lesions were more prominent in the ventricular walls and the septum. The fact that the infiltrates were predominant in the animals sacrificed at a shorter time after first inoculation and that fibrosis was more severe in those sacrificed at a longer time suggests that there is a progression of the infiltrative lesions to fibrosis, with a leukocytic activity indicative of a chronic phase. These lesions are similar to those described in human chronic Chagas' disease. This would demonstrate that this model is useful in evaluating a progress in the knowledge of the pathogenesis which is still a controversial issue, immunology, immunogenesis and chemotherapeutic agents of the chronic and indeterminate phases of this disease.

Trypanosoma cruzi: chemotherapy with benznidazole in mice inoculated with strains from Paraná State and from different endemic areas of Brazil

Strains of Trypanosoma cruzi from different geographical areas have shown different levels of susceptibility to trypanocidal drugs. The susceptibility in vivo to benznidazole was investigated in eighteen strains of T. cruzi. Twelve were isolated from chronic chagasic patients from different Chagas’ disease endemic areas. The other six strains were isolated from the northwestern region of Paraná state; two of them from patients three from triatomines (Triatoma sordida) and one from wild reservoir (Didelphis sp.). To test drug the infected mice were divided into two groups of twenty. One group was treated with benznidazole for twenty consecutive days and the other group was used as untreated control. The treatment began after detection of the infection by direct blood examination or haemoculture. The control of cure was done through haemoculture and indirect immunofluorescence test. The drug eliminated the inflammatory lesions of the skeletal muscle of mice considered cured and from the heart of most of them. Moreover, the inflammatory lesions were reduced in treated but not cured animals. The T. cruzi strains studied showed a gradient of drug susceptibility that varied from 0% to 100%. Ten strains were considered sensitive to the treatment (61 to 100% of cure), one strain was partially sensitive (50% of cure) and seven strains were considered resistant to the treatment (0 to 40% of cure). This variation was observed both in strains of T. cruzi isolated from domestic and sylvatic cycles

Estudo sobre triatomíneos e reservatórios silvestres de Trypanosoma cruzi no estado do Paraná, Sul do Brasil. Resultados preliminares

Em localidades da zona rural de cinco municípios da região noroeste do Paraná adultos e ninfas de Triatoma sordida foram capturados em 21 (41,2%) de 51 unidades domiciliares pesquisadas. Foram capturados 154 exemplares de T. sordida e 2 Panstrongylus megistus. De 135 exemplares examinados 58 (43,0%) apresentavam o Trypanosoma tipo cruzi. Constatou-se também a infecção em 57,1% (4/7) dos gambás (Didelphis sp) examinados. O peridomicílio apresentou-se mais infestado que o intradomicílio, sendo a casa de madeira abandonada a construção mais freqüente (34,7% do total investigado) e com maior taxa de infestação (53,9%). Os dados mostram um elevado índice de infestação da zona rural por espécies secundárias de triatomíneos com altas taxas de infecção por flagelados do tipo T. cruzi, em uma área endêmica para a doença de Chagas já em fase de vigilância epidemiológica.