Experimental infection of T4 Acanthamoeba genotype determines the pathogenic potential.

T4 is the Acanthamoeba genotype most related to cases of granulomatous amoebic encephalitis (GAE) in immunocompromised patients and of keratitis in contact lens wearers. The determination of the pathogenic potential of Acanthamoeba clinical and environmental isolates using experimental models is extremely important to elucidate the capacity of free-living organisms to establish and cause disease in hosts. The aim of this study was to compare and evaluate the histopathology and culture between two different routes of experimental infection of T4 Acanthamoeba isolated from environmental and clinical source in mice (intracranial and intraperitoneal). Swiss isogenic healthy mice were inoculated with 10(4) trophozoites by intracranial (IC) and intraperitoneal (IP) routes and observed during 21 days. The brains from animals inoculated by the IC route were collected and from the animals of the IP inoculation group, the brains, livers, kidneys, spleens, and lungs were removed. The organs were prepared and appropriately divided to be evaluated with histopathology and culture. There was no significant difference between the inoculation routes in terms of isolates recovery (χ(2) = 0.09; p = 0.76). In the IC group, isolate recovery rate was significantly higher in histopathology than the one achieved by culture (χ(2) = 6.45; p < 0.01). Experimental infection revealed that all isolates inoculated could be considered invasive because it was possible to recover evolutive forms of Acanthamoeba in both routes. This work represents the first in vivo pathogenicity assay of primary isolation source in Central region of Brazil showing in vivo pathogenicity and hematogenous spread capacity of these protozoa, improving the knowledge on free-living amoebae isolates.

[Trypanosoma rangeli parasite-vector-vertebrate interactions and their relationship to the systematics and epidemiology of American trypanosomiasis]. / Interacción tripanosoma-vector-vertebrado y su relación con la sistemática y la epidemiología de la tripanosomiasis Americana.

INTRODUCTION: Trypanosoma rangeli is a species of trypanosome second to T. cruzi, that is infective to humans in Latin America. Variability in the biological, biochemical and molecular characteristics between different isolates isolates of this parasite have been recorded. OBJECTIVE: Morphological and molecular characteristics were recorded from strains of T. rangeli that were isolated from different species of Rhodnius and maintained in different vertebrate species. MATERIALS AND METHODS: Nineteen strains of T. rangeli were isolated from R. prolixus, R. pallescens and R. colombiensis in Colombia, R. ecuadoriensis in Peru and R. pallescens in Panama. Polymorphism of blood trypomastigotes in ICR mice was evaluated and pleomorphism of P53 strain of T. rangeli KP1(-) inoculated in mouse, marsupial and canine was studied. RAPD analysis (randomly amplified polymorphic DNA analysis) of 12 strains isolated from four species of Rhodnius was performed. RESULT: Based on the total length of blood trypomastigotes, three discrete groups were observed. The P53 strain showed significant differences in the size of blood trypomastigotes in mouse, marsupial and canine. RAPD analysis showed that the strains segregated into two branches corresponding to strains of T. rangeli KP1(+) and T. rangeli KP1(-). All strains of T. rangeli KP1(-) clustered according to the species of Rhodnius from which they were isolated. CONCLUSION: These data reveal, for the first time, a close association amongst T. rangeli strains and Rhodnius species, confirming that each species of Rhodnius transmits to vertebrate hosts a parasite population with clear phenotypic and genotypic differences. This is further evidence that supports the concept of clonal evolution of these parasites.

Susceptibility of different Rhodnius species (Hemiptera, Reduviidae, Triatominae) to a Brazilian strain of Trypanosoma rangeli (SC58/KP1-).

INTRODUCTION: Specific host-parasite associations have been detected experimentally and suggest that triatomines of the genus Rhodnius act as biological filters in the transmission of Trypanosoma rangeli . OBJECTIVE: To analyze the susceptibility of four Rhodnius species ( Rhodnius robustus , Rhodnius neglectus , Rhodnius nasutus and Rhodnius pictipes ) to a Brazilian strain of T. rangeli (SC-58/KP1-). MATERIALS AND METHODS: We selected t hirty nymphs of each species, which were fed on blood infected with T. rangeli . Periodically, samples of feces and hemolymph were analyzed. Triatomines with T. rangeli in their hemolymph were fed on mice to check for transmission by bites. Later, the triatomines were dissected to confirm salivary gland infection. RESULTS: Specimens of R. pictipes showed higher rates of intestinal infection compared to the other three species. Epimastigotes and trypomastigotes were detected in hemolymph of four species; however, parasitism was lower in the species of the R. robustus lineage. Rhodnius robustus and R. neglectus specimens did not transmit T. rangeli by bite; after dissection, their glands were not infected. Only one specimen of R. nasutus and two of R. pictipes transmitted the parasite by bite. The rate of salivary gland infection was 16% for R. pictipes and 4% for R. nasutus . CONCLUSIONS: Both infectivity (intestinal, hemolymphatic and glandular) and transmission of T. rangeli (SC58/KP1-) were greater and more efficient in R. pictipes. These results reinforce the hypothesis that these triatomines may act as biological filters in the transmission of T. rangeli .

Multiplex Real-Time PCR Assay Using TaqMan Probes for the Identification of Trypanosoma cruzi DTUs in Biological and Clinical Samples.

BACKGROUND: Trypanosoma cruzi has been classified into six Discrete Typing Units (DTUs), designated as TcI-TcVI. In order to effectively use this standardized nomenclature, a reproducible genotyping strategy is imperative. Several typing schemes have been developed with variable levels of complexity, selectivity and analytical sensitivity. Most of them can be only applied to cultured stocks. In this context, we aimed to develop a multiplex Real-Time PCR method to identify the six T. cruzi DTUs using TaqMan probes (MTq-PCR). METHODS/PRINCIPAL FINDINGS: The MTq-PCR has been evaluated in 39 cultured stocks and 307 biological samples from vectors, reservoirs and patients from different geographical regions and transmission cycles in comparison with a multi-locus conventional PCR algorithm. The MTq-PCR was inclusive for laboratory stocks and natural isolates and sensitive for direct typing of different biological samples from vectors, reservoirs and patients with acute, congenital infection or Chagas reactivation. The first round SL-IR MTq-PCR detected 1 fg DNA/reaction tube of TcI, TcII and TcIII and 1 pg DNA/reaction tube of TcIV, TcV and TcVI reference strains. The MTq-PCR was able to characterize DTUs in 83% of triatomine and 96% of reservoir samples that had been typed by conventional PCR methods. Regarding clinical samples, 100% of those derived from acute infected patients, 62.5% from congenitally infected children and 50% from patients with clinical reactivation could be genotyped. Sensitivity for direct typing of blood samples from chronic Chagas disease patients (32.8% from asymptomatic and 22.2% from symptomatic patients) and mixed infections was lower than that of the conventional PCR algorithm. CONCLUSIONS/SIGNIFICANCE: Typing is resolved after a single or a second round of Real-Time PCR, depending on the DTU. This format reduces carryover contamination and is amenable to quantification, automation and kit production.

Domestic Rhodnius ecuadoriensis (Hemiptera, Reduviidae) infestation in Northern Peru: a comparative trial of detection methods during a six-month follow-up.

Two passive methods in the assessment of intradomiciliary infestation by Rhodnius ecuadoriensis were tested: (i) the Gomes Nu ez sensor box (GN), (ii) sheets of white typing paper and (iii) one active timed manual method. The study was carried out in the Alto Chicama River Valley, Province of Gran Chim , Department of La Libertad. The study design consisted of an initial searching of triatomines inside of the domestic environment by the manual capture active procedure (man/hour) covering all the studied houses. Then, matched pairs of GN boxes and paper sheets were simultaneously installed in the bedrooms of 207 households distributed in 19 localities. A comparative prospective trial of these passive detection devices were monitored at 2, 4 and, finally 6 months follow-up. Parasitological Trypanosoma rangeli and/or T. cruzi infections were investigated in two houses with high level of infestation by R. ecuadoriensis. 16.9% of the 207 households investigated by an initial active manual method were infested with R. ecuadoriensis. The proportion of infested houses fluctuated from 6.2 to 55.5% amongst the 19 localities investigated. T. rangeli natural infection was detected in R. ecuadoriensis specimens collected in two households. Parasite rates in the bugs ranged from 16.6 to 21.7% respectively. The most striking fact was an average rate of salivary gland infection ranging from 7.4 to 8.3%. At the end of the sixth month period, a cumulative incidence of 31.4% of positive GN boxes against 15.9% for paper sheets was recorded. All three methods combined detected domestic infestation in 129 (62.3%) of the 207 houses studied in the 19 localities. The range of houses infested varies from 6.7% to 92.9%. In areas with low bug density infestation rates, the methodology experienced in our studies, seems to be the best choice for investigations on domestic R. ecuadoriensis populations.

Enzootic transmission of Trypanosoma cruzi and T. rangeli in the Federal District of Brazil.

The Federal District of Brazil (DF) lies within the Cerrado biome, where open shrubland (savannas) is interspersed with riverside gallery forests and permanent swamps (veredas). Trypanosoma cruzi-infected native triatomines occur in the area, but the enzootic transmission of trypanosomatids remains poorly characterized. A parasitological survey involving sylvatic triatomines (166 Rhodnius neglectus collected from Mauritia flexuosa palms) and small mammals (98 marsupials and 70 rodents, totaling 18 species) was conducted in 18 sites (mainly gallery forests and veredas) of the DF. Parasites were isolated, morphologically identified, and characterized by PCR of nuclear (mini-exon gene) and kinetoplast DNA (kDNA). Six R. neglectus, seven Didelphis albiventris and one Akodon cursor were infected by trypanosomes; wild reservoir infection is documented for the first time in the DF. kDNA PCR detected T. cruzi in five R. neglectus and mini-exon gene PCR revealed T. cruzi I in isolates from D. albiventris. Parasites infecting one bug yielded T. rangeli KP1+ kDNA amplicons. In spite of the occurrence of T. cruzi-infected D. albiventris (an important wild and peridomestic reservoir) and R. neglectus (a secondary vector displaying synanthropic behavior), a low-risk of human Chagas disease transmission could be expected in the DF, considering the low prevalence infection recorded in this work. The detection of T. rangeli KP1+ associated with R. neglectus in the DF widens the known range of this parasite in Brazil and reinforces the hypothesis of adaptation of T. rangeli populations (KP1+ and KP1-) to distinct evolutionary Rhodnius lineages.

[Spatial distribution of Triatominae populations (Hemiptera: Reduviidae) in Mauritia flexuosa palm trees in Federal District of Brazil]. / Distribuição espacial de populações de triatomíneos (Hemiptera: Reduviidae) em palmeiras da espécie Mauritia flexuosa no Distrito Federal, Brasil.

To determine and analyze the distribution of Triatominae sylvatic populations in the Federal District of Brazil, 150 Mauritia flexuosa palm trees were sampled in six veredas of different landscapes (sylvatic, rural and peri-urban) in the rainy season. Triatomines were morphologically identified and grouped by sex and nymphal stage and were examined to verify infection by Trypanosoma cruzi and Trypanosoma rangeli. Twenty eight (18.6%) palm trees were infested by Rhodnius neglectus and fourteen (9.5%) by Psammolestes tertius. The frequency of Triatominae in palm trees with and without nests was significantly different being higher in palm trees with bird and mammal nests in the crown. The higher average number of insects/palm tree was observed in rural areas with estimates of up to 838 insects/hectare. The species age makeup presented a different pattern, with nymphs predominant for Rhodnius neglectus and adults predominant for Psammolestes tertius. Also, many Rhodnius neglectus eggs were collected, which indicates a reproductive event in February 2003. Among the nests found in palm trees, that of the Phacellodomus ruber (Furnariidae) bird had the greatest abundance of Triatominae, occurring on 42% of palm trees. The relative abundance of Rhodnius neglectus and Psammolestes tertius was greater in rural areas which contained higher number of nests in palm trees and lesser density of palms per hectare. None of the 96 triatomines examined were infected by Trypanosoma cruzi or Trypanosoma rangeli, despite our finding of Rhodnius neglectus infection by those parasites in former studies.

Sequence analysis of the spliced-leader intergenic region (SL-IR) and random amplified polymorphic DNA (RAPD) of Trypanosoma rangeli strains isolated from Rhodnius ecuadoriensis, R. colombiensis, R. pallescens and R. prolixus suggests a degree of co-evolution between parasites and vectors.

Spliced leader intergenic region (SL-IR) sequences from 23 Trypanosoma rangeli strains isolated from the salivary glands of Rhodnius colombiensis, R. ecuadoriensis, R. pallescens and R. prolixus and two human strains revealed the existence of 4 genotypes with CA, GT, TA, ATT and GTAT microsatellite repeats and the presence of insertions/deletions (INDEL) and single nucleotide polymorphism (SNP) characterizing each genotype. The strains isolated from the same vector species or the same Rhodnius evolutionary line presented the same genotypes, even in cases where strains had been isolated from vectors captured in geographically distant regions. The dendrogram constructed from the SL-IR sequences separated all of them into two main groups, one with the genotypes isolated from R. prolixus and the other group containing three well defined sub-groups with the genotypes isolated from R. pallescens, R. colombiensis and R. ecuadoriensis. Random amplified polymorphic DNA (RAPD) analysis showed the same two main groups and sub-groups supporting strict T. rangeli genotypes' association with Rhodnius species. Combined with other studies, these results suggest a possible co-evolutionary association between T. rangeli genotypes and their vectors.

Susceptibility of different Rhodnius species (Hemiptera, Reduviidae, Triatominae) to a Brazilian strain of Trypanosoma rangeli (SC58/KP1-) / Sensibilidad de diferentes especies de Rhodnius (Hemiptera, Reduviidae, Triatominae) a una cepa brasileña de Trypanosoma rangeli (SC58/KP1-)

Introduction: Specific host-parasite a ssociations have been detected experimentally and suggest that triatomines of the genus Rhodnius act as biological filters in the transmission of Trypanosoma rangeli . Objective: To analyze the susceptibility of four Rhodnius species ( Rhodnius robustus , Rhodnius neglectus , Rhodnius nasutus and Rhodnius pictipes ) to a Brazilian strain of T. rangeli (SC-58/KP1-). Materials and methods: We selected t hirty nymphs of each species, which were fed on blood infected with T. rangeli . Periodically, samples of feces and hemolymph were analyzed. Triatomines with T. rangeli in their hemolymph were fed on mice to check for transmission by bites. Later, the triatomines were dissected to confirm salivary gland infection. Results: Specimens of R. pictipes showed higher rates of intestinal infection compared to the other three species. Epimastigotes and trypomastigotes were detected in hemolymph of four species; however, parasitism was lower in the species of the R. robustus lineage. Rhodnius robustus and R. neglectus specimens did not transmit T. rangeli by bite; after dissection, their glands were not infected. Only one specimen of R. nasutus and two of R. pictipes transmitted the parasite by bite. The rate of salivary gland infection was 16% for R. pictipes and 4% for R. nasutus . Conclusions: Both infectivity (intestinal, hemolymphatic and glandular) and transmission of T. rangeli (SC58/KP1-) were greater and more efficient in R. pictipes. These results reinforce the hypothesis that these triatomines may act as biological filters in the transmission of T. rangeli .

Introducción. Se han detectado asociaciones biológicas huésped-parásito específicas que sugieren que los triatominos del género Rhodnius podrían actuar como filtros biológicos en la transmisión de Trypanosoma rangeli . Objetivo. Estudiar la sensibilidad de cuatro especies de Rhodnius ( Rhodnius robustus , Rhodnius neglectus , Rhodnius nasutus y Rhodnius p ictipes ) frente a la cepa de T. rangeli (SC-58/KP1-). Materiales y métodos. Se seleccionaron treinta ninfas de cada especie después de xenodiagnóstico artificial en sangre infectada con T. rangeli. Se examinaron periódicamente m uestras de heces y hemolinfa. Los insectos con hemolinfas infectadas fueron alimentados en ratones a fin de comprobar la transmisión por picadura y posteriormente disecados para confirmar la infección de las glándulas salivales . Resultados . En Rhodnius pictipes se encontró un mayor porcentaje de infección intestinal que en las otras especies . Se detectaron epimastigotes y tripomastigotes en la hemolinfa de las cuatro especies , y se encontró que el parasitismo fue menor en las especies del linaje R. robustus . Rhodnius robustus y R. neglectus no transmitían T. rangeli a ratones por picadura: después de la disección , sus glándulas no estaban infectadas. Solo un espécimen de R. nasutus y dos de R. pictipes transmitieron el parásito por la picadura . La tasa de infección glandular fue de 16 % para R. pictipes y de 4 % para R. nasutus . Conclusiones . La capacidad infecciosa ( hemolinfática, intestinal y glandular ) y la transmisión de T. rangeli (SC-58/KP1-) fueron mayores y más eficientes en R. pictipes . Estos resultados refuerzan la hipótesis de que estos triatominos actúan como filtros biológicos en la transmisión de T. rangeli .

Adaptabilidad de cepas argentinas de Trypanosoma cruzi en Dipetalogaster maxima (Uhler, 1894) / Adaptability of Argentine strains of Trypanosoma cruzi in Dipetalogaster maxima

Dipetalogaster maxima habita el sur de la península de Baja California, México. La adaptabilidad de las cepas argentinas de Trypanosoma cruzi a vectores propios de otras latitudes podría tener importancia epidemiológica y para su uso en xenodiagnóstico. El objetivo principal del presente estudio fue investigar si existía buena adaptación de cepas de T. cruzi (circulantes en Santiago del Estero, Argentina) en D. maxima, comparativamente con Triatoma infestans. Se utilizaron ninfas I de D. maxima y ninfas III de T. infestans, criadas en laboratorio. Se realizaron diez xenodiagnósticos sobre pacientes chagásicos crónicos no tratados, con serología positiva para cada especie, en paralelo. El peso promedio de sangre ingerida por cada ejemplar fue 61.4 mg para T. infestans y 63.8 mg para D. maxima. A los 30 días, la materia fecal de los insectos fue examinada al microscopio óptico. Se encontró que el 30% de los ejemplares de T. infestans y el 20% de los de D. maxima estaban infectados. Todos los pacientes que mostraron positividad en el xenodiagnóstico con D. maxima también fueron positivos con T. infestans. Comparados los resultados de infectividad, existió significación estadística válida para afirmar que las cepas de T. cruzi argentinas estudiadas se adaptaban a D. maxima. Consideramos importante este primer registro de infectividad de D. maxima con T. cruzi de pacientes chagásicos crónicos de la Argentina debido a la adaptabilidad significativa demostrada, ya que posibilitaría su utilización para xenodiagnóstico. Además, y a pesar del hábitat y la distribución actual de D.maxima, podría tener repercusiones epidemiológicas futuras, como consecuencia de los procesos de globalización y cambios climáticos que se observan en el planeta.

Distribuição espacial de populações de triatomíneos (Hemiptera: Reduviidae) em palmeiras da espécie Mauritia flexuosa no Distrito Federal, Brasil / Spatial distribution of Triatominae populations (Hemiptera: Reduviidae) in Mauritia flexuosa palm trees in Federal District of Brazil

Para determinar e analisar a distribuição espacial de populações silvestres de triatomíneos no Distrito Federal, Brasil, foram amostradas 150 palmeiras da espécie Mauritia flexuosa em seis veredas de diferentes paisagens (silvestre, rural e periurbana) na estação chuvosa. Os triatomíneos foram identificados morfologicamente, separados por sexo e estadio ninfal e examinados para verificar infecção por Trypanosoma cruzi e Trypanosoma rangeli. Vinte e oito (18,6 por cento) palmeiras estavam infestadas por Rhodnius neglectus e 14 (9,5 por cento) por Psammolestes tertius. A freqüência dessas espécies em palmeiras com e sem ninhos foi significativamente diferente, sendo superior nas palmeiras com ninhos de aves e mamíferos na copa. O maior número médio de insetos/palmeira foi observado nas áreas rurais, com estimativas de até 838 insetos/hectare. A composição etária das espécies apresentou um padrão diferente, Rhodnius neglectus com predomínio de ninfas e Psammolestes tertius com predomínio de adultos, sendo que muitos ovos de Rhodnius neglectus foram coletados indicando um evento reprodutivo em fevereiro de 2003. Entre os ninhos encontrados nas palmeiras, o do pássaro Phacellodomus ruber (Furnariidae) apresentou as maiores abundâncias de triatomíneos, ocorrendo em 42 por cento das palmeiras. A abundância relativa de Rhodnius neglectus e Psammolestes tertius foi maior em áreas rurais que possuíram maior número de ninhos nas palmeiras e menor densidade de palmeiras por hectare. Nenhum dos 96 triatomíneos examinados estava infectado por Trypanosoma cruzi ou Trypanosoma rangeli, apesar de encontrarmos infecção de Rhodnius neglectus por esses parasitas em estudos anteriores.

Ultrastructural observations on trypanosoma herpetosoma rangeli in the salivary glands of rhodnius ecuadoriensis, hemiptera, reduvidae

Fue realizado un estudio ultraestructural de las glándulas salivares del triatomino rhodnius ecuadoriensis experimentalmente infectado por el tripanosomatídeo, trypanosoma (herpetosoma) rangeli, con la finalidad de documentar los aspectos básicos de la infección glandular por este flagelado. Gran número de formas epimastigotes fueron encontradas en la hemolinfa que baña la parte externa de las glándulas en el hemoceloma. Algunos flagelados parecían intimamente asociados a la membrana basal del epitelio glandular y varios de ellos parecían estar iniciando el proceso de su invasión, rompiendo la lámina basal con el flagelo. En el interior de las células glandulares observamos que, T. rangeli se presentaba en la forma de esteromastigotes, células redondeadas rodeadas externamente por un flagelo, que envolvía apretadamente el parásito. En la luz glandular también observamos gran número de epimastigotes mezclados con los productos de la secreción glandular, el material presentó apenas pocas formas paramastigotes y trypomastigotes identificadas de manera inquívoca. El significado de los paramastigotes en el ciclo evolutivo del T. rangeli aguarda futuros estudios

Domestic Rhodnius ecuadoriensis (Hemiptera, Reduviidae) infestation in Northern Peru: a comparative trial of detection methods during a six-month follow-up

Two passive methods in the assessment of intradomiciliary infestation by Rhodnius ecuadoriensis were tested: (i) the Gomes Nuñez sensor box (GN), (ii) sheets of white typing paper and (iii) one active timed manual method. The study was carried out in the Alto Chicama River Valley, Province of Gran Chimú, Department of La Libertad. The study design consisted of an initial searching of triatomines inside of the domestic environment by the manual capture active procedure (man/hour) covering all the studied houses. Then, matched pairs of GN boxes and paper sheets were simultaneously installed in the bedrooms of 207 households distributed in 19 localities. A comparative prospective trial of these passive detection devices were monitored at 2, 4 and, finally 6 months follow-up. Parasitological Trypanosoma rangeli and/or T. cruzi infections were investigated in two houses with high level of infestation by R. ecuadoriensis. 16.9 percent of the 207 households investigated by an initial active manual method were infested with R. ecuadoriensis. The proportion of infested houses fluctuated from 6.2 to 55.5 percent amongst the 19 localities investigated. T. rangeli natural infection was detected in R. ecuadoriensis specimens collected in two households. Parasite rates in the bugs ranged from 16.6 to 21.7 percent respectively. The most striking fact was an average rate of salivary gland infection ranging from 7.4 to 8.3 percent. At the end of the sixth month period, a cumulative incidence of 31.4 percent of positive GN boxes against 15.9 percent for paper sheets was recorded. All three methods combined detected domestic infestation in 129 (62.3 percent) of the 207 houses studied in the 19 localities. The range of houses infested varies from 6.7 percent to 92.9 percent. In areas with low bug density infestation rates, the methodology experienced in our studies, seems to be the best choice for investigations on domestic R. ecuadoriensis populations

Clonal variation within a mucosal isolate derived from a patient with Leishmania (Viannia) braziliensis infection

No transcurso de um periodo de 5 anos foram estudados 3 isolados de um paciente com leishmaniose mucosa recidivante causada pela Leishmania (Viannia) braziliensis e 7 clones de um desses isolados. Este estudo foi feito pela analise dos serodemas e zimodemas. Os resultados indicaram a ocorrencia de variacoes fenotipicas clonais. Oito marcadores isoenzimaticos demonstraram diferencas nos padroes eletroforeticos em Acetato de Celulose (AC), bem como em camada fina de amido. Da mesma forma foram constatadas diferencas em um painel de anticorpos monoclonais especificos e subespecificos. Nossas observacoes indicam ainda que a leishmania (Viannia) braziliensis esta composta por subpopulacoes de parasitas com caracteristicas bioquimicas e antigenicas peculiares.

The triatomines of northern Peru, with emphasis on the ecology and infection by Trypanosomes of Rhodnius ecuadoriensis (Triatominae)

Information on the distribution and synanthropic behaviour of triatomines is essential for Chagas disease vector control. This work summarises such information from northern Peru, and presents new data on Rhodnius ecuadoriensis - an important local vector infesting 10-35 percent of dwellings in some zones. Three species are strongly synanthropic and may be suitable targets for chemical control of domestic/peridomestic bug populations. Panstrongylus herreri, the main domestic vector in the area, is probably present in sylvatic ecotopes in the Marañón river system. R. ecuadoriensis and Triatoma dimidiata seem exclusively domestic; biogeographical and ecological data suggest they might have spread in association with humans in northern Peru. Confirmation of this hypothesis would result in a local eradication strategy being recommended. Presence of trypanosome natural infection was assessed in 257 R. ecuadoriensis; Trypanosoma rangeli was detected in 4 percent of bugs. Six further triatomine species are potential disease vectors in the region (T. carrioni, P. chinai, P. rufotuberculatus, P. geniculatus, R. pictipes, and R. robustus), whilst Eratyrus mucronatus, E. cuspidatus, Cavernicola pilosa, Hermanlentia matsunoi, and Belminus peruvianus have little or no epidemiological significance. A strong community-based entomological surveillance system and collaboration with Ecuadorian public health authorities and researchers are recommended

Enzootic transmission of Trypanosoma cruzi and T. rangeli in the Federal District of Brazil

O Distrito Federal (DF) do Brasil está localizado no bioma Cerrado, um complexo de fisionomias savânicas incluindo matas de galeria e campos úmidos permanentes (veredas). Triatomíneos silvestres infectados por Trypanosoma cruzi ocorrem na área, mas a transmissão enzoótica de tripanossomatídeos permanece insuficientemente caracterizada. Um estudo parasitológico envolvendo triatomíneos silvestres (166 Rhodnius neglectus coletados em palmeiras da espécie Mauritia flexuosa) e pequenos mamíferos (98 marsupiais e 70 roedores, totalizando 18 espécies) foi conduzido em 18 áreas, principalmente matas de galeria e veredas. Os parasitas foram isolados, identificados morfologicamente e caracterizados por PCR do DNA do cinetoplasto (kDNA) e núcleo (gene mini-exon). Seis R. neglectus, sete Didelphis albiventris e um Akodon cursor estavam infectados por tripanossomatídeos; a infecção em reservatórios silvestres é documentada pela primeira vez no DF. O PCR do kDNA detectou T. cruzi em cinco R. neglectus e o PCR do gene mini-exon revelou T. cruzi I nos isolados de D. albiventris. Um dos insetos mostrou estar infectado por T. rangeli KP1+. Apesar da ocorrência de D. albiventris (um importante reservatório silvestre e peridoméstico) e R. neglectus (um vetor secundário capaz de invadir domicílios) infectados por T. cruzi, um baixo risco de transmissão da doença de Chagas humana seria esperado no DF, considerando a baixa prevalência da infecção apresentada neste trabalho. A evidência molecular apresentada neste trabalho confirma a circulação de T. rangeli KP1+ com R. neglectus como vetor, amplia a distribuição geográfica deste parasita no Brasil e reforça a hipótese de adaptação de populações de T. rangeli (KP1+ e KP1-) a diferentes linhagens evolutivas de espécies de Rhodnius.

Biogeography of Triatominae (Hemiptera: Reduviidae) in Ecuador: implications for the design of control strategies

Chagas disease control strategies strongly depend on the triatomine vector species involved in Trypanosoma cruzi transmission within each area. Here we report the results of the identification of specimens belonging to various species of Triatominae captured in Ecuador (15 species from 17 provinces) and deposited in the entomological collections of the Catholic University of Ecuador (Quito), Instituto Oswaldo Cruz (Brazil), the Natural History Museum London (UK), the London School of Hygiene and Tropical Medicine (UK), the National Institute of Hygiene (Quito), and the Vozandes Hospital (Quito). A critical review of published information and new field records are presented. We analysed these data in relation to the life zones where triatomines occur (11 life zones, excluding those over 2,200 m altitude), and provide biogeographical maps for each species. These records are discussed in terms of epidemiological significance and design of control strategies. Findings relevant to the control of the main vector species are emphasised. Different lines of evidence suggest that Triatoma dimidiata is not native to Ecuador-Peru, and that synanthropic populations of Rhodnius ecuadoriensis in southern Ecuador-northern Peru might be isolated from their sylvatic conspecifics. Local eradication of T. dimidiata and these R. ecuadoriensis populations might therefore be attainable. However, the presence of a wide variety of native species indicates the necessity for a strong longitudinal surveillance system

Leishmania mexicana: aspectos taxonomicos y rareza de la infeccion humana en Tres Bracos, Bahia, Brasil. / Leishmania mexicana: taxonomic aspects and rarity of human infection in Tres Bracos, Bahia, Brazil

Del estudio de 51 stocks de Leishmania aislados de pacientes humanos de leishmaniasis cutaneomucosa en Tres Bracos, Bahia, Brasil, los autores describen, en detalle, el analisis de los dos unicos stocks de L. mexicana, identificando uno de ellos como L. mexicana amazonensis. El otro aislado permanece en posicion taxonomica no definida pues considerandosele como un miembro de L. mexicana, encuentranse dificultades para su identificacion subespecifica. Evaluan tambien los parametros biologicos e isoenzimaticos y discutem el papel de los anticuerpos monoclonales en la tipificacion de estos stocks. Los autores remarcan la rareza de la transmision de parasitos del complejo L. mexicana en esta region, aun cuando estudien epidemiologicamente por mais de 8 anos la infeccion humana