[Show abstract][Hide abstract] ABSTRACT: A total of 179 individuals with acute Chagas disease mainly transmitted by oral source, from Pará and Amapá State, Amazonian, Brazil were included during the period from 1988 to 2005. Blood samples were used to survey peripheral blood for T. cruzi hemoparasites by quantitative buffy coat (QBC), indirect xenodiagnosis, blood culture and serology to detection of total IgM and anti-T. cruzi IgG antibodies by indirect immunofluorescence assay (IFA) and indirect hemagglutination assay (HA). All assays were performed pre-treatment (0 days) and repeated 35 (±7) and 68 (±6) days after the initiation of treatment with benznidazol and every 6 months while remained seropositive. The endpoint of collection was performed in 2005. Total medium period of follow-up per person was 5.6 years. Also, a blood sample was collected from 72 randomly chosen treated patients to perform polimerase chain reaction (PCR) method. Proportions of subjects with negative or positive serology according to the number of years after treatment were compared. In the endpoint of follow-up we found 47 patients (26.7%) serologically negative, therefore considered cured and 5 (2.7%) exhibited mild cardiac Chagas disease. Other 132 patients had persistent positive serologic tests. The PCR carried out in 72 individuals was positive in 9.8%. Added, there was evidence of therapeutic failure immediately following treatment, as demonstrated by xenodiagnosis and blood culture methods in 2.3% and 3.5% of cases, respectively. There was a strong evidence of antibody clearing in the fourth year after treatment and continuous decrease of antibody titers. Authors suggest that control programs should apply operational researches with new drug interventions four years after the acute phase for those treated patients with persistently positive serology.
PLoS ONE 05/2013; 8(5):e64450. · 3.53 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Trypanosoma cruzi and Trypanosoma rangeli are human-infective blood parasites, largely restricted to Central and South America. They also infect a wide range of wild and domestic mammals and are transmitted by a numerous species of triatomine bugs. There are significant overlaps in the host and geographical ranges of both species. The two species consist of a number of distinct phylogenetic lineages. A range of PCR-based techniques have been developed to differentiate between these species and to assign their isolates into lineages. However, the existence of at least six and five lineages within T. cruzi and T. rangeli, respectively, makes identification of the full range of isolates difficult and time consuming. Here we have applied fluorescent fragment length barcoding (FFLB) to the problem of identifying and genotyping T. cruzi, T. rangeli and other South American trypanosomes. This technique discriminates species on the basis of length polymorphism of regions of the rDNA locus. FFLB was able to differentiate many trypanosome species known from South American mammals: T. cruzi cruzi, T. cruzi marinkellei, T. dionisii-like, T. evansi, T. lewisi, T. rangeli, T. theileri and T. vivax. Furthermore, all five T. rangeli lineages and many T. cruzi lineages could be identified, except the hybrid lineages TcV and TcVI that could not be distinguished from lineages III and II respectively. This method also allowed identification of mixed infections of T. cruzi and T. rangeli lineages in naturally infected triatomine bugs. The ability of FFLB to genotype multiple lineages of T. cruzi and T. rangeli together with other trypanosome species, using the same primer sets is an advantage over other currently available techniques. Overall, these results demonstrate that FFLB is a useful method for species diagnosis, genotyping and understanding the epidemiology of American trypanosomes.
Infection, genetics and evolution: journal of molecular epidemiology and evolutionary genetics in infectious diseases 10/2010; 11(1):44-51. · 3.22 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Acute Chagas disease (ACD) is caused by Trypanosoma cruzi. ACD outbreaks due to probable oral transmission occur regularly in small family gatherings that are exposed to contaminated foods. We studied two cohorts of residents on islands in the Breves and Bagre municipalities, in July and August 2007, to identify risk factors of transmission and to recommend preventative measures. Of the 25 cases identified in both cohorts, 13 (52%) were men, and the most frequent symptoms were fever (96%),asthenia (80%), myalgia (76%), abdominal pain (64%), retro-orbital pain, headaches and asthma (52%). We recommend detailed investigation of future outbreaks and other studies to better understand and control oral transmission of T. cruzi.
Tropical Doctor 10/2009; 39(4):231-2. · 0.53 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: We characterized 28 new isolates of Trypanosoma cruzi IIc (TCIIc) of mammals and triatomines from Northern to Southern Brazil, confirming the widespread distribution of this lineage. Phylogenetic analyses using cytochrome b and SSU rDNA sequences clearly separated TCIIc from TCIIa according to terrestrial and arboreal ecotopes of their preferential mammalian hosts and vectors. TCIIc was more closely related to TCIId/e, followed by TCIIa, and separated by large distances from TCIIb and TCI. Despite being indistinguishable by traditional genotyping and generally being assigned to Z3, we provide evidence that TCIIa from South America and TCIIa from North America correspond to independent lineages that circulate in distinct hosts and ecological niches. Armadillos, terrestrial didelphids and rodents, and domestic dogs were found infected by TCIIc in Brazil. We believe that, in Brazil, this is the first description of TCIIc from rodents and domestic dogs. Terrestrial triatomines of genera Panstrongylus and Triatoma were confirmed as vectors of TCIIc. Together, habitat, mammalian host and vector association corroborated the link between TCIIc and terrestrial transmission cycles/ecological niches. Analysis of ITS1 rDNA sequences disclosed clusters of TCIIc isolates in accordance with their geographic origin, independent of their host species.
Infection, genetics and evolution: journal of molecular epidemiology and evolutionary genetics in infectious diseases 08/2009; 9(6):1265-74. · 3.22 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Trypanosoma cruzi is the most important parasitic infection in Latin America and is also genetically highly diverse, with at least six discrete typing units (DTUs) reported: Tc I, IIa, IIb, IIc, IId, and IIe. However, the current six-genotype classification is likely to be a poor reflection of the total genetic diversity present in this undeniably ancient parasite. To determine whether epidemiologically important information is "hidden" at the sub-DTU level, we developed a 48-marker panel of polymorphic microsatellite loci to investigate population structure among 135 samples from across the geographic distribution of TcI. This DTU is the major cause of resurgent human disease in northern South America but also occurs in silvatic triatomine vectors and mammalian reservoir hosts throughout the continent. Based on a total dataset of 12,329 alleles, we demonstrate that silvatic TcI populations are extraordinarily genetically diverse, show spatial structuring on a continental scale, and have undergone recent biogeographic expansion into the southern United States of America. Conversely, the majority of human strains sampled are restricted to two distinct groups characterised by a considerable reduction in genetic diversity with respect to isolates from silvatic sources. In Venezuela, most human isolates showed little identity with known local silvatic strains, despite frequent invasion of the domestic setting by infected adult vectors. Multilocus linkage indices indicate predominantly clonal parasite propagation among all populations. However, excess homozygosity among silvatic strains and raised heterozygosity among domestic populations suggest that some level of genetic recombination cannot be ruled out. The epidemiological significance of these findings is discussed.
[Show abstract][Hide abstract] ABSTRACT: An outbreak of Chagas disease occurred in Mazagão, Amapá, Brazilian Amazon in 1996. Seventeen of 26 inhabitants presented symptoms compatible with acute Chagas disease and were submitted to parasitological and serological tests. All 17 were positive in at least one parasitological test and 11 were also IgM or IgG anti-Trypanosoma cruzi positive. The nine asymptomatic patients were negative for parasites and one was positive for IgG anti-T. cruzi. Sixty-eight triatomines were captured (66 Rhodnius pictipes; two Panstrongylus geniculatus); 45 were infected with T. cruzi (43 R. pictipes; two P. geniculatus). Thirteen trypanosomatid strains were isolated: eight from humans and five from R. pictipes. Four were genotyped as T. cruzi I (two from humans; two from R. pictipes), seven as T. cruzi Z3 (six from humans; one from R. pictipes) and two as T. cruzi Z3 and T. rangeli (from R. pictipes). Treatment started for all patients leading to a decrease in parasitaemia in 16 during the follow-up period (6 months, 1, 5 and 7 years). All were serologically negative 7 years post-treatment. There was an overlap of genotypes in the same ecotope, raising the possibility of transmission through the oral route and the need for early therapeutic intervention for better patient management in the Brazilian Amazon.
Transactions of the Royal Society of Tropical Medicine and Hygiene 03/2009; 103(3):291-7. · 1.93 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The objective of this report is to describe treatment outcomes over a four-year period of patients with acute Chagas disease in the Amazon region of Brazil. An outbreak of Chagas disease in a low-income district of urban Belém, in September 2000, affected 11 people simultaneously, indicating the likelihood of indirect, oral transmission of Trypanosoma cruzi. Prior to treatment, patients underwent physical and clinical tests; blood samples were processed with immunofluorescence assay (IFA) and quantitative buffy coat (QBC). Following treatment with benznidazole, parasitological and serologic tests (artificial xenodiagnosis and blood culture for T. cruzi), electrocardiogram, and echocardiogram were administered at intervals over a four-year period. Four years after treatment for acute Chagas disease, all patients presented with negative parasitological tests and persistent IgG anti-T. cruzi antibodies with lowered titers; three patients presented electrocardiogram abnormalities consistent with chronic Chagas disease or sequel of acute disease. The satisfactory response to treatment and relevance of serial parasitological examinations of patients with acute Chagas disease are discussed.
Revista Panamericana de Salud Pública 02/2009; 25(1):77-83. · 0.85 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: In this study, we provide phylogenetic and biogeographic evidence that the Trypanosoma cruzi lineages T. cruzi I (TCI) and T. cruzi IIa (TCIIa) circulate amongst non-human primates in Brazilian Amazonia, and are transmitted by Rhodnius species in overlapping arboreal transmission cycles, sporadically infecting humans. TCI presented higher prevalence rates, and no lineages other than TCI and TCIIa were found in this study in wild monkeys and Rhodnius from the Amazonian region. We characterised TCI and TCIIa from wild primates (16 TCI and five TCIIa), Rhodnius spp. (13 TCI and nine TCIIa), and humans with Chagas disease associated with oral transmission (14 TCI and five TCIIa) in Brazilian Amazonia. To our knowledge, TCIIa had not been associated with wild monkeys until now. Polymorphisms of ssrDNA, cytochrome b gene sequences and randomly amplified polymorphic DNA (RAPD) patterns clearly separated TCIIa from TCIIb-e and TCI lineages, and disclosed small intra-lineage polymorphisms amongst isolates from Amazonia. These data are important in understanding the complexity of the transmission cycles, genetic structure, and evolutionary history of T. cruzi populations circulating in Amazonia, and they contribute to both the unravelling of human infection routes and the pathological peculiarities of Chagas disease in this region.
International journal for parasitology 12/2008; 39(5):615-23. · 3.39 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Parasites of wild primates are important for conservation biology and human health due to their high potential to infect humans. In the Amazon region, non-human primates are commonly infected by Trypanosoma cruzi and T. rangeli, which are also infective to man and several mammals. This is the first survey of trypanosomiasis in a critically endangered species of tamarin, Saguinus bicolor (Callitrichidae), from the Brazilian Amazon Rainforest. Of the 96 free-ranging specimens of S. bicolor examined 45 (46.8%) yielded blood smears positive for trypanosomes. T. rangeli was detected in blood smears of 38 monkeys (39.6%) whereas T. cruzi was never detected. Seven animals (7.3%) presented trypanosomes of the subgenus Megatrypanum. Hemocultures detected 84 positive tamarins (87.5%). Seventy-two of 84 (85.7%) were morphologically diagnosed as T. rangeli and 3 (3.1%) as T. cruzi. Nine tamarins (9.4%) yielded mixed cultures of these two species, which after successive passages generated six cultures exclusively of T. cruzi and two of T. rangeli, with only one culture remaining mixed. Of the 72 cultures positive for T. rangeli, 62 remained as established cultures and were genotyped: 8 were assigned to phylogenetic lineage A (12.9%) and 54 to lineage B (87.1%). Ten established cultures of T. cruzi were genotyped as TCI lineage (100%). Transmission of both trypanosome species, their potential risk to this endangered species and the role of wild primates as reservoirs for trypanosomes infective to humans are discussed.
[Show abstract][Hide abstract] ABSTRACT: Two hundred and thirty-three cases of the acute phase of Chagas disease, from Pará, Amapá and Maranhão, were observed between 1988 and 2005. One hundred and sixty were studied retrospectively from 1988 to 2002 and seventy-three were prospectively followed up from 2003 to 2005. Among the cases studied, 78.5% (183/233) formed part of outbreaks, probably due to oral transmission (affecting a mean of 4 individuals), and 21.5% (50/233) were isolated cases. Cases were taken to be acute if they presented positive direct parasitological tests (fresh blood, thick drop or Quantitative Buffy Coat, QBC) and/or positive anti Trypanosoma cruzi IgM. Xenodiagnosis was also performed on 224 patients and blood culturing on 213. All the patients had clinical and epidemiological evaluations. The most frequent clinical manifestations were fever (100%), headache (92.3%), myalgia (84.1%), pallor (67%), dyspnea (58.4%), swelling of the legs (57.9%), facial edema (57.5%), abdominal pain (44.3%), myocarditis (39.9%) and exanthema (27%). The electrocardiogram showed abnormalities of ventricular repolarization in 38.5%, low QRS voltage in 15.4%, left-axis deviation in 11.5%, ventricular ectopic beats in 5.8%, bradycardia in 5.8%, tachycardia in 5.8%, right branch block in 4.8% and atrial fibrillation in 4.8%. The most frequently observed abnormality on the echocardiogram was pericardial effusion, in 46.2% of the cases. Thirteen (5.6%) patients died: ten (76.9%) of them due to cardiovascular involvement, two due to digestive complications and one due to indeterminate causes.
Revista da Sociedade Brasileira de Medicina Tropical 01/2008; 41(6):602-14. · 0.93 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The isolation of biological clones of Trypanosoma cruzi by microscopically dispensing individual organisms or by serial dilution is laborious and time consuming. The inability to resolve mixed T. cruzi infections, from vectors and hosts, and to isolate clones of slow growing genotypes by efficient plating on solid media, has hindered characterisation studies and downstream applications. We have devised and validated a sensitive, solid medium plating technique for rapid in vitro isolation of clones representative of all the recognised T. cruzi lineages (TCI, TCIIa-e), including the slow growing strain CANIII (TC IIa) and Trypanosoma rangeli, with high plating efficiencies. Furthermore, the method is effective for the isolation of clones directly from silvatic triatomine bugs and from experimentally infected mice harbouring mixed infections, allowing resolution of multiclonal infections from varied sources.
International Journal for Parasitology 02/2007; 37(1):111-20. · 3.40 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Four cases of serious cardiac attacks by autochthonous Trypanosoma cruzi infection from the Brazilian Amazon are reported; three of them occurred in micro-epidemic episodes. The manifestations included sudden fever, myalgia, dyspnea and signs of heart failure. Diagnosis was confirmed by specific exams, especially QBC (Quantitative Buffy Coat) and natural xenodiagnosis. Despite treatment with benznidazol, three patients died with serious myocarditis, renal failure and cardiac tamponade. The authors call attention to the emergence of this disease and reveal a previously unknown pathogenicity of T. cruzi strains in this area, added to a non-usual transmission form.
[Show abstract][Hide abstract] ABSTRACT: The genus Panstrongylus includes 14 species widely distributed from Mexico to Argentina, some of them with great epidemiological significance as vectors of Chagas disease. We study the karyotype and the male meiotic process of Panstrongylus chinai, P. geniculatus, P. herreri, P. lignarius, P. megistus, P. rufotuberculatus and P. tupynambai. All species present the same sex mechanism (X(1)X(2)Y in males and X(1)X(1)X(2)X(2) in females) and they also have 20 autosomes, with the exception of P. megistus that only presents 18 autosomes. The analysis of C-banding patterns and meiotic chromosome behaviour show a great level of variability allowing the identification of three clearly differentiated groups. In the first group, we only include P. megistus because of its unusual number of autosomes. The second group includes P. chinai, P. herreri, P. lignarius and P. rufotuberculatus. Their autosomes present terminal heterochromatic regions that appear scattered throughout the nucleus and associated with the sex chromosomes. Actually, P. herreri and P. lignarius can be considered cytogenetically identical. Our results are in agreement with morphological, ecological and molecular data indicating that they should be regarded as the same species. The third group only includes P. tupynambai that shows autosomes without C-positive regions. Panstrongylus geniculatus shares characters will all the three groups. Its karyotypic features are extremely polymorphic depending on their geographic origin. Some populations do not show any heterochromatic regions, while others exhibit few or several heterochromatic blocks. The chromosomal variability observed, together with its wide distribution and phenetic variability, suggest that P. geniculatus is a species complex comprising at least two distinct species. Considering the entire subfamily, the level of cytogenetic variation in Panstrongylus is lower than that observed in Triatoma but considerably more than that of Rhodnius, which is a very homogenous genus in terms of chromosome appearance and behaviour. This would endorse the closer relationship between Panstrongylus and Triatoma, and their divergence from Rhodnius, in accordance with current tribal classification.
Infection Genetics and Evolution 11/2002; 2(1):47-56. · 3.26 Impact Factor