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BACKGROUND: Chagas disease (CD), caused by Trypanosoma cruzi, poses a major global public health challenge. Although vector-borne transmission is the primary mode of infection, oral transmission is increasingly concerning. METHODS: This study utilized long-amplicon-based sequencing (long-ABS), focusing on the 18S rRNA gene, to explore T. cruzi's genetic diversity and transmission dynamics during an acute CD outbreak in Colombia, an area without domestic infestation. RESULTS: Analyzing samples from five patients and five T. cruzi-positive marsupial samples, we identified coinfections between T. cruzi and Trypanosoma rangeli, mixed T. cruzi DTUs, suggesting possible links between human and marsupial T. cruzi infections. Coexistence of TcI, TcIV and T. rangeli suggests marsupial secretions as the possible source of T. cruzi transmission. Our investigation revealed diversity loss in DTUs TcIV and T. rangeli in humans after infection and in marsupial samples after culture. CONCLUSION: These findings provide significant insights into T. cruzi dynamics, crucial for implementing control and prevention strategies.
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Doença de Chagas , Surtos de Doenças , Variação Genética , Sequenciamento de Nucleotídeos em Larga Escala , Marsupiais , RNA Ribossômico 18S , Trypanosoma cruzi , Doença de Chagas/transmissão , Doença de Chagas/epidemiologia , Doença de Chagas/parasitologia , Trypanosoma cruzi/genética , Trypanosoma cruzi/isolamento & purificação , Humanos , Animais , Marsupiais/parasitologia , RNA Ribossômico 18S/genética , Colômbia/epidemiologia , Masculino , Coinfecção/epidemiologia , Coinfecção/parasitologia , Coinfecção/transmissão , Trypanosoma rangeli/genética , Feminino , Adulto , DNA de Protozoário/genéticaRESUMO
We assessed the diversity of triatomines, the rates of natural infection, and the discrete typing units (DTUs) of Trypanosoma cruzi isolated from them in two municipalities in the state of Sergipe, Brazil. Active searches for triatomines were conducted in the peridomicily and wild enviroments of 10 villages within the two municipalities. Triatomines were taxonomically identified and their feces were extracted using the abdominal compression method. Parasite detection was performed using optical microscopy. For Trypanosoma cruzi genotyping via PCR-FFLB, 151 samples of the subspecies Triatoma brasiliensis macromelasoma and Triatoma brasiliensis were isolated from both municipalities. In total, 505 triatomines were collected, with Triatoma brasiliensis macromelasoma being the most frequent species (58.81 %). Triatoma b. brasiliensis was the only species in both peridomestic and wild environments. Regarding the other species, T. pseudomaculata was found only in the peridomestic environment; and T. b. macromelasoma and Psammolestes tertius were found in the wild environment. Three Discrete Typing Units were identified: TcI (87.51 %) detected in T. b. brasiliensis and T. b. macromelasoma, TcI+TcIII (10.41 %) in T. b. macromelasoma, and TcI+Trypanosoma rangeli (2.08 %) in T. b. macromelasoma. It is concluded that T. b. macromelasoma is the species collected most frequently in the studied region and the one that presents the highest rates of natural infection, highlighting its epidemiological importance for the vectorial transmission of Chagas disease in Sergipe.
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Doença de Chagas , Genótipo , Insetos Vetores , Triatoma , Trypanosoma cruzi , Animais , Brasil , Trypanosoma cruzi/genética , Trypanosoma cruzi/classificação , Trypanosoma cruzi/isolamento & purificação , Doença de Chagas/parasitologia , Doença de Chagas/transmissão , Doença de Chagas/epidemiologia , Triatoma/parasitologia , Triatoma/classificação , Insetos Vetores/parasitologia , Insetos Vetores/classificação , Fezes/parasitologia , HumanosRESUMO
BACKGROUND: Chagas disease is caused by Trypanosoma cruzi, whose genetic structure is divided into six discrete typing units (DTUs) known as TcI-TcVI. In the Yucatan Peninsula, Mexico, information regarding the DTUs circulating in wild mammals is scarce, while this is important knowledge for our understanding of T. cruzi transmission dynamics. METHODS: In the current study, we sampled wild mammals in a sylvatic site of the Yucatan Peninsula and assessed their infection with T. cruzi by PCR. Then, for infected mammals, we amplified and sequenced nuclear and mitochondrial T. cruzi genetic markers for DTU identification. RESULTS: In total, we captured 99 mammals belonging to the orders Chiroptera, Rodentia and Didelphimorphia. The prevalence of infection with T. cruzi was 9% (9/99; 95% CI [5, 16]), and we identified TcI in a Jamaican fruit bat, Artibeus jamaicensis. Moreover, we fortuitously identified Trypanosoma dionisii in another Jamaican fruit bat and detected an unidentified Trypanosoma species in a third specimen. While the latter discoveries were not expected because we used primers designed for T. cruzi, this study is the first to report the identification of T. dionisii in a bat from Yucatan, Mexico, adding to a recent first report of T. dionisii in bats from Veracruz, and first report of this Trypanosoma species in Mexico. CONCLUSION: Further research is needed to enhance our knowledge of T. cruzi DTUs and Trypanosoma diversity circulating in wildlife in Southeastern Mexico.
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Doença de Chagas , Quirópteros , Trypanosoma cruzi , Animais , México/epidemiologia , Quirópteros/parasitologia , Trypanosoma cruzi/genética , Trypanosoma cruzi/isolamento & purificação , Doença de Chagas/veterinária , Doença de Chagas/epidemiologia , Doença de Chagas/transmissão , Reação em Cadeia da Polimerase , DNA de Protozoário , Prevalência , Trypanosoma/isolamento & purificação , Trypanosoma/genética , Trypanosoma/classificação , Roedores/parasitologiaRESUMO
Chagas disease, caused by Trypanosoma cruzi, remains a serious public health problem worldwide. The parasite was subdivided into six distinct genetic groups, called "discrete typing units" (DTUs), from TcI to TcVI. Several studies have indicated that the heterogeneity of T. cruzi species directly affects the diversity of clinical manifestations of Chagas disease, control, diagnosis performance, and susceptibility to treatment. Thus, this review aims to describe how T. cruzi genetic diversity influences the biology of the parasite and/or clinical parameters in humans. Regarding the geographic dispersion of T. cruzi, evident differences were observed in the distribution of DTUs in distinct areas. For example, TcII is the main DTU detected in Brazilian patients from the central and southeastern regions, where there are also registers of TcVI as a secondary T. cruzi DTU. An important aspect observed in previous studies is that the genetic variability of T. cruzi can impact parasite infectivity, reproduction, and differentiation in the vectors. It has been proposed that T. cruzi DTU influences the host immune response and affects disease progression. Genetic aspects of the parasite play an important role in determining which host tissues will be infected, thus heavily influencing Chagas disease's pathogenesis. Several teams have investigated the correlation between T. cruzi DTU and the reactivation of Chagas disease. In agreement with these data, it is reasonable to suppose that the immunological condition of the patient, whether or not associated with the reactivation of the T. cruzi infection and the parasite strain, may have an important role in the pathogenesis of Chagas disease. In this context, understanding the genetics of T. cruzi and its biological and clinical implications will provide new knowledge that may contribute to additional strategies in the diagnosis and clinical outcome follow-up of patients with Chagas disease, in addition to the reactivation of immunocompromised patients infected with T. cruzi.
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Doença de Chagas , Variação Genética , Trypanosoma cruzi , Trypanosoma cruzi/genética , Humanos , Doença de Chagas/imunologia , Doença de Chagas/parasitologia , Animais , Interações Hospedeiro-Parasita/genética , Interações Hospedeiro-Parasita/imunologiaRESUMO
Background: The Trypanosoma cruzi parasite is the causal agent of Chagas disease, recognized by the World Health Organization as a neglected tropical disease. Currently there are seven discrete typing units (DTUs) of T. cruzi distributed in America, but there are still gaps about its distribution in some endemic regions. Materials and Methods: Seventeen units isolated from Chiapas and Oaxaca in Mexico were identified by amplification of the C-5 sterol desaturase gene. Results: Three DTUs of T. cruzi, TcI (6), TcII (10), and TcIV (1) were detected by comparing polymorphic sites in specific regions. Conclusions: New DTUs are reported for both states, where TcII was the most common DTU. The genetic characterization of the isolates can help to understand the epidemiology of Chagas disease.
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Benznidazole (Bz) is the recommended drug for the treatment of Chagas disease; however, its efficacy may vary according to the sensitivity of Trypanosoma cruzi strains to the drug and host immune background. The study evaluated the immune response of peripheral blood mononuclear cells (PBMC) that were infected in vitro with the Colombian strain (Col) and treated with Bz. The co-cultures were incubated for 24 h, 5 and 10 days, where cytokine dosage was performed in the supernatant and evaluation of the cells for CD28+ and CTLA-4+ molecules in CD4+ and CD8+ lymphocytes, and CD80+ , CD86+ and HLA-DR+ in CD14+ cells. The results showed that Col induced a strong inflammatory response, with an increase in IFN-γ and TNF early in the infection (24 h), however, from 5 days of infection on, TNF production declined, and IL-10 production increased, which may be associated with a control mechanism of the exacerbated inflammatory response. The Bz treatment did not significantly alter the frequencies of the phenotypes evaluated both T cell subsets and CD14+ cells. Therefore, this study reinforces the need for typing the patient's strain to guide therapy and promote individualized treatment protocols due to the heterogeneous genetic background among T. cruzi strains.
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Doença de Chagas , Nitroimidazóis , Tripanossomicidas , Trypanosoma cruzi , Humanos , Leucócitos Mononucleares , Colômbia , Nitroimidazóis/farmacologia , Nitroimidazóis/uso terapêutico , Doença de Chagas/tratamento farmacológico , Tripanossomicidas/farmacologia , Tripanossomicidas/uso terapêuticoRESUMO
ABSTRACT Chagas disease (CD) is a neglected tropical disease caused by Trypanosoma cruzi and is genetically classified in six discrete typing units (DTUs). The isolates reported in Mexico are generally associated with DTU I. We presented a case of a prolonged cutaneous lesion in a Mexican man, caused by DTU II in coinfection with Bacillus velezensis and Corynebacterium sp. The patient assessment included a complete clinical history, physical exam, laboratory tests, and a skin biopsy. In the facial tissues, intracellular parasites were revealed. The PCR tests were positive for T. cruzi in tissue and blood samples. DNA satellite sequencing was correlated with the DTU II. The initial serological tests reported negative results. However, four months later, two serological tests reported positive results. These exams were performed in different health centers. Mexico is considered an endemic area for CD; nevertheless, this is just the second cutaneous case associated with a DTU different from DTU-I noted in this country. From an ecological point of view, this fact suggests a geographical expansion of DTU II and an association with atypical skin manifestations. Further studies should be conducted to understand this exciting association between DTU-II and prolonged cutaneous expression in humans.
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Parasites are important components of the immense n-dimensional trophic network that connects all living beings because they, among others, forge biodiversity and deeply influence ecological evolution and host behavior. In this sense, the influence of Trypanosomatidae remains unknown. The aim of this study was to determine trypanosomatid infection and richness in rats, opossums, and dogs in the semiarid Caatinga biome. We submitted DNA samples from trypanosomatids obtained through axenic cultures of the blood of these mammals to mini exon multiplex-PCR, Sanger, and next-generation sequencing targeting the 18S rDNA gene. Phylogenetic analyses were performed to identify genetic diversity in the Trypanosomatidae family. Shannon, Simpson, equability, and beta-diversity indices were calculated per location and per mammalian host. Dogs were surveyed for trypanosomatid infection through hemocultures and serological assays. The examined mammal species of this area of the Caatinga biome exhibited an enormous trypanosomatid species/genotypes richness. Ten denoised Operational Taxonomic Units (ZOTUs), including three species (Trypanosoma cruzi, Trypanosoma rangeli and Crithidia mellificae) and one Trypanosoma sp. five genotypes/lineages (T. cruzi DTU TcI, TcII, and TcIV; T. rangeli A and B) and four DTU TcI haplotypes (ZOTU1, ZOTU2, ZOTU5, and ZOTU10 merged), as well as 13 Amplicon Sequence Variants (ASVs), including five species (T. cruzi, T. rangeli, C. mellificae, Trypanosoma dionisii, and Trypanosoma lainsoni), five genotypes/lineages (same as the ZOTUs) and six DTU TcI haplotypes (ASV, ASV1, ASV2, ASV3, ASV5 and ASV13), were identified in single and mixed infections. We observed that trypanosomatids present a broad host spectrum given that species related to a single host are found in other mammals from different taxa. Concomitant infections between trypanosomatids and new host-parasite relationships have been reported, and this immense diversity in mammals raised questions, such as how this can influence the course of the infection in these animals and its transmissibility. Dogs demonstrated a high infection rate by T. cruzi as observed by positive serological results (92% in 2005 and 76% in 2007). The absence of positive parasitological tests confirmed their poor infectivity potential but their importance as sentinel hosts of T. cruzi transmission.
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Doença de Chagas , Trypanosomatina , Animais , Brasil/epidemiologia , Cães , Ecossistema , Gambás , Filogenia , RatosRESUMO
Trypanosoma cruzi is the causative agent of Chagas disease, a devastating parasitic disease endemic to Central and South America, Mexico, and the USA. We characterized the genetic diversity of Trypanosoma cruzi circulating in five triatomine species (Triatoma gerstaeckeri, T. lecticularia, T.indictiva, T. sanguisuga and T. recurva) collected in Texas and Southern Arizona using multilocus sequence typing (MLST) with four single-copy loci (cytochrome oxidase subunit II- NADH dehydrogensase subunit 1 region (COII-ND1), mismatch-repair class 2 (MSH2), dihydrofolate reductase-thymidylate synthase (DHFR-TS) and a nuclear gene with ID TcCLB.506529.310). All T. cruzi variants fall in two main genetic lineages: 75% of the samples corresponded to T. cruzi Discrete Typing Unit (DTU) I (TcI), and 25% to a North American specific lineage previously labelled TcIV-USA. Phylogenetic and sequence divergence analyses of our new data plus all previously published sequence data from those four loci collected in the USA, show that TcIV-USA is significantly different from any other previously defined T. cruzi DTUs. The significant level of genetic divergence between TcIV-USA and other T. cruzi DTUs should lead to an increased focus on understanding the epidemiological importance of this DTU, as well as its geographical range and pathogenicity in humans and domestic animals. Our findings further corroborate the fact that there is a high genetic diversity of the parasite in North America and emphasize the need for appropriate surveillance and vector control programs for Chagas disease in southern USA and Mexico.
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Doença de Chagas , Trypanosoma cruzi , Animais , Insetos Vetores/parasitologia , Tipagem de Sequências Multilocus , Filogenia , Sudoeste dos Estados Unidos/epidemiologia , Texas/epidemiologia , Trypanosoma cruzi/genéticaRESUMO
Background: Chagas disease (CD), caused by the protozoan Trypanosoma cruzi, is considered a public health problem in Latin America. In Colombia, it affects more than 437,000 inhabitants, mainly in Casanare, an endemic region with eco-epidemiological characteristics that favor its transmission. The objective of this study was to describe the clinical and epidemiological characteristics of the cases of acute CD in Casanare, eastern Colombia, in the period 2012-2020. Methods: In the present study, 103 medical records of confirmed cases of acute CD were reviewed. The departmental/national incidence and fatality were compared by year; the climatological data of mean temperature, relative humidity, and precipitation per year were reviewed and plotted at IDEAM (Colombian Meteorology Institute) concerning the number of cases of acute CD per month, and it was compared with the frequency of triatomines collected in infested houses by community surveillance. Univariate, bivariate, and multivariate analyses were performed, comparing symptoms and signs according to transmission routes, complications, and age groups. Results: The incidence was 3.16 cases per 100,000 inhabitants, and the fatality rate was 20% in the study period. The most frequent symptoms included: fever 98.1%, myalgia 62.1%, arthralgia 60.2%, and headache 49.5%. There were significant differences in the frequency of myalgia, abdominal pain, and periorbital edema in oral transmission. The main complications were pericardial effusion, myocarditis, and heart failure in the group over 18 years of age. In Casanare, TcI Discrete Typing Unit (DTU) has mainly been identified in humans, triatomines, and reservoirs such as opossums and dogs and TcBat in bats. An increase in the number of acute CD cases was evidenced in March, a period when precipitation increases due to the beginning of the rainy season. Conclusions: The results corroborate the symptomatic heterogeneity of the acute phase of CD, which delays treatment, triggering possible clinical complications. In endemic regions, clinical suspicion, diagnostic capacity, detection, and surveillance programs should be strengthened, including intersectoral public health policies for their prevention and control.
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Strains of Trypanosoma cruzi, etiological agent of Chagas disease, are classified into different discrete typing units that may present distinct dynamics of infection and susceptibility to benznidazole (BZ) treatment. Mice that were orally inoculated with T. cruzi IV strains exhibited a more intense course of infection compared with intraperitoneally inoculated mice, reflected by higher parasite loads. We evaluated the efficacy of BZ treatment in Swiss mice that were inoculated with T. cruzi IV strains from the Western Brazilian Amazon. The mice were orally (OR) or intraperitoneally (IP) inoculated with 2 × 106 culture-derived metacyclic trypomastigotes of the AM14, AM16, AM64, and AM69 strains of T. cruzi that were obtained from two outbreaks of orally acquired acute Chagas disease in the state of Amazonas, Brazil. The animals were treated with BZ (100 mg/kg/day for 20 days). Fresh blood examination, hemoculture, conventional and quantitative real-time polymerase chain reaction were performed to monitor the therapeutic effects of BZ. Significant reductions in five of 24 parameters of parasitemia and parasite load were found in different tissues in the OR group, indicating worse response to BZ treatment compared with the IP group, in which significant reductions in nine of those 24 parameters were observed. The cure rates in the OR groups ranged from 18.2% (1/11) to 75.0% (9/12) and in the IP groups from 58.3% (7/12) to 91.7% (11/12), for the AM14 and AM69 strains, respectively. These findings indicate that treatment with BZ had fewer beneficial effects with regard to reducing parasitemia and parasite load in different tissues of mice that were OR inoculated with four TcIV strains compared with IP inoculation. Therefore, the route of infection with T. cruzi should be considered when evaluating the therapeutic efficacy of BZ in patients with Chagas disease.
Assuntos
Doença de Chagas/parasitologia , Nitroimidazóis/uso terapêutico , Tripanossomicidas/uso terapêutico , Trypanosoma cruzi/classificação , Parede Abdominal/parasitologia , Animais , Brasil/epidemiologia , Doença de Chagas/tratamento farmacológico , Doença de Chagas/epidemiologia , Esôfago/parasitologia , Coração/parasitologia , Camundongos , Nitroimidazóis/farmacologia , Carga Parasitária , Parasitemia/tratamento farmacológico , Parasitemia/epidemiologia , Parasitemia/parasitologia , Estômago/parasitologia , Tripanossomicidas/farmacologia , Trypanosoma cruzi/efeitos dos fármacosRESUMO
Background: Trypanosoma cruzi has a high rate of biological and genetic variability, and its population structure is divided into seven distinct genetic groups (TcI-TcVI and Tcbat). Due to immigration, Chagas disease (ChD), caused by T. cruzi, has become a serious global health problem including in Europe. Therefore, the aim of this study was to evaluate the existence of genetic variability within discrete typing unit (DTU) TcV of T. cruzi in Bolivian patients with chronic ChD residing in Barcelona, Spain. Methods: The DNA was extracted from the peripheral blood of 27 patients infected with T. cruzi DTU TcV and the fragments of the genetic material were amplificated through the low stringency single primer-polymerase chain reaction (LSSP-PCR). The data generated after amplification were submitted to bioinformatics analysis. Results: Of the 27 patients evaluated in the study, 8/27 (29.6%) were male and 19/27 (70.4%) female, 17/27 (62.9%) were previously classified with the indeterminate clinical form of Chagas disease and 10/27 (37.1%) with Chagas cardiomyopathy. The LSSP-PCR detected 432 band fragments from 80 to 1,500 bp. The unweighted pair-group method analysis and principal coordinated analysis data demonstrated the existence of three distinct genetic groups with moderate-high rates of intraspecific genetic variability/diversity that had shared parasite's alleles in patients with the indeterminate and cardiomyopathy forms of ChD. Conclusions: This study demonstrated the existence of a moderate to high rate of intra-DTU TcV variability in T. cruzi. Certain alleles of the parasite were associated with the absence of clinical manifestations in patients harboring the indeterminate form of ChD. These results support the need to search for increasingly specific targets in the genome of T. cruzi to be correlated with its main biological properties and clinical features in patients with chronic ChD.
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Trypanosoma cruzi, as other kinetoplastids, has a complex mechanism of editing of mitochondrial mRNAs that requires guide RNAs (gRNAs) coded in DNA minicircles in the kinetoplast. There are many variations on this mechanism among species. mRNA editing and gRNA repertoires are almost unknown in T. cruzi. Here, gRNAs were inferred based on deep-sequenced minicircle hypervariable regions (mHVRs) and editing cascades were rebuilt in strains belonging to the six main T. cruzi lineages. Inferred gRNAs were clustered according to their sequence similarity to constitute gRNA classes. Extreme diversity of gRNA classes was observed, which implied highly divergent gRNA repertoires among different lineages, even within some lineages. In addition, a variable gRNA class redundancy (i.e., different gRNA classes editing the same mRNA region) was detected among strains. Some strains had upon four times more gRNA classes than others. Such variations in redundancy affected gRNA classes of all mRNAs in a concerted way, i.e., there are correlated variations in the number of gRNAs classes editing each mRNA. Interestingly, cascades were incomplete for components of the respiratory complex I in several strains. Finally, gRNA classes of different strains may potentially edit mitochondrial mRNAs from other lineages in the same way as they edit their own mitochondrial mRNAs, which is a prerequisite for biparental inheritance of minicircle in hybrids. We propose that genetic exchange and biparental inheritance of minicircles combined with minicircle drift due to (partial) random segregation of minicircles during kDNA replication is a suitable hypothesis to explain the divergences among strains and the high levels of gRNA redundancy in some strains. In addition, our results support that the complex I may not be required in some stages in the life cycle as previously shown and that linkage (in the same minicircle) of gRNAs that edit different mRNAs may prevent gRNA class lost in such stage.
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RNA Guia de Cinetoplastídeos , Trypanosoma brucei brucei , Trypanosoma cruzi , Sequência de Bases , DNA de Cinetoplasto , RNA Guia de Cinetoplastídeos/genética , Trypanosoma brucei brucei/genética , Trypanosoma cruzi/genéticaRESUMO
Trypanosoma cruzi is the etiologic agent of American trypanosomiasis and can infect humans and different species of domestic and wild animals. The marsupials are important wild reservoirs of T. cruzi, aiding in the maintenance of this agent in sylvatic and peri-domestic environments. The objective of this study was to report the parasitological and clinicopathological findings of a natural infection by T. cruzi in one specimen of Philander opossum that originated from the Brazilian Amazon. The animal was captured in a forest fragment near a rural community with reports of human Chagas disease. T. cruzi infection was diagnosed by blood smear examinations, blood culture, scent glands secretion culture, histopathological examination, and nested-PCR. Positive samples were subjected to PCR to characterize the discrete typing units (DTUs) of T. cruzi. Characteristic trypomastigotes of T. cruzi were observed in the blood smear, and spheromastigotes, epimastigotes, and trypomastigotes were visualized in the cultures. Non-suppurative myocarditis associated with amastigote clusters was the principal histopathological finding. DNA from T. cruzi was detected in samples of blood, blood cultures, scent glands secretion cultures, cardiac muscles, and the spleen. The TcI and the TcII/V/VI group DTUs were detected in blood culture and scent glands secretion cultures. Infection by T. cruzi can cause myocarditis in P. opossum and DTUs TcI and TcII/V/VI group mixed infection can be detected in the acute phase. P. opossum can be a source of infection for triatomine vectors and has the potential source for direct transmission of T. cruzi by secretions from the scent glands. These data are important to improve the understanding of the complex enzootic transmission cycle of T. cruzi in the Brazilian Amazon.
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Doença de Chagas/veterinária , Gambás , Trypanosoma cruzi/isolamento & purificação , Animais , Doença de Chagas/parasitologia , Doença de Chagas/patologia , Coração/parasitologia , Masculino , Miocárdio/patologia , Glândulas Odoríferas/parasitologia , Glândulas Odoríferas/patologiaRESUMO
BACKGROUND: Chagas disease is caused by the haemoflagellate protozoan Trypanosoma cruzi. Currently, T. cruzi recognizes seven discrete typing units (DTUs): TcI to TcVI and Tcbat. The genetic diversity of T. cruzi is suspected to influence the clinical outcome. Acute clinical manifestations, which include myocarditis and meningoencephalitis, are sometimes fatal; occur most frequently in children and in immunocompromised individuals. Acute disease is often overlooked, leading to a poor prognosis. CASE PRESENTATION: A 38-year-old man from a subtropical area of the Andes mountains of Ecuador was hospitalized after 3 weeks of evolution with high fever, chills, an enlarged liver, spleen, and lymph nodes, as well as facial edema. ECG changes were also observed. T. cruzi was identified in blood smears, culture and amplification of DNA by PCR. Tests for anti-T. cruzi IgG and IgM and HIV were negative. Molecular typing by restriction fragment length polymorphism (PCR-RFLP) determined the parasite to DTU TcI. In the absence of a timely anti-T. cruzi medication, the patient died. CONCLUSIONS: This is a case of severe pathogenicity and the virulence of a DTU TcI strain in an adult patient. The severe acute Chagas disease was probably overlooked due to limited awareness and its low incidence. Our findings suggest that T. cruzi DTU TcI strains circulating in Ecuador are capable of causing fatal acute disease. Early diagnosis and prompt treatment is of paramount importance to avoid fatalities in acute infections.
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Doença de Chagas/etiologia , Trypanosoma cruzi/genética , Trypanosoma cruzi/patogenicidade , Adulto , Doença de Chagas/parasitologia , Equador , Variação Genética , Humanos , Masculino , Tipagem Molecular , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , Trypanosoma cruzi/classificaçãoRESUMO
Trypanosoma cruzi, the etiological agent of Chagas disease, was initially classified into 6 Discrete Typing Units (DTUs). The hybrid DTUs TcV and TcVI are the most frequent in domestic transmission cycles throughout the Southern Cone countries of South America. Here, we genotyped parasite isolates from human residents in Pampa del Indio municipality, Chaco, to further characterize the structure of T. cruzi populations, and to assess the degree of overlapping between the domestic and sylvatic transmission cycles. Artificial xenodiagnostic tests were performed to blood samples from 125â¯T. cruzi-seropositive people (age range, 3-70â¯years) who represented 14.3% of all seropositive residents identified. Parasites were obtained from feces of T. cruzi-infected Triatoma infestans examined 30 or 60â¯days after blood-feeding, and grown in vitro. The cultured parasites were genotyped by means of two PCR-based protocols. DTUs were determined from 39 (31%) patients residing in 28 dwellings. The only DTUs identified were TcV (92%) and TcVI (8-36%). Households with more than one parasite isolate consistently displayed the same DTU. Further sequencing of a fragment of the TcMK gene from selected samples argue against the occurrence of mixed TcV-TcVI infections in the study population. Sequencing data revealed an unexpected degree of genetic variability within TcV including two apparently robust subgroups of isolates. Our results for human residents confirm the predominance of hybrid lineages (TcV and to a much lesser extent TcVI) and the absence of sylvatic genotypes (TcI and TcIII) in (peri)domestic transmission cycles in the Argentinean Chaco area. 245 words.
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Doença de Chagas/epidemiologia , Doença de Chagas/parasitologia , DNA de Protozoário , Hibridização Genética , Trypanosoma cruzi/genética , Argentina/epidemiologia , Doença de Chagas/diagnóstico , Doença de Chagas/transmissão , Genótipo , Geografia Médica , Humanos , Epidemiologia Molecular , Mutação , Polimorfismo de Nucleotídeo Único , Vigilância da População , Trypanosoma cruzi/isolamento & purificação , XenodiagnósticoRESUMO
Chagas disease is a major public health problem in Latin America and has spread to other countries due to immigration of infected persons. 10-30% of patients with chronic Chagas disease will develop cardiomyopathy. Chagas cardiomyopathy is the worst form of the disease, due to its high morbidity and mortality. Because of its prognostic value and adequate medical monitoring, it is very important to identify infected people who could develop Chagas cardiomyopathy. The aim of this study was to determine if discrete typing units (DTUs) of Trypanosoma cruzi are related to the presence of heart disease in patients with chronic Chagas disease. A total of 86 untreated patients, 41 with cardiomyopathy and 45 without heart involvement were submitted to clinical study. Electrocardiograms and echocardiograms were performed on the group of cardiopaths, in which all important known causes of cardiomyopathy were discarded. Sinus bradycardia and prolonged QTc interval were the most frequent electrocardiographic alterations and patients were classified in group I (46%) and group II (54%) of New York Hearth Association. In all cases real-time PCR genotyping assays were performed. In the group with cardiomyopathy, the most frequent DTU was TcI (56.1%), followed by TcII (19.5%). Mixed infections TcIâ¯+â¯TcII were observed in 7.3% of the patients. In the group without cardiac pathologies, TcI and TcII were found at similar rates (28.9 and 31.1%, respectively) and mixed infections TcIâ¯+â¯TcII in 17.8% of the cases. TcIII and TcIV were not detected in any sample. Taken together, our data indicate that chronic Chagas cardiomyopathy in Chile can be caused by strains belonging to TcI and TcII.
Assuntos
Cardiomiopatia Chagásica/parasitologia , Genótipo , Tipagem Molecular , Trypanosoma cruzi/genética , Adulto , Idoso , Doença de Chagas/parasitologia , Chile , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase em Tempo Real , Adulto JovemRESUMO
INTRODUCTION: Phylogenetic relationships between different lineages of Trypanosoma cruzi, the agent of Chagas disease, have been controversial for several years. However, recent phylogenetic and phylogenomic analyses clarified the nuclear relationships among such lineages. However, incongruence between nuclear and kinetoplast DNA phylogenies has emerged as a new challenge. This incongruence implies several events of mitochondrial introgression at evolutionary level. However, the mechanism that gave origin to introgressed lineages is unknown. Here, I will review and discuss how maxicircles of the kinetoplast were horizontally and vertically transferred between different lineages of T. cruzi. CONCLUSION: Finally, I will discuss what we know - and what we don't - about the kDNA transference and inheritance in the context of sexual reproduction in this parasite.
RESUMO
Abstract INTRODUCTION: Trypanosoma cruzi, the etiologic agent of Chagas disease, is widely distributed in nature, circulating between triatomine bugs and sylvatic mammals, and has large genetic diversity. Both the vector species and the genetic lineages of T. cruzi present a varied geographical distribution. This study aimed to verify the influence of sympatry in the interaction of T. cruzi with triatomines. Methods: The behavior of the strains PR2256 (T. cruzi II) and AM14 (T. cruzi IV) was studied in Triatoma sordida (TS) and Rhodnius robustus (RR). Eleven fifth-stage nymphs were fed by artificial xenodiagnosis with 5.6 × 103 blood trypomastigotes/0.1mL of each T. cruzi strain. Every 20 days, their excreta were examined for up to 100 days, and every 30 days, the intestinal content was examined for up to 120 days, by parasitological (fresh examination and differential count with Giemsa-stained smears) and molecular (PCR) methods. Rates of infectivity, metacyclogenesis and mortality, and mean number of parasites per insect and of excreted parasites were determined. RESULTS: Sympatric groups RR+AM14 and TS+PR2256 showed higher values of the four parameters, except for mortality rate, which was higher (27.3%) in the TS+AM14 group. General infectivity was 72.7%, which was mainly proven by PCR, showing the following decreasing order: RR+AM14 (100%), TS+PR2256 (81.8%), RR+PR2256 (72.7%) and TS+AM14 (36.4%). CONCLUSIONS: Our working hypothesis was confirmed once higher infectivity and vector capacity (flagellate production and elimination of infective metacyclic forms) were recorded in the groups that contained sympatric T. cruzi lineages and triatomine species.
Assuntos
Humanos , Animais , Vetores Artrópodes/fisiologia , Rhodnius/fisiologia , Triatoma/fisiologia , Trypanosoma cruzi/fisiologia , Simpatria , Vetores Artrópodes/genética , Vetores Artrópodes/patogenicidade , Rhodnius/genética , Rhodnius/patogenicidade , Especificidade da Espécie , Fatores de Tempo , Triatoma/genética , Triatoma/patogenicidade , Trypanosoma cruzi/genética , Trypanosoma cruzi/patogenicidade , Sangue/parasitologia , Brasil , Reação em Cadeia da Polimerase , Doença de Chagas/parasitologia , Doença de Chagas/transmissão , Xenodiagnóstico/métodos , Interações Hospedeiro-Parasita/fisiologia , Intestinos/parasitologia , CamundongosRESUMO
Los trabajos realizados de trampeo y captura de triatominos (que son los insectos vectores de la enfermedad de Chagas) en zonas silvestres de los alrededores de la localidad de Huayhuasi, municipio de Mecapaca en el departamento de La Paz. Se colocaron 50 trampas en lugares estratégicos escogidos de acuerdo a las características del área de estudio. De las 50 trampas, en 9 trampas se lograron atrapar a los triatominos, lo que representa el 4.5% de infestación de la zona de estudio. Se capturaron a 13 insectos (7 ninfas de diferentes estadíos y 6 adultos). Las características morfológicas de los insectos fueron las típicas del Triatoma infestans. En el examen en fresco de las deyecciones de los 6 especímenes adultos capturados, en 4 de ellos se detectó presencia del parásito Trypanosoma cruzi, lo cual demuestra un 67% de infección, porcentaje bastante significativo en consideración a la cercanía de estos lugares infestados a las zonas urbanas del municipio de Mecapaca. Los parásitos que los especímenes portaban, fueron aislados y cultivados en medio de cultivo LIT (Liver Infusion Tryptose) suplementado con 10% de suero bovino fetal e incubados en estufa a 26°C. El análisis del ADN extraído de los parásitos, ha revelado la DTU (Discrete Typing Unit) correspondiente al linaje TcI.
Trapping and capture of triatomines (which are the insect's vectors of Chagas' disease) were carried out in wild areas around the town of Huayhuasi, municipality of Mecapaca, in the department of La Paz. 50 traps were placed in strategic places chosen according to the characteristics already established. Of the 50 traps, nine traps managed to trap the triatomines, representing 4.5% of infestation in the study area. 13 insects (7 nymphs of different stages and 6 adults) were captured. The morphological characteristics of the insects were typical of Triatoma infestans. In the fresh examination of the excrement of the six captured adult specimens, in 4 of them the presence of the parasite Trypanosoma cruzi was detected, which shows a 67% infection, a very significant percentage considering the proximity of these infested places to the Urban areas of the municipality of Mecapaca. The parasites that the specimens carried were isolated and cultured in LIT (Liver Infusion Triptose) medium supplemented with 10% fetal bovine serum and incubated in an oven at 26 °C. Analysis of the DNA extracted from the parasites has revealed the DTU corresponding to the Tc I lineage.