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1. A Novel Genotype and First Record of Trypanosoma lainsoni in Argentina

Author: Anahí G. Díaz, Paula G. Ragone, Fanny Rusman, Noelia Floridia-Yapur, Rubén M. Barquez, M. Mónica Díaz, Nicolás Tomasini, and Patricio Diosque
Subjects: Trypanosoma lainsoni, Leopardus geoffroyi, Calomys spp., 18S rDNA genes, gGAPDH genes, morphology, Medicine
Abstract: Trypanosomes are a group of parasitic flagellates with medical and veterinary importance. Despite many species having been described in this genus, little is known about many of them. Here, we report a genetic and morphological characterization of trypanosomatids isolated from wild mammals from the Argentine Chaco region. Parasites were morphologically and ultrastructurally characterized by light microscopy and transmission electron microscopy. Additionally, 18s rRNA and gGAPDH genes were sequenced and analyzed using maximum likelihood and Bayesian inference. Morphological characterization showed clear characteristics associated with the Trypanosoma genus. The genetic characterization demonstrates that the studied isolates have identical sequences and a pairwise identity of 99% with Trypanosoma lainsoni, which belongs to the clade of lizards and snakes/rodents and marsupials. To date, this species had only been found in the Amazon region. Our finding represents the second report of T. lainsoni and the first record for the Chaco region. Furthermore, we ultrastructurally described for the first time the species. Finally, the host range of T. lainsoni was expanded (Leopardus geoffroyi, Carenivora, Felidae; and Calomys sp., Rodentia, Cricetidae), showing a wide host range for this species.
Published: 2020
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2. Evidence of hybridization, mitochondrial introgression and biparental inheritance of the kDNA minicircles in Trypanosoma cruzi I.

Author: Fanny Rusman, Noelia Floridia-Yapur, Paula G Ragone, Patricio Diosque, and Nicolás Tomasini
Subjects: Arctic medicine. Tropical medicine, RC955-962, Public aspects of medicine, RA1-1270
Abstract: BACKGROUND:Genetic exchange in Trypanosoma cruzi is controversial not only in relation to its frequency, but also to its mechanism. Parasexual genetic exchange has been proposed based on laboratory hybrids, but population genomics strongly suggests meiosis in T. cruzi. In addition, mitochondrial introgression has been reported several times in natural isolates although its mechanism is not fully understood yet. Moreover, hybrid T. cruzi DTUs (TcV and TcVI) have inherited at least part of the kinetoplastic DNA (kDNA = mitochondrial DNA) from both parents. METHODOLOGY/PRINCIPAL FINDINGS:In order to address such topics, we sequenced and analyzed fourteen nuclear DNA fragments and three kDNA maxicircle genes in three TcI stocks which are natural clones potentially involved in events of genetic exchange. We also deep-sequenced (a total of 6,146,686 paired-end reads) the minicircle hypervariable region (mHVR) of the kDNA in such three strains. In addition, we analyzed the DNA content by flow cytometry to address cell ploidy. We observed that most polymorphic sites in nuclear loci showed a hybrid pattern in one cloned strain and the other two cloned strains were compatible as parental strains (or nearly related to the true parents). The three clones had almost the same ploidy and the DNA content was similar to the reference strain Sylvio (a nearly diploid strain). Despite maxicircle genes evolve faster than nuclear housekeeping ones, we detected no polymorphisms in the sequence of three maxicircle genes showing mito-nuclear discordance. Lastly, the hybrid stock shared 66% of its mHVR clusters with one putative parent and 47% with the other one; in contrast, the putative parental stocks shared less than 30% of the mHVR clusters between them. CONCLUSIONS/SIGNIFICANCE:The results suggest a reductive division, a natural hybridization, biparental inheritance of the minicircles in the hybrid and maxicircle introgression. The models including such phenomena and explaining the relationships between these three clones are discussed.
Published: 2020
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3. Elucidating diversity in the class composition of the minicircle hypervariable region of Trypanosoma cruzi: New perspectives on typing and kDNA inheritance.

Author: Fanny Rusman, Nicolás Tomasini, Noelia-Floridia Yapur, Andrea F Puebla, Paula G Ragone, and Patricio Diosque
Subjects: Arctic medicine. Tropical medicine, RC955-962, Public aspects of medicine, RA1-1270
Abstract: BACKGROUND:Trypanosoma cruzi, the protozoan causative of Chagas disease, is classified into six main Discrete Typing Units (DTUs): TcI-TcVI. This parasite has around 105 copies of the minicircle hypervariable region (mHVR) in their kinetoplastic DNA (kDNA). The genetic diversity of the mHVR is virtually unknown. However, cross-hybridization assays using mHVRs showed hybridization only between isolates belonging to the same genetic group. Nowadays there is no methodologic approach with a good sensibility, specificity and reproducibility for direct typing on biological samples. Due to its high copy number and apparently high diversity, mHVR becomes a good target for typing. METHODOLOGY/PRINCIPAL FINDINGS:Around 22 million reads, obtained by amplicon sequencing of the mHVR, were analyzed for nine strains belonging to six T. cruzi DTUs. The number and diversity of mHVR clusters was variable among DTUs and even within a DTU. However, strains of the same DTU shared more mHVR clusters than strains of different DTUs and clustered together. In addition, hybrid DTUs (TcV and TcVI) shared similar percentages (1.9-3.4%) of mHVR clusters with their parentals (TcII and TcIII). Conversely, just 0.2% of clusters were shared between TcII and TcIII suggesting biparental inheritance of the kDNA in hybrids. Sequencing at low depth (20,000-40,000 reads) also revealed 95% of the mHVR clusters for each of the analyzed strains. Finally, the method revealed good correlation in cluster identity and abundance between different replications of the experiment (r = 0.999). CONCLUSIONS/SIGNIFICANCE:Our work sheds light on the sequence diversity of mHVRs at intra and inter-DTU level. The mHVR amplicon sequencing workflow described here is a reproducible technique, that allows multiplexed analysis of hundreds of strains and results promissory for direct typing on biological samples in a future. In addition, such approach may help to gain knowledge on the mechanisms of the minicircle evolution and phylogenetic relationships among strains.
Published: 2019
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4. Experimental evidence of biological interactions among different isolates of Trypanosoma cruzi from the Chaco Region.

Author: Paula G Ragone, Cecilia Pérez Brandán, Mercedes Monje Rumi, Nicolás Tomasini, Juan J Lauthier, Rubén O Cimino, Alejandro Uncos, Federico Ramos, Anahí M Alberti D'Amato, Miguel A Basombrío, and Patricio Diosque
Subjects: Medicine, Science
Abstract: Many infectious diseases arise from co-infections or re-infections with more than one genotype of the same pathogen. These mixed infections could alter host fitness, the severity of symptoms, success in pathogen transmission and the epidemiology of the disease. Trypanosoma cruzi, the etiological agent of Chagas disease, exhibits a high biological variability often correlated with its genetic diversity. Here, we developed an experimental approach in order to evaluate biological interaction between three T. cruzi isolates belonging to different Discrete Typing Units (DTUs TcIII, TcV and TcVI). These isolates were obtained from a restricted geographical area in the Chaco Region. Different mixed infections involving combinations of two isolates (TcIII + TcV, TcIII + TcVI and TcV + TcVI) were studied in a mouse model. The parameters evaluated were number of parasites circulating in peripheral blood, histopathology and genetic characterization of each DTU in different tissues by DNA hybridization probes. We found a predominance of TcVI isolate in blood and tissues respect to TcIII and TcV; and a decrease of the inflammatory response in heart when the damage of mice infected with TcVI and TcIII + TcVI mixture were compared. In addition, simultaneous presence of two isolates in the same tissue was not detected. Our results show that biological interactions between isolates with different biological behaviors lead to changes in their biological properties. The occurrence of interactions among different genotypes of T. cruzi observed in our mouse model suggests that these phenomena could also occur in natural cycles in the Chaco Region.
Published: 2015
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5. TcTASV Antigens of Trypanosoma cruzi: Utility for Diagnosis and High Accuracy as Biomarkers of Treatment Efficacy in Pediatric Patients

Author: Rubén O. Cimino, Mercedes Monje-Rumi, Anahí A. Alberti D’Amato, Daniel O. Sánchez, José Fernando Gil, Valeria Tekiel, Nicolás Tomasini, Noelia Floridia-Yapur, Patricio Diosque, Julio R. Nasser, Juan José Lauthier, and Paula G. Ragone
Subjects: 030231 tropical medicine, CHAGAS, Diagnostic accuracy, Antibody level, Strongyloides stercoralis, Serology, purl.org/becyt/ford/3.3 [https], 03 medical and health sciences, SEROLOGIA, FAMILAI MULTIGENICA, 0302 clinical medicine, Antigen, Virology, Parasite hosting, Medicine, Trypanosoma cruzi, TcTASV, biology, business.industry, ANTIGENOS SUPERFICIE, biology.organism_classification, Treatment efficacy, Infectious Diseases, Immunology, DIAGNOSTICO, SEGUIMIENTO, ELISA, purl.org/becyt/ford/3 [https], Parasitology, business
Abstract: The discovery and characterization of novel parasite antigens to improve the diagnosis of Trypanosoma cruzi by serological methods and for accurate and rapid follow-up of treatment efficiency are still needed. TcTASV is a T. cruzi-specific multigene family, whose products are expressed on the parasite stages present in the vertebrate host. In a previous work, a mix of antigens from subfamilies TcTASV-A and TcTASV-C (Mix A + C) was sensitive and specific to identify dogs with active infection of high epidemiological relevance. Here, TcTASV-A and TcTASV-C were assayed separately as well as together (Mix A + C) in an ELISA format on human samples. The Mix A + C presented moderate sensitivity (78%) but high diagnostic accuracy with a 100% of specificity, evaluated on healthy, leishmaniasic, and Strongyloides stercoralis infected patients. Moreover, antibody levels of pediatric patients showed-2 years posttreatment-diminished reactivity against the Mix A + C (P < 0.0001), pointing TcTASV antigens as promising tools for treatment follow-up. Fil: Floridia Yapur, Noelia Aldana del Rosario. Universidad Nacional de Salta. Facultad de Ciencias Naturales; Argentina. Universidad Nacional de Salta. Sede Regional Orán. Instituto de Investigación de Enfermedades Tropicales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta; Argentina Fil: Monje Rumi, Maria Mercedes. Universidad Nacional de Salta. Facultad de Ciencias Naturales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Patología Experimental. Universidad Nacional de Salta. Facultad de Ciencias de la Salud. Instituto de Patología Experimental; Argentina Fil: Ragone, Paula Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Patología Experimental. Universidad Nacional de Salta. Facultad de Ciencias de la Salud. Instituto de Patología Experimental; Argentina Fil: Lauthier, Juan José. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Patología Experimental. Universidad Nacional de Salta. Facultad de Ciencias de la Salud. Instituto de Patología Experimental; Argentina Fil: Tomasini, Nicolás. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Patología Experimental. Universidad Nacional de Salta. Facultad de Ciencias de la Salud. Instituto de Patología Experimental; Argentina Fil: D'Amato, Anahí Alberti. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Patología Experimental. Universidad Nacional de Salta. Facultad de Ciencias de la Salud. Instituto de Patología Experimental; Argentina Fil: Diosque, Patricio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Patología Experimental. Universidad Nacional de Salta. Facultad de Ciencias de la Salud. Instituto de Patología Experimental; Argentina Fil: Cimino, Rubén Oscar. Universidad Nacional de Salta. Sede Regional Orán. Instituto de Investigación de Enfermedades Tropicales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta; Argentina Fil: Gil, José Fernando. Universidad Nacional de Salta. Sede Regional Orán. Instituto de Investigación de Enfermedades Tropicales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Investigaciones en Energía no Convencional. Universidad Nacional de Salta. Facultad de Ciencias Exactas. Departamento de Física. Instituto de Investigaciones en Energía no Convencional; Argentina Fil: Sanchez, Daniel Oscar. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. - Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Biotecnológicas; Argentina Fil: Nasser, Julio Rubén. Universidad Nacional de Salta. Facultad de Ciencias Naturales; Argentina Fil: Tekiel, Valeria Sonia. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. - Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Biotecnológicas; Argentina
Published: 2019
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6. Evidence of hybridization, mitochondrial introgression and biparental inheritance of the kDNA minicircles in Trypanosoma cruzi I

Author: Paula G. Ragone, Noelia Floridia-Yapur, Fanny Rusman, Patricio Diosque, and Nicolás Tomasini
Subjects: 0301 basic medicine, Introgression, Genes, Protozoan, RC955-962, Biochemistry, purl.org/becyt/ford/1 [https], Spectrum Analysis Techniques, 0302 clinical medicine, Arctic medicine. Tropical medicine, Genotype, Cell Cycle and Cell Division, Energy-Producing Organelles, Protozoans, Genetics, KDNA, education.field_of_study, Chromosome Biology, DNA, Kinetoplast, Database and informatics methods, Sequence analysis, High-Throughput Nucleotide Sequencing, Eukaryota, Bioquímica y Biología Molecular, Flow Cytometry, Mitochondria, Nuclear DNA, Meiosis, Infectious Diseases, Cell Processes, Spectrophotometry, TCI, Cytophotometry, Cellular Structures and Organelles, Public aspects of medicine, RA1-1270, CIENCIAS NATURALES Y EXACTAS, HYBRIDIZATION, Research Article, Trypanosoma, Mitochondrial DNA, Evolutionary Processes, Bioinformatics, Trypanosoma cruzi, 030231 tropical medicine, Population, Genomics, Bioenergetics, Biology, Minicircle, DNA sequencing, Ciencias Biológicas, 03 medical and health sciences, BIPARENTAL INHERITANCE, education, purl.org/becyt/ford/1.6 [https], Gene, DNA sequence analysis, Evolutionary Biology, Ploidies, Organisms, Public Health, Environmental and Occupational Health, Biology and Life Sciences, Sequence Analysis, DNA, Cell Biology, TRYPANOSOMA CRUZI, DNA, Protozoan, Parasitic Protozoans, MITOCHONDRIAL INTROGRESSION, Hypervariable region, Research and analysis methods, Kinetoplasts, 030104 developmental biology, Genetic Loci, Hybridization, Genetic
Abstract: Background Genetic exchange in Trypanosoma cruzi is controversial not only in relation to its frequency, but also to its mechanism. Parasexual genetic exchange has been proposed based on laboratory hybrids, but population genomics strongly suggests meiosis in T. cruzi. In addition, mitochondrial introgression has been reported several times in natural isolates although its mechanism is not fully understood yet. Moreover, hybrid T. cruzi DTUs (TcV and TcVI) have inherited at least part of the kinetoplastic DNA (kDNA = mitochondrial DNA) from both parents. Methodology/Principal findings In order to address such topics, we sequenced and analyzed fourteen nuclear DNA fragments and three kDNA maxicircle genes in three TcI stocks which are natural clones potentially involved in events of genetic exchange. We also deep-sequenced (a total of 6,146,686 paired-end reads) the minicircle hypervariable region (mHVR) of the kDNA in such three strains. In addition, we analyzed the DNA content by flow cytometry to address cell ploidy. We observed that most polymorphic sites in nuclear loci showed a hybrid pattern in one cloned strain and the other two cloned strains were compatible as parental strains (or nearly related to the true parents). The three clones had almost the same ploidy and the DNA content was similar to the reference strain Sylvio (a nearly diploid strain). Despite maxicircle genes evolve faster than nuclear housekeeping ones, we detected no polymorphisms in the sequence of three maxicircle genes showing mito-nuclear discordance. Lastly, the hybrid stock shared 66% of its mHVR clusters with one putative parent and 47% with the other one; in contrast, the putative parental stocks shared less than 30% of the mHVR clusters between them. Conclusions/significance The results suggest a reductive division, a natural hybridization, biparental inheritance of the minicircles in the hybrid and maxicircle introgression. The models including such phenomena and explaining the relationships between these three clones are discussed., Author summary Chagas disease, an important public health problem in Latin America, is caused by the parasite Trypanosoma cruzi. Despite being a widely studied parasite, several questions on the biology of genetic exchange remain unanswered. Population genomic studies have inferred meiosis in T. cruzi, but this cellular division mechanism has not been observed in laboratory yet. In addition, previous results suggest that mitochondrial DNA (called kDNA) may be inherited from both parents in hybrids. Here, we analyzed a hybrid strain and its potential parents to address the mechanisms of genetic exchange at nuclear and mitochondrial levels. We observed that the hybrid strain had heterozygous patterns and DNA content compatible with a meiosis event. Also, we observed that the evolutionary histories of nuclear DNA and kDNA maxicircles were discordant and that the three strains shared identical DNA sequences. Mitochondrial introgression of maxicircle DNA from one genotype to another may explain this observation. In addition, we demonstrated that the hybrid strain shared kDNA minicircles with both parental strains. Our results suggest that hybridization implied meiosis and biparental inheritance of the kDNA. Further research is required to address such phenomena in detail.
Published: 2020

7. Potential association of Trypanosoma cruzi DTUs TcV and TcVI with the digestive form of Chagas disease

Author: S. Nuñez, C.M. Pérez Brandán, María M. Monje-Rumi, Patricio Diosque, Paula G. Ragone, Nicolás Tomasini, María Paola Zago, Noelia Floridia-Yapur, and N. Barrientos
Subjects: 0301 basic medicine, Microbiology (medical), Chagas disease, Adult, Male, Adolescent, DTUS, Trypanosoma cruzi, 030106 microbiology, Argentina, Biology, Microbiology, Asymptomatic, law.invention, 03 medical and health sciences, purl.org/becyt/ford/3.3 [https], Young Adult, law, parasitic diseases, Genetics, medicine, Prevalence, Humans, Blood culture, Chagas Disease, Typing, Molecular Biology, Ecology, Evolution, Behavior and Systematics, Polymerase chain reaction, Aged, Aged, 80 and over, ARGENTINA, medicine.diagnostic_test, CHAGAS DISEASE, DNA, Protozoan, Middle Aged, medicine.disease, biology.organism_classification, Virology, 030104 developmental biology, Infectious Diseases, DNA BLOT, CLINICAL PICTURES, purl.org/becyt/ford/3 [https], Female, medicine.symptom, Mixed infection
Abstract: The relationship among genetic diversity of Trypanosoma cruzi and clinical forms of Chagas disease remain elusive. In order to assess the possible association between different T. cruzi Discrete Typing Units (DTUs) and the clinical pictures of the disease, 205 chronic patients from Salta province, Argentina, were analysed. One hundred and twenty-two of these patients were clinically categorized as: cardiac 38.5% (47/122), digestive 15% (18/122), cardio-digestive 16% (20/122) and asymptomatic 30% (37/122). From each patient, blood samples were taken for both, Polymerase Chain Reaction (PCR) targeting kDNA and blood culture analyses. The presence of T. cruzi kDNA was detected in 43% (88/205) of the patients. T. cruzi DTUs were identified in 74% (65/88) of the kDNA positive patients by PCR-hybridization using specific probes. We detected the presence of DTUs TcI, TcII, TcV and TcVI. Single infections (i.e. presence of only one DTU in the sample) were detected in 38.64% of the samples (34/88), while mixed infections were 35.23% (31/88). TcV was the most prevalent DTU (60.3%- 53/88). The association analyses showed, for the first time to the best of our knowledge, that TcV and TcVI were associated with the digestive form of Chagas Disease (Fisher p = .0001). Fil: Monje Rumi, Maria Mercedes. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Patología Experimental. Universidad Nacional de Salta. Facultad de Ciencias de la Salud. Instituto de Patología Experimental; Argentina Fil: Floridia Yapur, Noelia Aldana del Rosario. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Patología Experimental. Universidad Nacional de Salta. Facultad de Ciencias de la Salud. Instituto de Patología Experimental; Argentina Fil: Zago, María Paola. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Patología Experimental. Universidad Nacional de Salta. Facultad de Ciencias de la Salud. Instituto de Patología Experimental; Argentina Fil: Ragone, Paula Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Patología Experimental. Universidad Nacional de Salta. Facultad de Ciencias de la Salud. Instituto de Patología Experimental; Argentina Fil: Pérez Brandán, Jimena María. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Patología Experimental. Universidad Nacional de Salta. Facultad de Ciencias de la Salud. Instituto de Patología Experimental; Argentina Fil: Nuñez, Sonia. Gobierno de la Provincia de Salta. Hospital San Bernardo.; Argentina Fil: Barrientos, Natalia. Gobierno de la Provincia de Salta. Hospital San Bernardo.; Argentina Fil: Tomasini, Nicolás. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Patología Experimental. Universidad Nacional de Salta. Facultad de Ciencias de la Salud. Instituto de Patología Experimental; Argentina Fil: Diosque, Patricio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Patología Experimental. Universidad Nacional de Salta. Facultad de Ciencias de la Salud. Instituto de Patología Experimental; Argentina
Published: 2019

8. The TcTASV proteins are novel promising antigens to detect activeTrypanosoma cruziinfection in dogs

Author: Juan José Lauthier, Paula G. Ragone, Jorge Diego Marco, Noelia Floridia-Yapur, Rubén O. Cimino, A. M. Alberti d'Amato, Daniel O. Sánchez, M. Monje Rumi, Nicolás Tomasini, Valeria Tekiel, Paola Andrea Barroso, Julio R. Nasser, and Patricio Diosque
Subjects: 0301 basic medicine, Chagas disease, Subfamily, Protein family, Otras Ciencias Biológicas, Trypanosoma cruzi, 030106 microbiology, 030231 tropical medicine, Antibodies, Protozoan, Antigens, Protozoan, Enzyme-Linked Immunosorbent Assay, Sensitivity and Specificity, law.invention, Ciencias Biológicas, Mice, 03 medical and health sciences, Dogs, 0302 clinical medicine, Antigen, law, medicine, Animals, Parasite hosting, Chagas Disease, Dog Diseases, Gene, IMMUNODIAGNOSIS, TCTASV ANTIGENS, biology, TRYPANOSOMA CRUZI, biology.organism_classification, medicine.disease, Virology, DOGS, Infectious Diseases, Recombinant DNA, Animal Science and Zoology, Parasitology, CIENCIAS NATURALES Y EXACTAS
Abstract: In regions where Chagas disease is endemic, canine Trypanosoma cruzi infection is highly correlated with the risk of transmission of the parasite to humans. Herein we evaluated the novel TcTASV protein family (subfamilies A, B, C), differentially expressed in bloodstream trypomastigotes, for the detection of naturally infected dogs. A gene of each TcTASV subfamily was cloned and expressed. Indirect enzyme-linked immunosorbent assays (ELISA) were developed using recombinant antigens individually or mixed together. Our results showed that dogs with active T. cruzi infection differentially reacted against the TcTASV-C subfamily. The use of both TcTASV-C plus TcTASV-A proteins (Mix A+C-ELISA) enhanced the reactivity of sera from dogs with active infection, detecting 94% of the evaluated samples. These findings agree with our previous observations, where the infected animals exhibited a quick anti-TcTASV-C antibody response, coincident with the beginning of parasitaemia, in a murine model of the disease. Results obtained in the present work prove that the Mix A+C-ELISA is a specific, simple and cheap technique to be applied in endemic areas in screening studies. The Mix A+C-ELISA could help to differentially detect canine hosts with active infection and therefore with high impact in the risk of transmission to humans. Fil: Floridia Yapur, Noelia Aldana del Rosario. Universidad Nacional de Salta. Facultad de Cs.naturales. Escuela de Biología. Cátedra de Química Biologica; Argentina. Universidad Nacional de Salta. Sede Regional Orán. Instituto de Investigación de Enfermedades Tropicales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Monje Rumi, Maria Mercedes. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Patología Experimental. Universidad Nacional de Salta. Facultad de Ciencias de la Salud. Instituto de Patología Experimental; Argentina Fil: Ragone, Paula Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Patología Experimental. Universidad Nacional de Salta. Facultad de Ciencias de la Salud. Instituto de Patología Experimental; Argentina Fil: Lauthier, Juan José. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Patología Experimental. Universidad Nacional de Salta. Facultad de Ciencias de la Salud. Instituto de Patología Experimental; Argentina Fil: Tomasini, Nicolás. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Patología Experimental. Universidad Nacional de Salta. Facultad de Ciencias de la Salud. Instituto de Patología Experimental; Argentina Fil: Alberti D'amato, Anahí Maitén. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Patología Experimental. Universidad Nacional de Salta. Facultad de Ciencias de la Salud. Instituto de Patología Experimental; Argentina Fil: Diosque, Patricio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Patología Experimental. Universidad Nacional de Salta. Facultad de Ciencias de la Salud. Instituto de Patología Experimental; Argentina Fil: Cimino, R.. Universidad Nacional de Salta. Sede Regional Oran. Instituto de Investigación de Enfermedades Tropicales; Argentina Fil: Marco, Jorge Diego. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Patología Experimental. Universidad Nacional de Salta. Facultad de Ciencias de la Salud. Instituto de Patología Experimental; Argentina Fil: Barroso, Paola Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Patología Experimental. Universidad Nacional de Salta. Facultad de Ciencias de la Salud. Instituto de Patología Experimental; Argentina Fil: Sanchez, Daniel Oscar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; Argentina Fil: Nasser, Julio Rubén. Universidad Nacional de Salta. Sede Regional Oran. Instituto de Investigación de Enfermedades Tropicales; Argentina Fil: Tekiel, Valeria Sonia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; Argentina
Published: 2016
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9. A Novel Genotype and First Record of Trypanosoma lainsoni in Argentina

Author: Fanny Rusman, M. Mónica Díaz, Rubén M. Barquez, Noelia Floridia-Yapur, Anahí G Díaz, Paula G. Ragone, Nicolás Tomasini, and Patricio Diosque
Subjects: Microbiology (medical), 18S RDNA GENES, 18S rDNA genes, lcsh:Medicine, Leopardus geoffroyi, Article, 18S ribosomal RNA, purl.org/becyt/ford/1 [https], gGAPDH genes, Genus, Calomys spp, morphology, transmission electron microscopy, parasitic diseases, Genotype, GGAPDH GENES, Immunology and Allergy, purl.org/becyt/ford/1.6 [https], Clade, Molecular Biology, Gene, Trypanosoma lainsoni, General Immunology and Microbiology, biology, lcsh:R, biology.organism_classification, TRANSMISSION ELECTRON MICROSCOPY, Infectious Diseases, Evolutionary biology, CALOMYS SPP, TRYPANOSOMA LAINSONI, Trypanosoma, MORPHOLOGY, LEOPARDUS GEOFFROYI, Cricetidae
Abstract: Trypanosomes are a group of parasitic flagellates with medical and veterinary importance. Despite many species having been described in this genus, little is known about many of them. Here, we report a genetic and morphological characterization of trypanosomatids isolated from wild mammals from the Argentine Chaco region. Parasites were morphologically and ultrastructurally characterized by light microscopy and transmission electron microscopy. Additionally, 18s rRNA and gGAPDH genes were sequenced and analyzed using maximum likelihood and Bayesian inference. Morphological characterization showed clear characteristics associated with the Trypanosoma genus. The genetic characterization demonstrates that the studied isolates have identical sequences and a pairwise identity of 99% with Trypanosoma lainsoni, which belongs to the clade of lizards and snakes/rodents and marsupials. To date, this species had only been found in the Amazon region. Our finding represents the second report of T. lainsoni and the first record for the Chaco region. Furthermore, we ultrastructurally described for the first time the species. Finally, the host range of T. lainsoni was expanded (Leopardus geoffroyi, Carenivora, Felidae, and Calomys sp., Rodentia, Cricetidae), showing a wide host range for this species.
Published: 2020
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10. Trypanosoma cruzi diversity in the Gran Chaco: Mixed infections and differential host distribution of TcV and TcVI

Author: Patricio Diosque, Juan José Lauthier, Paula G. Ragone, Nicolás Tomasini, Cecilia Pérez Brandán, María M. Monje-Rumi, Viviana Orellana, Miguel A. Basombrío, Rubén O. Cimino, and Anahí M. Alberti D’Amato
Subjects: Adult, Male, Rural Population, Microbiology (medical), CIENCIAS MÉDICAS Y DE LA SALUD, Adolescent, Genotype, Trypanosoma cruzi, Argentina, Ciencias de la Salud, Biology, Microbiology, Young Adult, Dogs, parasitic diseases, Genetics, Epidemiología, Animals, Humans, Parasite hosting, Parasitología, Chagas Disease, Dog Diseases, Typing, Child, Molecular Biology, Ecology, Evolution, Behavior and Systematics, DOMESTIC CYCLE, ARGENTINA, Coinfection, Host (biology), Hybridization probe, DNA, Protozoan, Middle Aged, biology.organism_classification, Virology, PREVALENCE, PCR, Cross-Sectional Studies, Infectious Diseases, HYBRIDIZATION, Mixed infection
Abstract: The transmission cycles of Trypanosoma cruzi in the Gran Chaco are complex networks involving domestic and wild components, whose interrelationships are not well understood. Knowing the circuit of transmission of the different Discrete Typing Units (DTUs) of T. cruzi in the complex environment of the Chaco region is relevant to understanding how the different components (reservoirs, vectors, ecotopes) interact. In the present study we identified the DTUs infecting humans and dogs in two rural areas of the Gran Chaco in Argentina, using molecular methods which avoid parasite culture. Blood samples of humans and dogs were typified by PCR-DNA blotting and hybridization assays with five specific DNA probes (TcI, TcII, TcIII, TcV and TcVI). PCR analyses were performed on seropositive human and dog samples and showed the presence of T. cruzi DNA in 41.7% (98/235) and 53% (35/66) samples, respectively. The identification of infective DTUs was determined in 83.6% (82/98) and 91.4% (32/35) in human and dog samples, respectively. Single infections (36.7% – 36/98) and a previously not detected high proportion of mixed infections (47.9% – 47/98) were found. In a 15.3% (15/98) of samples the infecting DTU was not identified. Among the single infections TcV was the most prevalent DTU (30.6% – 30/98) in human samples; while TcVI (42.8% – 15/35) showed the highest prevalence in dog samples. TcV/TcVI was the most prevalent mixed infection in humans (32.6% – 32/98); and TcI/TcVI (14.3% – 5/35) in dogs. Significant associations between TcV with humans and TcVI with dogs were detected. For the first time, the presence of TcIII was detected in humans from this region. The occurrence of one human infected whit TcIII (a principally wild DTU) could be suggested the emergence of this, in domestic cycles in the Gran Chaco. Fil: Monje Rumi, Maria Mercedes. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Salta. Instituto de Patología Experimental; Argentina. Universidad Nacional de Salta; Argentina Fil: Perez Brandan, Cecilia Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Salta. Instituto de Patología Experimental; Argentina. Universidad Nacional de Salta; Argentina Fil: Ragone, Paula Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Salta. Instituto de Patología Experimental; Argentina. Universidad Nacional de Salta; Argentina Fil: Tomasini, Nicolás. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Salta. Instituto de Patología Experimental; Argentina. Universidad Nacional de Salta; Argentina Fil: Lauthier, Juan José. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Salta. Instituto de Patología Experimental; Argentina. Universidad Nacional de Salta; Argentina Fil: Alberti D'Amato, Anahi M.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Salta. Instituto de Patología Experimental; Argentina. Universidad Nacional de Salta; Argentina Fil: Cimino, Ruben O.. Universidad Nacional de Salta. Facultad de Cs.naturales. Escuela de Biologia. Catedra de Quimica Biologica; Argentina. Universidad Nacional de Salta. Sede Regional Oran. Instituto de Investigación de Enfermedades Tropicales; Argentina Fil: Orellana, Viviana. Universidad Nacional de Salta. Facultad de Ciencias de la Salud; Argentina Fil: Basombrio, Miguel Angel Manuel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Salta. Instituto de Patología Experimental; Argentina Fil: Diosque, Patricio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Salta. Instituto de Patología Experimental; Argentina. Universidad Nacional de Salta; Argentina
Published: 2015
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11. Can Triatoma virus inhibit infection of Trypanosoma cruzi (Chagas, 1909) in Triatoma infestans (Klug)? A cross infection and co-infection study

Author: Soledad Ceccarelli, María Laura Susevich, Paula G. Ragone, Gerardo Anibal Marti, María Gabriela Echeverría, Jorge E. Rabinovich, Agustín Balsalobre, and Patricio Diosque
Subjects: 0301 basic medicine, Chagas disease, Trypanosoma cruzi, Otras Ciencias Biológicas, 030231 tropical medicine, DICISTROVIRIDAE, Microbiology, Ciencias Biológicas, 03 medical and health sciences, 0302 clinical medicine, parasitic diseases, Triatoma infestans, medicine, Animals, Chagas Disease, Triatoma, ASSOCIATED MICROORGANISMS, PROTOZOAN, Triatominae, Ecology, Evolution, Behavior and Systematics, Cross Infection, biology, Coinfection, Host (biology), biology.organism_classification, medicine.disease, Virology, 030104 developmental biology, Viral replication, CIENCIAS NATURALES Y EXACTAS
Abstract: Triatoma virus occurs infecting Triatominae in the wild (Argentina) and in insectaries (Brazil). Pathogenicity of Triatoma virus has been demonstrated in laboratory; accidental infections in insectaries produce high insect mortality. When more than one microorganism enters the same host, the biological interaction among them differs greatly depending on the nature and the infection order of the co-existing species of microorganisms. We studied the possible interactions between Triatoma virus (TrV) and Trypanosoma cruzi (the etiological agent of Chagas disease) in three different situations: (i) when Triatoma virus is inoculated into an insect host (Triatoma infestans) previously infected with T. cruzi, (ii) when T. cruzi is inoculated into T. infestans previously infected with TrV, and (iii) when TrV and T. cruzi are inoculated simultaneously into the same T. infestans individual. Trypanosoma cruzi infection was found in 57% of insects in the control group for T. cruzi, whereas 85% of insects with previous TrV infection were infected with T. cruzi. TrV infection was found in 78.7% of insects in the control group for TrV, whereas insects previously infected with T. cruzi showed 90% infection with TrV. A total of 67.9% of insects presented simultaneous infection with both types of microorganism. Our results suggest that TrV infection could increase adhesion of T. cruzi to the intestinal cells of triatomines, but presence of T. cruzi in intestinal cells would not increase the possibility of entry of TrV into cells. Although this study cannot explain the mechanism through which TrV facilitates the infection of triatomines with T. cruzi, we conclude that after TrV replication, changes at cellular level should occur that increase the adhesion of T. cruzi. Fil: Marti, Gerardo Anibal. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Estudios Parasitológicos y de Vectores. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Centro de Estudios Parasitológicos y de Vectores; Argentina Fil: Ragone, Paula Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Patología Experimental. Universidad Nacional de Salta. Facultad de Ciencias de la Salud. Instituto de Patología Experimental; Argentina Fil: Balsalobre, Agustin. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Estudios Parasitológicos y de Vectores. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Centro de Estudios Parasitológicos y de Vectores; Argentina Fil: Ceccarelli, Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Estudios Parasitológicos y de Vectores. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Centro de Estudios Parasitológicos y de Vectores; Argentina Fil: Susevich, Maria Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Estudios Parasitológicos y de Vectores. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Centro de Estudios Parasitológicos y de Vectores; Argentina Fil: Diosque, Patricio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Patología Experimental. Universidad Nacional de Salta. Facultad de Ciencias de la Salud. Instituto de Patología Experimental; Argentina Fil: Echeverria, Maria Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Departamento de Microbiología. Cátedra de Virología; Argentina Fil: Rabinovich, Jorge Eduardo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Estudios Parasitológicos y de Vectores. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Centro de Estudios Parasitológicos y de Vectores; Argentina
Published: 2017

12. Epidemiological modeling of Trypanosoma cruzi: Low stercorarian transmission and failure of host adaptive immunity explain the frequency of mixed infections in humans

Author: Juan P. Aparicio, Paula G. Ragone, Sébastien Gourbière, Nicolás Tomasini, and Patricio Diosque
Subjects: 0301 basic medicine, Mixed infections, Disease, Pathogenesis, Adaptive Immunity, Pathology and Laboratory Medicine, Systems Science, purl.org/becyt/ford/1 [https], 0302 clinical medicine, Agent-Based Modeling, Epidemiology, Genotype, Medicine and Health Sciences, Immune Response, lcsh:QH301-705.5, Protozoans, Mammals, Ecology, Transmission (medicine), Coinfection, Simulation and Modeling, purl.org/becyt/ford/1.2 [https], Acquired immune system, Infectious Diseases, Computational Theory and Mathematics, Modeling and Simulation, Host-Pathogen Interactions, Physical Sciences, Vertebrates, Algorithms, CIENCIAS NATURALES Y EXACTAS, Research Article, Neglected Tropical Diseases, Chagas disease, medicine.medical_specialty, Trypanosoma, Computer and Information Sciences, Trypanosoma cruzi, 030231 tropical medicine, Immunology, Humans transmission, Biology, Research and Analysis Methods, Models, Biological, Epidemiological modeling, 03 medical and health sciences, Cellular and Molecular Neuroscience, Immune system, Dogs, Genetics, medicine, Parasitic Diseases, Animals, Humans, Chagas Disease, Molecular Biology, Ecology, Evolution, Behavior and Systematics, Protozoan Infections, Organisms, Computational Biology, Biology and Life Sciences, biology.organism_classification, medicine.disease, Tropical Diseases, Virology, Parasitic Protozoans, Insect Vectors, Ciencias de la Computación, Vector-Borne Diseases, 030104 developmental biology, lcsh:Biology (General), Ciencias de la Computación e Información, Amniotes, Mathematics
Abstract: People living in areas with active vector-borne transmission of Chagas disease have multiple contacts with its causative agent, Trypanosoma cruzi. Reinfections by T. cruzi are possible at least in animal models leading to lower or even hardly detectable parasitaemia. In humans, although reinfections are thought to have major public health implications by increasing the risk of chronic manifestations of the disease, there is little quantitative knowledge about their frequency and the timing of parasite re-inoculation in the course of the disease. Here, we implemented stochastic agent-based models i) to estimate the rate of re-inoculation in humans and ii) to assess how frequent are reinfections during the acute and chronic stages of the disease according to alternative hypotheses on the adaptive immune response following a primary infection. By using a hybrid genetic algorithm, the models were fitted to epidemiological data of Argentinean rural villages where mixed infections by different genotypes of T. cruzi reach 56% in humans. To explain this percentage, the best model predicted 0.032 (0.008–0.042) annual reinfections per individual with 98.4% of them occurring in the chronic phase. In addition, the parasite escapes to the adaptive immune response mounted after the primary infection in at least 20% of the events of re-inoculation. With these low annual rates, the risks of reinfection during the typically long chronic stage of the disease stand around 14% (4%-18%) and 60% (21%-70%) after 5 and 30 years, with most individuals being re-infected 1–3 times overall. These low rates are better explained by the weak efficiency of the stercorarian mode of transmission than a highly efficient adaptive immune response. Those estimates are of particular interest for vaccine development and for our understanding of the higher risk of chronic disease manifestations suffered by infected people living in endemic areas., Author summary Chagas disease is caused by a unicellular parasite mainly transmitted by blood-feeding bugs. The disease starts with an acute phase when the parasite multiplies in tissues. The immune system is then activated and almost clears the parasites before the host enters the chronic phase of the disease. As hosts remain exposed to the vectors, secondary contacts may result in re-inoculation with the parasite. The re-inoculated parasites then have to face an adaptive immune response that may prevent reinfection. There is little quantitative knowledge about the frequency of such re-inoculation and the efficiency of the immune system to prevent them. Since reinfections can lead to people carrying two or more strains of the parasite, we can potentially infer on the frequency of reinfection from the detection of such mixed infections. We fitted agent-based models to field data to estimate the frequency of reinfection in humans and its timing after primary infection. Our models predicted that most reinfections occur during the chronic phase as the adaptive immune response fails to prevent them in 20% of re-inoculations. Such a rate of failure is particularly important as reinfections are thought to increase the risk of heart damage in the chronic phase of the disease.
Published: 2017

13. Biological behavior of different Trypanosoma cruzi isolates circulating in an endemic area for Chagas disease in the Gran Chaco region of Argentina

Author: Angel M. Padilla, Nélida Marcela Romero, Cecilia Pérez Brandán, Mercedes Monje Rumi, Miguel A. Basombrío, Patricio Diosque, Nicolás Tomasini, Anahí M. Alberti D’Amato, Marcela Portelli, Rubén O. Cimino, Juan José Lauthier, Julio R. Nasser, and Paula G. Ragone
Subjects: Chagas disease, Male, Biological Properties, CIENCIAS MÉDICAS Y DE LA SALUD, Endemic Diseases, Veterinary (miscellaneous), Trypanosoma cruzi, 030231 tropical medicine, Argentina, Antibodies, Protozoan, Biology, Natural Isolates, Serology, 03 medical and health sciences, Mice, 0302 clinical medicine, Genetic variation, medicine, Parasite hosting, Animals, Humans, Chagas Disease, Typing, Muscle, Skeletal, 030304 developmental biology, Trypanosoma Cruzi, 0303 health sciences, Myocardium, Genetic Variation, Heart, DNA, Protozoan, Bioquímica y Biología Molecular, medicine.disease, biology.organism_classification, Virology, DNA Fingerprinting, 3. Good health, Enzymes, Mice, Inbred C57BL, Disease Models, Animal, Medicina Básica, Infectious Diseases, Blood, DNA profiling, Insect Science, Multilocus sequence typing, Parasitology
Abstract: The biological behavior of the different . Trypanosoma cruzi strains is still unclear and the importance of exploring the relevance of these differences in natural isolates is of great significance. Herein we describe the biological behavior of four . T. cruzi isolates circulating sympatrically in a restricted geographic area in Argentina endemic for Chagas Disease. These isolates were characterized as belonging to the Discrete Typing Units (DTUs) TcI, TcIII, TcV and TcVI as shown by Multilocus Enzyme Electrophoresis and Multilocus Sequence Typing. In order to study the natural behavior of the different isolates and to preserve their natural properties, we developed a vector transmission model that allows their maintenance in the laboratory. The model consisted of serial passages of these parasites between insect vectors and mice. Vector-derived parasite forms were then inoculated in C57BL/6J mice and number of parasite in peripheral blood, serological response and histological damage in acute and chronic phases of the infection were measured. Parasites from DTUs TcI, TcIII and TcVI were detected by direct fresh blood examination, while TcV parasites could only be detected by Polimerase Chain Reaction. No significant difference in the anti-. T. cruzi antibody response was found during the chronic phase of infection, except for mice infected with TcV parasites where no antibodies could be detected. Histological sections showed that TcI isolate produced more damage in skeletal muscle while TcVI induced more inflammation in the heart. This work shows differential biological behavior among different parasite isolates obtained from the same cycle of transmission, permitting the opportunity to formulate future hypotheses of clinical and epidemiological importance. Fil: Ragone, Paula Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Patología Experimental. Universidad Nacional de Salta. Facultad de Ciencias de la Salud. Instituto de Patología Experimental; Argentina Fil: Pérez Brandan, Cecilia María. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Patología Experimental. Universidad Nacional de Salta. Facultad de Ciencias de la Salud. Instituto de Patología Experimental; Argentina Fil: Padilla, Angel M.. University of Georgia; Estados Unidos Fil: Monje Rumi, Maria Mercedes. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Patología Experimental. Universidad Nacional de Salta. Facultad de Ciencias de la Salud. Instituto de Patología Experimental; Argentina Fil: Lauthier, Juan José. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Patología Experimental. Universidad Nacional de Salta. Facultad de Ciencias de la Salud. Instituto de Patología Experimental; Argentina Fil: Alberti D'amato, Anahí Maitén. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Patología Experimental. Universidad Nacional de Salta. Facultad de Ciencias de la Salud. Instituto de Patología Experimental; Argentina Fil: Tomasini, Nicolás. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Patología Experimental. Universidad Nacional de Salta. Facultad de Ciencias de la Salud. Instituto de Patología Experimental; Argentina Fil: Cimino, Rubén Oscar. Universidad Nacional de Salta. Facultad de Cs.naturales. Escuela de Biología; Argentina Fil: Romero, Nélida M.. Universidad Nacional de Salta. Facultad de Cs.naturales. Escuela de Biología; Argentina Fil: Portelli, Marcela. Universidad Nacional de Salta. Facultad de Cs.naturales. Escuela de Biología; Argentina Fil: Nasser, Julio Rubén. Universidad Nacional de Salta. Facultad de Cs.naturales. Escuela de Biología; Argentina Fil: Basombrío, Miguel Ángel Manuel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Patología Experimental. Universidad Nacional de Salta. Facultad de Ciencias de la Salud. Instituto de Patología Experimental; Argentina Fil: Diosque, Patricio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Patología Experimental. Universidad Nacional de Salta. Facultad de Ciencias de la Salud. Instituto de Patología Experimental; Argentina
Published: 2012
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14. Interest and limitations of Spliced Leader Intergenic Region sequences for analyzing Trypanosoma cruzi I phylogenetic diversity in the Argentinean Chaco

Author: Maria Mercedes Monje Rumi, Nicolás Tomasini, Alejandra Falla, Cecilia Pérez Brandán, Anahí A. Alberti D’Amato, Alejandro G. Schijman, Felipe Guhl, Paula G. Ragone, Christian Barnabé, Juan José Lauthier, Michel Tibayrenc, Patricio Diosque, Carolina Cura, Claudia Herrera, and Miguel A. Basombrío
Subjects: RNA, Spliced Leader, Microbiology (medical), Genotype, Sequence analysis, Trypanosoma cruzi, Molecular Sequence Data, 030231 tropical medicine, Argentina, Biology, Polymerase Chain Reaction, Microbiology, 03 medical and health sciences, 0302 clinical medicine, Intergenic region, Phylogenetics, Genetics, Humans, Chagas Disease, Gene conversion, Molecular Biology, Phylogeny, Ecology, Evolution, Behavior and Systematics, 030304 developmental biology, 0303 health sciences, Geography, Molecular epidemiology, Haplotype, Genetic Variation, Bayes Theorem, Sequence Analysis, DNA, DNA, Protozoan, Phylogenetic diversity, Infectious Diseases, Haplotypes, Microsatellite, DNA, Intergenic, Microsatellite Repeats, Multilocus Sequence Typing
Abstract: Internal and geographical clustering within Trypanosoma cruzi I (TcI) has been recently revealed by using Multilocus Microsatellite Typing and sequencing of the Spliced-Leader Intergenic Region (SL-IR). In the present work, 14 isolates and 11 laboratory-cloned stocks obtained from a geographically restricted area in Chaco Province, Argentina, were analyzed by PCR and sequencing of SL-IR. We were able to differentiate 8 different genotypes that clustered into 4 groups. One of these groups was classified within the formerly described haplotype A and another one within the recently described SL-IR group E. Both were phylogenetically well-supported. In contrast, none of the stocks from the Chaco province were grouped within the cluster previously named haplotype D despite the fact that they shared a similar microsatellite motif in the SL-IR. No evidence of recombination or gene conversion within these stocks was found. On the other hand, multiple ambiguous alignments in the microsatellite region of SL-IR, affecting the tree topology and relationships among groups were detected. Finally, since there are multiple copies of the SL-IR, and they are arranged in tandem, we discuss how molecular processes affecting this kind of sequences could mislead phylogenetic inference.
Published: 2011
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15. Immuno-enzymatic evaluation of the recombinant TSSA-II protein of Trypanosoma cruzi in dogs and human sera: a tool for epidemiological studies

Author: Iván Sergio Marcipar, Paula G. Ragone, Juan José Lauthier, A. M. Alberti d'Amato, Marisa Juarez, Viviana Orellana, Rubén O. Cimino, M. Monje Rumi, José Fernando Gil, Silvana P. Cajal, Rosa Zacca, Norma Acosta, Alejandro J. Krolewiecki, I. R. López Quiroga, Patricio Diosque, and Julio R. Nasser
Subjects: Chagas disease, Trypanosoma cruzi, 030231 tropical medicine, Argentina, Antibodies, Protozoan, Antigens, Protozoan, Enzyme-Linked Immunosorbent Assay, Sensitivity and Specificity, law.invention, 03 medical and health sciences, 0302 clinical medicine, Dogs, Antigen, Cutaneous leishmaniasis, law, Seroepidemiologic Studies, parasitic diseases, medicine, Animals, Humans, Chagas Disease, Dog Diseases, 030304 developmental biology, chemistry.chemical_classification, 0303 health sciences, biology, biology.organism_classification, medicine.disease, Virology, Recombinant Proteins, 3. Good health, Infectious Diseases, Enzyme, chemistry, Antigens, Surface, biology.protein, Recombinant DNA, Animal Science and Zoology, Parasitology, Antibody, Xenodiagnosis
Abstract: SUMMARYThe rTSSA-II (recombinant Trypomastigote Small Surface II) antigen was evaluated by ELISA to detect anti-Trypanosoma cruziantibodies in sera from naturally infected dogs and humans. For this evaluation ELISA-rTSSA-II was standardized and groups were classified according to the results obtained through xenodiagnosis, ELISA and PCR. Sensitivity (Se), Specificity (Sp), Kappa index (KI) and area under curve (AUC) were determined. The Se was determined by using 14 sera from dogs infected withT. cruziVI (TcVI) whereas Sp was determined by using 95 non-chagasic sera by xenodiagnosis, ELISA-Homogenate and PCR. The performance of ELISA-rTSSA-II in dog sera was high (AUC=0·93 and KI=0·91). The Se was 92·85% (1 false negative) and Sp was 100%. Two sera from dogs infected with TcI and 1 with TcIII were negative. For patients infected withT. cruzi, reactivity was 87·8% (36/41), there was only 1 indeterminate, and Sp was 100%. Fifty-four sera from non-chagasic and 68 sera from patients with cutaneous leishmaniasis did not react with rTSS-II. ELISA-rTSSA-II showed a high performance when studying sera from naturally infected dogs and it also presented 100% Sp. This assay could be an important tool to carry out sero-epidemiological surveys on the prevalence ofT. cruzicirculating lineages in the region.
Published: 2011
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16. Evaluation of recombinant antigens of Trypanosoma cruzi to diagnose infection in naturally infected dogs from Chaco region, Argentina

Author: Noelia Floridia-Yapur, M. Monje Rumi, Julio R. Nasser, Nicolás Tomasini, I. Lopez-Quiroga, Paula G. Ragone, Iván Sergio Marcipar, A. M. Alberti d'Amato, Patricio Diosque, A. F. Vega-Benedetti, Rubén O. Cimino, and Juan José Lauthier
Subjects: dogs, Trypanosoma cruzi, Immunology, Argentina, Positive control, Antigens, Protozoan, Enzyme-Linked Immunosorbent Assay, Biology, Sensitivity and Specificity, law.invention, Kappa index, Dogs, Antigen, law, parasitic diseases, Animals, Chagas Disease, Dog Diseases, Antigens, Area under the curve, recombinant antigens, purl.org/becyt/ford/3.1 [https], biology.organism_classification, Virology, Recombinant Proteins, Recombinant DNA, Parasitology, ELISA, purl.org/becyt/ford/3 [https], Xenodiagnosis
Abstract: Dogs are considered the main mammal reservoir of Trypanosoma cruzi in domiciliary environments. Consequently, accurate detection of T. cruzi infection in canine populations is epidemiologically relevant. Here we analyzed the utility of the T. cruzi recombinant antigens FRA, SAPA, CP1, Ag1 and a SAPA/TSSA VI mixture, in an ELISA format. We used a positive control group of sera obtained from 38 dogs from the Chaco region in Argentina with positive Homogenate-ELISA reaction, all of them also positive by xenodiagnosis and/or PCR. The negative group included 19 dogs from a non-endemic area. Sensitivity, specificity, Area Under the Curve (AUC) of the Receiver-Operating Charactheristic (ROC) curve and Kappa index were obtained in order to compare the diagnostic efficiency of the tests. The SAPA/TSSA VI had the highest performance, with a sensitivity of 94.7% and an AUC ROC of 0.99 that indicates high accuracy. Among individual antigens, SAPA-ELISA yielded the highest sensitivity (86.8%) and AUC ROC (0.96), whereas FRA-ELISA was the least efficient test (sensitivity=36.8%; AUC ROC=0.53). Our results showed that the use of SAPA/TSSA VI in ELISA assays could be a useful tool to study dogs naturally infected with T. cruzi in endemic areas. Fil: Floridia Yapur, Noelia Aldana del Rosario. Universidad Nacional de Salta. Sede Regional Orán. Instituto de Investigación de Enfermedades Tropicales; Argentina. Universidad Nacional de Salta. Facultad de Ciencias Naturales. Escuela de Biología. Cátedra de Química Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta; Argentina Fil: Vega Benedetti, Ana Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Patología Experimental. Universidad Nacional de Salta. Facultad de Ciencias de la Salud. Instituto de Patología Experimental; Argentina Fil: Monje Rumi, Maria Mercedes. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Patología Experimental. Universidad Nacional de Salta. Facultad de Ciencias de la Salud. Instituto de Patología Experimental; Argentina Fil: Ragone, Paula Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Patología Experimental. Universidad Nacional de Salta. Facultad de Ciencias de la Salud. Instituto de Patología Experimental; Argentina Fil: Lauthier, Juan José. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Patología Experimental. Universidad Nacional de Salta. Facultad de Ciencias de la Salud. Instituto de Patología Experimental; Argentina Fil: Tomasini, Nicolás. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Patología Experimental. Universidad Nacional de Salta. Facultad de Ciencias de la Salud. Instituto de Patología Experimental; Argentina Fil: Alberti D'amato, Anahí Maitén. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Patología Experimental. Universidad Nacional de Salta. Facultad de Ciencias de la Salud. Instituto de Patología Experimental; Argentina Fil: López Quiroga, Inés Raquel. Universidad Nacional de Salta. Facultad de Ciencias Naturales. Escuela de Biología. Cátedra de Química Biológica; Argentina. Universidad Nacional de Salta. Sede Regional Orán. Instituto de Investigación de Enfermedades Tropicales; Argentina Fil: Diosque, Patricio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Patología Experimental. Universidad Nacional de Salta. Facultad de Ciencias de la Salud. Instituto de Patología Experimental; Argentina Fil: Marcipar, Iván Sergio. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Laboratorio de Tecnología Inmunológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta; Argentina Fil: Nasser, Julio Rubén. Universidad Nacional de Salta. Sede Regional Orán. Instituto de Investigación de Enfermedades Tropicales; Argentina. Universidad Nacional de Salta. Facultad de Ciencias Naturales. Escuela de Biología. Cátedra de Química Biológica; Argentina Fil: Cimino, Rubén Oscar. Universidad Nacional de Salta. Sede Regional Orán. Instituto de Investigación de Enfermedades Tropicales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta; Argentina. Universidad Nacional de Salta. Facultad de Ciencias Naturales. Escuela de Biología. Cátedra de Química Biológica; Argentina
Published: 2014
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17. Optimized multilocus sequence typing (MLST) scheme for Trypanosoma cruzi

Author: Nicolás Tomasini, Michel Tibayrenc, Paula G. Ragone, Cecilia Pérez Brandán, Maria Mercedes Monje Rumi, Juan José Lauthier, Christian Barnabé, Martin S. Llewellyn, Patricio Diosque, Anahí Maitén Alberti-D'Amato, Michael A. Miles, Louisa A. Messenger, Matthew Yeo, and Michael D. Lewis
Subjects: CIENCIAS MÉDICAS Y DE LA SALUD, lcsh:Arctic medicine. Tropical medicine, Genotype, Sequence analysis, lcsh:RC955-962, Trypanosoma cruzi, 030231 tropical medicine, SCHEME OF OPTIMIZATION, Ciencias de la Salud, Biology, Microbiology, 03 medical and health sciences, purl.org/becyt/ford/3.3 [https], 0302 clinical medicine, Medicine and Health Sciences, Parasitic Diseases, Chagas Disease, Typing, Parasite Evolution, 030304 developmental biology, Genetics, Whole genome sequencing, 0303 health sciences, Genetic diversity, Protozoan Infections, Strain (biology), lcsh:Public aspects of medicine, Public Health, Environmental and Occupational Health, Biology and Life Sciences, lcsh:RA1-1270, TRYPANOSOMA CRUZI, Amplicon Size, 3. Good health, Enfermedades Infecciosas, Infectious Diseases, Multilocus sequence typing, GENETIC DIVERSITY, purl.org/becyt/ford/3 [https], Parasitology, MULTILOCUS SEQUENCE TYPING, Research Article, Multilocus Sequence Typing
Abstract: Trypanosoma cruzi, the aetiological agent of Chagas disease possess extensive genetic diversity. This has led to the development of a plethora of molecular typing methods for the identification of both the known major genetic lineages and for more fine scale characterization of different multilocus genotypes within these major lineages. Whole genome sequencing applied to large sample sizes is not currently viable and multilocus enzyme electrophoresis, the previous gold standard for T. cruzi typing, is laborious and time consuming. In the present work, we present an optimized Multilocus Sequence Typing (MLST) scheme, based on the combined analysis of two recently proposed MLST approaches. Here, thirteen concatenated gene fragments were applied to a panel of T. cruzi reference strains encompassing all known genetic lineages. Concatenation of 13 fragments allowed assignment of all strains to the predicted Discrete Typing Units (DTUs), or near-clades, with the exception of one strain that was an outlier for TcV, due to apparent loss of heterozygosity in one fragment. Monophyly for all DTUs, along with robust bootstrap support, was restored when this fragment was subsequently excluded from the analysis. All possible combinations of loci were assessed against predefined criteria with the objective of selecting the most appropriate combination of between two and twelve fragments, for an optimized MLST scheme. The optimum combination consisted of 7 loci and discriminated between all reference strains in the panel, with the majority supported by robust bootstrap values. Additionally, a reduced panel of just 4 gene fragments displayed high bootstrap values for DTU assignment and discriminated 21 out of 25 genotypes. We propose that the seven-fragment MLST scheme could be used as a gold standard for T. cruzi typing, against which other typing approaches, particularly single locus approaches or systematic PCR assays based on amplicon size, could be compared., Author Summary The single-celled parasite Trypanosoma cruzi occurs in mammals and insect vectors in the Americas. When transmitted to humans it causes Chagas disease (American trypanosomiasis) a major public health problem. T. cruzi is genetically diverse and currently split into six groups, known as TcI to TcVI. Multilocus sequence typing (MLST) is a method used for studying the population structure and diversity of pathogens and involves sequencing DNA of several different genes and comparing the sequences between isolates. Here, we assess 13 T. cruzi genes and select the best combination for diversity studies. Outputs reveal that a combination of 7 genes can be used for both lineage assignment and high resolution studies of genetic diversity, and a reduced combination of four loci for lineage assignment. Application of MLST for assigning field isolates of T. cruzi to genetic groups and for detailed investigation of diversity provides a valuable approach to understanding the taxonomy, population structure, genetics, ecology and epidemiology of this important human pathogen.
Published: 2014

18. Preponderant clonal evolution of Trypanosoma cruzi I from Argentinean Chaco revealed by Multilocus Sequence Typing (MLST)

Author: Paula G. Ragone, Nicolás Tomasini, Juan José Lauthier, Patricio Diosque, Anahí M. Alberti D’Amato, Miguel A. Basombrío, Maria Mercedes Monje Rumi, and Cecilia Pérez Brandán
Subjects: RNA, Spliced Leader, Microbiology (medical), Lineage (genetic), CIENCIAS MÉDICAS Y DE LA SALUD, Genotype, Trypanosoma cruzi, Molecular Sequence Data, Argentina, Ciencias de la Salud, Microbiology, Linkage Disequilibrium, Evolution, Molecular, Intergenic region, Genetics, Humans, Chagas Disease, Molecular Biology, Alleles, Phylogeny, Ecology, Evolution, Behavior and Systematics, Polymorphism, Genetic, biology, Genetic Variation, biology.organism_classification, INTERCAMBIO GENÉTICO, RAPD, Enfermedades Infecciosas, Infectious Diseases, Genetic Loci, Genetic marker, TCI, Genetic structure, Multilocus sequence typing, DNA, Intergenic, ENFERMEDAD DE CHAGAS, Multilocus Sequence Typing, MLST
Abstract: Trypanosoma cruzi has been historically classified as a species with preponderant clonal evolution (PCE). However, with the advent of highly polymorphic markers and studies at geographically reduced scales,the PCE in T. cruzi was challenged. In fact, some studies have suggested that recombination in T. cruzi lineage I (TcI) is much more frequent than previously believed. Further analyses of TcI populations from different geographical regions of Latin America are needed to examine this hypothesis. In the present study,we contribute to this topic by analyzing the population structure of TcI from a restricted geographical area in the Chaco region, Argentina. We analyzed TcI isolates from different hosts and vectors using a Multilocus Sequence Typing (MLST) approach. These isolates were previously characterized by sequencing the spliced leader intergenic region (SL-IR). Low levels of incongruence and well-supported clusters for MLST dataset were obtained from the analyses. Moreover, high linkage disequilibrium was found and five repeated and overrepresented genotypes were detected. In addition, a good correspondence between SL-IR and MLST was observed which is expected under PCE. However, recombination is not ruled out because five out of 28 pairs of loci were incompatible with strict clonality and one possible genetic exchange event was detected. Overall, our results represent evidence of PCE in TcI from the study area.Finally, considering our findings we discuss the scenario for the genetic structure of TcI. Fil: Tomasini, Nicolás. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Salta. Instituto de Patología Experimental; Argentina. Universidad Nacional de Salta; Argentina Fil: Lauthier, Juan José. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Salta. Instituto de Patología Experimental; Argentina. Universidad Nacional de Salta; Argentina Fil: Monje Rumi, Maria Mercedes. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Salta. Instituto de Patología Experimental; Argentina. Universidad Nacional de Salta; Argentina Fil: Ragone, Paula Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Salta. Instituto de Patología Experimental; Argentina. Universidad Nacional de Salta; Argentina Fil: Alberti D'amato, Anahi M.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Salta. Instituto de Patología Experimental; Argentina. Universidad Nacional de Salta; Argentina Fil: Perez Brandan, Cecilia Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Salta. Instituto de Patología Experimental; Argentina. Universidad Nacional de Salta; Argentina Fil: Basombrio, Miguel Angel Manuel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Salta. Instituto de Patología Experimental; Argentina. Universidad Nacional de Salta; Argentina Fil: Diosque, Patricio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Salta. Instituto de Patología Experimental; Argentina. Universidad Nacional de Salta; Argentina
Published: 2014

19. Benznidazole treatment in chronic children infected with Trypanosoma cruzi: serological and molecular follow-up of patients and identification of Discrete Typing Units

Author: Julio R. Nasser, Rubén O. Cimino, Viviana Orellana, Nicolás Tomasini, Miguel A. Basombrío, Juan José Lauthier, José Fernando Gil, C.D. Lacunza, Patricio Diosque, A. M. Alberti d'Amato, C.M. Pérez Brandán, M. Monje Rumi, and Paula G. Ragone
Subjects: Chagas disease, Male, Rural Population, Adolescent, Genotype, Veterinary (miscellaneous), Trypanosoma cruzi, Antiprotozoal Agents, Argentina, Antibodies, Protozoan, Serology, medicine, Humans, Chagas Disease, Typing, Seroconversion, Child, biology, DNA, Kinetoplast, Nucleic Acid Hybridization, DNA, Protozoan, biology.organism_classification, medicine.disease, Virology, Hypervariable region, Infectious Diseases, Treatment Outcome, Benznidazole, Nitroimidazoles, Insect Science, Child, Preschool, Chronic Disease, Parasitology, Female, medicine.drug, Follow-Up Studies
Abstract: A total of 221 children from two rural settlements in Northeast Argentina were examined for T. cruzi infection. Blood samples were taken for serology tests and PCR assays. In addition, T. cruzi Discrete Typing Units (DTUs) were determined by hybridization with specific DNA probes of the minicircle hypervariable regions (mHVR). Serological results indicated that 26% (57/215) were reactive against T. cruzi antigens. PCR analyses were performed on seropositive samples showing presence of parasite DNA in 31 out of 53 samples (58.5%). All seropositive children underwent specific chemotherapy with Benznidazole (5mg/kg/day) for a period of two months and were monitored two and five years after treatment. Overall the treatment was well tolerated and low side effects were observed. Serological conversion was observed at two years post -treatment in one child form Pampa Avila and at five years in two children from Tres Estacas. However, at the end of the follow-up period, T. cruzi DNA could not be detected by PCR in samples from treated children, except in two cases. In addition, the results of hybridizations with specific DNA probes showed that DTU TcV was detected in 68% (21/31), TcVI in 7% (2/31) and TcV/VI in 3% (1/31) of the samples. Altogether, results of the follow-up of treated children showed a low rate of seroconversion; however trend toward seroconversion was evident at five years post-treatment. On the other hand, detection of T. cruzi DNA by PCR significantly decreased after Benznidazole treatment. The existence of data regarding serological and molecular follow-ups from controlled studies in the Chaco Region will be important for future treatment efforts against T. cruzi infection in this region. The results obtained in the present study represent a contribution in this regard.
Published: 2012

20. Experimental Evidence of Biological Interactions among Different Isolates of Trypanosoma cruzi from the Chaco Region

Author: Alejandro Uncos, Paula G. Ragone, Federico Ramos, Nicolás Tomasini, Anahí M. Alberti D´Amato, Miguel A. Basombrío, Rubén O. Cimino, Juan José Lauthier, Mercedes Monje Rumi, Cecilia Pérez Brandán, and Patricio Diosque
Subjects: Chagas disease, CIENCIAS MÉDICAS Y DE LA SALUD, Genotype, BIOLOGICAL INTERACTIONS, Trypanosoma cruzi, Science, Ciencias de la Salud, Parasitemia, Biology, Mice, purl.org/becyt/ford/3.3 [https], medicine, Animals, Humans, Chagas Disease, ISOLATES, Typing, Pathogen, Inflammation, Genetic diversity, Multidisciplinary, Genetic Variation, Heart, TRYPANOSOMA CRUZI, medicine.disease, biology.organism_classification, Virology, Biological interaction, Enfermedades Infecciosas, Immunology, Medicine, purl.org/becyt/ford/3 [https], Research Article
Abstract: Many infectious diseases arise from co-infections or re-infections with more than one genotype of the same pathogen. These mixed infections could alter host fitness, the severity of symptoms, success in pathogen transmission and the epidemiology of the disease. Trypanosoma cruzi, the etiological agent of Chagas disease, exhibits a high biological variability often correlated with its genetic diversity. Here, we developed an experimental approach in order to evaluate biological interaction between three T. cruzi isolates belonging to different Discrete Typing Units (DTUs TcIII, TcV and TcVI). These isolates were obtained from a restricted geographical area in the Chaco Region. Different mixed infections involving combinations of two isolates (TcIII + TcV, TcIII + TcVI and TcV + TcVI) were studied in a mouse model. The parameters evaluated were number of parasites circulating in peripheral blood, histopathology and genetic characterization of each DTU in different tissues by DNA hybridization probes. We found a predominance of TcVI isolate in blood and tissues respect to TcIII and TcV; and a decrease of the inflammatory response in heart when the damage of mice infected with TcVI and TcIII + TcVI mixture were compared. In addition, simultaneous presence of two isolates in the same tissue was not detected. Our results show that biological interactions between isolates with different biological behaviors lead to changes in their biological properties. The occurrence of interactions among different genotypes of T. cruzi observed in our mouse model suggests that these phenomena could also occur in natural cycles in the Chaco Region. Fil: Ragone, Paula Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Salta. Instituto de Patología Experimental; Argentina. Universidad Nacional de Salta; Argentina Fil: Perez Brandan, Cecilia Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Salta. Instituto de Patología Experimental; Argentina. Universidad Nacional de Salta; Argentina Fil: Monje Rumi, Maria Mercedes. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Salta. Instituto de Patología Experimental; Argentina. Universidad Nacional de Salta; Argentina Fil: Tomasini, Nicolás. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Salta. Instituto de Patología Experimental; Argentina. Universidad Nacional de Salta; Argentina Fil: Lauthier, Juan José. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Salta. Instituto de Patología Experimental; Argentina. Universidad Nacional de Salta; Argentina Fil: Cimino, Ruben O.. Universidad Nacional de Salta. Facultad de Cs.naturales. Escuela de Biologia. Catedra de Quimica Biologica; Argentina Fil: Uncos, Delfor Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Salta. Instituto de Patología Experimental; Argentina Fil: Ramos, Federico. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Salta. Instituto de Patología Experimental; Argentina Fil: Alberti D'amato, Anahi M.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Salta. Instituto de Patología Experimental; Argentina. Universidad Nacional de Salta; Argentina Fil: Basombrio, Miguel Angel Manuel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Salta. Instituto de Patología Experimental; Argentina Fil: Diosque, Patricio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Salta. Instituto de Patología Experimental; Argentina. Universidad Nacional de Salta; Argentina
Published: 2015
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20 results on '"Paula G. Ragone"'

1. A Novel Genotype and First Record of Trypanosoma lainsoni in Argentina

2. Evidence of hybridization, mitochondrial introgression and biparental inheritance of the kDNA minicircles in Trypanosoma cruzi I.

3. Elucidating diversity in the class composition of the minicircle hypervariable region of Trypanosoma cruzi: New perspectives on typing and kDNA inheritance.

4. Experimental evidence of biological interactions among different isolates of Trypanosoma cruzi from the Chaco Region.

5. TcTASV Antigens of Trypanosoma cruzi: Utility for Diagnosis and High Accuracy as Biomarkers of Treatment Efficacy in Pediatric Patients

6. Evidence of hybridization, mitochondrial introgression and biparental inheritance of the kDNA minicircles in Trypanosoma cruzi I

7. Potential association of Trypanosoma cruzi DTUs TcV and TcVI with the digestive form of Chagas disease

8. The TcTASV proteins are novel promising antigens to detect activeTrypanosoma cruziinfection in dogs

9. A Novel Genotype and First Record of Trypanosoma lainsoni in Argentina

10. Trypanosoma cruzi diversity in the Gran Chaco: Mixed infections and differential host distribution of TcV and TcVI

11. Can Triatoma virus inhibit infection of Trypanosoma cruzi (Chagas, 1909) in Triatoma infestans (Klug)? A cross infection and co-infection study

12. Epidemiological modeling of Trypanosoma cruzi: Low stercorarian transmission and failure of host adaptive immunity explain the frequency of mixed infections in humans

13. Biological behavior of different Trypanosoma cruzi isolates circulating in an endemic area for Chagas disease in the Gran Chaco region of Argentina

14. Interest and limitations of Spliced Leader Intergenic Region sequences for analyzing Trypanosoma cruzi I phylogenetic diversity in the Argentinean Chaco

15. Immuno-enzymatic evaluation of the recombinant TSSA-II protein of Trypanosoma cruzi in dogs and human sera: a tool for epidemiological studies

16. Evaluation of recombinant antigens of Trypanosoma cruzi to diagnose infection in naturally infected dogs from Chaco region, Argentina

17. Optimized multilocus sequence typing (MLST) scheme for Trypanosoma cruzi

18. Preponderant clonal evolution of Trypanosoma cruzi I from Argentinean Chaco revealed by Multilocus Sequence Typing (MLST)

19. Benznidazole treatment in chronic children infected with Trypanosoma cruzi: serological and molecular follow-up of patients and identification of Discrete Typing Units

20. Experimental Evidence of Biological Interactions among Different Isolates of Trypanosoma cruzi from the Chaco Region

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