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.

First record of Trypanosoma sp. (Protozoa: Kinetoplastida) in tuvira (Gymnotus aff. inaequilabiatus) in the Pantanal wetland, Mato Grosso do Sul State, Brazil

O objetivo do presente estudo foi reportar a infecção por Trypanosoma sp. em tuviras (Gymnotus aff. inaequilabiatus) oriundas do Pantanal Sul-mato-grossense, Brasil. Dez peixes provenientes do rio Paraguai, Pantanal Sul-mato-grossense, foram avaliados quanto à presença de hemoflagelados. Tripomastigotas de Trypanosoma sp. foram observados nas extensões sanguíneas de três peixes (30% de prevalência), e algumas formas encontravam-se em divisão. Por meio do exame a fresco e da centrifugação do sangue em capilar de hematócrito como métodos para diagnóstico, a taxa de prevalência foi de 80%. Esse é o primeiro relato de Trypanosoma sp. em tuviras no Brasil.

First record of Trypanosoma sp. (Protozoa: Kinetoplastida) in tuvira (Gymnotus aff. inaequilabiatus) in the Pantanal wetland, Mato Grosso do Sul State, Brazil

O objetivo do presente estudo foi reportar a infecção por Trypanosoma sp. em tuviras (Gymnotus aff. inaequilabiatus) oriundas do Pantanal Sul-mato-grossense, Brasil. Dez peixes provenientes do rio Paraguai, Pantanal Sul-mato-grossense, foram avaliados quanto à presença de hemoflagelados. Tripomastigotas de Trypanosoma sp. foram observados nas extensões sanguíneas de três peixes (30% de prevalência), e algumas formas encontravam-se em divisão. Por meio do exame a fresco e da centrifugação do sangue em capilar de hematócrito como métodos para diagnóstico, a taxa de prevalência foi de 80%. Esse é o primeiro relato de Trypanosoma sp. em tuviras no Brasil.

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.

Hemoprotozoa of freshwater turtles in Queensland

Blood smears from 27 turtles (15 Emydura signata, nine Elseya latisternum, and three Chelodina longicollis) from southeastern Queensland (Australia) were examined for infections by hemoprotozoan parasites between January and June 1999. Infections were found in 26 (96%) of the turtles. Twenty five (93%) were infected with the adeleorin coccidian Haemogregarina clelandi, eight (30%) with the hemosporidian Haemoproteus chelodinae, 11 (41%) with the kinetoplastid flagellate Trypanosoma chelodinae, and eight (30%) with a novel Trypanosoma sp. Despite the high prevalence and intensity of infections, there was no evidence of clinical disease in any of the turtles.

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.

Effect of management strategies on the performance of ruminant livestock production systems and the safety of plant and animal products in the city of Sikasso, Mali

The regional population growth in West Africa, and especially its urban centers, will bring about new and critical challenges for urban development policy, especially in terms of ensuring food security and providing employment for the growing population. (Peri-) urban livestock and vegetable production systems, which can contribute significantly to these endeavours, are limited by various constraints, amongst them limited access to expensive production factors and their (in)efficient use. To achieve sustainable production systems with low consumer health risks, that can meet the urban increased demand, this doctoral thesis determined nutrient use efficiencies in representative (peri-) urban livestock production systems in three West African cities, and investigated potential health risks for consumers ensuing from there. The field study, which was conducted during July 2007 to December 2009, undertook a comparative analysis of (peri-) urban livestock production strategies across 210 livestock keeping households (HH) in the three West African cities of Kano/Nigeria (84 HH), Bobo Dioulasso/Burkina Faso (63 HH) and Sikasso/Mali (63 HH). These livestock enterprises were belonging to the following three farm types: commercial gardening plus field crops and livestock (cGCL; 88 HH), commercial livestock plus subsistence field cropping (cLsC; 109 HH) and commercial gardening plus semi-commercial livestock (cGscL; 13 HH) which had been classified in a preceding study; they represented the diversity of (peri-) urban livestock production systems in West Africa. In the study on the efficiency of ruminant livestock production, lactating cowsand sheep herd units were differentiated based on whether feed supplements were offered to the animals at the homestead (Go: grazing only; Gsf: mainly grazing plus some supplement feeding). Inflows and outflows of nutrients were quantified in these herds during 18 months, and the effects of seasonal variations in nutrient availability on animals’ productivity and reproductive performance was determined in Sikasso. To assess the safety of animal products and vegetables, contamination sources of irrigated lettuce and milk with microbiological contaminants, and of tomato and cabbage with pesticide residues in (peri-) urban agriculture systems of Bobo Dioulasso and Sikasso were characterized at three occasions in 2009. Samples of irrigation water, organic fertilizer and ix lettuce were collected in 6 gardens, and samples of cabbage and tomato in 12 gardens; raw and curdled milk were sampled in 6 dairy herds. Information on health risks for consumers of such foodstuffs was obtained from 11 health centers in Sikasso. In (peri-) urban livestock production systems, sheep and goats dominated (P<0.001) in Kano compared to Bobo Dioulasso and Sikasso, while cattle and poultry were more frequent (P<0.001) in Bobo Dioulasso and Sikasso than in Kano. Across cities, ruminant feeding relied on grazing and homestead supplementation with fresh grasses, crop residues, cereal brans and cotton seed cake; cereal grains and brans were the major ingredients of poultry feeds. There was little association of gardens and livestock; likewise field cropping and livestock were rarely integrated. No relation existed between the education of the HH head and the adoption of improved management practices (P>0.05), but the proportion of HH heads with a long-term experience in (peri-) urban agriculture was higher in Kano and in Bobo Dioulasso than in Sikasso (P<0.001). Cattle and sheep fetched highest market prices in Kano; unit prices for goats and chicken were highest in Sikasso. Animal inflow, outflow and dairy herd growth rates were significantly higher (P<0.05) in the Gsf than in the Go cattle herds. Maize bran and cottonseed expeller were the main feeds offered to Gsf cows as dry-season supplement, while Gsf sheep received maize bran, fresh grasses and cowpea pods. The short periodic transhumance of Go dairy cows help them maintaining their live weight, whereas Gsf cows lost weight during the dry season despite supplement feeding at a rate of 1506 g dry matter per cow and day, resulting in low productivity and reproductive performance. The daily live weight gains of calves and lambs, respectively, were low and not significantly different between the Go and the Gsf system. However, the average live weight gains of lambs were significantly higher in the dry season (P<0.05) than in the rainy season because of the high pressure of gastrointestinal parasites and of Trypanosoma sp. In consequence, 47% of the sheep leaving the Go and Gsf herds died due to diseases during the study period. Thermo-tolerant coliforms and Escherichia coli contamination levels of irrigation water significantly exceeded WHO recommendations for the unrestricted irrigation of vegetables consumed raw. Microbial contamination levels of lettuce at the farm gate and the market place in Bobo Dioulasso and at the farm gate in Sikasso were higher than at the market place in Sikasso (P<0.05). Pesticide residues were detected in only one cabbage and one tomato sample and were below the maximum residue limit for consumption. Counts of thermo-tolerant coliforms and Escherichia coli were higher in curdled than in raw milk (P<0.05). From 2006 to x 2009, cases of diarrhea/vomiting and typhoid fever had increased by 11% and 48%, respectively, in Sikasso. For ensuring economically successful and ecologically viable (peri-) urban livestock husbandry and food safety of (peri-) urban foodstuffs of animal and plant origin, the dissemination and adoption of improved feeding practices, livestock healthcare and dung management are key. In addition, measures fostering the safety of animal products and vegetables including the appropriate use of wastewater in (peri-) urban agriculture, restriction to approve vegetable pesticides and the respect of their latency periods, and passing and enforcement of safety laws is required. Finally, the incorporation of environmentally sound (peri-) urban agriculture in urban planning by policy makers, public and private extension agencies and the urban farmers themselves is of utmost importance. To enable an efficient (peri-) urban livestock production in the future, research should concentrate on cost-effective feeding systems that allow meeting the animals’ requirement for production and reproduction. Thereby focus should be laid on the use of crop-residues and leguminous forages. The improvement of the milk production potential through crossbreeding of local cattle breeds with exotic breeds known for their high milk yield might be an accompanying option, but it needs careful supervision to prevent the loss of the local trypanotolerant purebreds.

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.