Biological aspects of the Trypanosoma cruzi (Dm28c clone) intermediate form, between epimastigote and trypomastigote, obtained in modified liver infusion tryptose (LIT) medium

Chagas disease, caused by the protozoan Trypanosoma cruzi, involves immunomediated processes. Canova (CA) is a homeopathic treatment indicated in the diseases in which the immune system is depressed. This study evaluated the Random Amplification of Polymorphic DNA (RAPD) profile of T. cruzi under the influence of CA and Benznidazole (BZ). Mice infected with the genetic lineage of T. cruzi II (Y strain) were divided into 4 groups:Infected animals treated with saline solution (control group); treated with CA; treated with BZ; treated with CA and BZ combined.Treatment was given at the 5th–25th days of infection (D5–25). The parasites were isolated by haemoculture in Liver Infusion Tryptose (LIT) medium: at D5 (before treatment), D13, 15 and 25 (during treatment) and D55 and 295 (after treatment). DNA was extracted from the mass of parasites. RAPD was done with the primers λgt11-F, M13F-40 and L15996, the amplified products were eletrophoresed through a 4% polyacrylamide gel. Data were analyzed by the coefficient of similarity using the DNA-POP program.163 markers were identified, 5 of them monomorphic. CA did not act against the parasites when used alone. The RAPD profiles of parasites treated with BZ and CA + BZ were different from those in the control group and in the group treated with CA. The actions of the CA and BZ were different and the action of BZ was different from the action of CA + BZ. These data suggest that CA may interact with BZ. The differences in the RAPD profile of the Y strain of T. cruzi produced by BZ, CA + BZ and the natural course of the infection suggest selection/suppression of populations.

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Role for a P-type H+-ATPase in the acidification of the endocytic pathway of Trypanosoma cruzi

Biochemical Journal ◽

10.1042/bj20051319 ◽

2005 ◽

Vol 392 (3) ◽

pp. 467-474 ◽

Cited By ~ 33

Author(s):

Mauricio Vieira ◽

Peter Rohloff ◽

Shuhong Luo ◽

Narcisa L. Cunha-E-Silva ◽

Wanderley De Souza ◽

...

Keyword(s):

Plasma Membrane ◽

Trypanosoma Cruzi ◽

Subcellular Fractionation ◽

Etiologic Agent ◽

Proton Pumping ◽

Endocytic Pathway ◽

Immunogold Electron Microscopy ◽

Intermediate Form ◽

Confocal Immunofluorescence Microscopy ◽

P Type

Previous studies in Trypanosoma cruzi, the etiologic agent of Chagas disease, have resulted in the cloning and sequencing of a pair of tandemly linked genes (TcHA1 and TcHA2) that encode P (phospho-intermediate form)-type H+-ATPases with homology to fungal and plant proton-pumping ATPases. In the present study, we demonstrate that these pumps are present in the plasma membrane and intracellular compartments of three different stages of T. cruzi. The main intracellular compartment containing these ATPases in epimastigotes was identified as the reservosome. This identification was achieved by immunofluorescence assays and immunoelectron microscopy showing their co-localization with cruzipain, and by subcellular fractionation and detection of their activity. ATP-dependent proton transport by isolated reservosomes was sensitive to vanadate and insensitive to bafilomycin A1, which is in agreement with the localization of P-type H+-ATPases in these organelles. Analysis by confocal immunofluorescence microscopy revealed that epitope–tagged TcHA1-Ty1 and TcHA2-Ty1 gene products are localized in the reservosomes, whereas the TcHA1-Ty1 gene product is additionally present in the plasma membrane. Immunogold electron microscopy showed the presence of the H+-ATPases in other compartments of the endocytic pathway such as the cytostome and endosomal vesicles, suggesting that in contrast with most cells investigated until now, the endocytic pathway of T. cruzi is acidified by a P-type H+-ATPase.

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Trypanosoma cruzi: Correlations of Biological Aspects of the Life Cycle in Mice and Triatomines

Memórias do Instituto Oswaldo Cruz ◽

10.1590/s0074-02761999000300021 ◽

1999 ◽

Vol 94 (3) ◽

pp. 397-402 ◽

Cited By ~ 8

Author(s):

Valdirene S Lima ◽

Regina HR Mangia ◽

João C Carreira ◽

Renato S Marchevsky ◽

Ana M Jansen

Keyword(s):

Life Cycle ◽

Trypanosoma Cruzi ◽

Biological Aspects

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Semisolid liver infusion tryptose supplemented with human urine allows growth and isolation of Trypanosoma cruzi and Trypanosoma rangeli clonal lineages

Revista da Sociedade Brasileira de Medicina Tropical ◽

10.1590/0037-8682-0190-2015 ◽

2016 ◽

Vol 49 (3) ◽

pp. 369-372 ◽

Cited By ~ 2

Author(s):

Emanuella Francisco Fajardo ◽

Marlene Cabrine-Santos ◽

Keila Adriana Magalhães Ferreira ◽

Eliane Lages-Silva ◽

Luis Eduardo Ramírez ◽

...

Keyword(s):

Trypanosoma Cruzi ◽

Human Urine ◽

Trypanosoma Rangeli ◽

Liver Infusion Tryptose

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Biological aspects of the DM28C clone of Trypanosoma cruzi after metacylogenesis in chemically defined media

Memórias do Instituto Oswaldo Cruz ◽

10.1590/s0074-02761988000100016 ◽

1988 ◽

Vol 83 (1) ◽

pp. 123-133 ◽

Cited By ~ 149

Author(s):

Victor T. Contreras ◽

Tania C. de Araújo-Jorge ◽

Myrna C. Bonaldo ◽

Neide Thomaz ◽

Helena S. Barbosa ◽

...

Keyword(s):

Trypanosoma Cruzi ◽

Biological Properties ◽

Biological Characterization ◽

Phagocytic Cells ◽

Biological Behaviour ◽

Metacyclic Trypomastigotes ◽

Bona Fide ◽

Biological Aspects

The biological characterization of the Trypanosoma cruzi clone Dm 28c in terms of its growth in LIT medium, cell-cycle, infectivity to mice and interaction with professional and non-professional phagocytic cells shows that it behaves as a bona fide T. cruzi representant. The biological properties of this myotropic clone do not change according to the origin of the trypomastigote forms (i. e., from triatomines, infected mice, cell-culture or from the chemically defined TAUP and TAU3AAG media). In addition Dm 28c metacyclic trypomastigotes from TAU3AAG medium display a high infectivity level to fibroblasts and muscle cells. Experiments on binding of cationized ferritin to trypomastigotes surface show the existence of cap-like structures of ferritin in regions near the kinetoplast. However the nature and role of these anionic sites remain to be determined. The results indicate that metacyclic trypomastigotes from Dm 28c clone obtained under chemically defined conditions reproduce the biological behaviour of T. cruzi, rendering this system very suitable for the study of cell-parasite interactions and for the isolation of trypanosome relevant macromolecules.

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Antibody dependent cell-mediated cytotoxicity against Trypanosoma cruzi

Parasitology ◽

10.1017/s0031182000051866 ◽

1977 ◽

Vol 75 (3) ◽

pp. 317-323 ◽

Cited By ~ 22

Author(s):

I. A. Abrahamsohn ◽

W. D. Da Silva

Keyword(s):

Trypanosoma Cruzi ◽

Normal Mouse ◽

Defined Medium ◽

Lag Phase ◽

Percentage Reduction ◽

Splenic Lymphocytes ◽

Dependent Cell ◽

Liver Infusion Tryptose ◽

Per Se

SummaryThis paper describes in vitro antibody dependent cytotoxicity against Trypanosoma cruzi epimastigotes by normal mouse splenic lymphocytes. Cytotoxicity was expressed as the percentage reduction in the number of motile parasites upon incubation with lymphocytes at 37 °C in a denned medium. Failure of the non-motile parasites to regain motility and their ensuring degeneration at 28 °C in liver infusion tryptose (LIT) medium confirmed loss of motility as a criterion of cytotoxicity. Incubation of T. cruzi at 37 °C for 18 h in a defined medium per se did not interfere with motility but was followed by a lag phase of the growth curve in LIT medium at 28 °C. The lag phase was prolonged for T. cruzi which had previously been incubated at 37 °C in the absence of cells.

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New achievements on biological aspects of copper complexes Casiopeínas®: Interaction with DNA and proteins and anti-Trypanosoma cruzi activity

Journal of Inorganic Biochemistry ◽

10.1016/j.jinorgbio.2012.01.010 ◽

2012 ◽

Vol 109 ◽

pp. 49-56 ◽

Cited By ~ 35

Author(s):

Lorena Becco ◽

Alejandra Rodríguez ◽

María Elena Bravo ◽

María José Prieto ◽

Lena Ruiz-Azuara ◽

...

Keyword(s):

Trypanosoma Cruzi ◽

Copper Complexes ◽

Biological Aspects

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Sensibilidad in vitro a benznidazol, nifurtimox y posaconazol de cepas de Trypanosoma cruzi de Paraguay

Biomédica ◽

10.7705/biomedica.5187 ◽

2020 ◽

Vol 40 (4) ◽

pp. 749-763

Author(s):

Nidia Acosta ◽

Gloria Yaluff ◽

Elsa López ◽

Christopher Bobadilla ◽

Analía Ramírez ◽

...

Keyword(s):

Trypanosoma Cruzi ◽

Liver Infusion Tryptose

Introducción. Trypanosoma cruzi, agente causal de la enfermedad de Chagas, exhibe una sustancial heterogeneidad fenotípica y genotípica que puede influir en las variaciones epidemiológicas y clínicas de la enfermedad, así como en la sensibilidad a los fármacos utilizados en el tratamiento.Objetivo. Evaluar la sensibilidad in vitro al benznidazol, el nifurtimox y el posaconazol de 40 cepas clonadas de T. cruzi de Paraguay, con distintos genotipos, huéspedes y localidades de origen.Materiales y métodos. En su estado epimastigote, los parásitos se incubaron en medio de cultivo LIT (Liver Infusion Tryptose) con diferentes concentraciones de cada fármaco en ensayos por triplicado. El grado de sensibilidad se estimó a partir de las concentraciones inhibitorias del 50 y el 90% (IC50 e IC90) y se obtuvieron los valores promedio y la desviación estándar de cada cepa y fármaco. La significación estadística entre grupos se determinó mediante análisis de varianzas con el test no paramétrico de Wilcoxon/Kruskal-Wallis y valores de p<0,05.Resultados. Se observó un amplio rango de respuesta a los fármacos. Se identificaron dos grupos de parásitos (A y B) con diferencias significativas en la sensibilidad al benznidazol (p<0,0001), y tres grupos (A, B, C) en cuanto a la sensibilidad al nifurtimox y el posaconazol (p<0,0001).Conclusiones. En general, las cepas fueron más sensibles al nifurtimox que al benznidazol y el posaconazol. Estas diferencias evidencian la heterogeneidad de las poblaciones de T. cruzi que circulan en Paraguay, lo que debe considerarse en el tratamiento y el seguimiento de las personas afectadas.

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Protocol optimization for the detection of Trypanosoma cruzi DNA in açai (Euterpe oleraceae) pulp

Acta Amazonica ◽

10.1590/1809-4392201903426 ◽

2021 ◽

Vol 51 (1) ◽

pp. 79-84

Author(s):

Gabrielle Virgínia Ferreira CARDOSO ◽

Andrey Carlos do Sacramento de OLIVEIRA ◽

Andréia Silva da SILVA ◽

Marcos Clécio de Lemos SILVA ◽

Joelson Sousa LIMA ◽

...

Keyword(s):

Trypanosoma Cruzi ◽

Detection Threshold ◽

Chain Reaction ◽

Pcr Assay ◽

Accurate Identification ◽

Oral Transmission ◽

Protocol Optimization ◽

Optimized Protocol ◽

Liver Infusion Tryptose ◽

Polymerase Chain

ABSTRACT Chagas disease, caused by the protozoan Trypanosoma cruzi, has often been linked to oral transmission through açai consumption. Molecular methods that allow fast and accurate identification of the pathogen are important for the detection of the presence of the parasite in this food. This study aimed to optimize polymerase chain reaction (PCR)-based detection of T. cruzi DNA in açai pulp. Several dilutions of T. cruzi DTU TcI trypomastigote forms were cultured in liver infusion tryptose (LIT) medium. Trypanosoma cruzi DNA was extracted from the cells and subjected to PCR. Subsequently, culture dilutions were added to açai pulp to evaluate the detection threshold of the optimized PCR assay. We demonstrate that our assay can detect T. cruzi DNA in açai pulp at a concentration of 1.08 × 10-10 ng µL-1. We conclude that our optimized protocol is effective and can be used as an important tool for the detection of T. cruzi contamination in açaí.

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Hemoculture and Polymerase Chain Reaction Using Primers TCZ1/TCZ2 for the Diagnosis of Canine and Feline Trypanosomiasis

ISRN Veterinary Science ◽

10.5402/2012/419378 ◽

2012 ◽

Vol 2012 ◽

pp. 1-6 ◽

Cited By ~ 3

Author(s):

Luciano José Eloy ◽

Simone Baldini Lucheis

Keyword(s):

Polymerase Chain Reaction ◽

Trypanosoma Cruzi ◽

Chain Reaction ◽

Blood Samples ◽

Transmission Cycle ◽

Epidemiologic Surveillance ◽

Parasite Detection ◽

Liver Infusion Tryptose ◽

Polymerase Chain

Introduction. American trypanosomiasis, also known as Chagas disease, is a zoonosis caused by Trypanosoma cruzi (T. cruzi). Dogs and cats participate actively in this parasite's transmission cycle. This study aimed at evaluating the occurrence of T. cruzi in dogs and cats from Botucatu, SP, Brazil, as well as at evaluating the technique of hemoculture in LIT (liver infusion tryptose) medium by polymerase chain reaction (PCR). Methods. Blood samples were collected from 50 dogs and 50 cats in Botucatu-SP, Brazil. For hemoculture, the samples were inoculated in LIT medium, and readings were performed for four months. Upon completion of such period, all the hemocultures were processed for parasitic DNA extraction. The PCR reactions were performed by using primers TCZ1/TCZ2. Results. Ten dogs and ten cats (20%) were positive to PCR, and four dogs and three cats (7%) were positive to hemoculture. Only in a one cat sample (1%) there was confirmation of positive hemoculture by PCR for T. cruzi. Conclusions. Results showed that PCR was a suitable tool for the confirmation of the parasite detection in hemoculture samples, and that dogs and cats from Botucatu, SP, Brazil, are maintaining the role of household reservoirs of T. cruzi, which reinforces the need for constant epidemiologic surveillance for this zoonosis.

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