ABSTRACT
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.
Subject(s)
Acanthamoeba/genetics , Acanthamoeba/pathogenicity , Amebiasis/parasitology , Keratitis/parasitology , Acanthamoeba/classification , Acanthamoeba/physiology , Acanthamoeba Keratitis/parasitology , Amebiasis/pathology , Animals , Brain/parasitology , Brazil , Female , Genotype , Humans , Keratitis/pathology , Male , Mice , Trophozoites/physiology , VirulenceABSTRACT
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 .
Subject(s)
Rhodnius/parasitology , Trypanosoma rangeli/physiology , Animals , Brazil , Female , Host-Parasite Interactions , Male , MiceABSTRACT
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 .
Subject(s)
Animals , Female , Male , Mice , Rhodnius/parasitology , Trypanosoma rangeli/physiology , Brazil , Host-Parasite InteractionsABSTRACT
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.
Subject(s)
Humans , Animals , Insect Vectors , Marsupialia , Rhodnius , Rodentia , Trypanosoma , Trypanosomiasis , Brazil , Chagas Disease , Disease Reservoirs , DNA, Kinetoplast , DNA, Protozoan , Polymerase Chain Reaction , Prevalence , Trypanosoma , Trypanosoma cruzi , TrypanosomiasisABSTRACT
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.
Subject(s)
Arecaceae/parasitology , Insect Vectors/classification , Triatominae/classification , Animals , Brazil , Female , Insect Vectors/physiology , Male , Population Density , Seasons , Triatominae/physiologyABSTRACT
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.
Subject(s)
Animals , Female , Male , Insect Vectors , Triatominae , Brazil , Insect Vectors , Population Density , Population Dynamics , Rural Population , Seasons , Triatominae , Urban PopulationABSTRACT
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.
Subject(s)
Insect Vectors/classification , Marsupialia/parasitology , Rhodnius/parasitology , Rodentia/parasitology , Trypanosoma/isolation & purification , Trypanosomiasis/veterinary , Animals , Brazil/epidemiology , Chagas Disease/epidemiology , Chagas Disease/transmission , DNA, Kinetoplast/analysis , DNA, Protozoan/analysis , Disease Reservoirs , Humans , Polymerase Chain Reaction , Prevalence , Trypanosoma/classification , Trypanosoma/genetics , Trypanosoma cruzi/isolation & purification , Trypanosomiasis/epidemiology , Trypanosomiasis/transmissionABSTRACT
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.
Subject(s)
Entomology/instrumentation , Insect Control/instrumentation , Insect Vectors , Rhodnius , Animals , Female , Housing , Humans , Male , Matched-Pair Analysis , Peru , Population DensityABSTRACT
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
Subject(s)
Animals , Humans , Male , Female , Entomology , Insect Control , Insect Vectors , Rhodnius , Housing , Matched-Pair Analysis , Peru , Population DensityABSTRACT
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.
