Ecological diversity of Trypanosoma cruzi transmission in the Amazon basin. The main scenaries in the Brazilian Amazon.

The ecological diversity of Trypanosoma cruzi transmission in the Brazilian Amazon region is directly interlinked with the parasite's extensive reservoir, composed of 33 species of wild mammals within the following orders: Marsupialia, Chiroptera, Rodentia, Xenarthra, Carnivora and Primates; and of 16 species of wild triatomines, of which ten may be infected with T. cruzi. Four scenarios for the diversity of T. cruzi transmission in the Brazilian Amazon region are evident: (i) T. cruzi transmission between vectors and wild mammals, which is characterized as a wild enzooty encompassing the entire Amazon basin; (ii) accidental T. cruzi transmission from vectors and wild mammals to humans, when they invade the wild ecotope or when these vectors and wild mammals invade human homes; (iii) occupational Chagas disease among piassava (Leopoldinia piassaba) palm fiber gatherers, transmitted by the vector Rhodnius brethesi, for which these palm trees are the specific ecotope; (IV) oral T. cruzi transmission to humans through food contamination, particularly in juices from plants such as assai, which today is considered to be endemic in the Brazilian Amazon region, with more than 1500 cases notified.

Molecular epidemiology of American trypanosomiasis in Brazil based on dimorphisms of rRNA and mini-exon gene sequences.

American trypanosomiasis is transmitted in nature via a sylvatic cycle, where Trypanosoma cruzi interacts with wild triatomines and mammalian reservoirs, or via a domestic cycle where the parasite comes into contact with humans through domiciliated triatomines. The pool of T. cruzi isolates consists of sub-populations presenting a broad genetic diversity. In contrast to the heterogeneity suggested by isoenzyme analysis, PCR amplification of sequences from the 24S alpha rRNA gene and from the non-transcribed spacer of the mini-exon gene indicated dimorphism among T. cruzi isolates, which enabled the definition of two major parasite lineages. In the present study, 157 T. cruzi isolates obtained from humans, triatomines and sylvatic mammalian reservoirs from 12 Brazilian states were analysed by the 24S alpha RNA and mini-exon typing approaches. The stocks were classified into the two proposed lineages and according to the domestic or sylvatic cycle of the parasite. Data presented provide evidence for a strong association of T. cruzi lineage 1 with the domestic cycle, while in the sylvatic cycle both lineages circulate equally. Molecular typing of human parasite isolates from three well-characterised endemic regions of Chagas disease (Minas Gerais, Paraiba and Piaui) and from Amazonas State, where T. cruzi is enzootic, suggests that in some endemic areas in Brazil there is a preferential linkage between both cycles mediated by lineage-1 stocks.

Brazilian isolates of Trypanosoma cruzi from humans and triatomines classified into two lineages using mini-exon and ribosomal RNA sequences.

Traditional molecular and biochemical methods, such as schizodeme analysis, karyotyping, DNA fingerprinting, and enzyme electrophoretic profiles, have shown a large variability among Trypanosoma cruzi isolates. In contrast to those results, polymerase chain reaction (PCR) amplification of sequences from the 24S alpha ribosomal RNA gene and from the mini-exon gene nontranscribed spacer indicated a dimorphism among T. cruzi isolates, which enabled the definition of two major parasite lineages. In the present study, 86 T. cruzi field stocks (68 isolated from humans with defined presentations of Chagas' disease and 18 from triatomines) derived from four Brazilian geographic areas were typed by the PCR assay based on the DNA sequences of the mini-exon and 24S alpha rRNA genes. These stocks were ordered into the two major T. cruzi lineages. Lineage 1 was associated mainly with human isolates and lineage 2 with the sylvatic cycle of the parasite.

Spotted fever in Brazil: a seroepidemiological study and description of clinical cases in an endemic area in the state of São Paulo.

During 1985-1995, illnesses clinically and epidemiologically compatible with Brazilian spotted fever were identified in 17 patients in the county of Pedreira, in the state of São Paulo, Brazil. Spotted-fever group rickettsial infection was confirmed by serology and/or immunostaining of tissues in 10 of these patients. Immunostaining confirmed infection in a 37-year-old pregnant patient, although rickettsial antigens were not demonstrable in the tissues of the fetus. A serosurvey was conducted in four localities in the county to determine the prevalence of subclinical or asymptomatic infections with spotted fever group rickettsiae. Five hundred and twenty-five blood samples were tested by an indirect immunofluorescence assay for antibodies reactive with Rickettsia rickettsii. Twenty-two (4.2%) of these samples demonstrated titers > or = 1:64. The results indicate that Brazilian spotted fever is endemic within this region of Brazil.

Chagas' disease in the Brazilian Amazon. II. A serological survey.

A serological survey, involving indirect immunofluorescence testing of blood sera samples, was carried out on the residents of one in every five dwellings in the town of Barcelos (in the northern part of the State of Amazonas, on the right bank of the Rio Negro, 490 Km from Manaus by river) and on the rural populations of the villages of Piloto and Marará (also on the right bank of the Rio Negro, 30 minutes by boat from Barcelos). A total of 710 sera samples were tested, 628 from the resident population in the town of Barcelos, 35 from Piloto and 47 from Marará. The tests were carried out using human anti-gammaglobulin type IgG (Biolab) and antigen from formolized culture of T. cruzi Y strain. The sera were serially diluted from 1:40 to 1:320 in PBS 7.2. Of the 710 samples examined 89 (12.5%) were positive for anti-T.cruzi antibodies: 2 of these (2.2%) at a dilution of 1:320; 12 (13.4%) at 1:160; 38 (42.6%) at 1:80; and the remainder at 1:40, giving a median serological dilution of 1:80. The following questions are discussed: the high serological prevalence for Chagas' infection found in our survey; the possibility of serological cross-reactions; the need for confirmatory tests for the positives reactions; and the strong correlation between our results and preliminary epidemiological data (such as the level of human contact with wild triatominae, know locally as "Piaçava's lice". We draw attention to the isolation by xenodiagnosis of one strain of T.cruzi from a patient with positive serology for Chagas' infection.

Chagas' disease in the Brazilian Amazon. I--A short review.

At least eighteen species of triatominae have been found in the Brazilian Amazon, nine of them naturally infected with Trypanosoma cruzi or "cruzi-like" trypanosomes and associated with numerous wild reservoirs. Despite the small number of human cases of Chagas' disease described to date in the Brazilian Amazon the risk that the disease will become endemic in this area is increasing for the following reasons: a) uncontrolled deforestation and colonization altering the ecological balance between reservoir hosts and wild vectors; b) the adaptation of reservoir hosts of T. cruzi and wild vectors to peripheral and intradomiciliary areas, as the sole feeding alternative; c) migration of infected human population from endemic areas, accompanied by domestic reservoir hosts (dogs and cats) or accidentally carrying in their baggage vectors already adapted to the domestic habitat. In short, risks that Chagas' disease will become endemic to the Amazon appear to be linked to the transposition of the wild cycle to the domestic cycle in that area or to transfer of the domestic cycle from endemic areas to the Amazon.

Chagas' disease in the Brazilian Amazon. III. A cross-sectional study (1).

Two serological surveys for Chagas' infection were carried out, in 1991 and 1993, respectively, using a conglomerate family samples from the residents in the town of Barcelos (in the northern part of the State of Amazonas, on the right bank of the Rio Negro, 490 Km up-river from Manaus), using indirect immunofluorescent tests for anti-T. cruzi antibodies. In the first survey (1991), 628 blood samples from the residents of 142 dwellings were tested, showing positive in 12.7% for anti-T. cruzi antibodies and in 1993 an other 658 samples from residents of 171 dwellings showed positive in 13.7% of the tests, thus confirming the previous results. From 170 individuals with positive serology for T. cruzi antibodies, 112 (66%) were interviewed and submitted to electrocardiographic and clinical examinations; 82 (73.2%) of them gave consent for xenodiagnosis. From the 112 interviewed 52 (46.4%) recognized the triatomines as "piaçavas' lice", 48 (42.8%) knew the bugs from their work places being gatherers of piaçava fibers in rural areas and 19 (16.9%) said that have been bitten by bugs in their huts. Only 2 (2.4%) of 82 xenodiagnosis applied were positive for T. cruzi and 9 (8%) of the ECG had alterations compatible with Chagas' disease.

Epidemiological aspects of the Brazilian spotted fever: serological survey of dogs and horses in an endemic area in the State of São Paulo, Brazil.

In order to obtain information on Brazilian spotted fever, a study in domestic animals was performed in the County of Pedreira, State of São Paulo, Brazil, where 17 human cases had been notified. Serum samples obtained from animals were tested by indirect immunofluorescence for detectable antibodies to spotted fever-group rickettsiae. Seropositivity was revealed in 12 (36.4%) of 33 dogs and seven (77.8%) of nine horses from the endemic area. For comparison, blood samples from dogs and horses from non endemic area were tested and four (12.9%) of 31 dogs and three (27.3%) of 11 horses were positive. The highest titers of antibodies by IFA (IgG > or = 1:1024) were found only in three dogs and six horses from endemic area. The results suggest that dogs as horses may serve as environmental sentinels for establishing the prevalence of foci of spotted fever in Brazil.

[The evolution of chronic chagasic cardiopathy. I. The influence of parasitemia]. / Evolução da cardiopatia chagásica crônica. I--Influência da parasitemia.

During eight years (1982-1990) the evolution of chronic chagasic cardiopathy and its relation to parasitemia was evaluated in 279 patients, 85 men and 194 women, studied by resting electrocardiography and xenodiagnosis. All patients were residents in Virgem da Lapa, State of Minas Gerais, Brazil and their ages varied from 7 to 76 years (average 42.6 y). According to the results of the electrocardiograms the evolution of chagasic cardiopathy was classified as a) unchanged--when there was no change of the initial pattern off the ECG, b) progressive--when there was deterioration of the ECG pattern and c) regressive-when there was normalization or regression of the ECG alterations. Regarding xenodiagnosis 120 were considered with positive parasitemia, one or more xenodiagnoses positive, and 159 with negative parasitemia--all xenodiagnoses negative. The results showed: a) chagasic cardiopathy unchanged in 172 (61.6%) patients, b) progressive in 99 (35.5%) patients and c) regressive in 8 (2.9%). There was no relation between the evolution of chagasic cardiopathy and parasitemia. Independent from parasitemia, the cardiopathy was progressive according to the age of the patients and significantly greater in males. In conclusion we can state that evolution of chronic chagasic cardiopathy is associated with the age and with the male sex, but not with parasitemia, and this may suggest that parasitemia is not related to the development of the chronic chagasic cardiopathy.

[Human populations attacked by wild Triatominae in the Amazonas: a new form of transmission of Chagas disease?]. / Ataque de populações humanas por triatomíneos silvestres no Amazonas: uma nova forma de transmissao da infecção chagásica?

An "attack" of wild triatomines (Rhodnius brethesi) to piaçaca workers (Leopoldinia piaçaba) is confirmed in the locality of Acuquaia, at Padauari river, affluent of Rio Negro in the municipality of Barcelos, State of Amazonas, Brazil. A serological prevalence of 12.5% for T. cruzi antibodies in human population, in the city of Barcelos, has already been described in a previous paper. A strong association between the serological positivity and the population contact with wild triatomines, known in the area as "piaçava's lice", was verified.

[Chagas disease in Virgem da Lapa, Minas Gerais, Brazil. IV. Clinical and epidemiological aspects of left ventricular aneurism]. / Doença de Chagas em virgem da Lapa, Minas Gerais, Brasil. IV. Aspectos clínicos e epidemiológicos do aneurisma ventricular esquerdo.

The study aimed at the evaluation of the clinical and epidemiological characteristics of the aneurysm found in the left ventricle in chronic Chagas' disease patients. Three hundred, eighty eight people (298 chagasic patients and 90 randomly selected healthy individuals) were submitted to echocardiography. The ventricular function was assessed in the M mode by calculating the fraction of ejection, and in the bidimensional mode by analyzing he global systolic function. Segmental contractility was evaluated according to the method described by American Society of Echocardiography. Aneurysm of the left ventricle was diagnosed in 58 (18.8%) patients, all from the chagasic population. From these, 38 (12.7%) were found in the apical segment; 10 (3.4%) in the interventricular septum; and 2 (0.7%) each in the posterior wall; the inferior wall; apico-septal; and inferior-posterior. We could not observe any significant difference for the aneurysm frequencies in relation to age group, gender and race, and no association between aneurysm and arterial hypertension could be made. Of the 56 individuals presenting aneurysm, 55 (98.2%) were symptomatic with predominant palpitations; 53 (94.6%) showed an aberrant ECG with predominant ventricular extra-systoles followed by changes in conduction; and 34 (60.%) showed an impairment of the ventricular function, regardless of the affected segment. In view of these results we consider the apical aneurysm of the left ventricle as a marker of Chagas' disease and as an indicator of high morbidity of the human T. cruzi infection in Virgem da Lapa.

[Parasitemia in chronic chagasic patients evaluated using the index of infected Triatoma in xenodiagnosis]. / Parasitemia em pacientes chagásicos crônicos avaliada pelo índice de triatomíneos infectados no xenodiagnóstico.

As part of a pre-treatment study of chronic Chagas infections, the parasitemia of 206 patients (85 men and 121 women, aged 7 to 80 y) from Virgem da Lapa, Minas Gerais State Brazil, was evaluated by three xenodiagnoses per patient during a one year period. Each time, 40 3rd or 4th instar nymphs of Triatoma infestans were applied. The parasitemia was arbitrarily classified as: not detected (when all nymphs were negative), low (when the number of infected nymphs was less than 2%), medium (when it was higher than 2% and up to 7%) and high (when higher than 7%). The parasitemia was not detected in 105 (51%) of the patients, and was considered low in 55 (26.7%), medium in 27 (13.1%) and high in 19 (9.2%). There was no significant differences in levels of parasitemia in relation to sex or age, but the high parasitemia was more frequent among the patients with chagasic cardiomyopathy. Persistent parasitemia (all three xenodiagnoses positive) was observed in 100% of the patients with high parasitemia, in 22.2% with medium and in none with low parasitemia.

[Development of chronic human chagas cardiopathy in the hinterland of the Paraíba State, Brazil, in a 4.5 year period]. / Evolução da cardiopatia chagásica crônica humana no Sertão do Estado da Paraíba, Brasil, no período de 4,5 anos.

Two sectional studies about chronic Chagas' disease were performed at a 4.5 year interval, involving the urban populations of Agua Branca, Catingueira, Emas, Imaculada, Mãe D'Agua, Olho D'Agua, Piancó and São José de Caiana counties in the Sertão region of the State of Paraíba. The evolution of heart disease was evaluated in 125 matched pairs of chronic chagasic and non-chagasic patients of the same sex, age and county of origin through electrocardiograms (ECG) at rest. The following evaluation criteria were considered: unchanged - no change in the original ECG pattern; progressive-changes in ECG pattern from normal to abnormal or deterioration of ECG abnormalities; and ECG normalization. In chagasic patients evolution of the disease was unchanged in 101 (80.8%), progressive in 13 (10.4%) and ECG normalization in 11 (8.8%), while those observed in non-chagasic patients were respectively values 117 (93.6%), 6 (4.8%) and 2%) patients. Findings indicate that the share of Chagas-linked etiological component affecting the development of chronic Chagas cardiopathy was 5.9% with an estimated annual average of 1.3%. There was no significant difference in the frequency of progressive disease between the sexes either in the chagasic or in the non-chagasic group. On the other hand, progression of heart disease occurred earlier among chagasic patients. Lethality caused by heart disease was 1.6% (2 cases) in the chagasic group and zero in the non-chagasic group during the period under survey. These morbidity and mortality rates were significantly lower than those found in endemic areas such as Virgem da Lapa and Pains-Iguatama in the State of Minas Gerais and can probably be attributed to the weaker pathogenic it of human infection by Trypanosoma cruzi in the Paraíba hinterland.

[Comparative controlled study on the use of benznidazole, nifurtimox and placebo, in the chronic form of Chagas' disease, in a field area with interrupted transmission. I. Preliminary evaluation]. / Estudo comparativo controloado com emprego de benznidazole, nifurtimox e placebo, na forma crônica da doença de Chagas; em uma área de campo com transmissão interrompida. I. Avalição preliminar.

A controlled clinical trial was carried out to evaluate the therapeutic efficacy and tolerance of nifurtimox and benznidazole in patients with chronic Chagas' disease. All patients had immunofluorescence and complement fixation reactions positives for T. cruzi antibodies and at least two xenodiagnoses positives in three performed before treatment, and they were submitted to clinical examinations, ECG and X-ray of the heart and esophagus. Of 77 patients studied, 27 were treated with nifurtimox and 26 with benznidazole in the dosage of 5 m/kg/day for 30 consecutive days, and 24 received a placebo in tablets similar to benznidazole. From the 77 patients, 64 (83.1%) accomplished the treatment: 23 (88.4%) with benznidazole, 19 (70.3%) with nifurtimox and 22 (91.6%) with placebo. The patients were evaluated, clinically, serologically and parasitologically (six xenodiagnoses within one year after treatment). The benznidazole group showed only 1.8% of positive xenodiagnoses post-treatment, the nifurtimox 9.6% and the placebo 34.3%. All serologic reactions continued positive and there were no clinical, ECG or X-ray changes one year after treatment.

Epidemiological aspects of the Brazilian spotted fever: seasonal activity of ticks collected in an endemic area in São Paulo, Brazil.

Ticks were collected from vegetation and animals at monthly intervals during one year (1993-1994) in an endemic area of Brazilian spotted fever in the Country of Pedreira, State of São Paulo. Six species of ticks were identified Amblyomma cajennense, Amblyomma cooperi, Amblyomma triste, Anocentor nitens, Rhipicephalus sanguineus and Boophilus microplus. Only the first species was sufficiently numerous to permit a quantitative study with seasonal activity, although the distribution and source of capture of other species were observed and are reported. This information is correlated with the epidemiology of tick-borne rickettsiosis.

[Xenodiagnosis in chronic Chagas' disease. I. The sensitivity of Panstrongylus megistus and Triatoma infestans]. / Xenodiagnóstico na doença de Chagas crônica. I--Sensibilidade de Panstrongylus megistus e Triatoma infestans.

From January 1986 to February 1994, 563 xenodiagnosis (XD) were applied in 563 chronic chagasic patients from different areas of Brazil; 292 were women and 271 were men between 6 and 89 years (average: 41.4 +/- 14.7 years). To each XD 40 nymphs on the 4th stage were used: 20 from Panstrongylus megistus (Pm) and 20 from Triatoma infestans (Ti) in fast, during at least 14 days. The exam in each nymph was made 45 days after being applied on the patient, by observation in optical microscopy of the drugs and/or the grinded from the digestive tube. The results are: a) 205 (36.4%) positive XD, including 85 (15.1%) due only nymphs of Pm, 44 (7.8%) Ti and 76 (13.5%) Pm and Ti; b) positively in 4.9% of the nymphs from Pm and in 3.0% of the Ti nymphs examined. These results' analysis showed that the Pm nymphs were more sensitive that Ti's ones to the infection by Trypanosoma cruzi, increasing considerably the xenopositivity, independently from birthplace, sex or age of the patients. These results point out that to increase the efficacy of XD in chronic Chagas' disease, the exam must have more than a species of triatomine with different sensibilities to the T. cruzi infection, and in case of using one species on XD, Pm must substitute Ti.

[Morbidity of Chagas disease in areas of Sertão da Paraiba and Caatinga do Piauí]. / Morbidade da doença de Chagas em áreas do Sertão da Paraíba e da Caatinga do Piauí.

A clinical and electrocardiographic case control study was carried out with 186 pairs of persons with positive and negative serology for T. cruzi infection from the Sertão Paraíba and in 200 seropositive cases from the region of Caatinga in the State of Piauí, North-eastern Brazil. The predominant clinical manifestations in seropositive cases in both areas were: palpitations, dyspnea on effort, precordial pain, dysphagia, odynophagia, pyrosis and intestinal constipation. The EKG abnormalities rates suggestive of chronic chagasic cardiopathy were respectively in Paraíba and Piauí: AV block 3.8% and 2%, RBBB III 6.4% and 7%, RBBB III+ LAB 10.7% and 10.5%, and multifocal extrasystoles 2.7% and 3%. Xenodiagnosis in a sample of 54 seropositive individuals in the Sert-ao of Para-iba and in 120 in the Caatinga of Piauí was revealed 13% and 34% positive; PCR tests in a sample of 47 seropositives in Paraíba and 101 in Piauí revealed positives in 44.6% and 59.5% respectively. Blood culture in LIT media of 101 seropositive cases from the Caatinga of Piauí was positive for T. cruzi in 25.7%. A triatomine survey carried out in a sample of 132 domiciles and peridomiciles in the Sertão of Paraíba and in a sample of 159 in the Caatinga of Piauí showed the following results: In Paraíba, 16 specimens of T. brasiliensis, not infected with T. cruzi, were captured. In Piauí, 750 triatomines were captured, of these 625 were examined: 49 were T. pseudomaculata, not infected with T. cruzi (19 in peridomiciles and 30 in the domiciles), and 576 were T. brasiliensis (371 in the domiciles and 205 in the peridomiciles) and of this latter specie 32 (5.5%) were infected with T. cruzi (31 in the domiciles and one in the peridomicile).

[Cross-sectional study of intestinal parasites and Chagas' disease in the Municipality of Novo Airão, State of Amazonas, Brazil]. / Estudo das parasitoses intestinais e da infecção chagásica no Município de Novo Airão, Estado do Amazonas, Brasil.

A cross-sectional study was performed on the residents of one in every ten households in the town of Novo Airão, in the northern meso-region of the State of Amazonas, 250 kilometers from Manaus by riverboat. A family cluster sample of 89 dwellings was studied. A stool sample was requested from each of the inhabitants for examination using the Lutz sedimentation and Baermann-Moraes-Coutinho techniques, and blood was taken by venous puncture for Trypanosoma cruzi Elisa antibody testing and immunofluorescence. From a total of 316 stool samples, 87.6% had one or more parasites: Ascaris lumbricoides (35.1%), Entamoeba histolytica (29.1%), Giardia lamblia (17.4%), and other parasites with lower prevalence rates. These results were directly correlated with lack of sanitation and clean water supply. Of the 346 sera examined, 16 (4.6%) were reactive to T. cruzi antibodies, but only three showed a correlation between this result and human contact with wild triatomines, known locally as "piassava lice".

[Epidemiological, social, and sanitary aspects in an area of the Rio Negro, State of Amazonas, with special reference to intestinal parasites and Chagas' disease]. / Aspectos epidemiológicos, sociais e sanitários de uma área no Rio Negro, estado do Amazonas, com especial referência às parasitoses intestinais e à infecção chagásica.

A cross-sectional study was carried out on the residents of one in every four dwellings in the town of Barcelos (in the northern part of the State of Amazonas, on the right bank of the Rio Negro, 490 kilometers from Manaus by river), in order to evaluate social and sanitary conditions and specific indicators for intestinal parasites and Chagas' infection. During the survey, two questionnaires were applied, a household one to evaluate social and sanitary aspects, and an individual one, for social and epidemiological evaluation of the population conditions. A conglomerate family sample of 171 dwellings was studied. From each of the 658 habitants, a sample was requested for stool examination by Lutz sedimentation and Baermann-Moraes-Coutinho techniques modified by Willcox & Coura (1989), and blood was collected in filter paper for immunofluorescence test by Camargo (1966) and Souza & Camargo (1966) methods modified by Petana & Willcox (1975). The stool examination showed 69.4% of samples with one or more parasites. Ascaris lumbricoides was predominant with 51% of positivity and Entamoeba histolytica, although surveyed by a non-specific method, was present in 19.7%. Surprisingly, 20.1% of the 658 sera samples were reactive for T. cruzi antibodies at a dilution of 1:20 and 13.7% at 1:40. There was a strong correlation between this result and the level of human contact with wild triatomines, known locally as "piasava lice", and we succeeded in isolating by xenodiagnosis one strain of T. cruzi from one patient, a sixty-one-year old man (n. 209 -1), a native of the area, with positive serology for Chagas' disease and who worked in agriculture and transporting piasava and was very familiar with "piasava lice".