Ecological diversity of Trypanosoma cruzi transmission in the Amazon basin. The main scenaries in the Brazilian Amazon.

The ecological diversity of Trypanosoma cruzi transmission in the Brazilian Amazon region is directly interlinked with the parasite's extensive reservoir, composed of 33 species of wild mammals within the following orders: Marsupialia, Chiroptera, Rodentia, Xenarthra, Carnivora and Primates; and of 16 species of wild triatomines, of which ten may be infected with T. cruzi. Four scenarios for the diversity of T. cruzi transmission in the Brazilian Amazon region are evident: (i) T. cruzi transmission between vectors and wild mammals, which is characterized as a wild enzooty encompassing the entire Amazon basin; (ii) accidental T. cruzi transmission from vectors and wild mammals to humans, when they invade the wild ecotope or when these vectors and wild mammals invade human homes; (iii) occupational Chagas disease among piassava (Leopoldinia piassaba) palm fiber gatherers, transmitted by the vector Rhodnius brethesi, for which these palm trees are the specific ecotope; (IV) oral T. cruzi transmission to humans through food contamination, particularly in juices from plants such as assai, which today is considered to be endemic in the Brazilian Amazon region, with more than 1500 cases notified.

Comparative phylogeography of Trypanosoma rangeli and Rhodnius (Hemiptera: Reduviidae) supports a long coexistence of parasite lineages and their sympatric vectors.

To make reliable interpretations about evolutionary relationships between Trypanosoma rangeli lineages and their insect vectors (triatomine bugs of the genus Rhodnius) and, thus, about the determinant factors of lineage segregation within T. rangeli, we compared phylogenies of parasite isolates and vector species. Sixty-one T. rangeli isolates from invertebrate and vertebrate hosts were initially evaluated in terms of polymorphism of the spliced-leader gene (SL). Further analysis based on SL and SSUrRNA sequences from 33 selected isolates, representative of the overall phylogenetic diversity and geographical range of T. rangeli, supported four phylogenetic lineages within this species. By comparing the phylogeny of Rhodnius species with that inferred for T. rangeli isolates and through analysis of the geographical range of the isolates, we showed that there is a very significant overlap in the distribution of Rhodnius species and T. rangeli lineages. Congruence between phylogeographical analysis of both T. rangeli lineages and complexes of Rhodnius species are consistent with the hypothesis of a long coexistence of parasites and their vectors, with lineage divergence associated with sympatric species of Rhodnius apparently without association with particular vertebrate hosts. Separation of T. rangeli isolates from vectors of distinct complexes living in sympatry favours the absence of gene flow between the lineages and suggests evolution of T. rangeli lineages in independent transmission cycles, probably associated to specific Rhodnius spp. ecotopes. A polymerase chain reaction assay based on SL intergenic sequences was developed for simultaneous identification and lineage genotyping of T. rangeli in epidemiological surveys.

Phylogeny, taxonomy and grouping of Trypanosoma rangeli isolates from man, triatomines and sylvatic mammals from widespread geographical origin based on SSU and ITS ribosomal sequences.

Phylogenetic relationships among Trypanosoma rangeli isolates from man, wild mammals and triatomine bugs from widespread geographical origin were inferred by comparison of the small subunit of ribosomal gene sequences. The phylogenetic trees indicated that the subgenus Herpetosoma is polyphyletic and strongly supported division of this group into two monophyletic lineages, one made up of T. rangeli, T. rangeli-like and allied species and other consisting of T. lewisi and related taxa. Based on phylogenetic analysis, morphology, behaviour in vertebrate and invertebrate hosts and epidemiology we propose: a) the validation of Herpetosoma as a taxon comprised only for species of group lewisi and the maintenance of T. lewisi as the type-species of this subgenus; b) the classification of T. rangeli, T. rangeli-like and allied species into a 'T. rangeli-clade' more closely related to Schizotrypanum than to T. lewisi or T. brucei. The phylogenetic tree disclosed at least 4 groups within the clade T. rangeli, all confirmed by polymorphism of the internal transcribed spacer, thus conferring for the first time phylogenetic support to groups of T. rangeli and corroborating the high complexity of this taxon. Grouping was independent of their mammalian host-species and geographical origin, indicating that other factors are determining this segregation.

Epidemiological and control aspects of schistosomiasis in Brazilian endemic areas.

The present work analyzes the epidemiology of schistosomiasis in Brazil, its expansion, the attempts to control the disease, and the overall difficulties. The authors present the distribution of schistosomiasis intermediary hosts in Brazil, the migration routes of the human population, and disease distribution in highly and lowly endemic areas and isolated foci. They also analyze the controlling programs developed from 1977 to 2002, indicating the prevalence evolution and the reduction of disease morbi-mortality. In addition, the authors also evaluate controlling methods and conclude that: (a) no isolated method is able to control schistosomiasis, and every controlling program should consider the need of a multidisciplinary application of existing methods; (b) in long term, basic sanitation, potable water supply, as well as sanitary education, and community effective participation are important for infection control; (c) in short term, specific treatment at endemic areas, associated with control of intermediary hosts at epidemiologically important foci, are extremely relevant for controlling disease morbidity, although not enough for interrupting infection transmission.

Epidemiological and control aspects of schistosomiasis in Brazilian endemic areas

The present work analyzes the epidemiology of schistosomiasis in Brazil, its expansion, the attempts to control the disease, and the overall difficulties. The authors present the distribution of schistosomiasis intermediary hosts in Brazil, the migration routes of the human population, and disease distribution in highly and lowly endemic areas and isolated foci. They also analyze the controlling programs developed from 1977 to 2002, indicating the prevalence evolution and the reduction of disease morbi-mortality. In addition, the authors also evaluate controlling methods and conclude that: (a) no isolated method is able to control schistosomiasis, and every controlling program should consider the need of a multidisciplinary application of existing methods; (b) in long term, basic sanitation, potable water supply, as well as sanitary education, and community effective participation are important for infection control; (c) in short term, specific treatment at endemic areas, associated with control of intermediary hosts at epidemiologically important foci, are extremely relevant for controlling disease morbidity, although not enough for interrupting infection transmission.

Randomly amplified polymorphic DNA analysis of Trypanosoma rangeli and allied species from human, monkeys and other sylvatic mammals of the Brazilian Amazon disclosed a new group and a species-specific marker.

We characterized 14 trypanosome isolates from sylvatic mammals (9 from primates, 1 from sloth, 2 from anteaters and 2 from opossum) plus 2 human isolates of Brazilian Amazon. These isolates were proven to be Trypanosoma rangeli by detection of metacyclic trypomastigotes in the salivary glands of triatomines and by a specific PCR assay. Polymorphism determined by randomly amplified polymorphic DNA (RAPD) revealed that most (12) of the Brazilian T. rangeli isolates from the Amazon differed from those of other geographical regions, thus constituting a new group of T. rangeli. Four Brazilian isolates clustered together with a previously described group (A) that was described as being composed of isolates from Colombia and Venezuela. Isolates from Panama and El Salvador form another group. The isolate from Southern Brazil did not cluster to any of the above-mentioned groups. This is the first study that assesses the genetic relationship of a large number of isolates from wild mammals, especially from non-human primates. A randomly-amplified DNA fragment (Tra625) exclusive to T. rangeli was used to develop a PCR assay able to detect all T. rangeli groups.

Morphologic, karyotypic, and molecular evidence of a new form of Chiropotes (primates, pitheciinae).

Morphologic, karyotypic, and molecular analyses were carried out in 25 specimens of a distinct morph of Chiropotes (henceforth termed Chiropotes sp.) obtained from a number of localities in the Brazilian Amazon. Pelage coloration clearly distinguishes the collected specimens and all other known species of this genus. A distinct karyotype was described for Chiropotes sp. It differs from C. satanas chiropotes by two pericentric inversions, and from C. satanas utahicki by three, which suggests that these taxa are reproductively isolated. Morphometric analyses did not show significant differentiation between these Chiropotes taxa. Molecular analyses confirmed the monophyly of the subfamily Pitheciinae and genera Chiropotes, Cacajao, and Pithecia (the latter appearing as the most basal lineage of the pithecine clade). The genetic distances between C. s. utahicki and Chiropotes sp. from Rio Negro were greater than those between three recognized species of Pithecia, but smaller than those between Cacajao calvus and Cacajao melanocephalus. The most appropriate name for Chiropotes sp. from Rio Negro is C. israelita. This species, C. s. chiropotes, and C. s. utahicki are allopatric. Pelage coloration, karyotype, and molecular analysis strongly indicate that C. chiropotes, C. utahicki, and Chiropotes israelita deserve species status.

In vivo and in vitro metacyclogenesis tests of two strains of Trypanosoma cruzi in the triatomine vectors Triatoma pseudomaculata and Rhodnius neglectus: short/long-term and comparative study.

Metacyclogenesis of Trypanosoma cruzi of the Y and Berenice strains was studied in Triatoma pseudomaculata and Rhodnius neglectus. Results in vivo showed a higher production of metacyclic trypomastigotes in R. neglectus' digestive tube than in T. pseudomaculata. In vitro experiments were also carried out in order to compare the behavior of culture forms of T. cruzi incubated in extracts of different compartments (stomach, intestine, and rectum) of the digestive tract of both species of triatomines. A higher percentage of metacyclic trypomastigotes for both parasite strains, Y and Berenice, was detected in the rectum extract of R. neglectus in comparison to that from T. pseudomaculata. The same results were obtained with in vitro experiments, using parasites incubated in urine from each of those vectors. The adhesion of parasites to the incubated rectum epithelial cells was also compared. In incubations with the Y strain no significant differences were detected between the two triatomine species but, however, with the Berenice strain the mean percentage of cells with adhered parasites was higher in R. neglectus than in T. pseudomaculata.

Two main clusters within Trypanosoma cruzi zymodeme 3 are defined by distinct regions of the ribosomal RNA cistron.

Trypanosoma cruzi is currently classified into 2 major phylogenetic lineages, T. cruzi I and II, that correlate with the formerly described zymodeme 1 and 2, respectively. Another isoenzymic group (zymodeme 3-Z3) was also described. In this study, we analysed the genetic diversity among Z3 isolates of the Brazilian Amazon by restriction fragment length polymorphism of the intergenic transcribed spacers (ITSs) of the ribosomal RNA cistron and the size of the divergent domain D7 of the 24Salpha rRNA gene. DNAs from 12 T. cruzi Z3 isolates obtained from humans (2), Panstrongylus geniculatus (1), and Rhodnius brethesi (9) were submitted to PCR amplification of the ITSs plus the 5.8S rDNA. The PCR products were digested with 4 distinct endonucleases and the profiles analysed by a numerical methodology. The phenetic dendrogram revealed a clear dichotomy in the Z3 group, defining 2 groups that were named Z3-A and Z3-B. Dimorphism was also found in the band sizes of the amplified D7 divergent domain of the 24Salpha rDNA, which showed a perfect correlation with the ITSs clustering. The organization of the ribosomal cistron was investigated by Southern blotting and shown to be conserved in the genome of the 2 Z3 groups. This study shows that the rDNA cistron allows the definition of 2 distinct subclusters in Z3 isolates.

The genetic diversity of Brazilian Trypanosoma cruzi isolates and the phylogenetic positioning of zymodeme 3, based on the internal transcribed spacer of the ribosomal gene.

There is a high level of genetic diversity among the clinical isolates of the taxon Trypanosoma cruzi. The results of recent studies on well defined gene loci have revealed that T. cruzi can be divided into two major phylogenetic lineages, designated T. cruzi I and T. cruzi II. Further intra-lineage polymorphisms were explored, in clinical isolates from five representative regions of Brazil, using the internal transcribed spacers that flank the 5.8S ribosomal RNA gene. These regions were amplified in a PCR and the products were digested with six restriction enzymes before being subjected to RFLP analysis in polyacrylamide gels. The results were used to construct a complex evolutionary tree, in which the isolates were clustered into two major phylogenetic lineages with a low level of similarity. To investigate the evolutionary relationships between the isolates, the PCR products were cloned and sequenced. The results of the subsequent phylogenetic analysis confirmed the presence of the two major groups of T. cruzi and showed that zymodeme 3, a third iso-enzymatic sub-group, is closer to T. cruzi I than to T. cruzi II.

Spotted fever in Brazil: a seroepidemiological study and description of clinical cases in an endemic area in the state of São Paulo.

During 1985-1995, illnesses clinically and epidemiologically compatible with Brazilian spotted fever were identified in 17 patients in the county of Pedreira, in the state of São Paulo, Brazil. Spotted-fever group rickettsial infection was confirmed by serology and/or immunostaining of tissues in 10 of these patients. Immunostaining confirmed infection in a 37-year-old pregnant patient, although rickettsial antigens were not demonstrable in the tissues of the fetus. A serosurvey was conducted in four localities in the county to determine the prevalence of subclinical or asymptomatic infections with spotted fever group rickettsiae. Five hundred and twenty-five blood samples were tested by an indirect immunofluorescence assay for antibodies reactive with Rickettsia rickettsii. Twenty-two (4.2%) of these samples demonstrated titers > or = 1:64. The results indicate that Brazilian spotted fever is endemic within this region of Brazil.

Fungal flora of the digestive tract of 5 species of triatomines vectors of Trypanosoma cruzi, Chagas 1909.

A study of the mycobiota in the digestive tract of 5 important species of triatomines, Triatoma brasiliensis, T infestans, T. sordida, T. pseudomaculata and T. vitticeps, was made. The digestive tracts of 164 adults and 535 nymphs of those triatomines were studied and 393 fungal strains were isolated. The genera with the greatest number of species were Penicillium (19 species), Aspergillus (17 species) and Acremonium (5 species) and the most frequent species, in decreasing order, were Penicillium corylophilum, Aspergillus niger, Penicillium felluttanum, Cladosporium herbarum, Penicillium waksmanii, Aspergillus awamori and Paecilomyces variotii. Among the isolated fungi, we found species that are recognized as entomopathogenic and pathogenic for humans and animals.

[Cross-sectional study of intestinal parasites and Chagas' disease in the Municipality of Novo Airão, State of Amazonas, Brazil]. / Estudo das parasitoses intestinais e da infecção chagásica no Município de Novo Airão, Estado do Amazonas, Brasil.

A cross-sectional study was performed on the residents of one in every ten households in the town of Novo Airão, in the northern meso-region of the State of Amazonas, 250 kilometers from Manaus by riverboat. A family cluster sample of 89 dwellings was studied. A stool sample was requested from each of the inhabitants for examination using the Lutz sedimentation and Baermann-Moraes-Coutinho techniques, and blood was taken by venous puncture for Trypanosoma cruzi Elisa antibody testing and immunofluorescence. From a total of 316 stool samples, 87.6% had one or more parasites: Ascaris lumbricoides (35.1%), Entamoeba histolytica (29.1%), Giardia lamblia (17.4%), and other parasites with lower prevalence rates. These results were directly correlated with lack of sanitation and clean water supply. Of the 346 sera examined, 16 (4.6%) were reactive to T. cruzi antibodies, but only three showed a correlation between this result and human contact with wild triatomines, known locally as "piassava lice".