Marli Maria Lima

Epidemiology of Chagas disease in Jaguaruana, Ceará, Brazil. I. Presence of triatomines and index of Trypanosoma cruzi infection in four localities of a rural area

In order to assay the triatomine infestation and domiciliation in the rural area of Jaguaruana district, state of Ceará, Brazil, we studied, from November 2000 to April 2002, 4 localities comprising 158 domiciles as a whole, with an average of 4 inhabitants/house, who are dwelling in there for more than 7 years. Most houses have tile-covered roofs and the walls built with plaster-covered bricks (57%), followed by bricks without plaster (33%), and mud walls (7.5%). A total of 3082 triatomines were captured from different locations, according to the following capture plan: (a). intradomiciles: 238 Triatoma brasiliensis, 6 T. pseudomaculata, 9 Rhodnius nasutus, and 2 Panstrongylus lutzi; (b). peridomiciles (annexes): 2069 T. brasiliensis, 223 T. pseudomaculata, 121 R. nasutus, and 1 P. lutzi; (c). wild, in carnauba palms (Copernicia prunifera): 413 R. nasutus. From the captured triatomines, 1773 (57.5%) were examined. The natural index of Trypanosoma cruzi infection ranged from 10.8% to 30.2% (average of 17%), depending on the species and the location from where the triatomines were captured.

Aspects of peridomiciliary ecotopes in rural areas of Northeastern Brazil associated to triatomine (Hemiptera, Reduviidae) infestation, vectors of Chagas disease

Artificial ecotopes of 121 peridomiciliary environments in four rural localities in the state of Ceará, Brazil, were studied and the type of material of the ecotopes was identified as triatomine infestation. Two thousand two hundred and four Triatoma brasiliensis Neiva, 340 Triatoma pseudomaculata Corrêa and Espínola, 121 Rhodnius nasutus Stall, and 5 Panstrongylus lutzi (Neiva and Pinto) were captured. Out of the 323 ecotopes found (X = 2.0 +/- 1.8 per dwelling) - such as pigpens, henhouses, corrals, perches, dovecotes, piles of roofing tiles, bricks, wood, and straw - 30.3% were infested by triatomines in all different developmental stages, including eggs. A substantial number of triatomines were found in perches, however the largest infestation took place in roofing materials used in the construction of goat/sheep corrals, henhouses, and pigpens, where 98% of them were captured: 1372 triatomines were located in the roofing tile covers, 285 in the straw, 187 in the perches, 77 in the coverings of roofing tiles and straw, and 13 in the straw and wood Among all the different pile of materials, roofing tiles were the most infested (50%) followed by bricks (38.9%) and woods (36.1%). T. brasiliensis colonized mainly brick piles (chi-squared = 16.539; p < 0.05) and roofing tiles (chi-squared = 5,090.58; p < 0.05); T. pseudomaculata preferred wood perches (chi-squared = 472.39; p < 0.05) and woodpiles (chi-squared = 126.0 p < 0.05), and R. nasutus was principally found in roofing straw (chi-squared = 384.43; p < 0.05). These findings suggest that triatomines tend to colonize peridomiciliary ecotopes similar to their original habitats.

Morphological aspects of the larval instars of Chrysomya albiceps (Diptera, Calliphoridae) reared in the laboratory

In order to study the morphology of young Chrysomya albiceps forms, newly hatched larvae were collected at 2 hr intervals, during the first 56 hr; after this time the collection was made at 12 hr intervals. For identification and drawing, larvae were placed between a slide and a coverslip. The cephalopharyngeal skeletons along with the first and last segments were cut off for observation of their structures and spiracles. The larvae present microspines, which are distributed randomly throughout the 12 segments of the body surface; the cephalopharyngeal skeleton varies in shape and extent of sclerotization according to larval instar; the second and third instars have relatively long processes (tubercles) on the dorsal, lateral and ventral surfaces, with microspine circles on the terminal portion. Guimaraes et al. (1978) were the first researchers who recorded the presence of Chrysomya albiceps (Wiedemann, 1819), Chrysomya megacephala (Fabricius, 1974) and Chrysomya putoria (Wiedemann, 1818) in Brazil. According to them, these species were probably introduced in the southern region of Brazil by ships coming from Angola, Africa. These Diptera are considered important from the sanitary standpoint, since they may be mechanical vector of enteropathogens for humans as well as cause facultative myiasis. Today, they are widely distributed throughout the neotropical region (Prado & Guimaraes 1982, Baumgartner 1988), and have drawn the attention of several researchers. However, only the last two species have been well studied in terms of biology and taxonomy of the adults. The taxonomy of C. albiceps is still quite unknown since it was long thought to be indistinguishable from those of C. rufifacies (Macquart, 1842) (Patton 1922, Zumpt 1965, Kurahashi 1981, Richard & Gerrish 1983), a species that has not yet been recorded in Brazil. However, more accurate observations have shown some morphological aspects of the third instar of both species that allow their differentiation (Erzinclioglu 1987, Tantawi & Greenberg 1993). During the 70’s the two species were geographically isolated, C. albiceps being restricted to Africa and C. rufifacies to regions of Eastern Asia and Australia, although occupying the same biological niches in their respective natural biotopes

Cultivation-independent methods reveal differences among bacterial gut microbiota in triatomine vectors of Chagas disease

Background Chagas disease is a trypanosomiasis whose agent is the protozoan parasite Trypanosoma cruzi, which is transmitted to humans by hematophagous bugs known as triatomines. Even though insecticide treatments allow effective control of these bugs in most Latin American countries where Chagas disease is endemic, the disease still affects a large proportion of the population of South America. The features of the disease in humans have been extensively studied, and the genome of the parasite has been sequenced, but no effective drug is yet available to treat Chagas disease. The digestive tract of the insect vectors in which T. cruzi develops has been much less well investigated than blood from its human hosts and constitutes a dynamic environment with very different conditions. Thus, we investigated the composition of the predominant bacterial species of the microbiota in insect vectors from Rhodnius, Triatoma, Panstrongylus and Dipetalogaster genera. Methodology/Principal Findings Microbiota of triatomine guts were investigated using cultivation-independent methods, i.e., phylogenetic analysis of 16s rDNA using denaturing gradient gel electrophoresis (DGGE) and cloned-based sequencing. The Chao index showed that the diversity of bacterial species in triatomine guts is low, comprising fewer than 20 predominant species, and that these species vary between insect species. The analyses showed that Serratia predominates in Rhodnius, Arsenophonus predominates in Triatoma and Panstrongylus, while Candidatus Rohrkolberia predominates in Dipetalogaster. Conclusions/Significance The microbiota of triatomine guts represents one of the factors that may interfere with T. cruzi transmission and virulence in humans. The knowledge of its composition according to insect species is important for designing measures of biological control for T. cruzi. We found that the predominant species of the bacterial microbiota in triatomines form a group of low complexity whose structure differs according to the vector genus.

Investigation of Chagas disease in four periurban areas in northeastern Brazil: epidemiologic survey in man, vectors, non-human hosts and reservoirs

Chagas disease was investigated in four periurban areas of Ceará state, northeastern Brazil, through serological, parasitological and molecular methods in humans, reservoirs and vectors. A cross-sectional survey revealed a seroprevalence rate of 1.2% (13/1076 residents, six also proving positive through PCR). Trypanosoma cruzi infection was not detected in children under 10 years old. Triatoma pseudomaculata prevailed in the peridomiciles: 63 specimens, 69% (34/49) infected with trypanosomatids. Rhodnius nasutus was captured in Copernicia prunifera palm trees (n=280; 25.0% infected with trypanosomatids) and inside dwellings (n=8, all uninfected). Trypanosoma cruzi seropositive reservoirs, represented by Didelphis albiventris (n=27), Rattus rattus (n=24), Thrichomys laurentius (n=2), Mus musculus (n=1) and Monodelphis domestica (n=1), were identified. Among domestic dogs (n=96) seroprevalence reached 21.9%. Miniexon multiplex PCR assays characterized TcI in triatomines. Both TcI and TcII were detected in wild mammal hosts. We conclude that Trypanosoma cruzi circulates within a domestic zoonotic cycle, requiring continuous surveillance. Insecticide application to domiciles does not appear to prevent continuous reintroduction of wild triatomine specimens, presenting a challenge to authorities involved in Chagas disease control.

Epidemiologia da doença de Chagas em quatro localidades rurais de Jaguaruana, Estado do Ceará: soroprevalência da infecção, parasitemia e aspectos clínicos

A cross-sectional study on Chagas disease that examined the populations of four localities (no = 541 inhabitants) in the municipality of Jaguaruana, State of Ceara, showed seroprevalence of Chagas infection of 3.1%, as assessed by indirect immunofluorescence, indirect hemagglutination and ELISA tests. The rate was higher among adults over 50 years old, without any difference in relation to sex. Positive parasitemia was found in 11.8% (2/17) of the seropositive individuals by means of indirect xenodiagnosis and in 75% (9/12) by means of the polymerase chain reaction (p < 0.05). Cardiopathy was found by means of anamnesis, physical examination and resting electrocardiogram in 41% (7/17) of the seropositive individuals and in 11.8% (2/17) of the seronegative controls (p < 0.05). Analysis of these results showed that the prevalences of positive parasitemia and chronic Chagas cardiopathy were similar to those in the Caatinga area of Piaui and greater than in the Sertao area of Paraiba, although all these areas historically presented Triatoma brasiliensis and Triatoma pseudomaculata as the primary vectors responsible for Chagas infection transmission.

Characterization of the microbiota in the guts of Triatoma brasiliensis and Triatoma pseudomaculata infected by Trypanosoma cruzi in natural conditions using culture independent methods

BackgroundChagas disease is caused by Trypanosoma cruzi, which is transmitted by triatomine vectors. The northeastern region of Brazil is endemic for Chagas disease and has the largest diversity of triatomine species. T. cruzi development in its triatomine vector depends on diverse factors, including the composition of bacterial gut microbiota.MethodsWe characterized the triatomines captured in the municipality of Russas (Ceará) by sequencing the cytochrome c oxidase subunit I (COI) gene. The composition of the bacterial community in the gut of peridomestic Triatoma brasiliensis and Triatoma pseudomaculata was investigated using culture independent methods based on the amplification of the 16S rRNA gene by polymerase chain reaction (PCR), denaturing gradient gel electrophoresis (DGGE), DNA fragment cloning, Sanger sequencing and 454 pyrosequencing. Additionally, we identified TcI and TcII types of T. cruzi by sequencing amplicons from the gut metagenomic DNA with primers for the mini-exon gene.ResultsTriatomines collected in the peridomestic ecotopes were diagnosed as T. pseudomaculata and T. brasiliensis by comparing their COI sequence with GenBank. The rate of infection by T. cruzi in adult triatomines reached 80% for T. pseudomaculata and 90% for T. brasiliensis. According to the DNA sequences from the DGGE bands, the triatomine gut microbiota was primarily composed of Proteobacteria and Actinobacteria. However, Firmicutes and Bacteroidetes were also detected, although in much lower proportions. Serratia was the main genus, as it was encountered in all samples analyzed by DGGE and 454 pyrosequencing. Members of Corynebacterinae, a suborder of the Actinomycetales, formed the next most important group. The cloning and sequencing of full-length 16S rRNA genes confirmed the presence of Serratia marcescens, Dietzia sp., Gordonia terrae, Corynebacterium stationis and Corynebacterium glutamicum.ConclusionsThe study of the bacterial microbiota in the triatomine gut has gained increased attention because of the possible role it may play in the epidemiology of Chagas disease by competing with T. cruzi. Culture independent methods have shown that the bacterial composition of the microbiota in the guts of peridomestic triatomines is made up by only few bacterial species.

Could the bug Triatoma sherlocki be vectoring Chagas disease in small mining communities in Bahia, Brazil?

Searches for Chagas disease vectors were performed at the type locality from which Triatoma sherlocki Papa et al. (Hemiptera: Reduviidae: Triatominae) was described in the municipality of Gentio do Ouro, in the state of Bahia, Brazil, and in a small artisan quarry‐mining community ∼13 km distant in a remote area of the same municipality. The latter site represents a new locality record for this species. Adults, nymphs and exuviae of T. sherlocki were found in 21% of human dwellings, indicating that the species is in the process of domiciliation. Prevalence of Trypanosoma cruzi infection in collected bugs was 10.8%. Simple predictive approaches based on environmental similarity were used to identify additional sites likely suitable for this species. The approach successfully predicted an additional five sites for the species in surrounding landscapes. Ecological and entomological indicators were combined to discuss whether this scenario likely represents an isolated case or an emerging public health problem.

Is Rhodnius nasutus (Hemiptera; Reduviidae) changing its habitat as a consequence of human activity?

Rhodnius nasutus, a triatomine species autochthonous in the Brazilian Northeast, is primarily associated with the Copernicia prunifera palm tree (Carnauba). For the first time, the colonization of this triatomine in another tree species is reported. To investigate the existence of an infected triatomine focus located in a periurban area of the county of Jaguaruana, Ceará, situated in the Brazilian Northeast, where soil is greatly altered and natural vegetation scarce, an entomological survey was performed. During 2 consecutive days in August 2006 and 4 in December 2006, with the aid of live-bait traps, nine C. prunifera palms and seven Licania rigida trees (Oiticica), among other typical trees of the region, were sampled. In April 2007, 13 L. rigida trees and five C. prunifera palm trees were newly sampled. Considering the three investigation periods, a total of 20 R. nasutus specimens in C. prunifera and 52 in L. rigida were captured, in all developmental stages, 12.5 and 20.7%, respectively, harboring T. cruzi-like protozoa. The authors suggest that environmental damages are facilitating the ability of this species to colonize other trees besides palms.

On palms, bugs, and Chagas disease in the Americas

Palms are ubiquitous across Neotropical landscapes, from pristine forests or savannahs to large cities. Although palms provide useful ecosystem services, they also offer suitable habitat for triatomines and for Trypanosoma cruzi mammalian hosts. Wild triatomines often invade houses by flying from nearby palms, potentially leading to new cases of human Chagas disease. Understanding and predicting triatomine-palm associations and palm infestation probabilities is important for enhancing Chagas disease prevention in areas where palm-associated vectors transmit T. cruzi. We present a comprehensive overview of palm infestation by triatomines in the Americas, combining a thorough reanalysis of our published and unpublished records with an in-depth review of the literature. We use site-occupancy modeling (SOM) to examine infestation in 3590 palms sampled with non-destructive methods, and standard statistics to describe and compare infestation in 2940 palms sampled by felling-and-dissection. Thirty-eight palm species (18 genera) have been reported to be infested by ∼39 triatomine species (10 genera) from the USA to Argentina. Overall infestation varied from 49.1-55.3% (SOM) to 62.6-66.1% (dissection), with important heterogeneities among sub-regions and particularly among palm species. Large palms with complex crowns (e.g., Attalea butyracea, Acrocomia aculeata) and some medium-crowned palms (e.g., Copernicia, Butia) are often infested; in slender, small-crowned palms (e.g., Euterpe) triatomines associate with vertebrate nests. Palm infestation tends to be higher in rural settings, but urban palms can also be infested. Most Rhodnius species are probably true palm specialists, whereas Psammolestes, Eratyrus, Cavernicola, Panstrongylus, Triatoma, Alberprosenia, and some Bolboderini seem to use palms opportunistically. Palms provide extensive habitat for enzootic T. cruzi cycles and a critical link between wild cycles and transmission to humans. Unless effective means to reduce contact between people and palm-living triatomines are devised, palms will contribute to maintaining long-term and widespread, albeit possibly low-intensity, transmission of human Chagas disease.