João Carlos Araujo Carreira

STRUCTURAL VARIATION IN THE GLYCOINOSITOLPHOSPHOLIPIDS OF DIFFERENT STRAINS OF TRYPANOSOMA CRUZI

The structures of the glycoinositolphospholipids (GIPLs) from five strains of the protozoan parasiteTrypanosoma cruzi have been determined. Two series of structures were identified, all but one containing the same Man4(AEP)GlcN-Ins-PO4 core. Series 1 oligosaccharides are substituted at the third mannose distal to inositol (Man 3) by ethanolamine-phosphate or 2-aminoethylphosphonic acid, as are some glycosyl-phosphatidylinositol-protein anchors ofT. cruzi. The core can be further substituted by terminal (1–3)-linked β-galactofuranose units. In contrast, Series 2 oligosaccharides do not have additional phosphorus-containing groups attached to Man 3, the latter being substituted instead by a single side chain unit of β-galactofuranose. Series 1 oligosaccharides are present in all strains (G, G-645, Tulahuen CL, and Y) whereas Series 2 structures are present mainly in CL and Y strains. The lipid moiety in the GIPLs from the G, G-645 and Tulahuen strains is predominantly ceramide, as reported for the Y strain, whilst that from the CL strain is a mixture of ceramide and alkylacylglycerol species. The lipid moiety of the GIPLs, and probably also the phosphoinositol-oligosaccharide structures may play an important immunomodulatory role in infection byT. cruzi.

Leishmaniose em cães domésticos: aspectos epidemiológicos

In Barra de Guaratiba, an endemic area for American visceral leishmaniasis (AVL) in Rio de Janeiro, Brazil, control campaigns were unable to reduce canine infection rates. This difficulty prompted an in-depth study of dogs as a reservoir for Leishmania chagasi in the peri-urban environment through clinical and serological follow-up using the immunofluorescence and Western blot techniques. Recognition of 29 kDa and 32 kDa peptides by sera from dogs with proven L. chagasi infection was observed. Furthermore, only sera from symptomatic dogs recognized the 68.5 kDa antigen, so the latter should be considered a parameter for culling dogs from endemic areas. The WB technique proved to be more sensitive than IFA, since the 29 and 32 kDa peptide fractions were even recognized by sera from AVL seronegative dogs up to 8 months before IFA seroconversion. Proximity to wooded areas was an important risk factor for L. chagasi infection in dogs, possibly due to the presence of wild reservoirs.

Infection of a mammal by monogenetic insect trypanosomatids (Kinetoplastida, trypanosomatidae)

Monogenetic insect trypanosomatids of the genera Crithidia, Leptomonas and Herpetomonas, multiplied as in axenic cultures, for many months, in the lumen of the scent glands of the opossum Didelphis marsupialis. Specific antibodies were detected in the serum of the animals but there was no evidence of invasion of their tissues by the parasites.

Natural infection of Didelphis aurita (Mammalia: Marsupialia) with Leishmania infantum in Brazil

BackgroundThe opossum Didelphis have been considered as natural hosts of Leishmania parasites in the New World, suggesting an important role in the epidemiology of Visceral Leishmaniasis (VL). Among six extant species that belong to the genus Didelphis, only two (D. marsupialis and D. albiventris), have been mentioned as natural hosts of Leishmania infantum in Brazil and Colombia. In the present paper, it is reported for the first time, the observation of intracellular parasites (amastigotes) in tissues of Didelphis aurita naturally infected with Leishmania infantum in Brazil. We also discuss some aspects associated to the relationship between L. infantum and the geographical distribution of some species of the genus Didelphis.MethodsThe opossums studied were caught by wire traps (Tomahawk) in Barra de Guaratiba, a peri-urban area in Rio de Janeiro. The opossums were killed with an overdose of Thiopental sodium.At necropsy, macroscopic alterations were examined and samples from liver, spleen, lymph nodes, ear, abdominal skin, scent glands and bone marrow were collected for parasitological and molecular diagnoses.ResultsForty-eight opossums were captured in an AVL endemic region, 30 being caught in a mangrove area and eighteen animals in a forest area near to some residential-yards. Among the thirty opossums trapped in the mangrove area, all of them were negative by both imprint and sera samples assayed on Dipstick Tests, that is a test based on a combination of protein-A colloidal gold conjugate and rk39 Leishmania antigen to detect anti-Leishmania antibody in serum or plasma. At the macroscopic examination one out of eighteen opossums, caught close to the forest, presented alterations compatible with spleen hypertrophy and three were positive by Dipstick Tests (16.6%) and presented amastigotes in the spleen and in one of them, the parasites were also observed in a submandibular lymph node. Leishmania infantum infections were confirmed through dot blot hybridization using a L. infantum-specific biotinylated probe.ConclusionsIn the present paper we present the first report of amastigotes in the tissues of Didelphis aurita (Mammalia: Marsupialia) naturally infected with Leishmania infantum. We also attempt to claim the particular role of some opossum species as hosts of Leishmania infantum, contributing at least in part on the description of potential sylvatic reservoirs.

Trypanosoma cruzi: Correlations of Biological Aspects of the Life Cycle in Mice and Triatomines

The infection pattern in Swiss mice and Triatomine bugs (Rhodnius neglectus) of eleven clones and the original stock of a Trypanosoma cruzi isolate, derived from a naturally infected Didelphis marsupialis, were biochemically and biologically characterized. The clones and the original isolate were in the same zymodeme (Z1) except that two clones were found to be in zymodeme 2 when tested with G6PDH. Although infective, neither the original isolate nor the clones were highly virulent for the mice and lesions were only observed in mice infected with the original stock and one of the clones (F8). All clones and the original isolate infected bugs well while only the original isolate and clones E2 and F3 yielded high metacyclogenesis rates. An observed correlation between absence of lesions in the mammal host and high metacyclogenesis rates in the invertebrate host suggest a evolutionary trade off i.e. a fitness increase in one trait which is accompanied by a fitness reduction in a different one. Our results suggest that in a species as heterogeneous as T. cruzi, a cooperation effect among the subpopulations should be considered.

Mapping of the environmental contamination of Toxoplasma gondii by georeferencing isolates from chickens in an endemic area in Southeast Rio de Janeiro State, Brazil

The environmental contamination of Toxoplasma gondii in an endemic area in Brazil was mapped by georeferencing isolates from chickens in farms in the Southeast of the state of Rio de Janeiro. Tissue samples obtained from 153 adult chickens were analyzed by the mouse bioassay for T. gondii infection. These animals were reared free-range on 51 farms in the municipalities of Rio Bonito and Maricá. The ArcGIS kernel density estimator based on the frequency of T. gondii-positive chickens was used to map the environmental contamination with this parasite. A questionnaire was applied to obtain data on the presence and management of cats and the type of water consumed. Of the farms studied, 64.7% were found to be located in areas of low to medium presence of T. gondii, 27.5% in areas with a high or very high contamination level and 7.8% in non-contaminated areas. Additionally, 70.6% kept cats, 66.7% were near water sources and 45.0% were in or near dense vegetation. Humans used untreated water for drinking on 41.2% of the farms, while all animals were given untreated water. The intensity of environmental T. gondii contamination was significantly higher on farms situated at a distance >500 m from water sources (P=0.007) and near (≤500 m) dense vegetation (P=0.003). Taken together, the results indicate a high probability of T. gondii infection of humans and animals living on the farms studied. The kernel density estimator obtained based on the frequency of chickens testing positive for T. gondii in the mouse bioassay was useful to map environmental contamination with this parasite.

Canine Leishmaniasis in Brazil: Serological Follow-Up of a Dog Population in an Endemic Area of American Visceral Leishmaniasis

We performed a serological, clinical, and parasitological follow-up of a dog population in an endemic area of American Visceral Leishmaniasis estimated by indirect immunofluorescent assay (IFA) and western blot (WB). After twelve months, the results obtained from IFA demonstrated that 50% were seropositive and two serological profiles were observed: the first one ranging from 1/40 to 1/80 and the second ≥1/160. By WB, it was observed that the same percentage and sera from positive dogs presented the recognition of the peptides of 29 and 32 kDa up to 8 months before IFA serum conversion. Among the positive dogs, all the sera from symptomatic ones with tissue parasitism recognized the peptide of 68.5 kDa. Our results suggest the need of modifications in the control measures regarding the elimination of the dogs. They also corroborate the high sensitivity and specificity of western blot in the diagnosis of canine leishmaniasis, suggesting the possibility of its association with IFA.

Haementeria lutzi Pinto, 1920 (Hirudinea: Glossiphoniidae) as a putative Vector of Trypanosoma evansi (Kinetoplastida: Trypanosomatidae) in the Pantanal Matogrossense (MS, Brazil)

In the present study, it was shown under experimental conditions that Trypanosoma evansi could be mechanically transmitted to Rattus norvegicus by leeches (Haementeria lutzi). Additionally, we also described some aspects related to the behavior of the Trypanosoma evansi in the leeches after an infective blood feeding, as follows: a) 10 minutes after the parasites were ingested; they promptly progressed to the coelomic cavity. b) Approximately, from 10 to 30 minutes inside the gut, rounded and dividing forms together with stumpy and slender trypomastigotes showed a random dispersion. c) 24 hours after, the trypanosomes also invaded both, the salivary glands as well as the proboscis cells. Our results suggest that leeches of the species Haementeria lutzi could have some role as a probable alternative vector of Trypanosoma evansi at wetlands in Brazil.