Trypanosoma cruzi genotypes associated with domestic Triatoma sordida in Bolivia

Triatoma sordida is the second species of Triatominae considered of epidemiological significance in Bolivia. Associated with Triatoma infestans in various regions, it is as yet the only triatomine species established in human dwellings in localities of Velasco province, Department of Santa Cruz. This domestication is considered as primary. Flagellate parasites were detected in 16.2% of domiciliary T. sordida and the kDNA-PCR confirmed the presence of Trypanosoma cruzi. Frequencies of T. cruzi clonets 20 and 39, common clonets in Bolivian domestic cycle (T. infestans), were established by their direct detection in feces using PCR and hybridization. These clonets present low frequencies in T. sordida and synanthropic mammals. Forty-six stocks were isolated and analysed by multilocus enzyme electrophoresis (MLEE). The MLEE showed a higher clonal diversity than in T. infestans domestic cycle and the genotypes were clustered in the two principal lineages of T. cruzi. Within each lineage, a broad variability was observed. Mixture of genotypes was mostly observed in mammals. The large diversity of T. cruzi in this cycle should be related to its sylvatic origin. Moreover, the current limited sample of stocks suggests a lineage association with specific hosts.     

Low probability of transmission of Trypanosoma cruzi to humans by domiciliary Triatoma sordida in Bolivia

The role of Triatoma sordida in the domestic transmission of Trypanosoma cruzi was assessed in 7 rural localities in Velasco Province, Department of Santa Cruz, Bolivia. Tri. sordida, the only triatomine species identified in these localities, was found inside 58.0% of houses but not in large numbers (3.1 bugs per infested house on average). A total of 220 faecal samples from domiciliary bugs was examined microscopically and by the polymerase chain reaction for the presence of trypanosomes: 21.4% were infected. Analysis of blood meals of domiciliary Tri. sordida showed that humans were the commonest host (70.4%), followed by chickens and dogs. Four of 418 persons tested were seropositive for Tryp. cruzi. Only 2 of a second group of 62 persons living in dwellings infested by Tri. sordida were seropositive. Tryp. cruzi infection was demonstrated in dogs and domestic rats. Three other species of small mammals were found to be infected with trypanosomes. In our study area, domestic Tri. sordida are mainly incriminated in the transmission of Tryp. cruzi to synanthropic animals, whereas transmission to humans is very rare. The presence in houses of small populations of Tri. sordida infected with Tryp. cruzi is therefore currently insufficient for this insect to constitute a major epidemiological risk factor.     

Evaluation of vectors of Chagas' disease eradication programs in Chile by serological study of children under 10 years old

Chagas' disease is a parasitic zoonosis with high prevalence in Chile. It is distributed in rural and periurban section in the northern most seven out of thirteen regions in which the country is divided, and affects about 142,000 individuals dispersed in most of the 165 counties sited in the chagasic zone. Triatoma infestans--intradomiciliary species--is the main and practically exclusive vector of Trypanosoma cruzi. Diverse tools may be utilized in order to interrupt the domestic cycle of transmission of T. cruzi: health education, housing improvement and elimination of vectors by dwelling insecticide sprayings. This last resource has received priority in Chile in the last 12 years. To evaluate the effectivity of the programs for eliminating T. infestans a serological study for Chagas' disease--comprising 8,767 children less than 10 years old from 27 rural counties insecticide sprayed in the last 12 years--was carried out. A global total of 125 (1.4%) children resulted positive, figure significantly lower than 5.4% found in 1982-1990 in the same age group. Distribution by regions of positive individuals showed a decrease of prevalence in each of them: III Region, from 9.8 to 1.0%, IV Region, 7.2 to 2.0%, V Region, 5.2 to 1.9%, and Metropolitan Region, 1.4% to 0.6. Even though positive children have still been found in 46.7% of localities of the studied counties, it is possible to affirm that the vector control programs have been effective and must be maintained, and increased in those localities with T. cruzi infection in children under 10 years of age, with the general aim of eradicate Chagas' disease transmission in Chile before 2000.     

Current human and canine seroprevalence of Chagasic infection in San Pedro de Atacama County, II Region of Antofagasta, Chile, 1995

In order to assess the impact of a control program against Triatoma infestans launched in 1988, based on insecticide spraying of dwellings, a serological survey for chagasic infection was carried out during 1995 in three localities from San Pedro de Atacama County (22 degrees 55' South lat., 68 degrees 12' West long.), II Region of Antofagasta in northern Chile. Blood samples from 531 children and adolescents and 65 dogs were subjected to ELISA test and indirect immunofluorescent test for Chagas' disease respectively. Tests resulted positive in 12 (2.3%) persons, all above 5 years old, in contrast with the 16.8% serological positivity observed in 1985. Three (4.6%) dogs (two 0-12 months old) resulted positive. These results indicate that dwelling sprayings with long-term activity insecticides against T. infestans is a good tool to prevent new human infections with T. cruzi. However, active vector transmission among domestic animals (canines) could be recently acquired.     

Biological characterization of Trypanosoma cruzi stocks from Chilean insect vectors

Fifty-seven Trypanosoma cruzi stocks isolated from Triatoma infestans and Triatoma spinolai of the five different geographic endemic areas of Chile were studied by schizodeme and molecular karyotype analysis. Four different genotypes are found in the sylvatic T. spinolai vector and five in the T. infestans domiciliary vector. Of these genotypes, two common genotypes overlap on both transmission cycles exclusively in the extreme northern endemic areas of Chile. Metacyclic trypomastigotes obtained in vitro or cell-derived trypomastigotes proved to be infective in gamma-irradiated Balb/c mice for the study of the immune response and biological behavior. Of a total of 57 T. cruzi stocks obtained, 19 of them, representing all the different genotypes found in Chile, were tested on a murine experimental model and then fully studied. Female compared with male animals demonstrated greater resistance to Chagas disease with all the T. cruzi stocks tested. The immune response was assessed by lytic antibodies that were studied by the in vitro antibody-dependent complement-mediated lytic assay with the use of bloodstream trypomastigotes as target cells. In one unique parasite genotype the elicited lytic antibodies reacted in a genotype-specific manner, in contrast with lytic antibodies generated by other T. cruzi genotypes. Parasitemias were high, moderate, and low, with mortality ranges of 6-50%, 0-45%, and 0-10%, respectively. No association was found between specific infective genotypes and virulence or mortality. Independently of the T. cruzi strain studied, each population displayed a characteristic parasitemia curve and prepatent period. A considerable number of the parasite stocks proved to be mixed populations, according to molecular karyotype patterns obtained before and after differentiation and amplification of the parasites. This fact created difficulty in assessing the identity of the genotype really infective to mice.     

CMV infection and cytomegalovirus disease in immunodepressed patients

An active entomologic survey was conducted by a team of trained health workers in a rural area endemic for Chagas' disease in central Brazil. They used pyrethrum as a flushing agent and 4,232 houses were inspected for triatomine bugs both inside and in the immediate environs. Houses with Triatoma infestans or evidence of an established colony were identified and defined as infested houses (cases). The building and environmental characteristics of 161 randomly selected infested houses were compared with 161 matched, noninfested houses (controls) that were the shortest distance from the infested house. Domestic and peridomestic potential risk factors associated with house infestation by Triatoma infestans were assessed by logistic regression analysis. Incomplete house construction (odds ratio [OR] = 2.5, 95% confidence interval [CI] = 1.5-4.1) was confirmed as a risk factor related to the presence or evidence of Triatoma infestans in the dwellings. The study also disclosed a statistically significant association between the presence of rats (OR = 1.6, 95% CI = 1.1-2.6) and indoor crop storage (OR = 2.3, 95% CI = 1.1-5.2) and house infestation. Further experimental field studies using tagged rodents should be conducted to assess their epidemiologic role in the domestic chain of Trypanosoma cruzi transmission.     

Internalization of non-permeant materials into cultured mammalian cells by vortex-stirring in the presence of a high molecular weight polyacrylic acid: direct evidence of internalization and its dependence on the cell lines

BACKGROUND: Trypanosoma cruzi, the cause of Chagas' disease, is often transmitted by transfusion in Latin America. Previous studies showed that at least 1 in 1000 eligible blood donors at the Los Angeles County+University of Southern California (LAC+USC) Medical Center Blood Bank had specific antibodies to T. cruzi. In June 1993, serologic screening of prospective allogeneic donors at epidemiologic risk for T. cruzi infection was begun voluntarily. STUDY DESIGN AND METHODS: The risk of T. cruzi infection in all eligible donors was assessed by questionnaire. At-risk donors were screened serologically for antibodies to T. cruzi with an enzyme immunoassay, and confirmatory testing was done with a radioimmunoprecipitation assay. RESULTS: During the 29-month study period 1311 (39.5%) of 3320 donors were judged to be at risk for T. cruzi infection. Seven donors (1/475) were reactive by an enzyme immunoassay, and six of these seven (1/ 553) were positive in a radioimmunoprecipitation assay. All radioimmunoprecipitation assay-positive donors had been born in countries in which Chagas' disease is endemic. One person in this group had received a transfusion in his homeland. CONCLUSION: These results demonstrate that a substantive proportion of eligible blood donors at our institution have antibodies specific for T. cruzi and that a commercially available assay can be used to detect these antibodies. Our data suggest that the risk of transmission of T. cruzi by transfusion could be eliminated by serologic testing limited to persons born in or transfused in countries in which Chagas' disease is endemic.     

Constipation, laxative use, and colon cancer among middle-aged adults

Reported is an evaluation of 13 years of intervention by the Chagas Disease Control Programme in an endemic area (Montalvania) in the State of Minas Gerais, Brazil. The design used was an epidemiological panel study. The reduction of Trypanosoma cruzi infection rates was estimated from data collected on three separate occasions: a serological survey in 1975-80, a quasi-experimental study in 1987, and the present investigation. A random sample of 156 households was selected and blood samples were collected from 653 inhabitants. The data routinely collected by the control programme were analysed to correlate the results with the incidence of T. cruzi. The overall prevalence of infection was 2.3%; however, no participant under 14 years of age was found to have a positive serological test. The total reduction in T. cruzi infection rates in this area from the start of the programme's activities was estimated to be 83.5%. Cross-sectional comparisons for the age groups 2-6 years and 7-14 years indicated a 100% reduction in T. cruzi incidence rates; but cohort comparisons showed that 100% reduction was achieved only for the 2-6-years age group.     

Use of the restricted academic task in ADHD dose-response relationships

An expression genomic library of Trypanosoma cruzi (T. cruzi) constructed using pcDNA3 plasmid was used for the immunisation (25 micrograms) of Balb/c mice. Expression of T. cruzi antigens in the muscle of inoculated mice was detected by indirect immunofluorescence 7 days after immunisation. Specific IgG antibodies were significatively increased (P < 0.05) in animals that were reimmunized with 50 micrograms of the genomic library. An antigen specific lymphoproliferative response was detected in one animal of the group inoculated with one dose of the library.     

Variants of papillary carcinoma of the thyroid: experience at Asir Central Hospital

Twenty Trypanosoma cruzi stocks attributed to the 19/20, 32, and 39 major clones (Tibayrenc et al. 1986) were used to infect experimentally third instar larvae of Triatoma infestans. Three variables were considered: (i) percentage of infected insects; (ii) number of flagellates per insect (NFI); and (iii) percentage of metacyclic trypomastigotes per insect. Differences between the genotypes under study for all parameters considered were detected. These differences were statistically significant (P < 10(-3)), except between the 39 and 32 clonal genotypes for the NFI parameter. The correlation coefficient between the genetic distance and the biological parameters determined by the nonparametric Mantel's test was strongly significant (P < 10(-4)). Data obtained suggest clearly that populations of parasites belonging to the 19/20 genotype are more efficiently transmitted (high transmissibility genotype) by the vector than the 32 genotype (low transmissibility genotype), while the 39 genotype presents intermediary characteristic. Results confirm the working hypothesis that the subdivision of T. cruzi into discrete clonal lineages has an impact on the vectorial competence of T. infestans, the most important vector of the chagasic infection in South America, and that different clonal lineages do not exhibit the same vectorial transmissibility. This fact is relevant both for Chagas' disease epidemiology and for the use of xenodiagnosis.     

Systematics of Triatoma sordida, T. guasayana and T. patagonica (Hemiptera, Reduviidae)

Because of the relative epidemiological significance of Triatoma sordida, T. guasayana and T. patagonica, and the need to resolve doubts about their taxonomic validity, we report here a detailed taxonomic comparison of the three species using multivariate analysis of morphometric measures combined with comparisons of their genitalia and antennal structures. From the 17 metric variables studied, the length of the second segment of the rostrum and the anteocular length provided a discrimination function able to separate without error T. sordida from T. guasayana and T. patagonica. The multivariate discriminant functions classified T. guasayana and T. patagonica with an error of 2.44%. Comparison of the male genitalia of T. guasayana and T. sordida showed that there are minor differences in the articulatory apparatus, the median process of the pygophore, the phallosome support and the vesica, with bigger differences in the endosomal process and the phallosome. However, the already described male genitalia of T. patagonica is very similar to that of T. sordida. Analysis of antennal structure by scanning electron microscope showed that sensilla distribution around the pedicel is slightly different in the three species and sensilla density is highest in T. sordida and lowest in T. patagonica. The study showed that the three species form a closely related group. The results confirm the earlier classification of sordida and guasayana as separate species, but they raise some doubts about the taxonomic status of T. patagonica.     

Host preferences of Triatoma sordida

Triatoma sordida is a widespread vector of Trypanosoma cruzi, living in both sylvatic and domestic habitats. To see if Tri. sordida 'prefers' to feed from avian or mammalian hosts, two experiments were carried out. In the first experiment, in which fifth-instar nymphs were allowed free access to two unrestrained hosts, a guinea-pig and a pigeon, 98% of those feeding fed on the guinea-pig, and the mammalian bloodmeals were generally larger than the avian, with mean weights of 50.9 and 30.2 mg, respectively. In the second experiment, groups of nymphs were exposed to a single host (guinea-pig or pigeon) at two different densities (10 and 30 bugs/host). Whatever the density of the bugs, a much larger proportion of the bugs offered feeds on the guinea-pig took bloodmeals than of those offered the pigeon (55% v. 9%), and the bugs which fed on the mammal again generally took larger bloodmeals (mean weight = 50.9 mg) than the few which fed on the bird (mean weight = 12.8 mg; P < 0.0001). Bug density was found to have a greater effect on the size of the bloodmeals taken from pigeon than on those taken from guinea-pig. Triatoma sordida therefore does not appear to have a marked preference for avian blood; it is probably a generalist species that can invade a variety of habitats to exploit the range of available vertebrate hosts.     

Reducing the hindsight bias utilizing attorney closing arguments

Dogs, the main domestic reservoirs of Trypanosoma cruzi in the Argentine chaco, may be useful as sentinels of vector-mediated transmission of Trypanosoma cruzi in control programmes if canine infections acquired by all other routes could be excluded. In the present study, in the province of Santiago del Estero, Argentina, the dog populations in the neighbouring villages of Amamá, Trinidad, Mercedes, Villa Matilde and Pampa Pozo were studied immediately before spraying of residual insecticides (in 1992 in Amamá, Trinidad and Mercedes and 1993-1994 in the other villages) and twice during a subsequent programme of triatomine surveillance (in 1994 and 1996). Overall seropositivity for Trypanosoma cruzi infection (i.e. the proportion of dogs found positive by at least two different serological tests in any one survey) steadily declined from 65% (54/83) in 1992 to 39% (70/182) in 1994 and 15% (36/237) in 1996. No seroconversion was detected between the 1994 and 1996 surveys in any dog aged > 2 years. However, among the dogs aged < 2 years (which were born after virtual elimination of Triatoma infestans from the villages), seropositivity fell from 15% (12/81) in 1994 to 5% (6/131) in 1996. Thirteen demographic, behavioural and entomological variables were analysed by logistic multiple-regression analysis for their association with seropositivity in dogs born locally during the surveillance programme. The total number of Triatoma guasayana collected in the bedroom areas of the owner's house, having a mother which was seropositive for Trypanosoma cruzi, and the number of seropositive dogs with which it cohabited were all found to be significant predictors of seropositivity in any dog. The results are the first indication that Triatoma guasayana is the vector responsible for the new cases of Trypanosoma cruzi infection seen in dogs in domestic or peridomestic sites during the surveillance programme.     

Risk factors for Trypanosoma cruzi infection among children in central Brazil: a case-control study in vector control settings

This population-based case-control study was conducted in northern Goias State, central Brazil, in rural settings under vector control surveillance. One hundred forty-nine children seropositive for Trypanosoma cruzi antibodies, selected in a cross-sectional survey carried out in village schools, were compared with 298 seronegative classmate controls matched for age, sex, and place of residence. Information on potential environmental, familiar, and social economic risk factors for T. cruzi infection was collected during household visits, and interviews with parents and entomologic inspections of domestic and peridomestic environments were conducted. The presence of triatomines in dwellings or evidence of triatomine colonization was found to be statistically associated with seropositivity in children. The presence of exuviae and a report of triatomines indoors or outdoors by householders in the past were strong predictors of an infected child. Children from seropositive mothers had a 3.9-fold increase in the risk of having anti-T. cruzi antibodies after adjusting for the confounding variables, including triatomine capture, mother's age, and family size in multivariate analysis. Parent's report of vector presence showed a 97.7% sensitivity in identifying a dwelling with at least one seropositive child. The possibility of transplacental T. cruzi transmission and its implication for Chagas' disease control were considered.     

Relationships between neuroradiological and clinical features in apparently idiopathic hypopituitarism

Chagas disease vector insects Triatoma infestans and Rhodnius prolixus (fifteen stage III nymphs per 4 litre cage) were allowed to feed on anaesthetized mice for 1 h (control group), or on active non-anaesthetized mice (NAM) for 2, 4 or 8 h exposure. The bloodmeal size (weight increase) for both species was proportional to the duration of contact with NAM, due to ingestion of multiple small bloodmeals, up to 142% of control weight for T. infestans with 8 h exposure to NAM. The mean weight increase of T. infestans nymphs after 4 h contact with NAM was similar to that of the control group, whereas for R. prolixus, 8 h contact with NAM gave only 64% of the control value. For both species of insect, within 4 h of feeding, > 20% of the bloodmeal weight was lost by defaecation and diuresis. The proportions of unfed nymphs and mortality during 2 h contact with NAM were significantly higher for R. prolixus, demonstrating better exploitation of the host blood source by T. infestans, apparently because during blood-feeding the latter insect species caused less irritation to the host.     

Trypanosoma cruzi: effect of immunization on the risk of vector-delivered infection in guinea pigs

The protective effect of experimental immunization was studied in guinea pigs exposed to vectorial infection by Trypanosoma cruzi. Immunized animals received an inoculum of live-attenuated T. cruzi epimastigotes into a granuloma previously induced by Freund's complete adjuvant in the hind footpad. Seven days later, a delayed-type hypersensitivity reaction was triggered by reinjection of the parasites in the front footpad. The animals were then placed in Triatoma infestans-colonized corrals and exposed to vectorial T. cruzi transmission of the parasite for up to 200 days. The effectiveness of this immunizing protocol was controlled in terms of the number of bites necessary for infection (NBNI) in immunized as compared with control animals. Periodic entomological census allowed for the determination of vector biting and infection rates and the calculation of NBNI. Although this measurement was quite variable between yards, an overall average of 4,973 bites was enough to infect a control guinea pig in 4 separate experiments. The corresponding figure for the experimental group was 21,307 bites, implying that immunized animals could resist a 4.28-fold increase (range: 1.99-8.32) in the number of vector bites before becoming infected.     

Psychomotor development in children with triphalangeal thumbs. A preliminary study

A double-blind, randomized, clinical field trial was designed to test the efficacy and tolerance of a specific drug treatment in children in the indeterminate phase of infection by Trypanosoma cruzi. Children were treated with benznidazole at a dose of 5 mg/kg/day for 60 days or placebo and followed-up for 48 months. The treated children showed a significant decrease in geometric mean titers of antibodies against T. cruzi measured by indirect hemagglutination, indirect immunofluorescence, and ELISA. After a four year follow-up, 62% of the benznidazole-treated children and no placebo-treated child were seronegative for T. cruzi when tested by an ELISA using a T. cruzi flagellar calcium-binding protein (F29). Xenodiagnosis carried out after 48 months of follow-up was positive in 4.7% of the benznidazole-treated children and in 51.2% of the placebo-treated children. These results show the tolerance to and efficacy of benznidazole against T. cruzi in seropositive children six to 12 years of age. We used an early serologic marker of cure after treatment, consisting of a recombinant antigen implemented in a rapid, conventional serologic procedure.     

Immune information, self-organization and meaning

From January 1989 to April 1995, 465 specimens of Triatoma vitticeps were collected in the locality of Triunfo, 2nd District of Santa Maria Madalena municipal district, State of Rio de Janeiro. The bugs were found indoors by local residents with predominance of adults. The flight activity was high in hot months when the incidence in the domicile also increased. Two hundred and two bugs (111 alive and 91 dead) were examined for Trypanosoma cruzi infection. This was detected in 31 of the dead bugs (34%) and 88 (79%) of the live bugs examined. With a view to investigate the possible vertebrate hosts of the T. cruzi isolates, the blood of 122 mammals was examined through Giemsa-stained smears, hemocultures and xenodiagnosis. T. cruzi was detected in three specimens of Didelphis marsupialis and T. (M.) theileri was detected in one specimen of Bos taurus. The parasites were isolated from triatomine feces, xenoculture and hemoculture. No evidence of human infection was detected in 58 inhabitants examined, as evaluated by indirect imunofluorescence technique using T. cruzi epimastigotes as antigens. These results show that T. vitticeps is still a sylvatic species although nymphs have been found inside the domicile. Thus, an epidemiological vigilance is necessary to know the behaviour of this species following the continuous modifications promoted by the presence of man.     

Chagas disease: impact of Triatoma infestans control program in Alto del Carmen, Huasco Province, III Region Atacama, Chile

A control program of Triatoma infestans has been carried out in Alto del Carmen, an endemic chagasic rural county in the III Region, Chile. The program started in 1988 with an attack phase consisting in a masshouse spraying with residual insecticide, followed by an entomological surveillance phase with health education for community participation and vector detection in eventually reinfested houses. A yearly evaluation in 1992, 1993, 1994 and 1995 was carried out in order to determine the effectiveness of vector control activities. In 1992, 24.1% of dwellings was infested, whereas in 1993, 1994 and 1995 the infestation rates were 3.9%, 2.8% and 4.0% respectively. The similar infestation rates found in 1993, 1994 and 1995 suggest passive dispersion of triatomas from areas without surveillance. Additionally, in 1994, 110 (16.0%) out of 688 examined people resulted serologically positive. It is noteworthy that all of the children in the 0-4 year age group--born after the attack phase--resulted serologically negative. This fact may indicate the interruption of vectorial transmission of Chagas' disease in Alto del Carmen county. It is concluded that the control activities performed in the county constitute good strategies to the Nacional Program of Control of T. infestans, but for the success of such a program it is necessary to integrate the efforts of all endemic areas with an active community participation.