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Isolation of Paecilomyces lilacinus (Thom) Samson (Ascomycota: Hypocreales) from the Chagas disease vector, Triatoma infestans Klug (Hemiptera: Reduviidae) in an endemic area in Argentina.

A survey for entomopathogenic fungi of the Chagas disease vector Triatoma infestans was conducted in two provinces of Argentina from March-December 2003. Field-collected insects that died in the laboratory were individually maintained in moist chamber and incubated at 22 degrees C. Triatominae adults infected with the fungus Paecilomyces lilacinus were found at El Quebracho (27 degrees 34'S-64 degrees 31'W), Santiago del Estero province, Argentina, in December 2003. Paecilomyces lilacinus was cultured and isolated from infected insects in SDAY, PYG and MEA media. Pathogenicity tests were conducted and positive results were recorded. The median survival time (MST) of T. infestans exposed to a P. lilacinus conidial suspension was 12.8 days, and 100% mortality occurred at 30 days post-treatment. This is the first record of natural infection caused by P. lilacinus in T. infestans in the world.

Animals↗

Density-dependent timing of defaecation by Triatoma infestans.

Defaecation timing of the Triatominae vectors of Trypanosoma cruzi directly affects the transmission probability of Chagas disease to mammal hosts. Experimental studies with fifth instar nymphs of Triatoma infestans showed that defaecation time was negatively affected by blood meal size and positively affected by starvation period and bug initial weight. Since blood meal size and starvation period are both density-dependent, low density domestic populations of T. infestans would represent a higher transmission risk than high density populations. As low density populations could occur in recently reinfested houses after control using insecticide, vigilance activities should be reinforced to protect the human population at the highest risk of disease transmission.

Animals↗

Control of Chagas disease in Brazil.

Chagas disease (South American trypanosomiasis) is a chronic but often fatal disease endemic throughout much of Latin America. Serological surveys suggest around 24 million people seropositive for the causative agent, Trypanosoma cruzi (Fig. 1), with over 65 million living in the endemic areas and at risk to infection. In Brazil, over 25 million people are considered at risk, and control of the disease constitutes one of Brazil's public health priorities. Treatment or vaccination against T. cruzi is impossible at the public health level because suitable drugs or vaccines are not available. But it is well recognized that transmission can be interrupted by eliminating the domestic vectors - blood-sucking reduviid bugs of the subfamily Triatominae. In Brazil, eradication of Triatoma infestans - the major domestic vector of T. cruzi - is now seen as a feasible target by the Ministry of Health. However, although other domestic vectors can also be controlled, they will retain their sylvatic ecotopes from which they can reinvade houses. In this article, Joao Carlos Pinto Dias explains the current Brazilian policy, high-lighting the successful elimination of T. infestans from much of the southern part of the country.

Journal Article↗

Exploring the role of insect host factors in the dynamics of Trypanosoma cruzi-Rhodnius prolixus interactions.

Members of the subfamily Triatominae, family Reduviidae, comprise a large number of insect species of which some are vectors of Trypanosoma cruzi, the causative agent of Chagas' disease. This article outlines research on the process of transformation and the dynamics of developmental stages of Trypanosoma cruzi in the triatomine insect hosts. Special attention is given to the interactions of parasites with gut molecules, and the gut environment, and with host developmental physiology and intestinal organization. The vector insect's permissiveness to Trypanosoma cruzi, which develops in the vector gut, largely depends on the host nutritional state, the parasite strain, trypanolytic compounds, digestive enzymes, lectins, resident bacteria in the gut and the endocrine system of the insect vector. Finally, the mechanisms of these interactions and their significance for Trypanosoma cruzi transmission are discussed.

Animals↗

Nested clade and phylogeographic analyses of the Chagas disease vector Triatoma brasiliensis in Northeast Brazil.

Triatoma brasiliensis (Hemiptera: Reduviidae: Triatominae) is the most important Chagas disease vector in the semiarid areas of Northeast Brazil. We analyzed mitochondrial cytochrome b sequence variation among 136 individuals representing 16 populations from across the species' distribution. Neighbor-joining and parsimony tree-building methods were used in conjunction with nested clade analysis to describe the systematics and phylogeography of this species. Our results indicate that T. brasiliensis is composed of four genetically distinct chromatic forms (referred to as brasiliensis, macromelasoma, juazeiro, and melanica) that present inter-population divergence values (0.027-0.119, corrected K2-p) and a pattern of haplotype geographic distribution compatible with the existence of a species complex. As a consequence, such forms can be treated as isolated targets in vector control programs. We were unable to infer what is shaping the population structure of the brasiliensis form as we obtained mutually exclusive causes of structure, namely a barrier to gene flow caused by past population fragmentation, and isolation by distance between populations (which would permit gene flow). We found indication of mitochondrial DNA introgression occurring among forms in putative hybrid zones.

Animals↗

Trypanosoma cruzi genotypes associated with domestic Triatoma sordida in Bolivia.

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

Animals↗

Parity between kinetoplast DNA and mini-exon gene sequences supports either clonal evolution or speciation in Trypanosoma rangeli strains isolated from Rhodnius colombiensis, R. pallescens and R. prolixus in Colombia.

Trypanosoma rangeli are kinetoplastid protozoa which have been largely recognized and defined in several Latin American countries in relation to T. cruzi, because the two trypanosome species are frequently found in mixed infections in triatominae vectors, humans and a variety of wild and domestic mammals. We report the molecular characterization of 18 T. rangeli strains isolated from the salivary glands of naturally infected Rhodnius colombiensis, R. pallescens and R. prolixus by using two independent set of molecular markers. kDNA and mini-exon amplification indicated dimorphism within both DNA sequences: KP1, KP2 and KP3 or KP2 and KP3 products for kDNA mini-circles and 380 or 340bp products for the mini-exon. One of two associations was observed within individual strains: KP1, KP2 and KP3 kDNA products with the 340bp mini-exon product and the KP2 and KP3 kDNA products with the 380bp mini-exon product. Independent mitochondrial and nuclear molecular markers showed a clear division of T. rangeli into two major phylogenetic groups associated with specific vectors in Colombia and in other Latin America countries. These results support either clonal evolution or speciation in T. rangeli populations, probably derived as a secondary adaptation to their parasitic condition in triatomine vectors.

Animals↗

Peridomestic distribution of Triatoma garciabesi and Triatoma guasayana in north-west Argentina.

The Reduviid bugs Triatoma garciabesi Carcavallo et al. (Hemiptera: Reduviidae: Triatominae), previously known as T. sordida Ståhl in the semi-arid chaco region, and T. guasayana Wygodzinsky & Abalos, vectors of Trypanosoma cruzi Chagas (Kinetoplastida: Trypanosomatidae), were found to occupy wide but different ranges of ecotopes in the peridomestic environment. At Amamá and nearby rural villages in north-western Argentina, a combined total of 1233 specimens were collected from 325/2314 (14%) sites surveyed at 6-monthly intervals from November 1994 to November 1996. Triatoma garciabesi and T. guasayana displayed a significantly different distribution among peridomestic ecotopes. Triatoma garciabesi predominated over T. guasayana in the prevalence of infested sites, the number of colonies and the number of bugs collected. For T. garciabesi, the predominant ecotopes most likely to yield T. garciabesi repeatedly were the rugged bark of Prosopis alba or P. nigra (Fabaceae) trees, where chickens roosted, and chicken coops. For T. guasayana the main ecotopes were goat or sheep corrals, piled materials and orchard fences. Triatoma garciabesi and T. guasayana were collected concurrently from the same site on 9/2314 (0.4%) occasions, and on different dates at the same site on 12 (0.5%) occasions. The observed low frequency of mixed populations (< 1%) was not significantly different from that expected from a hypothesis of independence. Triatoma garciabesi clearly outnumbered T. guasayana in four of the nine mixed populations, none of which persisted as such. Neither T. garciabesi nor T. guasayana colonized human habitations, even in the absence of T. infestans (formerly the predominant domestic vector of T. cruzi in this area), a situation that apparently has not changed in the last 50 years in northern Argentina.

Animals↗

Sampling Rhodnius neglectus in Mauritia flexuosa palm trees: a field study in the Brazilian savanna.

Two sampling methods (manual capture and live-baited adhesive traps) were compared for collecting the bug Rhodnius neglectus Lent (Hemiptera: Reduviidae: Triatominae) from palm trees, Mauritia flexuosa L. (Arecaceae), in the savanna of Brasília DF. R. neglectus was found in 19/50 (38%) of palm trees sampled. The detection rate was much higher by visual inspection and manual capture (18/50=36%) than by our trapping method (5/50=10%), although one tree was found to be positive by trapping but not by manual capture. Bugs collected manually were mostly (146/154=95%) found among the dead organic material in palm crowns. In combination, these sampling techniques are useful for quick detection of triatomine bug infestation in palm trees, especially in areas of high ecological value where the palms should not be cut and dissected, but arboreal Rhodnius are suspected to transmit enzootic Trypanosoma cruzi that might represent a risk of causing human cases of Chagas disease.

Animals↗

Cryptic speciation in Triatoma sordida (Hemiptera:Reduviidae) from the Bolivian Chaco.

Eight natural Bolivian populations of two closely related species of Triatominae, Triatoma sordida and T. guasayana, were analysed by multilocus enzyme electrophoresis at 20 loci. Both species were readily separated and no natural hybrid was observed. Among the silvatic sample of T. sordida, strong departure from panmixia within and between loci was detected in two sites of the Chaco, suggesting two reproductively separate populations easily recognized at Idh2 and Mdh2 loci. Genetic distance between them was in agreement with the hypothesis of distinct species. However, the detection of 3% of putative hybrids suggested a recent evolutionary divergence.

Alleles↗

Domiciliation process of Rhodnius stali (Hemiptera: Reduviidae) in Alto Beni, La Paz, Bolivia.

We report a systematic collection of Triatominae inside houses and in the peridomestic environment of Alto Beni, department of La Paz, Bolivia. This area is free of Triatoma infestans and although we detected previously seropositivity for Trypanosoma cruzi, the Alto Beni region is not officially considered as endemic for Chagas disease. From 11 houses of five localities, we collected adults, nymphs and eggs of a Rhodnius species, which was confirmed by morphological and morphometric analysis as Rhodnius stali. This little-known species has long been confused with R. pictipes, and was originally described from museum specimens labelled as R. pictipes. Our data show that R. stali is able to establish colonies in domestic and peridomestic habitats in Bolivia, and it is probably the vector responsible for Chagas disease seropositivity observed in the indigenous population of Alto Beni.

Animals↗

Type of rectal contents and infectivity of domiciliary populations of Triatoma infestans (Hemiptera: Reduviidae) in Argentina.

Different types of rectal content in domiciliary populations of Triatoma infestans (Hemiptera: Reduviidae: Triatominae) were associated with differences in the degree of infection with Trypanosoma (Schizotrypanum) cruzi. Field studies were carried out in a region of Argentina (Cordoba) where Chagas' disease is endemic. The rectal contents of domiciliary T. infestans were classified by color (clear, yellow, or brown) and the number of infective metacyclic stages of T. cruzi. The seasonal percentage of triatomines with metacyclics did not vary significantly, except in adults, in which it was greatest in spring (November-December). The mean density of T. cruzi per microliter of rectal material varied during the year, according to instar and color of the rectal contents. Clear recta had most metacyclics. December (late spring) may be more important for vectorial transmission of T. cruzi, because there are more infective adults (788) with higher counts of rectal metacyclics.

Age Factors↗

Antennal phenotype of Triatoma dimidiata populations and its relationship with species of phyllosoma and protracta complexes.

Triatoma dimidiata (Latreille 1811) Reduviidae Triatominae is the main vector of Chagas disease in several countries of Latin America. As for other vector species, the characterization of T. dimidiata subpopulations within particular geographical regions or occupying different habitats could help in better planning of vector control actions. A first objective in this study was to evaluate the antennal phenotype as a phenetic marker to characterize populations of T. dimidiata collected in different geographic areas and domestic and sylvatic habitats. A second objective was to evaluate the phenetic relationships of T. dimidiata with other species of the phyllosoma complex: longipennis, pallidipennis, and phyllosoma. The antennal sensilla of T. dimidiata specimens collected in Mexico, Central America, and Colombia were analyzed and compared with the antennal sensilla of T. longipennis, T. pallidipennis, and T. phyllosoma. T. barberi was used as an outgroup in the analysis. For each specimen, the ventral side of the three distal segments of the antennae was drawn, identifying and counting four types of sensilla. In T. dimidiata, univariate and multivariate analysis showed differences between sexes, among populations collected in different habitats within the same region, and among populations collected in different geographic regions. Two types of antennal sensilla showed a latitudinal variation. Domestic specimens showed intermediate characteristics of the antennal phenotype, between sylvatic cave- and sylvatic forest-collected specimens. The antennal phenotypes show high similarities among T. pallidipennis, T. phyllosoma, and T. longipennis, with a better differentiation of T. pallidipennis. T. dimidiata is separated from the other members of the complex by a similar distance to T. barberi, of the protracta complex.

Animal Structures↗

A salivary nitrophorin (nitric-oxide-carrying hemoprotein) in the bedbug Cimex lectularius.

Salivary gland homogenate of the bedbug Cimex lectularius caused vasodilation of the preconstricted rabbit aortic ring in the absence of endothelium. Vasodilation was augmented in the presence of superoxide dismutase and inhibited in the presence of Methylene Blue. Utilization of the Griess reaction indicated the presence of reactive nitrogen equivalents of the order of 337 +/- 57 pg equivalent NO2- per pair of salivary glands (mean +/- S.E.M.; N = 3). Salivary gland homogenates have a nitrosyl-hemoprotein that releases nitric oxide in a pH-dependent manner. The fraction containing the NO-carrying hemoprotein, when separated by HPLC, caused vasodilation of the preconstricted rabbit aortic strip. Furthermore, the presence of a nitrosyl-hemoprotein in Cimex lectularius salivary gland was verified by electron paramagnetic resonance spectroscopy. It is proposed that, as in the case of Rhodnius prolixus (Triatominae), Cimex lectularius salivary glands contain a hemoprotein (nitrophorin) that carries NO from the glands to the host tissues. However, because Cimex lectularius and Rhodnius prolixus belong to different hemipteran families (Cimicidae and Reduvidae) and evolved independently to blood feeding, Cimex lectularius and Rhodnius prolixus nitrophorin may be a case of convergent evolution.

Animals↗

Morphometric variability in the Chagas' disease vector Rhodnius prolixus.

Morphometric variability was studied in six domestic Venezuelan populations of the blood-sucking bug Rhodnius prolixus Stal 1897 (Reduviidae, Triatominae) and in a sylvatic population identified as R. robustus Larrousse 1927. Evidence is here provided by both uni- and multifactorial analyses of extensive variation of morphological traits between the R. prolixus populations studied. Regardless the geographic or climatic environmental factor tested, none can be retained in a selective model accounting for the morphological variability observed. Moreover, the results failed to support any correlation between the morphological Mahalanobis' distances and geographical distances. The genetic relationships between these populations inferred from the present data, are more consistent with some demic structure, resulting from random genetic drift by founder effects, than with any alternative population genetic model. It is noteworthy that the range of variation of these morphological traits in R. prolixus includes the putative R. robustus population. Therefore, the species-specific status of R. robustus, at the very least the local Trujillo population studied, is questioned. In addition, a preliminary multifactorial analysis bearing on the three other Rhodnius relatives, R. pictipes Stal 1872, R. nasutus Stal 1859 and R. neglectus Lent 1954, confirmed the marked morphological differentiation of R. pictipes from all other species and showed a clear morphological differentiation of R. nasutus and R. neglectus both from one another and from R. prolixus.

Analysis of Variance↗

[Transfusional Chagas infection detected in the program for the control of Chagas disease in the state of São Paulo (Brazil)].

A system of surveillance for Chagas' disease aiming at a systematic investigation of the occurrence of triatominae in human dwellings in S. Paulo, Brazil was proposed. It included a serological survey of residents in house considered to be potential breeding places for blood-sucking triatomines. Serologically positive cases were observed to be distributed in age groups from 19 years of age upwards. Case-investigation revealed that the infection had been acquired either in S. Paulo in the past or recently in other States. A serologically positive (titre = 128 - IgG) case of an 8-year-old male child, was detected by the In direct Fluorescent Antibody Technique (IFAT). In S. Paulo State natural transmission is now of low probability. An epidemiological investigation disclosed the fact that this was a case of blood transfusion infection. The donor was found to be serologically positive (IFAT, titre = 1024 - IgG). His case-history was typical of vector transmitted infection. It is worthy of note that blood had been donated by this patient in four instances, without his condition having been diagnosed. The necessity of organizing an integrated Public Health Service to take more efficient care of such cases is stressed.

Adult↗

[Influence of temperature and humidity on the nymphal development of Rhodnius robustus].

OBJECTIVE: To assess the significant differences in the nymphal development of the Rhodnius robustus Larrousse, 1927 under different temperatures and humidity conditions. This is a species found in the northern region of Brazil (states of Acre, Amazonas, and Pará), Colombia, Equator, Peru and Venezuela. METHODS: Three groups of triatominae were kept under the following laboratory conditions: 33/40 (33+1 degree C and 40+/-5% of relative humidity - RH), 33/70 (33+/-1 degree C and 70+/-5% RH), and 28/70 (28+/-1 degree C and 70+/-5% RH). The incubation period of the eggs, developmental time of each stage, mortality percentage, number of bloodmeals, and the total amount of time from the egg hatching to adult ecdysis were observed. RESULTS: The shortest average time of nymphal development was observed in the 28/70 group, with following averages: 14.4, 17.3, 20.3, 22.8, and 40. Significant differences were observed in the embryonic development between the groups (p<0.01). For all groups, the number of bloodmeals had a gradual increase near the adult phase, except for the 3rd instar of the 33/70 group. The smallest mortality percentages were seen in the 28/70 group. CONCLUSIONS: High temperatures, regardless of the humidity, can accelerate the biological development cycle of R. robustus. However, these temperatures can impair the colonies' survival, preventing their maintenance in the laboratory.

Animals↗

[Acute Chagas' disease: transmission routes, clinical aspects and response to specific therapy in diagnosed cases in an urban center].

The authors report clinical features and therapeutic response of 24 outpatients with acute Chagas' disease, and 3 in the initial chronic phase, referred to the Clinic for Infectious and Parasitic Diseases of the FMUSP "Clínicas" Hospital between 1974 and 1987. The following transmission routes were involved: triatominae in 7 cases, blood transfusion in 9, kidney transplantation and/or blood transfusion in 4, accidental in 1, oral route in 3, probably breast feeding in 1, congenital or breast feeding in 1, and congenital or blood transfusion in 1. Six patients infected by triatominac acquired the disease between 1974 and 1980 and one in 1987. The blood transfusion infected patients acquired the disease in Greater São Paulo, seven of whom after 1983. The acute phase Chagas' disease was oligosymptomatic in 4 patients: three of such patients being immunocompromised by drugs or other diseases. Another two adult immunocompromised patients developed myocarditis and congestive heart failure. Clinical features were severe in 5 from 6 children under two years, irrespective of the transmission route. Evaluation of the acute phase patients treated with benznidazol (4-10 mg/kg/day) showed: therapeutic failure in 4/16 (25.0%); possible cure in 9/16 (53.2%) and inconclusive results in 3/16 (18.8%). The antibody and complement-mediated lysis reaction was in keeping with the xenodiagnosis in 18/22 cases, having shown negative results after treatment earlier than classical serological reactions. One aplastic anaemia patient receiving corticosteroid presented lymphoproliferative disease 6 years after being treated with benznidazol for acute Chagas' disease.

Adolescent↗