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Background: Triatomines are blood-sucking insects capable of transmitting Trypanosoma cruzi, the parasite that causes Chagas disease in humans. Vectorial transmission entails an infected triatomine feeding on a vertebrate host, release of triatomine infective dejections, and host infection by the entry of parasites through mucous membranes, skin abrasions, or the biting site; therefore, transmission to humans is related to the triatomine-human contact. In this cross-sectional study, we evaluated whether humans were detected in the diet of three sylvatic triatomine species (Mepraia parapatrica, Mepraia spinolai, and Triatoma infestans) present in the semiarid-Mediterranean ecosystem of Chile., Methods: We used triatomines collected from 32 sites across 1100 km, with an overall T. cruzi infection frequency of 47.1% (N = 4287 total specimens) by conventional PCR or qPCR. First, we amplified the vertebrate cytochrome b gene (cytb) from all DNA samples obtained from triatomine intestinal contents. Then, we sequenced cytb-positive PCR products in pools of 10-20 triatomines each, grouped by site. The filtered sequences were grouped into amplicon sequence variants (ASVs) with a minimum abundance of 100 reads. ASVs were identified by selecting the best BLASTn match against the NCBI nucleotide database., Results: Overall, 16 mammal (including human), 14 bird, and seven reptile species were identified in the diet of sylvatic triatomines. Humans were part of the diet of all analyzed triatomine species, and it was detected in 19 sites representing 12.19% of the sequences., Conclusions: Sylvatic triatomine species from Chile feed on a variety of vertebrate species; many of them are detected here for the first time in their diet. Our results highlight that the sylvatic triatomine-human contact is noteworthy. Education must be enforced for local inhabitants, workers, and tourists arriving in endemic areas to avoid or minimize the risk of exposure to Chagas disease vectors., (© 2023. The Author(s).)

It is not currently known which individuals with chronic Chagas disease (ChD) will develop cardiopathy in a determined period and which will be maintained asymptomatic with normal routine laboratory tests all their lives. The parasite burden is a factor that could explain this different evolution. The objective of this study was to quantify Trypanosoma cruzi burden by real-time PCR in blood (qPCR-B) and dejections of triatomines fed by xenodiagnosis (qPCR-XD) in 90 individuals with chronic ChD untreated, classified according to XD results and the presence or absence of cardiopathy. All individuals came from hyperendemic areas of Chile and participated in the study under Informed Consent. The standard qPCR curves for qPCR-B and qPCR-XD were elaborated with a mixture of known concentrations of T. cruzi strains, performing DNA serial dilutions (1/10) with a dynamic range between 10 5 and 10 -1 parasite equivalents/mL. The TaqMan Ⓡ detection system was applied in a Stratagene Mx3000P thermocycler (Agilent Technologies, USA) with cruzi 1 and cruzi 2 satellite primers. 22.2% and 15.6% of cases with cardiopathy or without cardiopathy were XD positive. There was no significant difference between the groups. The positivity of qPCR-B and qPCR-XD in the positive XD group was 82.35% and 100%, respectively, while in the negative XD group was 55.26% and 42.10%, respectively. A superior qPCR value in chronic ChD patients with and without cardiopathy was determined for qPCR in cases with positive XD and positive qPCR-XD. The receiver operating characteristic (ROC) curve analyses show better accuracy for detecting parasite burden (area under the curve, AUC) for qPCR-XD in comparison to qPCR-B. That is to say, major performance in DNA samples obtained of positive XD (gold standard for viable T. cruzi) detected and quantified by qPCR-XD. A high percentage of cases with XD and qPCR-XD positive (80-100%) have result concordant with qPCR-B. In absence of XD, future challenges are especially related to the low parasitic load of chronic ChD patients treated with trypanocidal drugs and post-therapy parasitological evaluations by qPCR-B. Finally, no statistically significant differences were found between presence or absence of cardiopathy and XD, qPCR-B or qPCR-XD., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Background: Trypanosoma cruzi is a protozoan parasite that is transmitted by triatomine vectors to mammals. It is classified in six discrete typing units (DTUs). In Chile, domestic vectorial transmission has been interrupted; however, the parasite is maintained in non-domestic foci. The aim of this study was to describe T. cruzi infection and DTU composition in mammals and triatomines from several non-domestic populations of North-Central Chile and to evaluate their spatio-temporal variations., Methodology/principal Findings: A total of 710 small mammals and 1140 triatomines captured in six localities during two study periods (summer/winter) of the same year were analyzed by conventional PCR to detect kDNA of T. cruzi. Positive samples were DNA blotted and hybridized with specific probes for detection of DTUs TcI, TcII, TcV, and TcVI. Infection status was modeled, and cluster analysis was performed in each locality. We detected 30.1% of overall infection in small mammals and 34.1% in triatomines, with higher rates in synanthropic mammals and in M. spinolai. We identified infecting DTUs in 45 mammals and 110 triatomines, present more commonly as single infections; the most frequent DTU detected was TcI. Differences in infection rates among species, localities and study periods were detected in small mammals, and between triatomine species; temporally, infection presented opposite patterns between mammals and triatomines. Infection clustering was frequent in vectors, and one locality exhibited half of the 21 clusters found., Conclusions/significance: We determined T. cruzi infection in natural host and vector populations simultaneously in a spatially widespread manner during two study periods. All captured species presented T. cruzi infection, showing spatial and temporal variations. Trypanosoma cruzi distribution can be clustered in space and time. These clusters may represent different spatial and temporal risks of transmission., Competing Interests: The authors have declared that no competing interests exist.

The etiologic agent of Chagas disease is Trypanosoma cruzi, a protozoan whose life cycle involves obligatory passage through vertebrate and invertebrate hosts in a series of stages. The aim of this study was to explore the transferability of mixed discrete typing units (DTUs) of T. cruzi present in chronic chagasic patients when passed through an invertebrate host during xenodiagnosis (XD) and then when transferred to axenic cultures to obtain T. cruzi isolates. DTUs of T. cruzi present in these two hosts and axenic cultures were identified by kDNA PCR amplification and subsequent hybridization with DTU-specific probes. Mixtures of Tc I, Tc II, Tc V and Tc VI DTUs were detected in blood samples. However as a result of XD and axenic cultures it was possible to identify mostly Tc V. We conclude that the transferability of an isolate of T.cruzi derived from mixed DTUs present in human blood depends upon the starved invertebrate host used for xenodiagnosis., (Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.)

The objective of this study was to investigate if there is specific host-parasite association in Chilean populations of Trypanosoma cruzi. For this purpose, two groups of parasites were analyzed, one from chronic chagasic patients, and the other from Triatoma infestans triatomines in three regions of the country. The first group consisted of four types of samples: parasites from peripheral blood of non-cardiopathic T. cruzi infected patients (NB); parasites from their corresponding xenodiagnosis (NX); parasites from peripheral blood of T. cruzi infected cardiopathic patients (CB) and parasites from their xenodiagnostics (CX). The T. infestans sample in turn was from three regions: III, V and M (Metropolitan). The genetic differentiation by the Fisher exact method, the lineage distribution of the samples, the molecular phylogeny and the frequency of multiclonality were analysed. The results show that not only are the groups of T. cruzi clones from Chagas disease patients and vectors genetically differentiated, but also all the sub-groups (NB, NX, CB and CX) from the III, V and M regions. The analysis of lineage distribution was concordant with the above results, because significant differences among the percentages of TcI, TcIII and hybrids (TcV or TcVI) were observed. The phylogenetic reconstruction with these Chilean T. cruzi samples was coherent with the above results because the four chagasic samples clustered together in a node with high bootstrap support, whereas the three triatomine samples (III, V and M) were located apart from that node. The topology of the tree including published T. cruzi clones and isolates was concordant with the known topology, which confirmed that the results presented here are correct and are not biased by experimental error. Taken together the results presented here are concordant with a specific host-parasite association between some Chilean T. cruzi populations.

Unlabelled: We evaluate the elimination of the microscopic stage of conventional xenodiagnosis (XD) to optimize the parasitological diagnosis of Trypanosoma cruzi in chronic Chagas disease. To this purpose we applied under informed consent two XD cages to 150 Chilean chronic chagasic patients. The fecal samples (FS) of the triatomines at 30, 60 and 90 days post feeding were divided into two parts: in one a microscopic search for mobile trypomastigote and/or epimastigote forms was performed. In the other part, DNA extraction-purification for PCR directed to the conserved region of kDNA minicircles of trypanosomes (PCR-XD), without previous microscopic observation was done. An XD was considered positive when at least one mobile T. cruzi parasite in any one of three periods of incubation was observed, whereas PCR-XD was considered positive when the 330 bp band specific for T. cruzi was detected. 25 of 26 cases with positive conventional XD were PCR-XD positive (concordance 96.2%), whereas 85 of 124 cases with negative conventional XD were positive by PCR-XD (68.5%). Human chromosome 12 detected by Real-time PCR used as exogenous internal control of PCR-XD reaction allowed to discounting of PCR inhibition and false negative in 40 cases with negative PCR-XD., Conclusion: PCR-XD performed without previous microscopic observation is a useful tool for detection of viable parasites with higher efficiency then conventional XD.

In Chile, the main vector of Chagas disease, Triatoma infestans, is under control after insecticide spraying. However, it has been found colonizing wild habitats. This study evaluated Trypanosoma cruzi infection of sylvatic and domiciliary T. infestans and identified their parasite genotypes. The sample studied was composed mainly of T. infestans sylvatic nymphs and domiciliary adults from a semi-urban area with human dwellings under vector control surveillance. Results showed prevalences of 57.7% in nymphs and 68.6% in adults. Hybridization tests showed a major T. cruzi lineage (TcI) circulating in sylvatic (93.3%) and domiciliary (100%) T. infestans. TcII, TcV, and TcVI were also detected, mainly in nymphs, suggesting differential adaptation of T. cruzi lineages among instars. We also discuss the origin of domiciliary individuals of T. infestans and the risk of human infection by triatomines of sylvatic foci that invade houses despite vector control programs.

In order to obtain more information about the population structure of Chilean Trypanosoma cruzi, and their genetic relationship with other Latino American counterparts, we performed the study of T. cruzi samples detected in the midgut content of Triatoma infestans insects from three endemic regions of Chile. The genetic characteristics of these samples were analysed using microsatellite markers and PCR conditions that allow the detection of predominant T. cruzi clones directly in triatomine midgut content. Population genetic analyses using the Fisher's exact method, analysis of molecular variance (AMOVA) and the determination of F(ST) showed that the northern T. cruzi population sample was genetically differentiated from the two southern population counterparts. Further analysis showed that the cause of this genetic differentiation was the asymmetrical distribution of TcIII T. cruzi predominant clones. Considering all triatomines from the three regions, the most frequent predominant lineages were TcIII (38%), followed by TcI (34%) and hybrid (8%). No TcII lineage was observed along the predominant T. cruzi clones. The best phylogenetic reconstruction using the shared allelic genetic distance was concordant with the population genetic analysis and tree topology previously described studying foreign samples. The correlation studies showed that the lineage TcIII from the III region was genetically differentiated from the other two, and this differentiation was correlated with geographical distance including Chilean and mainly Brazilian samples. It will be interesting to investigate whether this geographical structure may be related with different clinical manifestation of Chagas disease.

Objectives: This study compared three parasitological methods applied simultaneously in individuals with untreated chronic Chagas' disease in order to determine their individual and combined performances., Methods: From a total of 100 chronic chagasic patients from endemic areas of Chile, with informed consent, we extracted 2 mL of peripheral venous blood for PCR (PCR-B) and applied two xenodiagnosis (XD) boxes with seven uninfected Triatoma infestans nymphs each for microscopic examination and PCR of faecal samples of the triatomines fed on each patient (PCR-XD). The PCR-B and PCR-XD reactions were performed with oligonucleotides 121 and 122, which anneal to the four constant regions of the minicircles of Trypanosoma cruzi kinetoplasts. The 330 bp PCR product was analysed by electrophoresis in a 2% agarose gel and visualized by staining with ethidium bromide., Results: PCR-B detected T. cruzi in 58% of the cases, while PCR-XD proved to be more sensitive than XD (67% versus 14%, respectively) (P = 0.0001). There was no difference between the detection power of PCR-B and PCR-XD (P = 0.222). The percentage detected as positive was much greater when the three tests were considered (84%) (P = 0.00001)., Conclusions: The simultaneous application of more than one technique for the parasitological diagnosis of Chagas' disease in untreated individuals increases the possibility of detection of T. cruzi.

Chagas disease is one of the most important vector-borne diseases in Latin America. The disease, caused by the flagellate protozoan Trypanosoma cruzi, is commonly transmitted to humans by Triatoma infestans in South America. Using mitochondrial DNA sequences, we assessed alternative biogeographic scenarios of dispersal of T. infestans using coalescence simulations. We also assessed phylogeographic structure and spatial genetics of T. infestans in Chile. Two major routes of dispersal in southern South America were supported including a dual-origin of T. infestans in Chile. Phylogeographic analyses identified two primary clades with Chilean haplotypes partitioned into either a northern cluster with Peruvian and Bolivian haplotypes or a north-central cluster with Argentinean and Uruguayan haplotypes. The north-central clade is further divided into two subgroups. Domestic and sylvatic T. infestans in central Chile were not segregated in the phylogeographic reconstruction. Spatial genetic analyses show higher distances in northern Chile, congruent with the presence of two divergent lineages of T. infestans. Phylogenetic evidence does not unequivocally support the hypothesized Bolivian origin of T. infestans, so we discuss alternative scenarios., (Copyright © 2010 Elsevier B.V. All rights reserved.)

Triatoma infestans is one of the main domestic vectors of Chagas disease. Reports of wild habitat occurrences have recently increased. In Chile, after a successful elimination campaign of T. infestans domestic infestation, a sylvatic focus was reported in bromeliads in the metropolitan region. Here, we report a new focus of sylvatic T. infestans inhabiting rock piles in the Valparaíso region in central Chile. All T. infestans captured were nymphal instars living among the stones, which were inhabited by several mammal species, along with the sylvatic triatomine vector Mepraia spinolai. We found a prevalence of infection with Trypanosoma cruzi of 36.54% in T. infestans, similar to the previous report for sylvatic specimens from bromeliads. Sylvatic populations of T. infestans should be studied at different geographic scales to elucidate their role in the maintenance of the sylvatic transmission cycle of T. cruzi and their possible role in threatening the domestic elimination of this vector. This information should be used to re-design the control programs in Chile to avoid the re-establishment of the domestic cycle.

We report results of Trypanosoma cruzi infection and parasite genotypes in the wild Octodon degus and synantropic reservoir Rattus rattus from an endemic area with sylvatic Triatoma infestans as the only detected vector. The infection status was determined by hemi-nested PCR directed to minicircles DNA and genotyping by hybridization tests with a panel of five specific probes, including two probes for TcI subgroups (clones 19 and 20). O. degus was found infected with 13.3% and mainly with sublineage TcIId, and less with TcIIb and TcI. Meantime the synantropic R. rattus was found infected with 27.7% and mainly with TcI and much less with TcIId, TcIIb and TcIIe. The results are discussed to explain the distribution of T. cruzi genotypes between these two reservoirs and the importance of sylvatic foci of T. infestans allowing the permanence of the wild and peridomestic cycle of Chagas disease.

In the xenodiagnosis (XD) of American trypanosomiasis (Chagas disease), Trypanosoma cruzi in the triatomine bugs fed on the patient can now be detected using PCR (XD-PCR) as well as by microscopy (XD-M). In a study to compare XD-PCR with XD-M, triatomine bugs were fed on 50 cases of chronic American trypanosomiasis, of whom only 25 were ever found positive by XD-M. Overall, the bugs fed on 34 of the patients (all 25 cases found positive by XD-M and nine of the other patients) were found PCR-positive, giving a 330-bp fragment corresponding to part of the hyper variable region of the kinetoplast DNA of T. cruzi. Of the 25 patients who were ever found positive by XD-M, 20 gave bugs that were smear-positive on day 90 and a similar number (24; P=0.125) gave bugs that were PCR-positive at this time. On day 30, however, the bugs fed on only 11 of these 25 patients were found positive by microscopy, whereas 23 of these patients were found positive by XD-PCR (P=0.0016). Thus, not only was XD-PCR more sensitive than XD-M but it was also quicker, revealing more cases within 30 days than detected using XD-M over a period of 90 days.

We detected Trypanosoma cruzi in blood samples of the wild rodent Octodon degus by xenodiagnosis and a polymerase chain reaction (PCR) using the domestic and wild vectors of Chagas disease, Triatoma infestans and Mepraia spinolai, respectively. We captured 35 rodents and extracted DNA from blood samples and intestinal contents of vectors fed on O. degus. Our results indicate that the percentage of rodents naturally infected with T. cruzi depends on the biologic sample used for PCR and on the vector species for xenodiagnosis. The PCR with blood samples did not detect T. cruzi DNA, but the PCR with intestinal contents showed that both vectors were positive for T. cruzi. The PCR performed with M. spinolai intestinal contents detected four times more T. cruzi-positive O. degus than the PCR with Triatoma infestans intestinal contents (22.9% and 5.7%, respectively). We report the improvement of T. cruzi detection in sylvatic animals by a combination of PCR and xenodiagnosis using sylvatic vectors, especially in disease-endemic areas with low parasitemias in mammals.

Background: Insects of the subfamily triatominae are the biological vectors of Trypanosoma cruzi, the causal agent of Chagas disease., Aim: To search for wild colonies of triatomines in the Metropolitan Region of Chile., Material and Methods: Ad hoc traps were placed in two endemic zones of the Metropolitan Region of Chile, during 30 nights. The dejections of 16 T infestans and 43 M spinolai specimens were examined under the microscope, searching for live metacyclic trypomastigotes. A polymerase chain reaction (PCR) was performed in macerates of all insects looking for T cruzi DNA., Results: A total of 269 bugs were captured. Forty four were Triatoma infestans and 225 were Mepraia spinolai. They were not syntopic, since T infestans was restricted to a Southern zone (Calera de Tango) while M spinolai was only found in the Northern zone (Til-Til). Both species were found associated to terrestrial bromeliads (Puya sp) but M spinolai was also detected in stony grounds. Microscopic examination of dejections yielded a trypano-triatomine index of 56.3 and 32.6 for T infestans and M spinolai, respectively. PCR detected T cruzi DNA in 41 and 43% of T infestans and M spinolai specimens, respectively., Conclusions: The finding of T infestans in a wild habitat is noticeable. This is the first report of such phenomenon in Chile. The high infection rates with T cruzi, explains the maintenance of Chagas disease wild cycle in Chile.

Background: The great variability in the clinical presentation of Chagas disease may depend in part in the genetic variability of Trypanosoma cruzi populations., Aim: To compare prepatent period, parasitemia, mortality and histological lesions in mice infected with two populations of Trypanosoma cruzi isolated in Chile., Material and Methods: Two Trypanosoma cruzi populations, isolated from Chilean Triatomides and genetically characterized by kinetoplast restriction fragment DNA profiles, were compared. Two groups of 40 Balb/c mice were studied. Each mouse was inoculated with 10(4) trypomastigotes, of the V-121 and sp COMB 2 Trypanosoma cruzi populations. The prepatent period, parasitemia, mortality and histopathological lesions, at different evolutionary stages of infection were registered during 32 days., Results: Prepatency and mortality were similar in both groups of mice. However, parasitemia was significantly greater in mice inoculated with V-121 than those inoculated with sp COMB 2. Amastigote pseudocysts and inflammation were present only in skeletal muscle and myocardium in both groups of mice. The intensity of tissue involvement was associated to the level of parasitemia, therefore it was greater in mice inoculated with V-121 population., Conclusions: V-121 population of Trypanosoma cruzi caused a greater parasitemia than COMB 2, in inoculated mice.

In total, 61 Chilean isolates of Trypanosoma cruzi, were analysed using schizodeme and Southern analysis, using as probes the highly variable regions of minicircles from cloned parasites. Isolates were collected and amplified from domestic and wild triatomines, and from infected subjects in all the endemic areas of Chile. Three major parasite genotypes could be detected in the domestic transmission cycle, whilst 1 major T. cruzi genotype is circulating in the wild transmission cycle. Schizodeme analysis suggested that T. cruzi isolates are mixed populations, whereas the Southern analyses detected only 3 mixed isolates using 4 selected minicircle segments as probes.

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.

Many studies have attempted to assess the relative effects of different vectors of a disease on animal populations. To this end, three measures have been proposed: Vectorial efficiency, Vectorial capacity and recently Vectorial effectiveness (or Vectorial impact). In this study we relate these measures to derive some of their properties emphasising in the vectorial impact for its importance in both, population performance of parasites and the proportion of the prevalence of one parasite due to a given vector. We applied the quantitative expressions advanced in this study to a simple Chilean example with one parasite (Trypanosoma cruzi), two vectors (Triatoma infestans and Mepraia spinolai) and one animal population (humans: Chagas's disease).

Background: Triatoma spinolai is the only wild vector for Chagas disease in Chile and its epidemiological importance is being studied., Aim: To study the proportion of insects infected with Trypanosoma cruzi (tripano-triatomine index) in a zone with epidemiological risk in the Metropolitan Region of Chile., Methods: Four hundred ninety two specimens of Triatoma spinolai were collected in four sites of a quarry zone, 14 kilometers north of Santiago. Their maturity and the presence of Trypanosoma cruzi in their intestinal contents were determined., Results: Mean tripano-triatomine index was 26.02 +/- 2% (range 0 to 34% in different sites). The proportion of infected insects increased along with their maturity and 58% of adult specimens were infected. There was a seasonal variation of the proportion of infected specimens, being lower in March and June and higher in July and February., Conclusions: The studied zone has a potential epidemiological risk for the transmission of Chagas disease by Triatoma spinolai.

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.

Twenty-one Trypanosoma cruzi stocks isolated from Triatoma infestans and humans of the Chilean Andean highlands were studied at the genotypic level by schizodeme and molecular karyotype analyses, which allowed a clear distinction of the parasites from those hosts. A phenotypical characterization was performed by proteolytic activity after electrophoretic fractionation without discrimination among the stocks. Metacyclic trypomastigotes obtained in vitro proved to be infective to Swiss mice and the study of immune response and biological behavior was assessed. Of a total of 21 T. cruzi stocks, only 11 proved to be infective in mice due to difficulties in obtaining metacyclic trypomastigotes with the parasite populations isolated from humans. Western blot analysis revealed a complex immune response even in the first days postinfection with each T. cruzi strain studied. Antigenic recognition by each immune serum is characteristic, although several major and common antigens were detected. Lytic antibodies were studied by the in vitro complement-mediated lytic assay using purified metacyclic trypomastigotes as target cells. All the T. cruzi isolates tested induced lytic antibodies in this experimental model. Parasitemias were moderate and characteristic for each T. cruzi strain. Results are compared with metacyclic forms of the infective and pathogenic Tulahuen strain.

Triatoma spinolai is a wild species of triatomine existing exclusively in Chile, with scarce epidemiological importance in the transmission of Trypanosoma cruzi to man. It has been found in environments from 3000 meters down to the sea level. Exceptionally it has been found in human dwellings. In order to clarify the possibility of an adaptation of T. spinolai to human dwellings a study was undertaken in 50 rural-periurban dwellings from regions III, IV, V and Metropolitan in which this triatomine was found. The dwellings were classified in houses (35) and huts (15). The insect was trapped in 35 dwellings (24 houses and 11 huts) located in declivities (70%) and 15 dwellings (14 and 1) located in plains. This proportion is significantly higher than the average of 26.2% for dwellings located in declivities. It is concluded that the presence of T. spinolai in dwellings, more than a triatomine invasion is the result of the construction of huts or houses in the insect habitat.

Triatoma infestans is the principal vector for Chagas disease in Chile. Although there is enough epidemiological information about human infection, the effect of vector and host (animals and man) populations interrelations over Chagas disease prevalence has not been studied. This work studies a determinist mathematical model for the dynamics of Chagas disease transmission by Triatoma infestans, relating entomological population parameters with infection propagation. The effect of invasion of susceptible animal and human populations by infected triatomine insects and the stability of Chagas disease endemic was simulated. The Ro parameter was estimated. This is a gross appraiser of the infection reproductive rate that relates triatomine insect density, bite rate and mortality. The infected population at equilibrium was estimated. The resulting endemic was highly stable with a great amount of chronic and few acute cases. There is always an acute Chagas epidemic stage after a recent invasion. The need for a threshold triatomine insect population for the genesis of an endemic is proposed.

Chagas' disease is endemo-enzootic in periurban and rural sections of the first seven (18 degrees 30'-34 degrees 36' South lat.), out of the thirteen political-administrative regions of Chile. Permanent inhabited localities situated at different altitudes over the sea level (OSL) are classified in: coastal settlement (0-1,499 m OSL), intermediate settlement (1,500-3,000 m OSL) and altiplano settlement (> 3,000 m OSL). A total of 1,558 dwellings and their occupants from regions I and II were surveyed in 1982-1984 for different epidemiological Trypanosoma cruzi infection parameters. Hence, 261 dwellings, 189 domestic mammals and 517 people (ages 5- > or = 60 years) from the Altiplano settlement were studied. The results obtained were: 4(1.5%) and 2(0.8%) of dwellings infested with Triatoma infestans, according to occupants information and by observation of direct evidences respectively; 8(4.2%) animals (1 dog, 4 goats and 3 sheep) with a positive hemagglutination test (IHAT) for Chagas' disease; 21 people (4.1%) with a positive IHAT; 4(19.0%) and 3(14.3%), out of the 21 IHAT positive individuals with all types of electrocardiogram (EKG) abnormalities and EKG abnormalities suggestive of a chagasic etiology respectively, whereas 65(13.1%) and 15(3.0%), out of the 496 IHAT negative ones presented the same kinds of EKG abnormalities.(ABSTRACT TRUNCATED AT 250 WORDS)

Examination of 54 Triatoma infestans from a village near the European Southern Observatory La Silla in Chile and of 9 Triatoma spinolai from the territory of the observatory showed that 10 T. infestans were infected with trypanosomatids. Mice were infected with in vitro cultures initiated with five different trypanosomatid isolates and treated with the immunosuppressive drug cyclophosphamide to increase the parasitemia of the flagellates. Evidence of the presence of T. cruzi was provided by a comparative biometrical analysis of blood trypomastigotes and the occurrence of intracellular amastigotes. Three methods for further identification were used: examination of kDNA ultrastructure, disc electrophoresis of soluble proteins and the Aaptos papillata II lectin induced agglutination. We obtained the following results for all isolates: (1) presence of a central band of the kDNA; (2) T. cruzi specific double bands of the protein patterns; (3) positive reaction with Aaptos papillata II. No differences between the five isolates from Chile and T. cruzi or T. cruzi-like strains from other countries could be observed. Based on these results an infection of the bugs with T. rangeli and T. conorhini could be excluded.

Chagas disease probably affects over half a million people in Chile, principally in rural communities in the fertile valleys of the arid 'norte chico' region, north of Santiago. The main domestic vector is Triatoma infestans, but Triatoma spinolai, although mainly in rocky sylvatic and peridomestic ecotopes, also invades houses. Since the Spanish invasion in the sixteenth century, and particularly during the last 100 years, the endemic region has suffered an ecological breakdown, largely due to excessive timbering and over-grazing, which has led to a denuded landscape with severe loss of agricultural productivity. This breakdown, combined with uneconomically sized farms and poor marketing, exacerbates the poverty of the rural communities. As in other similar areas of Latin America, the combination of poverty and poor education discourages improvements in housing which would reduce the risk of vector-transmitted Chagas disease. This paper reviews the historical and ecological background of the endemic region of Chile, both as a basis for further work, and as a point of comparison with other endemic areas. The review attempts to show how the current status of Chagas disease is likely to be maintained through its association with poor quality housing, poverty and ecological degradation, drawing parallels with other endemic++ areas and suggesting ways by which the ecological damage might be reversed.

Fifty-three stocks of Trypanosoma cruzi were isolated in Chile, 13 from patients, 32 from the domestic triatomine vector Triatoma infestans, and 8 from the silvatic and peridomestic vector T. spinolai. The majority of isolates from triatomine bugs were made by the direct culture of infected faeces. The 53 stocks and a single clone were characterized by a combination of starch-gel and cellulose acetate enzyme electrophoresis. Three groups of T. cruzi stocks were apparent from either simple visual comparisons of isozyme profiles or numerical taxonomy. The groups were designated Chilean zymodeme (Z) 1, which was similar to Brazilian Z1, Chilean Z2a, similar to Brazilian Z2 and Chilean Z2b, similar to Bolivian Z2 and with prominent heterozygous isozyme profiles. Chilean Z1 was isolated only from T. spinolai colonizing farm walls inhabited by the rodent Octodon degus. Chilean Z2a and Z2b were both isolated from domestic T. infestans T. infestans and man, in some cases within the same household. Hardy-Weinberg equilibria were not found amongst a group of 22 stocks from a single locality and deviations from theoretical Hardy-Weinberg distributions were compatible with the absence of genetic exchange in the sampled population of T. cruzi.

Culture forms of 12 Chilean and 9 Bolivian Trypanosoma cruzi stocks were compared isoenzymatically by the following enzymes: non-specific esterase, phosphoglucomutase, glucose-6-phosphate dehydrogenase, glucosephosphate isomerase, and alcohol dehydrogenase. On the basis of the electrophoretic mobility of these enzymes the stocks were classified into two main groups. Ten Chilean stocks were characterized as group II; two stocks showed enzyme patterns of group I. In contrast, five Bolivian stocks were classified as belonging to group I, the other four to group II. The results show that the two groups of T. cruzi overlap in Triatoma infestans suggesting that both groups of T. cruzi are infective for man. The classification of stocks into two groups is discussed in the light of published results of Brazilian T. cruzi stocks. A strong association of groups with the transmission cycles as it seems to be in Brazil does not exist in Chile and Bolivia.