Trypanosomatids (Protozoa: Kinetoplastida) in three species of Armored Catfish from Mogi-Guaçu river, Pirassununga, São Paulo, Brazil / Tripanossomatídeos (Protozoa: Kinetoplastida) em três espécies de cascudos do rio Mogi-Guaçu, Pirassununga, São Paulo, Brasil

Abstract Trypanosome infections have been reported in several species of fish, in majority of cases described on the basis of morphological characteristics. Trypanosomes in fish are heteroxenous and transmitted by hirudineans. This study aims to evaluate the prevalence and density of infections by Trypanosoma sp. in blood from three species of catfish, Hypostomus regani, H. strigaticeps, H. albopunctatus, from the Mogi Guaçu River, Pirassununga, São Paulo, Brazil. Further, this study intends to characterize the Trypanosoma specimens found in the blood of these fish by morphological and molecular techniques. The trypanosomes overall prevalence observed was 47.6% with a general average density of 0.75 parasites/µl of blood. Hypostomus regani and Hypostomus strigaticeps showed a significant difference in prevalence. The average densities of parasites were not significantly different among the three fish species. Similar findings were observed for the monthly variations in densities. The parasites found in the three species of catfish studied showed similar morphological characteristics. The morphological data and the statistical analyses used in this study didn’t show the formation of groups. The analyses provided evidence of the presence of pleomorphisms in the trypanosomes found in the three studied fish.

Resumo Infecções por tripanossomas foram descritas em diversas espécies de peixes, sendo a maioria das descrições baseada nas características morfológicas. Tripanossomas de peixes são heteroxenos e transmitidos por hirudíneos. Este estudo tem como objetivo a avaliação da prevalência e densidade da infecção por Trypanosoma sp. no sangue de três espécies de cascudos, Hypostomus regani, H.strigaticeps, H.albopunctatus, do Rio Mogi-Guaçu, Pirassununga, São Paulo, Brasil. Além disso, pretende-se a caracterização das espécies de Trypanosoma encontradas no sangue desses peixes, por meio da utilização de técnicas morfólogicas e moleculares. A prevalência total de tripanossomas foi de 47,6%, e a densidade média foi de 0,75 parasitas/ µl de sangue. Hypostomus regani e Hypostomus strigaticeps apresentaram diferenças significativas em prevalência. As médias das densidades dos parasitas não apresentaram diferenças significativas entre as três espécies de peixes estudadas. O mesmo foi observado com a variação mensal das densidades. Os parasitas encontrados nas três espécies de cascudo possuíam características morfológicas semelhantes. As análises morfológicas e estatísticas obtidas neste estudo não mostraram a formação de grupos. As análises evidenciaram a presença de pleomorfismo dos tripanossomas encontrados nas três espécies de peixes estudados.

Trypanosoma evansi is alike to Trypanosoma brucei brucei in the subcellular localisation of glycolytic enzymes

Trypanosoma evansi, which causes surra, is descended from Trypanosoma brucei brucei, which causes nagana. Although both parasites are presumed to be metabolically similar, insufficient knowledge of T. evansi precludes a full comparison. Herein, we provide the first report on the subcellular localisation of the glycolytic enzymes in T. evansi, which is a alike to that of the bloodstream form (BSF) of T. b. brucei: (i) fructose-bisphosphate aldolase, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), hexokinase, phosphofructokinase, glucose-6-phosphate isomerase, phosphoglycerate kinase, triosephosphate isomerase (glycolytic enzymes) and glycerol-3-phosphate dehydrogenase (a glycolysis-auxiliary enzyme) in glycosomes, (ii) enolase, phosphoglycerate mutase, pyruvate kinase (glycolytic enzymes) and a GAPDH isoenzyme in the cytosol, (iii) malate dehydrogenase in cytosol and (iv) glucose-6-phosphate dehydrogenase in both glycosomes and the cytosol. Specific enzymatic activities also suggest that T. evansi is alike to the BSF of T. b. brucei in glycolytic flux, which is much faster than the pentose phosphate pathway flux, and in the involvement of cytosolic GAPDH in the NAD+/NADH balance. These similarities were expected based on the close phylogenetic relationship of both parasites.

Caracterización molecular de Trypanosoma vivax en ovinos naturalmente infectados en dos hatos de los municipios San Fernando y Biruaca estado Apure Venezuela / Molecular characterization of Trypanosoma vivax in naturally-infected sheep from two farms at San Fernando and Biruaca municipalities Apure state Venezuela

Este estudio tuvo como objetivo efectuar una caracterización molecular de Trypanosoma vivax en ovinos de dos hatos en los cuales estos rumiantes, conjuntamente con vacunos y búfalos de agua, comparten la misma área agroecológica, estableciendo el potencial papel de los ovinos como fuente de infección de tripanosomosis por T. vivax para los grandes rumiantes. La técnica de microcentrifugación capilar (TMC) fue usada para establecer el porcentaje de infecciones activas por tripanosomas existente en los animales evaluados. Se empleó un ensayo de reacción en cadena de la polimerasa (PCR) para confirmar la identificación de especie, mientras que un ensayo de PCR-RFLP (polimorfismo en la longitud de los fragmentos de restricción) permitió evaluar la variabilidad intraespecífica entre los aislados de T. vivax detectados en ovinos vs. aquellos provenientes de bóvidos (vacunos y búfalos de agua), colectados en la misma área de producción. De las 320 muestras de sangre de ovinos colectadas, la TMC detectó positividad en 11 (4,35 por ciento), lo que es de gran relevancia epidemiológica debido a la baja sensibilidad de esta metodología. Los resultados de PCR permitieron caracterizar a T. vivax como la especie presente en todas las infecciones activas detectadas. Todos los animales infectados mostraron un valor de hematocrito inferior (P<0,05) al registrado en animales no infectados (22,435 vs. 31,450). El ensayo de PCR-RFLP permitió observar la existencia de perfiles de restricción similares entre los aislados de T. vivax evaluados, sugiriéndose la ausencia de variación intraespecífica para el marcador molecular en estudio, independientemente del origen de hospedador del que provino la muestra (ovinos, vacunos, búfalos de agua). Estos resultados permiten sugerir que los aislados de T. vivax que infectan ovinos, vacunos y búfalos de agua en el área de estudio pudiesen estar estrechamente relacionados desde un punto de vista genético y, consecuentemente...

This study was made to achieve a molecular characterization of Trypanosoma vivax in two Venezuelan farms where both small ruminants (mainly ovines) and bovines (cattle and water buffaloes) share the same agroecological area. In addition, it was made to assess the role of sheep as source of T. vivax infection for cattle and buffalo herds. The microhematocrit centrifugation technique (MHC) was used to establish the percentage of current trypanosome infection. A PCR-based assay was used to confirm the species identification while a PCR-RFLP assay was used for studying intra-specific variation among T. vivax from sheep vs. those from other livestock from the same area. From 320 sheep blood samples, MHC detected 11 (4,35 percent) which is of remarkable epidemiological significance due to the low sensitivity of this method. Based on PCR results, T. vivax was characterized as the only species responsible for all sheep infections. All infected animals showed a lower packed cell volume value (P<0,05) when compared with the non-infected (22,435 vs. 31,450). The PCR-RFLP technique revealed similar profiles among T. vivax isolates suggesting a non intra-specific variation within the molecular marker amplified regardless the host (sheep water buffaloes or cattle). Thus, it was suggested that T. vivax infecting sheep, cattle, and buffaloes in the study area could be genetically closely related. These findings show that sheep may play an important role in the epidemiology of livestock trypanosomiasis in this area and they might be incorporated into therapeutic and preventive programs against livestock trypanosomiasis.

Informações sobre a biologia do conorhinus megitus Burm / Information on the biology of conorhinus megitus Burm

Revisões históricas aos avanços científicos para o controle da doença, o Simpósio Internacional Comemorativo do Centenário da Descoberta da Doença de Chagas (1909-2009).

Nova tripanossomíase humana: estudos sobre a morfologia e o ciclo evolutivo do Schizotrypanum n. gen., n. sp., agente etiológico de nova entidade mórbida do homem / New human trypanosomiasis: studies on the morphology and life cycle of Schizotrypanum No gen., n. sp. etiological agent of a new morbid entity of man

Revisões históricas aos avanços científicos para o controle da doença, o Simpósio Internacional Comemorativo do Centenário da Descoberta da Doença de Chagas (1909-2009).

Bovine Trypanosomosis in three districts of East Gojjam Zone bordering the Blue Nile River in Ethiopia

Background: Bovine trypanosomosis is a serious constraint to agricultural production in extensive areas of Ethiopia. Methodology: A cross-sectional study was conducted to determine the prevalence of bovine infection with trypanosomes and to identify the prevailed trypanosome species in three districts of the East Gojjam zone bordering the Blue Nile River from March 2005 to February 2006. Cattle from 9 different localities were checked using microscopical examination of wet blood smears; thin and stained bloodsmears; and by blood centrifugation followed by the examination of the resultant buffy coats. Result: Of the total 3;360 cattle investigated; 8.2(3.5; 11.6and 9.4from Dejen; Machakel and Baso-Liben districts respectively) were found to be infected with trypanosomes. Of the total 275 positive animals; 249 (90.5) appeared to be infected with Trypanosoma vivax; 11 (4) were infected with T. congolense; and 15 (5.5) were infected with mixed infection of T. vivax and T. congolense. The prevalence of infection with T. vivax was significantly higher than that of T. congolense (P 0.001). Taking 24-46as normal PCV value; the mean PCV for the trypanosome-infected cattle (22.09) was lower than those for the trypanosome-negative animals (26.03). Conclusion: Trypanosomosis is a disease of considerable importance to the major economic districts bordering the Blue Nile River of the East Gojjam zone; Ethiopia; given the disease's potential to threaten the health and productivity of cattle in this region

Predominance of Trypanosoma rangeli infection in children from a Chagas disease endemic area in the west-shore of the Panama canal

A total of 206 serum samples from children (3-14 years old) living in the Amador County (La Chorrera District, Province of Panama) were screened by indirect immunofluorescence antibody test (IFAT) for the presence of antibodies against Trypanosoma cruzi. Positive sera were confirmed by recombinant enzyme-linked immunosorbent assay (ELISA) and Western blot analysis. The presence of blood trypanosomes was investigated by hemoculture and subsequently identify by a duplex polymerase chain reaction (PCR) followed by dot blot hybridization. The results indicated a prevalence of 9.7 percent for trypanosome infections, a seroprevalence of 2.9 percent against T. cruzi and a predominance of T. rangeli infection (6.8 percent). The immunological and clinical implications of these findings are discussed.

Trypanosoma evansi isolated from capybara (Hidrochaeris hidrochaeris)

A study was conducted to determine the morphological and biometric characteristics of Trypanosoma isolated from 50 capybaras animals, raised in captivity in the Peruvian Amazon. Trypanosoma was found in 14 blood samples using the microhaematocrit, wide drop, and Giemsa-stain methods and T. evansi was identified through morphological details in all 14 positive samples (the subterminal kinetoplast, the developed undulating membrane, and a long free flagellum were used for the identification of the agent)

Specific antibody levels and antigenic recognition of Wistar rats inoculated with distinct isolates of Trypanosoma evansi

"Mal de Cadeiras", an enzootic disease caused by Trypanosoma evansi, is one of the most important trypanosomiases in the Brazilian Pantanal region. The disease affects mainly horses, which are widely used in extensive cattle production, an activity of greatest economical significance for the region. The parasite also infects sylvan (coatis and capybaras) and domestic (dogs) animals, respectively considered wild and domestic reservoirs of T. evansi. For a better understanding of the interaction of T. evansi with its rodent host, we evaluated the differences in the specific antibody level patterns and in the parasitic peptides recognition patterns of experimentally infected Wistar rats. The rats experimentally infected with T. evansi isolates obtained from coatis, dogs and horses were submitted to indirect immunofluorescence test (IgM e IgG) and Western blotting. The serological titers for IgM and IgG ranged between 1:40 and 1:160. The most recognized polypeptide profiles were in a range of 17 and 74 kDa. Our data suggest that the humoral immune response in Wistar rats is not sufficient for granting an effective control of T. evansi infections

Metalloproteases in Trypanosoma rangeli-infected Rhodnius prolixus

Protease activities in the haemolymph and fat body in a bloodsucking insect, Rhodnius prolixus, infected with Trypanosoma rangeli, were investigated. After SDS-polyacrylamide gel electrophoresis containing gelatin as substrate, analysis of zymograms performed on samples of different tissues of controls and insects inoculated or orally infected with short or long epimastigotes of T. rangeli, demonstrated distinct patterns of protease activities: (i) proteases were detected in the haemolymph of insects which were fed on, or inoculated with, short epimastigotes of T. rangeli (39 kDa and 33 kDa, respectively), but they were not observed in the fat body taken from these insects; (ii) protease was also presented in the fat bodies derived from naive insects or controls inoculated with sterile phosphate-saline buffer (49 kDa), but it was not detected in the haemolymph of these insects; (iii) no protease activity was observed in both haemolymph and fat bodies taken from insects inoculated with, or fed on, long epimastigotes of T. rangeli. Furthermore, in short epimastigotes of T. rangeli extracts, three bands of the protease activities with apparent molecular weights of 297, 198 and 95 kDa were detected while long epimastigotes preparation presented only two bands of protease activities with molecular weights of 297 and 198 kDa. The proteases from the insect infected with T. rangeli and controls belong to the class of either metalloproteases or metal-activated enzymes since they are inhibited by 1,10-phenanthroline. The significance of these proteases in the insects infected with short epimastigotes of T. rangeli is discussed in relation to the success of the establishment of infection of these parasites in its vector, R. prolixus

Revisión de los aspectos biológicos y diagnósticos del Trypanosoma (Herpetosoma) rangeli / Review of the biologic and diagnostic aspects of Trypanosoma (Herpetosoma) rangeli

Esta revisión tiene tres objetivos básicos: a) estimular aún más la investigación de esta prevalente infección humana. b) examinar el arsenal de técnicas diagnósticas disponíbles al momento y, las nuevas pruebas descritas recientemente. c) enfatizar el significado que tiene, el parasitismo por el Trypanosoma (Herpetosoma) rangeli, en las áreas endémicas de la Enfermedad de Chagas distribuidas en las Américas Central y del Sur. Trypanosoma rangeli y Trypanosoma cruzi son parásitos que circulan superponiéndose en muchas áreas de Latinoamérica utilizando prácticamente los mismos triatominos vectores. Una vasta gama de especies de mamíferos han sido encontradas infectadas naturalmente con T. rangeli en diversos países. Se revisa la biología del parasitismo y el ciclo biológico del tripanosoma haciendo énfasis en este último. Infecciones crónicas por T. rangeli en el hombre pueden, serológicamente, ser confundidas con las del T. cruzi. Ambas especies presentan antígenos comunes que provocan las conocidas reacciones serológicas cruzadas. Desafortunadamente, no conocemos la real distribución de las infecciones por el T. rangeli en la mayoría de las áreas mencionadas. Nuevos estudios epidemiológicos son necesarios, para examinar el problema de las infecciones humanas mixtas, por estos tripanosomas.

This review has three objectives: a) To stimulate further research of this prevalent human infection b) to examine the progress of current diagnostic techniques and c) to emphasise the significance of the flagellate parasite Trypanosoma (Herpetosoma) rangeli in Chagas' Disease endemic areas of South and Central America. Both Trypanosoma rangeli and Trypanosoma cruzi overlap in many of the areas of Latin America utilising the same triatomine vectors. Also a vast range of mammalian species have been found naturally infected with T. rangeli. The biology of the parasitism of T. rangeli is revised and emphasis is given regarding its biological cycle. T. cruzi and T. rangeli share common antigens and cross react serologically. Human infection in the chronic phase may be misdiagnosed as T. cruzi infection. Conventional and modern diagnostic and identification methods are discussed. Unfortunately we do not know the real distribution of T. rangeli infections in most areas and epidemiological studies to examine concomitant dual infections deserve further investigation.

Trypanosoma (Schizotrypanum) cruzi: histopathology in mice infected with strains isolated from Didelphis marsupialis from the Valley of Caracas (Venezuela)

The histopathological alterations produced in NMRI strain mice by isolates of Trypanosoma cruzi from Didelphis marsupialis captured near human dwellings in the valley of Caracas, Venezuela are described. The donor opossums showed pseudocysts and amastigotes and trypomastigotes only in the heart muscle, and few areas of discrete inflammations and lysis of some muscle cells. Mice were parasitized in the heart, skeletal muscle, jejunum, colon, liver, lung, urinary bladder, penis, seminal vesicle, prostate, pancreas, and brain. All the isolates produced histiolymphocyticinflammation, severe in cardiac and skeletal muscle, and light in the smooth muscle of the intestine and urinary bladder; certain of the isolates produced destruction of muscle fibers. These findings, together with the morphology and behavior reported in previous papers, suggest that the isolates from this mammal reservoir and from the local vector Panstrongylus geniculatus) belong to the same type. The possible venereal transmission through the parasitosis of the urogenital system is discussed. The necessity for characterization of strains of the parasite that have been isolated from areas of intense urbanization, where the ecological changes have reduced the number of host species, is emphasized; such studies may help to clarify the extreme heterogeneity of T. cruzi and the parasitoses it induces

A simplified and highly sensitive detection of Trypanosoma evansi by DNA amplification.

In Southeast Asia Trypanosoma evansi infection is a disease of economic importance since it affects the health of buffalo, cattle and swine. The acute stage symptoms include abortion, central nervous system disorder and even death, and in the chronic condition working capacity and productivity of the animals are affected. A polymerase chain reaction (PCR)-based detection technique has been developed with a sensitivity of 0.5 pg of parasite DNA or one single parasite in 10 microliters of blood samples which were allowed to clot and then boiled before DNA amplification. This permitted storage of blood collection at ambient temperature for at least one month. Phosphate-saline-glucose solution, normally used as trypanosome maintenance buffer, inhibited PCR. Although DNA primers used were derived from T. evansi specific sequence, amplification of the genome of T. brucei and T. equiperdum generated the same 227 bp fragment. This method should now make it possible to detect infections in livestock in the very early stages where microscope examination is equivocal and to monitor groups of animals after trypanocidal treatment.

Didelphis marsupialis: reservorio primario de tripanosomiasis americana urbana / Didelphis marsupialis: primary reservoirs of american urban trypanosomiasis

Se realizó un estudio sistemático utilizando ratones albinos de las características morfobiológicas de cepas de T. Cruzi aisladas de P. Geniculatus, adultos capturados en el interior de viviendas humanas de Caracas. Las cepas aisladas de estos reservorios invadieron en ratones albinos a los tejidos y visceras parasitadas por los aislados de P. Geniculatus

Estudo comparado do ciclo evolutivo de triatomíneos a duas temperaturas / Comparison of life cycles of triatominae according to changes in temperature

We studied the life cicle of several triatominae species: Dipetalogaster maximus (Uhler, 1894); Panstrongylus herreri Wygodzinsky, 1948; Panstrongylus megistus (Burmeister, 1835); Rhodnius ecuadoriensis Lent & Leon, 1958; Rhodnius nasutus Stal, 1859; Rhodnius neglectus Lent, 1954; Rhodnius pictipes Stal, 1872; Rhodnius prolixus Stal, 1859; Rhodnius robustus Larrousse, 1927; Triatoma brasiliensis Neiva, 1911; Triatoma infestans (Klug, 1834); Triatoma maculata (Erichson, 1848); Triatoma matogrossensis Leite & Barbosa, 1953; Triatoma platensis Neiva, 1913; Triatoma protracta (Uhler, 1894); Triatoma sordida (Stal, 1859); Triatoma tibiamaculata (Pinto, 1926) e Triatoma vitticeps (Stal, 1859) (Hemiptera, Reduviidae). The main purpose of the study was to obtain information to improve control measures specially in those peridomiciliar species. The experiments were performed in two climatized chambers, both with an humidity of 70 ± 5% and photoperiod of 12 hours. One was maintained at 25 ± 0,5ºC and the other at 30 ± 1ºC.

Trypanosoma (Herpetosoma) rangeli tejera, 1920: influence of host weight, size of inoculum and route of infection upon experimental parasitemia

Para estudiar la influecia de la edad del hospedador, del tamaño del inóculo y su ruta de adminsitración sobre la infección por Trypanosoma (Herpetosoma) rangeli, fueron inoculados i.p. 12 lotes de ratones albinos (cepa NMRI) de 6.0 g de peso con 25.0 - 2.5x10**6 metatripomastigotes/g obtenidos del medio LIY. Los inóculos mas bajos produjeron parasitemias bajas pero persistentes; los inóculos mas elevados originaron níveles altos de parasitemias que cayeron prontamente, sugiriendo la mobilización de mecanismos de resistencia a níveles diferentes de actividad. En otros experimentos, la inoculación i.p. y el usodeem%de em de ratones de 6.0 g dieron parasitemias mas elevadas que cuando se usaron inoculaciones s.c. o animales de 16.0 y 26.0 g. Estos resultados indican la necesidad deemplear una metodologúa uniforme cuando se investigue el posible carácter heterogénico del complejo de T. rangeli

Comportamiento de la infección y diferenciación morfologica entre trypanosoma cruzi y T. rangeli en el intestino del vector Rhodnius prolixus / Course of the infection and morphological differentiation between trypanosoma cruzi and T. rangeli in the intestine of the vector Rhodnius prolixus

A distribuiçäo de amastigotas, epimastigotas, esferomastigotas e tripomastigotas foi estudada no intestino de Rhodnius prolixus infectado com Trypanosoma cruzi (amostra Molino) e T. rangeli (amostra San Agustín). Medidas foram feitas de 500 cinetoplastos de organismos morfologicamente diferentes para verificar se é possível identificar as espécies usando esse caracter. A maioria dos tripomastigotas de cada espécie foi diferenciada pelo comprimento do corpo e o tamanho do cinetoplasto. Porém, em algumas infeccöes por T. rangeli, foi observado que menos de 1% dos epimastigotas curtos eram morfologicamente idênticos às mesmas formas de T. cruzi. Nesses casos o T. rangeli pode ser confundidos com T. cruzi, e que poderia ter graves implicaçöes

Trypanosoma (herpetosoma) rangeli Tejera, 1920: the course of infection in immunosuppressed mice

Las infecciones experimentales por Trypanosoma (Herpetosoma) rangeli Tejera, 1920 muestran típicamente un repentino aumento de la parasitemia la cual, igualmente, declina y desaparece de manera abrupta e irregular. La droga cyclofosfamida fue usada para inmunosuprimir ratones albinos de 6, 16 y 26g infectados i.p. con T. rangeli de cultivo LIT. Las curvas iniciales de parasitemia fueron similares en los animales tratados y controles, pero la droga extendió el período de parasitemia patente y la repetición del tratamiento reactivó la parasitemia críptica,. Por cuanto ha sido demostrado que T. rangeli realiza un ciclo intracelular mediante multiplicación de amastigotes, es sugerido que el aumento de la parasitemia y el mantenimiento de infecciones crípticas son debidos al ciclo tisular del parásito en el cual tripomastigotes "jóvenes" reinvaden los tejidos mientras que las formas "adultas", incapaces de penetrar, predominan en sangre durante los niveles más elevados de la parasitemia de donde pueden ser tomados por el insecto vector o eliminados más tarde por la respuesta inmune adquirida del hospedador; supresión mediada por drogas de esta respuesta podría prolongar la parasitemia