The population structure of Trypanosoma cruzi: expanded analysis of 54 strains using eight polymorphic CA-repeat microsatellites

Recently we cloned and sequenced the first eight Trypanosoma cruzi polymorphic microsatellite loci and studied 31 clones and strains to obtain valuable information about the population structure of the parasite. We have now studied 23 further strains, increasing from 11 to 31 the number of strains obtained from patients with chronic Chagas disease. This expanded set of 54 strains and clones analyzed with the eight microsatellites markers confirmed the previously observed diploidy, clonal population organization and very high polymorphism of T. cruzi. Moreover, this new study disclosed two new features of the population genetic structure of T. cruzi. The first was the discovery that, similarly to what we had previously shown for strains isolated from insect vectors, mammals and humans with acute disease, isolates from patients in the chronic phase of Chagas disease could also be multiclonal, albeit at a reduced proportion. Second, when we used parsimony to display the genetic relationship among the clonal lineages in an unrooted Wagner network we observed, like before, a good correlation of the tree topography with the classification in three clusters on the basis of single locus analysis of the ribosomal RNA genes. However, a significant new finding was that now the strains belonging to cluster 2 split in two distant sub-clusters. This observation suggests that the evolutionary history of T. cruzi may be more complex than we previously thought.

Haematophagy and cleptohaematophagy of Clerada apicicornis (Hemiptera: Lygaeidae), a potential biological control agent of Rhodnius prolixus (Hemiptera: Reduviidae)

Because of its ability to prey on Triatominae in rural houses, Clerada apicicornis has been suggested as a potential biological control agent of Rhodnius prolixus. It has also been suggested as a potential vector of mammalian trypanosomes such as Trypanosoma cruzi, because of its ability to take blood directly from mammals. To help resolve these conflicting ideas, we assessed the haematophagic behaviour of C. apicicornis by carrying out feeding trials on laboratory animals. Cleptohaematophagic behaviour was also assessed by allowing C. apicicornis to feed on R. prolixus previously engorged with avian blood. The low proportion of blood meals taken directly from laboratory animals indicates a facultative haematophagy in this species, whereas a greater proportion of nymphs and adults were able to obtain vertebrate blood by predation on engorged R. prolixus. The results suggest that C. apicicornis is unlikely to be effective as a biological control agent, but is also unlikely to have a significant role in the transmission of vertebrate pathogens.

Mepraia spinolai in the Southeastern Pacific Ocean Coast (Chile) - First insular record and feeding pattern on the Pan de Azúcar Island

In a field collection performed at Pan de Azúcar Island in Northern Chile, 95 specimens representing all instars of Mepraia spinolai were collected. The intestinal contents of 55 specimens were examined for Trypanosoma cruzi infection and were found to be negative. This is the first record of an insular habitat for M. spinolai, where the insects had fed mainly on seabirds (78%), some on marine mammals (5%), and some on reptiles (7%).

Monitoring the domiciliary and peridomiciliary invasion process of Triatoma rubrovaria in the State of Rio Grande do Sul, Brazil

The presence of Triatoma rubrovaria in Brazil has only been confirmed in the States of Paraná and Rio Grande do Sul (RS), where it is found naturally infected with Trypanosoma cruzi. In the wild environment it occurs in rocky habitats and has an eclectic diet, feeding from cockroaches, reptiles and mammals. Data from the Chagas Disease Control Program obtained by the Fundação Nacional de Saúde, between 1975 and 1997, indicate a growing domiciliary and peridomiciliary invasion of T. rubrovaria in RS, where it has become the most frequently Triatominae species captured in this state since the control of Triatoma infestans. In order to monitor this process, we analyzed collection data derived from 22 years of control campaigns against T. infestans. Collection data for triatomines from domestic habitats show an inverse relationship, with high numbers of T. infestans and low numbers of T. rubrovaria during 1976-1987, compared to the following ten years, 1986-1997, when the number of T. infestans dropped drastically and that of T. rubrovaria increased. There are no consistent indications of intradomiciliary colonization by T. rubrovaria, since only low numbers of nymphs have been captured in the intradomiciliary ecotopes. Nevertheless, this species appears to have preadaptive characteristics for anthropic ecotopes, and should be kept under constant epidemiological surveillance.

Description of males of Parabronema pecariae Ivaschkin, 1960 (Nematoda, Habronematoidea) parasitizing peccaries (Mammalia, Tayassuidae) in Brazil

Nematodes studied herein and identified as Parabronema pecariae were collected in 1936 in the States of Rio de Janeiro and Pará and in 1940 in the State of Mato Grosso do Sul, Brazil. This species was proposed, with basis on female specimens that had been described earlier as Parabronema sp. Although the presence of males of P. pecariae was previously reported in Brazil, their description was not provided. The present paper deals with the first complete morphometric data on male specimens of P. pecariae recovered from peccaries (Pecari tajacu and Tayassu pecari).

Studies on the virulence and attenuation of Trypanosoma cruzi using immunodeficient animals

Tissue invasion and pathology by Trypanosoma cruzi result from an interaction between parasite virulence and host immunity. Successive in vivo generations of the parasite select populations with increasing ability to invade the host. Conversely, prolonged in vitro selection of the parasite produces attenuated sublines with low infectivity for mammals. One such subline (TCC clone) has been extensively used in our laboratory as experimental vaccine and tested in comparative experiments with its virulent ancestor (TUL). The experiments here reviewed aimed at the use of immunodeficient mice for testing the infectivity of TCC parasites. It has not been possible to obtain virulent, revertant sublines by prolonged passaged in such mice.

Molecular karyotype and chromosomal localization of genes encoding ß-tubulin, cysteine proteinase, hsp 70 and actin in Trypanosoma rangeli

The molecular karyotype of nine Trypanosoma rangeli strains was analyzed by contour-clamped homogeneous electric field electrophoresis, followed by the chromosomal localization of ß-tubulin, cysteine proteinase, 70 kDa heat shock protein (hsp 70) and actin genes. The T. rangeli strains were isolated from either insects or mammals from El Salvador, Honduras, Venezuela, Colombia, Panama and southern Brazil. Also, T. cruzi CL-Brener clone was included for comparison. Despite the great similarity observed among strains from Brazil, the molecular karyotype of all T. rangeli strains analyzed revealed extensive chromosome polymorphism. In addition, it was possible to distinguish T. rangeli from T. cruzi by the chromosomal DNA electrophoresis pattern. The localization of ß-tubulin genes revealed differences among T. rangeli strains and confirmed the similarity between the isolates from Brazil. Hybridization assays using probes directed to the cysteine proteinase, hsp 70 and actin genes discriminated T. rangeli from T. cruzi, proving that these genes are useful molecular markers for the differential diagnosis between these two species. Numerical analysis based on the molecular karyotype data revealed a high degree of polymorphism among T. rangeli strains isolated from southern Brazil and strains isolated from Central and the northern South America. The T. cruzi reference strain was not clustered with any T. rangeli strain.

Human infection by Pseudoterranova decipiens (Nematoda, Anisakidae) in Chile: report of seven cases

From 1997 to 1999, we identified seven human cases of infection by fourth stage larvae of Pseudoterranova decipiens in Chile. All identified larvae were coughed up by the patients. Subjects were 10-55 years old; five were female. Some patients complained of coughing, expectoration, pharyngeal pain, nausea or anal and nasal pruritus. Larvae of three patients were coughed up from 36 h to 7 days after having eaten raw (cebiche or sushi) or lightly fried fish. P. decipiens has a marine life cycle. Infective third stage larva develop to adult stage in pinniped mammals. The nematode eggs are voided with the host faeces and develop and hatch releasing third stage larvae. Some crustaceans and fish act as hosts of third stage larvae. Man is an accidental host for third or fourth stage larvae.

Activity of Tabanids (Insecta: Diptera: Tabanidae) Attacking the Reptiles Caiman crocodilus (Linn.) (Alligatoridae) and Eunectes murinus (Linn.) (Boidae), in the Central Amazon, Brazil

Tabanid females are better known as hematophagous on man and other mammals, and linked to mechanical transmission of parasites. The association between tabanids and reptiles is poorly known, but has been gaining more corroboration through experiments and occasional observation in the tropics. The present study was conducted at a military base (CIGS/BI-2), situated 54 km from Manaus, Amazonas, in a small stream in a clearing (02°45'33"S; 59°51'03"W). Observations were made monthly, from April 1997 to March 1998, during two consecutive days. At the same time, other vertebrate animals were offered, including humans. However in this paper only data obtained on a common caiman, Caiman crocodilus (Linn.), and an anaconda, Eunectes murinus (Linn.), in diurnal observations from 05:30 a.m. to 18:30 p.m., will be discussed. A total of 254 tabanid specimens were collected, 40 from the anaconda and 214 from the caiman. Four tabanid species were recorded on these two reptiles: Stenotabanus cretatus Fairchild, S. bequaerti Rafael et al., Phaeotabanus nigriflavus (Kröber) and Tabanus occidentalis Linn. Diurnal activities showed species-specific patterns. The first three species occurred only in the dry season. T. occidentalis occurred during the whole observation period, and with increased frequency at the end of the dry season. We observed preferences for body area and related behavior of the host. Observations on the attack of tabanids on one dead caiman are also presented.

The opossum Didelphis virginiana as a synanthropic reservoir of Trypanosoma cruzi in Dzidzilché, Yucatán, México

In México, the role of mammals in the transmission cycle of Trypanosoma cruzi is poorly known. In the State of Yucatán, an endemic area of Chagas disease, both Didelphis virginiana and D. marsupialis occur sympatrically. However, until now, only the former species had been found infected with T. cruzi. To evaluate the role of D. virginiana in a peridomestic transmission, nine periods of capture-recapture were performed around the village of Dzidzilché, Yucatán. The sex, age, reproductive status, location, and presence of infection with T. cruzi were recorded for each opossum. The chromosome morphology was used to identify the opossum species. T. cruzi was identified by the presence of pseudocysts of amastigotes in cardiac muscle fibers of Balb/c mice inoculated with strains isolated from opossums. However, xenodiagnosis was the best diagnostic method. Triatoma dimidiata, the vector, were collected in and around the opossums' nests, and human dwellings; and were checked for T. cruzi. From 102 blood samples of D. virginiana examined 55 (53.9%) were positive to T. cruzi, the only two D. marsupialis captured were negative. Significant differences were found between infection, and both sex and reproductive condition. Eight out of 14 triatomines collected in peridomestic nests (57.1%), and 32 of 197 captured inside houses (16.3%) were found infected, suggesting a peridomestic transmission. The statistically high abundance of infected opossums and triatomines during the dry season (March to May) suggested the existence of a seasonality in the peridomestic transmission of T. cruzi in Dzidzilché.

Phorcotabanus cinereus (Wiedemann, 1821) (Diptera, Tabanidae), an ornithophilic species of Tabanid in Central Amazon, Brazil

In Central Amazon, Brazil, the tabanid Phorcotabanus cinereus (Wiedemann) was recorded attacking the native duck Cairina moschata (Linnaeus) (Anseriformes, Anatidae). The flight and behavior of the tabanid during the attacks and the host's defenses were videotaped and analyzed in slow motion. The tabanid was recorded flying rapidly around the heads of the ducks before landing. Landing always took place on the beak, and then the tabanid walked to the fleshy caruncle on the basal part of the beak to bite and feed. Firstly the duck defends itself through lateral harsh head movements, and then, when it is being bitten, it defends itself by rubbing its head on the body, or dipping the head into water, when swimming. If disturbed, the fly resumed the same pattern of flight as before and would generally try to land again on the same host and bite in the same place. This feeding activity was observed predominantly between 9:30 am and 4:30 pm and always in open areas, near aquatic environments, from June 1996 to January 1997, the dry season in Central Amazon. To test the attractiveness of other animals to P. cinereus, mammals, caimans and domestic and wild birds were placed in suitable habitat and the response of P. cinereus observed. P. cinereus did not attack these animals, suggesting that this species has a preference for ducks, which are plentiful in the region.

Experimental models of Schistosoma mansoni infection

Experimental models of Schistosoma mansoni infections in mammals have contributed greatly to our understanding of the pathology and pathogenesis of infection. We consider here hepatic and extrahepatic disease in models of acute and chronic infection. Experimental schistosome infections have also contributed more broadly to our understanding of granulomatous inflammation and our understanding of Th1 versus Th2 related inflammation and particularly to Th2-mediated fibrosis of the liver.

Host-ectoparasite specificity in a small mammal community in an area of Atlantic Rain Forest (Ilha Grande, State of Rio de Janeiro), Southeastern Brazil

The analyses of the ectoparasite species associated with a small mammal community on Ilha Grande, a coastal island in southern of the state of Rio de Janeiro, Brazil, evaluated the level of host-ectoparasite specificity. Was used the Jaccard index for qualitative data to analyse the similarity. The lowest value of similarity occurred between Proechimys iheringi and Marmosops incanus and between Sciurus aestuans and Nectomys squamipes (Cj = 0.08) and the highest between P. iheringi and Oxymycterus sp. (Cj = 0.33). This index showed a low value of similarity across the ectoparasite community. The only exception from this pattern of high host specificity occurred with P. iheringi and Oxymycterus sp., which shared five species of ectoparasites. The similarity values, for most of the cases, is smaller than 0.2.

Estudo morfométrico do gênero Panstrongylus Berg, 1879 (Hemiptera, Reduviidae, Triatominae)

The subfamily Triatominae is actually represented by 137 species distributed among 6 tribes and 19 genera. Within this subfamily, the genus Panstrongylus, Berg 1879, is composed by 13 species widespread in sylvatic, peridomestic, and domestic habitats of Neotropical regions. These species are vectors of Chagas disease and consequently are found associated with its main hosts, such as birds and mammals. Interest in species of this genus has been increasing in the last few years. Reports of several authors indicate these Triatominae to invade and colonize houses, increasing their epidemiological significance. Morphometry was used in this study to investigate correlations among possible closely related species. We measured 224 specimens among 13 species through a set of metric variables of the head. The results indicated that the genus Panstrongylus seems to be homogeneous since 10 of the 14 species were shown to be closely related.

Siphonaptera parasites of wild rodents and marsupials trapped in three mountain ranges of the Atlantic Forest in Southeastern Brazil

A study of the associations between small mammals and fleas was undertaken in three areas of the Atlantic Forest in Souhtheastern Brazil: Serra da Fartura, SP, Serra da Bocaina, SP, and Itatiaia, RJ. Trapping of small rodents and marsupials was done every 3 months during 2 years, from June 1999 to May 2001. A total 502 rodents (13 species) and 50 marsupials (7 species) were collected, and 185 hosts out of 552 (33.5%) captured in the traps were parasitized by 327 fleas belonging to 11 different species. New host records were determined for several flea species, and 5 significant associations between fleas and hosts were also found.