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.     

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.     

Domestic infestation by Triatoma infestans and prevalence of Trypanosoma cruzi seropositives in a rural area of the Argentinian northeast

An epidemiological study was carried out from April 1991 to December 1993 to obtain a general view of T. cruzi domestic transmission in rural areas of San Miguei Department, Corrientes. From 100 analyzed households, 50.0% was infested by T infestans (Klug, 1834) and 1.0% by T. sordida (Stal, 1859). Domiciliary colonization by T. sordida is reported for first time in Corrientes province. T. cruzi infection of T. infestans was 23.1%. Indirect hemagglutination and indirect immunofluorescence antibody tests were used for detection of anti T. cruzi antibodies in 388 human sera, 23.4% showed serological reactivity. An important high rate (12.9%) was observed in the age group of higher transmission risk. Seropositives percentages increased with age and reached 50.0% in 31-40 years old group. Domestic infestation by T. infestans, seropositive human prevalence to T. cruzi and householders precarious life conditions prove that this endemic disease is still a problem in the studied area.     

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.     

Clonal population structure of Colombian sylvatic Trypanosoma cruzi

Isoenzyme variability and evidence of genetic exchange were evaluated in 75 wild stocks of Trypanosoma cruzi obtained from different hosts from 5 geographical regions within the endemic area in Colombia. Cluster analysis of genetic variability was attempted. Thirty-three multilocus enzyme genotypes (clonets) were identified from 75 stocks, 27 of which clustered with zymodeme Z1 and 6 with zymodeme Z3. Two stocks isolated from human infections showed the potential risk to rural communities in Colombia. The stocks exhibited departures from Hardy-Weinberg expectations, including both fixed heterozygote and fixed homozygote demes, where both segregation and recombination were absent. To inspect for population subdivision that might falsely imply clonality in these stocks, Wright's F statistics were calculated. Theta values (Fst) were significantly different from 0 when 33 clonets, 27 Z1-like clonets, and 5 geographical subpopulations were compared; thus, a significant amount of divergence has occurred between and within them. In addition, linkage disequilibrium was detected for most possible pairwise comparisons of loci. In conclusion, the above results all support a scenario of long-term clonal evolution in Colombian sylvatic T. cruzi populations.     

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.     

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.     

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.     

Genetic variability and geographic differentiation among three species of Triatomine bugs (Hemiptera-Reduviidae)

Three species of triatomine bugs, Triatoma sordida, T. guasayana, and T. patagonica, were examined by cytogenetic (C-banded karyotypes and male meiotic process) and isoenzymatic studies. These three species, with different importance as Chagas' disease vectors, were found to be closely related according to their known ethologic, ecologic, and morphologic traits. Although they have the same diploid chromosome number (2n = 22 constituted by 20 autosomes and an XY male/XX female sex mechanism), each species has a distinct chromosomal behavior during male meiosis and a specific amount and localization of C-heterochromatic blocks. Moreover, these chromosome characteristics allowed us to differentiate two T. sordida populations. Isoenzymatic data confirmed the taxonomic status of the three species and together with our cytogenetic results questioned the species homogeneity of T. sordida.     

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.     

Beauveria bassiana (Hyphomycetes) as a possible agent for biological control of Chagas disease vectors

Beauveria bassiana (Balsamo) Vuillemin (isolate CG306) was tested on 3rd instars of 9 Triatoma spp., 4 Rhodnius spp., 2 Panstrongylus spp. and Dipetalogaster maxima (Uhler) at 25 degrees C and 50% RH. Quantitative sporulation of the fungus on cadavers was studied at 25 degrees C and 97% RH. Mortality, estimates of survival time, and conidial production on cadavers differed significantly among the genera and species tested. Panstrongylus herreri Wygodzinsky, Dipetalogaster maxima, Triatoma picturata Usinger, Rhodnius robustus Larrousse, Rhodnius prolixus St?l, Triatoma infestans (Klug), and Triatoma brasiliensis Neiva were most susceptible to fungal infection. Most conidia per cadaver were produced on Triatoma williami Galv?o, Souza & Lima and Triatoma lecticularia (St?l). Results indicate that B. bassiana (CG306) might be a candidate for control of important or potential vectors of Trypanosoma cruzi using the fungus as a bioinsecticide.     

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.     

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.     

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.     

Metric differences between silvatic and domestic Triatoma infestans (Heteroptera: Reduviidae) in Bolivia

One hundred and fifty-seven specimens of Bolivian Triatoma infestans (Klug 1834), including 44 from the silvatic focus at Cochabamba (Bolivia), were compared using morphometric characters of the head capsule. From these specimens, 10 silvatic and 28 domestic adults were also compared using additional morphometric characters of the membranous part of the hemelytra. Nonparametric univariate comparisons between specimens from silvatic and domestic ecotopes indicated the importance of the postocular region as a possible diagnostic character for nymphs and adults from the different ecotopes, and they detected wing differences in males. Populations became more distinct entities when head or wing characters were considered jointly in canonical variate analysis. Regardless of whether size variation was considered, canonical variate analysis generally showed greater significance for wing than for head features. These morphological differences between silvatic and domestic bugs, particularly unrelated to size differences, are interpreted to indicate incipient separation between silvatic and domestic populations that had not been detected by previous isoenzyme analyses, and suggest a reinterpretation of the epidemiological significance of silvatic populations of T. infestans in Bolivia.     

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.     

Genetic multilocus studies of different strains of Cryptococcus neoformans: taxonomy and genetic structure

The genotypes of 107 strains of Cryptococcus isolated from the environment or from patients from various geographical areas were determined by multilocus enzyme electrophoresis (MLEE). We analyzed the relationships between genotype structure and serotype and between genotype structure and strain origin. Twelve of the 14 enzyme-encoding loci studied were polymorphic, giving rise to 48 electrophoretic types. The genotypes of C. neoformans and C. laurentii were very similar. MLEE could not distinguish between these two pathogenic species. A correlation between the genetic multilocus structure and the origin of the sample (from the environment or patients) existed. A second analysis detected a correlation between genotype distribution and serotype. The second analysis considered three serotype groups (B, C, and A plus D plus A/D), proving that serotypes A, D, and A/D are closely related. MLEE is a useful epidemiological tool for improving our understanding of the biology of this fungus.     

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.     

Effect of fasting on the composition of the fat body lipid of Dipetalogaster maximus, Triatoma infestans and Panstrongylus megistus (Hemiptera: Reduviidae)

Modifications in content and lipid composition induced by fasting were examined in fat bodies from adults of Triatominae, Dipetalogaster maximus, Triatoma infestans and Panstrongylus megistus. With fasting, total lipid stores dropped approximately 50% for T. infestans and more than 70% for P. megistus. Total lipids analyzed by thin layer chromatography and fractionated by column chromatography on Unisil showed triacylglycerols as the main component in the three species, although P. megistus showed high levels of diacylglycerols (31-46%). Cholesterol amounted to 8-15%. In diacylglycerol fractions, C16:0, C18:1 and C18:0 fatty acids were detected; their ratio varied with species but it was not dependent on nutritional status. In triacylglycerol fractions C18:1 fatty acid was the major component at different times (48-68%); the ratio of monounsaturated to saturated in this fraction was 1.3, 2.6 and 1.2 for D. maximus, T. infestans and P. megistus respectively. The remarkable drop in lipid stores without noticeable changes in their relative composition would suggest that all types of lipid are used at similar rates. The higher content of diacylglycerols in P. megistus may be associated with the better flight performance of this species.     

Factors limiting the domestic density of Triatoma infestans in north-west Argentina: a longitudinal study

Reported are the environmental and demographic risk factors associated with the domestic infestation and density of Triatoma infestans in three heavily infested rural villages in Santiago del Estero Province, Argentina. In a one-factor unadjusted analysis, the number of T. infestans captured per person-hour was associated significantly and negatively with the use of domestic insecticides by householders, type of thatch used in the roofs and the age of the house; and positively with the following: degree of cracking of the indoor walls and presence of hens nesting indoors. In one model, using multiple linear regression and a backward stepwise elimination procedure, most of the variation in the overall abundance of T. infestans was explained by insecticide use and the presence of hens nesting indoors; in another model using the same procedure it was explained by insecticide use, bug density in 1988 and previous spraying with deltamethrin in 1985. Variations in bug density per capture stratum (household goods, beds, walls and roof) were explained by the bug density in other strata and by one or two of the following risk factors: hens nesting indoors, type of roof, presence of cracks in the walls and number of people living in the house. Bug density might be locally controlled by the availability of refuges in the roofs and walls, by the presence of hens nesting indoors and by the use of domestic insecticides. Certain local materials, such as a grass known as simbol, could be successfully used in rural housing improvement programmes aimed at reducing the availability of refuges for insects in the roof.