Prevention of electrocardiographic and histopathologic alterations in the murine model of Chagas' disease by preinoculation of an attenuated Trypanosoma cruzi strain

The effects of infection with Trypanosoma cruzi on the electrocardiographic tracings of mice were studied in 4.groups of animals: (1) normal; (2) infected with a pathogenic T. cruzi strain (TS COB); (3) immunized with 3 intraperitoneal inocula of 10(6) attenuated T. cruzi epimastigotes (TCC) and (4) immunized-infected, which sequentially received the treatments of groups 3 and 2. Infection and protection were confirmed by xenodiagnosis and histopathology. Isolated alterations such as extrasystolia, 1st degree atrioventricular block, arrhythmia and ST elevation were observed in normal as well as infected mice. However, tracings taken repeatedly on each mouse over a 293 day period revealed a set of alterations which were more frequently seen in infected (14/22) than in normal (4/27) animals (p = 0.00048). These alterations consisted of supraventricular tachycardia, sinus bradycardia and persisting, first degree AV blocks, often associated to pacemaker changes. Inoculation of attenuated T. cruzi (group 3) did not increase these alterations (2/27 mice) but significantly prevented their development after challenge with the pathogenic strain (1/19 versus 14/22 mice, p = 0.000095). Thus, preimmunization reduced not only parasitemia but also a pathogenic consequence of T. cruzi infection. This evidence is relevant for immunoprevention studies against Chagas' disease.

Eosinophil levels in the acute phase of experimental chagas' disease

Eosinophil dynamics, in bone marrow, blood and peritoneal exudate, of resistant C57B1/6 (C57) and susceptible A/Snell (A/Sn) mice was comparatively studied during the acute phase of infection by Trypanosoma cruzi Y strain. A decline was observed in bone marrow eosinophil levels in A/Sn, but not in C57 mice, soon after infection, those of the former remaining significantly below those of the latter up to the 4th day of infection. Bone marrow eosinophil levels of C57 mice declined subsequently to levels comparable to those of A/Sn mice, the number of these cells in this compartment remaining 50% those of non infected controls, in both strains, up to the end of the experiment on the 14th day of infection. The fluctuations in eosinophil levels in blood and peritoneal space were similar in both mice strains studied. Concomitantly with depletion of eosinophils in the marrow, depletion in blood and a marked rise of these cells in the peritoneal space, initial site of infection, occurred in both strains. The difference in eosinophil bone marrow levels, between C57 and A/Sn mice, observed in the first four days of infection, suggests a higher eosinopoiesis capacity of the former in this period, which might contribute to their higher resistance to T. cruzi infection.

Chronic myocardial damage in experimental T. cruzi infection of a new world primate, Cebus sp. monkey

Eighteen Cebus apella monkeys, (juvenile and adult of both sexes) were inoculated five years ago, with three Trypanosoma cruzi strains (CA1, n = 10; Colombian, n=4 and Tulahuen, n=4), either by conjunctival or intraperitoneal route, once or repeatedly. Parasitological, hematological, serological, enzymatic, radiographic, electro and echocardiographic findings have been peviously published15 and they are similar to those observed in human pathology. The most frequent electrocardiographic alteration was right branch bundle block. Six animals, chosen at random, were sacrificed. Those sacrificed 20 to 25 months post-first inoculation showed focal accumuli of leukocytes with myocytolysis. Foci of diffuse interstitial fibrosis with mild infiltrate of leukocytes among fibers were observed in the animals sacrificed 36 to 47 months post-inoculation. No parasites were seen. The lesions were more prominent in the ventricular walls and the septum. The fact that the infiltrates were predominant in the animals sacrificed at a shorter time after first inoculation and that fibrosis was more severe in those sacrificed at a longer time suggests that there is a progression of the infiltrative lesions to fibrosis, with a leukocytic activity indicative of a chronic phase. These lesions are similar to those described in human chronic Chagas' disease. This would demonstrate that this model is useful in evaluating a progress in the knowledge of the pathogenesis which is still a controversial issue, immunology, immunogenesis and chemotherapeutic agents of the chronic and indeterminate phases of this disease.

Possible oral transmission of acute Chagas' disease in Brazil

In October, 1986, 7 to 22 days after a meeting at a farm in Paraíba state, 26 individuals presented with a febrile illness associated with bilateral eyelid and lower limb edema, mild hepatosplenomegaly, lymphadenopathy and, occasionally a skin rash. A 11-year-old boy exhibited atrial premature complexes and a 74-year-old patient developed acute heart failure. In two patients hospitalized in São Paulo city, acute Chagas' disease was diagnosed by the demonstration of circulating Trypanosoma cruzi. At autopsy in a fatal case, acute Chagas' cardiomyopathy was demonstrated. Xenodiagnosis were positive in 9 out of 14 tested patients. A specific IgG immune response was found in all patients and specific IgM antibodies were identified in 20 out of 22 tested patients. A epidemiological survey showed the existence of Triatoma brasiliensis in the outbuildings of this farm, but none in the house where most of the guests stayed. A high rate of infection with Trypanosoma cruzi was found in opossums. These observations together with those related to the food consumed by the patients, lead the authors to suggest that the human infections resulted from oral contamination probably originating from naturally infected marsupials in the area or crushed infected bugs.

Lethality of triatomines (Hemiptera: Reduviiae), vectors of Chagas' disease, feeding on blood baits containing synthetic insecticides, under laboratory conditions

A laboratory study was conducted to test the toxicity of synthetic insecticides added to defibrinated sheep blood kept at room temperature and offered as food to the following triatomine species: Triatoma infestans, Panstrongylus megistus, Triatoma vitticeps, Triatoma pseudomaculata, Triatoma brasiliensis and Rhodnius prolixus. The insecticides used, at a concentration of 1g/l, were: HCH, DDT, Malathion and Trichlorfon, and the lethalithy observed at the end of a 7-day period varied according to the active principle of each. HCH was the most effective by the oral route, killing 100% of the insects, except P. megistus (95.7%) and T. pseudomaculata (94.1%). Trichlorfon killed the insects at rates ranging from 71.8% (T. vitticeps) to 98% (R. prolixus). Malathion was slightly less efficient, killing the insects at rates from 56.8% (T. vitticeps) to 97% (T.brasiliensis). DDT was the least effective, with a killing rate of 10% (T. vitticeps) to 75% (T.brasiliensis). Since the tests were performed at room temperature, we suggest that baits of this type should be tried for the control of triatomines in the field.

Embryology of Triatoma infestans (KLUG), (Hemiptera, Reduviidae), a Chagas' disease vector

This study reports the embryogenesis of T. infestans (Hemiptera, Reduviidae). Morphological parameters of growth sequences from oviposition until hatching (12-14 d 28ºC) were established. Five periods, as percent of time of development (TD), were characterized from oviposition until hatching. The most important morphological features were: 1) formation of blastoderm within 7% of TD; 2) germ band and gastrulation within 30% of TD; 3) nerve cord, limb budding, thoracic and abdominal segmentation and formation of body cavity within 50% of TD; 4) nervous system and blastokinesis end, and development of embryonic cuticle within 65% of TD; 5) differentiation of the mouth parts, fat body, and Malphigian tubules during final stage and completion of embryo at day 12 to day 14 around hatching. These signals were chosen as appropriate morphological parameters which should enable the evaluation of embryologic modifications due to the action/s of different insecticides

Lethal effect of a bait for Rhodnius prolixus (Hemiptera: Reduviidae), the vector of Chagas' disease, containing hexachlorocyclohexane (HCH), under laboratory conditions

The lethal effect of a bait containing an aqueous hexachlorocyclohexane (HCH) suspension at the concentration of 1g/l and maintained at room temperature was studied in the laboratory over a period of 12 weeks. The suspension was placed in a latex bag hanging inside a 1000-ml beaker tightly covered with nylon netting, and left there with no changes for 85 days. Sixteen groups of R. prolixas bugs, consisting on average of 30 specimens each, were successively exposed to the bait and observed at different intervals for one week each. The mortality rate was 100% for all groups, except for the 16th, whose mortality rate was 96.7%. As the groups succeeded one another, mortality started to occur more rapidly and was more marked at the 6- and 24-h intervals. Later tests respectively started at 6:00 a.m. and 6:00 p.m. showed that diurnal and nocturnal periodicity in the offer of food had no effect on mortality. First- and 2nd- instar nymphs and adults male were more sensitive and 5th- instar nymphs were more resistant to the active principle of the bait.

Possible risk factors for vertical transmission of Chagas' disease

The author emphasizes the importance of the congenital transmission of Chagas' disease and discusses the possible risk factors for transmission such as age, origin, obstetrical history and maternal form of disease. Exacerbation of infection during pregnancy is also considered as a possible risk factor for transmission. Besides, a relationship between the frequency of transmission and gestational age is presented. Concerning breast-feeding, the risk of transmission is directly related to the acute phase of maternal disease and bleeding nipples. The deleterious effects of chagasic infection on the fetus and newborn are also considered.

A critical evaluation of the expression of parasitemia in experimental Chagas' disease

This paper aims to study the best way to express the parasitemia of Trypanosoma cruzi's experimentally infected animals. Individual scores may have a great variability, not emphasized by the majority of the authors. A group of 50 rats infected with 1x10(6) trypomastigotes of T. cruzi Y strain was used and the parasitemia was estimated by BRENER' s method. The results showed that the median can avoid false results due to very high or low parasitemias but it does not have the mathematic properties necessary for analysis of variance. The comparison of the means of the original and transformed data, with their respective coefficients of variability (CV), showed that the logarithmic mean (Mlog) have the minor value of CV. Therefore, the Mlog is the best way to express the parasitemia when the data show great variability. The number of the animal for group did not affect the variability of data when the Mlog and CV were used.

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: 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 "Piacava'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.

Seroepidemiological and clinical study of Chagas' disease in Nicaragua

With the aim of determining the prevalence, immunological profile, and knowing the electrocardiographic alterations, a clinical and Seroepidemiological study of Chagas' disease was performed in three rural settlements located at the North, East and West of Nicaragua. Anti T. cruzi antibodies were searched by indirect immunofluorescence (IFI) and hemagglutination (IHA) in a total of 803 subjects. Seropositives and the same number of seronegatives, matched by age and sex, were included in a case-control design for the electrocardiographic assessment. Antibody prevalence was 13.1, 4.3 and 3.2% in the respective settlements. In the first two the immunological profile corresponds to that of an endemic zone of long standing, were transmission has decreased, and in the third the pattern is of a zone under control. Electrocardiographic changes compatible with Chagas' disease were found in seropositive individuals, but difference with control group was not statistically significant. It is concluded that the disease is endemic in the three settlements and the clinical aspect requires further evaluation, including additional cardiologic techniques.

Chagas' disease in the Brazilian Amazon: III. a cross-sectional sutdy

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.

Domestic vectors of Chagas' disease in three rural communities of Nicaragua

A triatomine survey was conducted in three rural settlements of Nicaragua (Santa Rosa, Quebrada Honda and Poneloya) where Chagas' disease is endemic, to determine rates of house infestation, evaluate the housing condition and to asess the performance of the María sensor box in detection of domestic vectors. A total of 184 households were selected and vectors were sought by the methods of timed manual capture and by sensor boxes. The sole vectors species found in this study was Triatoma dimidiata. Of the examined bugs 50, 60 and 33%, in the respective communities, were infected with T. cruzi. The rates of house infestation as determined by manual capture and sensor boxes were respectively, 48.3% and 54.2% in Santa Rosa, 29.8% and 51.2% in Quebrada Honda and in Poneloya 3.8 and 5.9% with significant difference between the methods in Quebrada Honda. When compared with the manual capture, the Maria sensor box detected vectors in 71.4% of positive houses in two of the communities but also was able to detect bugs in 39.3% and 41.1% of houses where manual capture had been negative. Housing condition was evaluated according to three structural parameters, in this way, in the first community 79.2% of houses were classified as bad, 20.8% as regular; in the second one 42.5% were bad and 57.5% regular, whereas in the third 62.5% of the houses were regular. Rates of infestation did not differ greatly between the different housing conditions. Our results show that the sensor box is as efficient as manual capture and could be implemented in our country.

Tumor-like lesion due to Chagas' disease in a patient with lymphocytic leukemia

A 73 year-old white male, living in the interior of the state of Mato Grosso do Sul, in central Bra­zil, after an initial diagnosis of sinusitis was transferred to the neurology service with a 3-day evolution of intracranial hypertension. Exams showed lymphocytic leukemia and a tumor-like lesion, either an expanding inflammatory process such as an abscess or a neoplasm. Treatment with Ceftriaxone and Decadron was started and intracranial hypertension was controlled. Methotrexate was injected on the occasion of the next puncture considering a possible leukemia infiltration. Flagellate forms of T. cruzi were observed in the CSF and treatment with Benznidazole was started. After 4 days the CSF presented fractionated forms of trypomastigotes. The protein level was 27%. Signs of intracranial hypertension ceased. Tomography and magnetic resonance images showed an important reduction of the tumor-like lesion. The clinical condition of the patient improved.