High Parasitic Loads Quantified in Sylvatic Triatoma melanica, a Chagas Disease Vector.

Triatoma melanica is a sylvatic vector species in Brazil. In We aimed to characterize the Trypanosoma cruzi discrete typing units (DTUs), the parasitic loads, and the blood meal sources of insects collected in rocky outcrops in rural areas in the state of Minas Gerais. An optical microscope (OM) and kDNA-PCR were used to examine natural infection by T. cruzi, and positive samples were genotyped by conventional multilocus PCR. Quantification of the T. cruzi load was performed using qPCR, and the blood meal sources were identified by Sanger sequencing the 12S rRNA gene. A total of 141 T. melanica were captured. Of these, ~55% (61/111) and ~91% (63/69) were positive by OM and KDNA-PCR, respectively. We genotyped ~89% (56/63) of the T. cruzi-positive triatomines, with TcI (~55%, 31/56) being the most prevalent DTU, followed by TcIII (~20%, 11/56) and TcII (~7%, 4/56). Only TcI+TcIII mixed infections were detected in 10 (~18%) specimens. A wide range of variation in the parasitic loads of T. melanica was observed, with an overall median value of 104 parasites/intestine, with females having higher T. cruzi loads than N2, N4, and N5. TcII showed lower parasitic loads compared to TcI and TcIII. The OM positive diagnosis odds ratio between T. cruzi infection when the parasite load is 107 compared to 103 was approximately 29.1. The most frequent blood meal source was Kerodon rupestris (~58%), followed by Thrichomys apereoides (~18%), Wiedomys cerradensis (~8%), Galactis cuja (~8%) and Gallus gallus (~8%). Our findings characterize biological and epidemiological aspects of the sylvatic population of T. melanica in the study area, highlighting the need to extend surveillance and control to this vector.

Avaliação do conhecimento sobre a doença de Chagas em escolares das zonas rural e urbana de municípios endêmicos em Minas Gerais / Assessment of knowledge about Chagas disease in schoolchildren in rural and urban areas of endemic municipalities in Minas Gerais state, Brazil

Resumo O objetivo deste estudo foi avaliar o conhecimento de escolares do ensino médio de três escolas localizadas em municípios mineiros endêmicos para a doença de Chagas e extrair informações relativas à sua percepção em relação aos aspectos gerais, biológicos, clínicos e epidemiológicos da doença, do vetor e controle entomológico da doença de Chagas, além de comparar o conhecimento dos participantes em função da localização das escolas (zona rural ou urbana). Para isso, aplicou-se um questionário semiestruturado a 68 alunos. Todos os participantes já ouviram falar a respeito da doença de Chagas, afirmando ter adquirido esse conhecimento, sobretudo, com familiares e escola. Além disso, muitos participantes demonstraram conhecer os hábitos dos vetores, tais como hematofagia e fontes alimentares. Não houve associação clara entre o perfil de urbanização e o conhecimento dos participantes. No entanto, alunos da zona urbana apresentaram menos conhecimento prático a respeito dos triatomíneos e aspectos relativos à transmissão da doença, em relação aos alunos da zona mais "ruralizada". Sugere-se o aprofundamento do tema nos currículos escolares do ensino médio e a necessidade de elaboração de ações de educação em saúde voltadas à vigilância entomológica da doença de Chagas nesses municípios e, provavelmente, em outras regiões do Brasil.

Abstract This study aimed to evaluate the knowledge of high school students from three schools located in endemic municipalities of Minas Gerais to Chagas disease and to extract information related to their perception regarding the general, biological, clinical and epidemiological aspects of the disease, vector and entomological control of Chagas disease, in addition to comparing the knowledge of participants according to the location of schools (rural or urban area). For this, a semi-structured questionnaire was applied to 68 students. All participants have heard about Chagas disease, stating that they have acquired this knowledge, especially with family members and school. In addition, many participants demonstrated to know the habits of vectors, such as hematophagy and dietary sources. There was no clear association between the urbanization profile and the participants' knowledge. However, students from the urban area presented less practical knowledge about triatoins and aspects related to the transmission of the disease, in relation to students in the most "ruralized" area. It is suggested the deepening of the theme in high school curricula and the need to develop health education actions aimed at entomological surveillance of Chagas disease in these municipalities and, probably, in other Brazilian regions.

Fast recovery of house infestation with Triatoma brasiliensis after residual insecticide spraying in a semiarid region of Northeastern Brazil.

The northeastern semiarid region stands out in the Brazilian context regarding the eco-epidemiology of Chagas disease, in which Triatoma brasiliensis is the main vector of Trypanosoma cruzi. Persistent house invasion threatens the relative levels of progress achieved over previous decades. We conducted an intervention trial with a five-year follow-up to assess the impacts of residual spraying with pyrethroid insecticides on house infestation with T. brasiliensis in 18 rural villages (242 houses) located in the Tauá, Ceará. House infestations were assessed by systematic manual searches for triatomines in every domestic and peridomestic habitat on five occasions. Triatomines were collected in peridomestic (57.5%), sylvatic (35.8%), and intradomiciliary (6.7%) habitats. The most important ecotopes of T. brasiliensis were containing roofing tiles, bricks or rocks (23.4% ± 9.1). Residual insecticide spraying substantially reduced baseline house infestation rates from 27.9% to 5.9% by 6 months post first spraying (MPS). The decline was substantially greater in intradomiciles (11.2% to 0.8%) than in peridomiciles (16.7% to 5%). The mean relative density of triatomines recovered its preintervention values at 14 MPS in intradomiciles, and in the main peridomestic ecotopes. The house infestation levels recorded at 14 MPS persisted thereafter despite all reinfested houses were selectively sprayed on every occasion. Overall average bug infection rates with T. cruzi in the five occasions were in intradomiciles (11.1%), peridomiciles (4.7%) and wild habitats (3.3%). In peridomicile T. cruzi infection rates decreased significantly at all stages after chemical intervention. In intradomicile, the only significant difference occurred at 20 MPS (7.7% to 30.8%). The vectorial capacity of T. brasiliensis, combined with its invasive potential from sylvatic sources and the limited effectiveness of chemical control in the harsh caatinga landscape, pose serious obstacles to the definite elimination of domestic transmission risks. Systematic vector surveillance supported by community participation and locally adapted environmental management measures are needed to reduce the risks of establishment of domestic transmission with T. cruzi in this region.

Triatoma brasiliensis Neiva, 1911: food sources and diversity of Trypanosoma cruzi in wild and artificial environments of the semiarid region of Ceará, northeastern Brazil.

BACKGROUND: Knowledge of triatomine food sources in different ecotopes enables the estimation of T. cruzi transmission risk in diverse environments, as well as its dynamics of dispersion and ecological niche. For Triatoma brasiliensis in the Caatinga, in the northeast of Brazil, seasonal differences influence feeding eclecticism and rates of T. cruzi infection. The objective of the present study was to monitor food sources and to characterize the populations of T. cruzi associated with T. brasiliensis in wild and domestic environments in the Caatinga of northeast Brazil. METHODS: A cross-sectional study based on a search for triatomines in wild and domestic environments, was undertaken at five different time periods from 2009 to 2015. Insects from 2015 were used for identification of food sources. Two universal primers, based on the conserved regions of the 12S rRNA locus, were used to amplify fragments of 215 bp. The content of the intestinal tract of triatomines was identified by a comparison between the sequences obtained and those deposited in the GenBank database, using BLAST. In triatomines with parasitological diagnosis of infection by trypanosomatids, xenoculture was performed for the isolation and characterization of strains, using cox2, the amplification of the SL-IL mini-exon intergenic spacer and the polymorphism of the D7 divergent domain of the gene 24αrDNA-LSU. RESULTS: Food sources were identified in 76.3% (213/279) T. brasiliensis specimens sampled in 2015. The most frequent sources in a total of 20 vertebrate species were: rodents (58%, 123/213), ruminants (30%, 64/213) and cats (6%, 12/213). A total of 49% (44/89) of the samples of T. cruzi isolated in the period from 2009 to 2015 were characterized: TcII (43%, 19/44), TcI (41%, 18/44) and TcIII (16%, 7/44). CONCLUSIONS: The feeding eclecticism of T. brasiliensis shows its importance in maintaining the transmission dynamics of T. cruzi, with evidence of intense circulation between anthropic and wild environments. Attention should be placed on the association among T. brasiliensis, rodents and ruminants, in addition to the presence of TcIII in the study region.

Description of an amazing nest of Caatinga Cachalote Pseudoseisura cristata infested by Psammolestes tertius in Tauá, state of Ceará, Northeastern Brazil

Psammolestes tertius Lent & Jurberg 1965 is a triatomine species part of the tribe Rhodniini. It is only wild and found in bird nests, mostly of the Furnariidae family. This work describes the amazing infestation of a nest of Pseudoseisura cristata in the municipality of Taua, Ceará. The nest harbored a colony with about 40 specimens of P. tertius in all evolutionary stages, all negative for trypanosomatids. A rodent of the species Wiedomys pyrrhorhinos was seen running away from the nest. The materials used to build the nest included the typical natural wigs, and various anthropogenic materials such as razor blades, shaver, fork, sunglasses, metal fragments, fragments of plastic objects, wire, barbed wire, plastic sandal straps, paper, plastic lid, pieces of plastic bags, gas cylinder label, large feathers and the exuviae of a snake.

Domestic, peridomestic and wild hosts in the transmission of Trypanosoma cruzi in the Caatinga area colonised by Triatoma brasiliensis.

The role played by different mammal species in the maintenance of Trypanosoma cruzi is not constant and varies in time and place. This study aimed to characterise the importance of domestic, wild and peridomestic hosts in the transmission of T. cruzi in Tauá, state of Ceará, Caatinga area, Brazil, with an emphasis on those environments colonised by Triatoma brasiliensis. Direct parasitological examinations were performed on insects and mammals, serologic tests were performed on household and outdoor mammals and multiplex polymerase chain reaction was used on wild mammals. Cytochrome b was used as a food source for wild insects. The serum prevalence in dogs was 38% (20/53), while in pigs it was 6% (2/34). The percentages of the most abundantly infected wild animals were as follows: Thrichomys laurentius 74% (83/112) and Kerodon rupestris 10% (11/112). Of the 749 triatomines collected in the household research, 49.3% (369/749) were positive for T. brasiliensis, while 6.8% were infected with T. cruzi (25/369). In captured animals, T. brasiliensis shares a natural environment with T. laurentius, K. rupestris, Didelphis albiventris, Monodelphis domestica, Galea spixii, Wiedomys pyrrhorhinos, Conepatus semistriatus and Mus musculus. In animals identified via their food source, T. brasiliensis shares a natural environment with G. spixii, K. rupestris, Capra hircus, Gallus gallus, Tropidurus oreadicus and Tupinambis merianae. The high prevalence of T. cruzi in household and peridomiciliar animals reinforces the narrow relationship between the enzootic cycle and humans in environments with T. brasiliensis and characterises it as ubiquitous.

Domestic, peridomestic and wild hosts in the transmission of Trypanosoma cruzi in the Caatinga area colonised by Triatoma brasiliensis

The role played by different mammal species in the maintenance of Trypanosoma cruzi is not constant and varies in time and place. This study aimed to characterise the importance of domestic, wild and peridomestic hosts in the transmission of T. cruzi in Tauá, state of Ceará, Caatinga area, Brazil, with an emphasis on those environments colonised by Triatoma brasiliensis. Direct parasitological examinations were performed on insects and mammals, serologic tests were performed on household and outdoor mammals and multiplex polymerase chain reaction was used on wild mammals. Cytochrome b was used as a food source for wild insects. The serum prevalence in dogs was 38% (20/53), while in pigs it was 6% (2/34). The percentages of the most abundantly infected wild animals were as follows: Thrichomys laurentius 74% (83/112) and Kerodon rupestris 10% (11/112). Of the 749 triatomines collected in the household research, 49.3% (369/749) were positive for T. brasiliensis, while 6.8% were infected with T. cruzi (25/369). In captured animals, T. brasiliensis shares a natural environment with T. laurentius, K. rupestris, Didelphis albiventris, Monodelphis domestica, Galea spixii, Wiedomys pyrrhorhinos, Conepatus semistriatus and Mus musculus. In animals identified via their food source, T. brasiliensis shares a natural environment with G. spixii, K. rupestris, Capra hircus, Gallus gallus, Tropidurus oreadicus and Tupinambis merianae. The high prevalence of T. cruzi in household and peridomiciliar animals reinforces the narrow relationship between the enzootic cycle and humans in environments with T. brasiliensis and characterises it as ubiquitous.

Population dynamics of Triatoma vitticeps (Stål, 1859) in Itanhomi, Minas Gerais, Brazil.

Taxonomic markers (head structure morphometry, isoenzymes and randon amplified polymorphism of DNA - RAPD) were used to understand the population dynamics of Triatoma vitticeps, predominant triatomine species in Itanhomi district, using samples obtained from domestic, peridomiciliary and sylvatic habitats. Morphometric analysis revealed sexual dimorphism within the three samples although specimens could not be separated according to the habitat in which they were captured. Forty-two bands were analyzed from RAPD profiles generated using four primers. A dendrogram constructed from Dice's similarity coefficient values showed that migration of the insects between the habitats has occurred, without structuring of populations. Moreover, the dendrogram obtained from the genetic distance values showed an important gene flow between the sylvatic and domestic habitats. No polymorphism was found in the electrophoretic mobility of proteins for the ten enzymes studied. Our results revealed movement of triatomines between the three habitats, suggesting that the presence of T. vitticeps in houses should not be ignored. As invasion of houses by sylvatic insects is frequent and the natural infection indices of this species are among the highest known, epidemiological vigilance studies may reveal possible changes in T. vitticeps behaviour which could present future risks to public health.

Population dynamics of Triatoma vitticeps (Stål, 1859) in Itanhomi, Minas Gerais, Brazil

Taxonomic markers (head structure morphometry, isoenzymes and randon amplified polymorphism of DNA - RAPD) were used to understand the population dynamics of Triatoma vitticeps, predominant triatomine species in Itanhomi district, using samples obtained from domestic, peridomiciliary and sylvatic habitats. Morphometric analysis revealed sexual dimorphism within the three samples although specimens could not be separated according to the habitat in which they were captured. Forty-two bands were analyzed from RAPD profiles generated using four primers. A dendrogram constructed from Dice's similarity coefficient values showed that migration of the insects between the habitats has occurred, without structuring of populations. Moreover, the dendrogram obtained from the genetic distance values showed an important gene flow between the sylvatic and domestic habitats. No polymorphism was found in the electrophoretic mobility of proteins for the ten enzymes studied. Our results revealed movement of triatomines between the three habitats, suggesting that the presence of T. vitticeps in houses should not be ignored. As invasion of houses by sylvatic insects is frequent and the natural infection indices of this species are among the highest known, epidemiological vigilance studies may reveal possible changes in T. vitticeps behaviour which could present future risks to public health.

Comparative developmental and susceptibility to insecticide of Bolivian and Brazilian populations of Triatoma infestans

The triatomine bug Triatoma infestans probably originated in Bolivia and dispersed passively over wide areas of South America, where it is the principal vector of Trypanosoma cruzi. In the region of its probable origin this species shows colonization in two different ecotopes, so that it may be encountered in sylvatic as well as in artificial habitats. The sylvatic colonization pattern is not observed in the rest of its range, where T. infestans is exclusive to man-made habitats. The objective of this study was to compare several aspects of two T. infestans populations, one from Minas Gerais (Brazil) and the other from the Cochabamba Valley (Bolivia), with a view to elucidate the factors associated with the different colonization patterns observed for this species. The differences between the developmental cycle, weight, capacity to ingest blood and mortality rate of first instar nymphs should indicate more fragility of Brazilian population that may be related to its elimination possibility

Characterization of Rhodnius neglectus from two regions of Brazil using isoenzymes, genitalia morphology and morphometry

Among the triatomines considered as secondary in the epidemiology of Chagas disease, Rhodnius neglectus is frequently captured in artificial ecotopes, especially peridomiciliary ones, rarely producing colonies indoors. Nevertheless, the presence of breeding colonies in houses was unquestionably demonstrated in some areas of the State of Goiás, Brazil. Previous isoenzyme comparisons of this species with morphologically close triatomines, such as R. prolixus,R. robustuts or R. nasutus, did not produce definitive conclusions because of doubt about the geographical origin of the R. neglectus. We present here, for the first time, the isoenzyme profile of topotypes of R. neglectus. In addition, wild caught specimens from the type locality, Uberaba (Minas Gerais, Brazil), were compared to wild caught specimens from Jaraguá (Goiás, Brazil), where R. neglectus is more frequently reported invading houses. We used isoenzyme, morphology and morphometry analysis. Neither morphological nor enzymatic differences were found between areas, but metric, size-related divergence was evidenced between them.

Variations of the external male genitalia in three populations of Triatoma infestans Klug, 1834

Triatoma infestans is the triatomine that presents the greatest dispersion area in South America. However, it is not known whether the original characteristics of this insect remained in its long dispersion process. The purpose of this work was to study comparatively the external male genitalia of insects from diferent populations of T. infestans, two from Brazil (Minas Gerais and Bahia) and one from Bolivia (Cochabamba Valley), and to investigate the correlation between the morphological and behavioral variations. Differences were observed in one of the structures of the external genitalia (endosoma process) that could be used to characterize the insects from the three populations studied.