Extra-tissular Schistosoma mansoni egg granulomata in the peritoneal cavity of mice

The presence of Schistosoma mansoni eggs surrounded by inflammatory cells were detected within the peritoneal cavity of experimentally infected mice. The histological and ultrastructural analysis revealed the predominantly macrophagic composition of these structures. The presence of epithelioid cells, macrophages in different stages of activation and the architectural pattern of the cells, characterize these structures as extra-tissular true granulomas. Granulomas much similar to those observed in the peritoneal cavity of infected mice were also detected after the intraperitoneal injection of viable eggs in non-infected mice. Collagen fibers were observed in between the inflammatory cells of granulomas obtained 10 weeks after infection and 48 hours after the injection of viable eggs into the peritoneal cavity. In later times of infection or injection the amount of collagen fibers increases resulting in a typical pattern of healed schistosoma egg granulomas. The possible influence of the immune response on the genesis of the granulomatous reaction as well as the influence of the vascularized connective tissue on this process is discussed.

Evaluation of antibody responses in american visceral leishmaniasis by ELISA and immunoblot

American visceral leishmaniasis (AVL) is an important disease among children of northeast Brazil. In order to characterize antibody responses during AVL, sera of hospitalized patients were analyzed by ELISA and Western blot using a Leishmania chagasi antigen preparation. The ELISA was positive (asorbance [greater than or equals to] 0.196) at a serum dilution of 1:1024 in all patients at presentation, and fell to ward control levels over the following year. Only one of 72 control subjects tested positive, and that donor had a sibling with AVL. Immunoblots of the patients' sera recognized multiple bands, the most frequent of which were at approximately 116 kDa, 70 kDa, and 26 kDa. Less frenquently observed were bands at approximately 93 kDa, 74 kDa, 62 kDa, 46 kDa and 32 kDa. The ELISA responses and patterns of banding were distinctive for AVL, and could be used to differentiate patients with AVL from those with Chagas' disease of cutaneous leishmaniasis. Sera from six AVL patients followed for up to six weeks after treatment identified no new bands. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) of surface iodinated parasite proteins showed one major band and four minor bands, whereas SDS-PAGE of biotinylated prarasite proteins revealed a banding pattern similar to those of patient sera. AVL appears to produce characteristic immunoblot patterns which can be used along with a sensitive screening ELISA to diagnose AVL.

A potential vector of Schistosoma mansoni in Uruguay

Susceptibily experiments were carried out with a Biomphalaria straminea-like planorbid snail (Biomphalaria aff. straminea, species inquirenda) from Espinillar, near Salto (Uruguay), in the area of the Salto Grande reservoir, exposed individually to 5 miracidia of Schistosoma mansoni (SJ2 and BH2 strains). Of 130 snails exposed to the SJ2 strain, originally infective to Biomphalaria tenagophila, 30 became infected (23%). The prepatent (precercaria) period ranged from 35 to 65 days. The cercarial output was irregular, following no definite pattern, varying from 138 to 76,075 per snail (daily average 4.3 to 447.5 and ending up with death. Three specimens that died, without having shed cercarie, on days 69 (2) and 80 after exposure to miracidia, had developing secondary sporocysts in their tissues, justifying the prospect of a longer precercarial period in these cases. In a control group of 120 B. teangophila, exposed to the SJ2 strain, 40 became infected, showing an infection rate (33.3%) not significantly different from that of the Espinillar snail (X [raised to the power of] 2 = 3.26). No cercarie were produced by any of the Espinilar snails exposed to miracidia of the BH2 strain, originally infective to Biomphalaria glabrata. Four specimens showed each a primary sporocyst in one tentacle, which disappeared between 15 and 25 days post-exposure, and two others died with immature, very slender sporocysts in their tissues on days 36 and 54. In a control group of 100 B. glabrata exposed to BH2 miracidia, 94 shed cercariae (94%) and 6 remained negative. Calculation of Frandsen's (1979a, b) TCP/100 index shows that "Espinillar Biomphalaria-SJ2 S. mansoni" is a vector-parasite "compatible" combination. Seeing that tenagophila-borne schistosomiasis is prevalent in Rio de Janeiro and São Paulo states and has recently spread sothwards to Santa Catarina state, and the range of B. tenagophila overlaps taht of the Espinillar Biomphalaria, the possibility of schistosomiais establishing itself in Uruguay, although not imminent, is not to be disregarded.

Experimental american trypanomiasis in rats: sympathetic denervation, parasitism and inflammatory process

Tissue parasitism, inflammatory process (histologic methods) and sympathetic denervation (glyoxylic acid-induced histofluorescence for demonstration of catecholamines) were studied in the heart (atrium and verntricle) and the submandibular gland of rats infected with the Y strain of Trypanosoma cruzi. In the heart paralleling intense parasitism and inflammatory process, the sympathetic denervation started at day 6 of infection and at the end of the acute phase (day 20) practically no varicose nerve terminals were found in both myocardium and vessels. In the submandibular gland, in spite of the rarity of anastigote pseudocysts and the scarcity of inflammatory foci, slight to moderate (days 13-15 of infection) or moderate to severe denervation (day 20) was found. At day 120 of infection both organs exhibited normal pattern of sympathetic innervation and only the heart showed some inflammatory foci and rare psudocysts (ventricle). Our data suggest the involvement of circulating factors in the sympathetic denervation phenomena but indicate that local inflammatory process is, at least, an aggravating factor.

Fonte alimentar e potencial vetorial de Triatoma vitticeps (Stal, 1859) com relação à doença de Chagas humana no estado do Espírito Santo, Brasil (Hemiptera, Reduviidae)

In paralel with several other epidemiologic and entomologic data of 19 Municipalities of Espírito Santo State, Brazil, the feeding pattern of 222 Triatoma vitticeps is studied through precipitin tests. Very high levels of natural infection with Trypanosoma cruzi are observed in adult insects, in contrast with the abscence or minimum degrees of infection among nymphs and human individuals. The precipitin tests showed the contact of the insects with multiple blood sources, chiefly human and birds, followed by rodents and marsupials. The data suggest that T. vitticeps in spite of being highly antropophilic, become infected by T. cruzi in sylvatic ambient and occasionally invade houses. The species doesn't seem to be - at least until now - a good vector in the domestic cycle of Chagas' disease. Several factors seem to be involved in this conclusion, mainly the low density of the insect in the houses, its hardness to coloniza them, its slowness concerning to suction and defecation and possibly its low susceptibility to different T. cruzi strains.

Two new species of Euphumosia from Papua New Guinea (Diptera: Calliphoridae)

Two new species of the calliphorid blow flies are described from Papua New Guinea under the names of Euphumosia lopesi and E. robertsi. Body coloration and mesonotal pattern are characteristic and differ from those of the closely allied species, E. evittata Togerson & James, 1967.

Human and dogs Toxocara canis infection in a poor neighborhood in Bogota

Prevalence of Toxocara canis antibodies was studied in a poor community of Bogotá, Colombia. Two-hundred-sevem patients, from both sexes and all age groups, were studied. Positive Elisa titers were found in 47.5% of the population, a high prevalence compared with reports from developed countries. T. canis ova were positive in 43.6% of fecal samples from dog puppies. An endemic pattern of the disease is described: socioeconomic status, weathers, pollution, poor hygiene and a significant population of infected dogs. Neither the physical examination nor Elisa titers could detect any case of T. canis disease.

The thermal stability of yellow fever vaccines

The assessment of yellow fever vaccine thermostability both in lyophilized form and after reconstitution were analyzed. Two commercial yellow fever vaccines were assayed for their thermal stability. Vaccines were exposed to test temperatures in the range of 8 (graus) C to 45 (graus) C. Residual infectivity was measured by a plaque assay using Vero cells. The titre values were used in an accelerated degradation test that follows the Arrhenius equation and the minimum immunizing dose was assumed to be 10 (ao cubo) particles forming unit (pfu)/dose. Some of the most relevant results include that (i) regular culture medium show the same degradation pattern of a reconstituted 17D-204 vaccine; (ii) reconstituted YF-17D-204 showed a predictable half life of more than six days if kept at 0 (graus) C; (iii) there are differences in thermostability between different products that are probably due to both presence of stabilizers in the preparation and the modernization in the vaccine production; (iv) it is important to establish a proper correlation between the mouse infectivity test and the plaque assay since the last appears to be more simple, economical, and practical for small laboratories to assess the potency of the vaccine, and (v) the accelerated degradation test appears to be the best procedure to quantify the thermostability of biological products.

Histological and ultrastructural studies of the marsupial Didelphis albiventris Peyer's patches

Differing from the studied Eutheria the white belly opossum Peyer"s patches do not present a conspicous dome. M cells are located in the inmer layer of bilaminal invaginations formed at the bottom of the villi. A great variation in the morphology of M cells was observed. The enterocytes located at the epithelial inner layer may present endocytic vesicles, and the microvilli are shorter tha the microvilli of enterocytes lining the small intestine. As these morphological aspects have been described to exist in the enterocytes of the lancet opossum small intstine it was surmised that the opossum Peyer's patches special epithelium could represent the persistence in adult animals of a cellular pattern established before the intestinal maturation had occurred.

The importance of the opossum (Didelphis albiventris) as a reservoir for Trypanosoma cruzi in Bambuí, Minas Gerais state

In a survey realized on the sylvatic and peridomestic environment at Bambuí county, Minas Gerais State, 44 (37.9%) out of 116 opossums (Didelphis albiventris) captured were found to be naturally infected with Trypanosoma cruzi. One handred and forty three parasite samples were obtanied from 43 infected opossums using simultaneously hemoculture, xenodiagnosis (Triatoma infestans, Panstrongylus megistus and Rhodnius neglectus) and examination of anal glands contents. The parasite samples were characterized according to six isoenzyme patterns. All samples, independently of the method of isolation, presented an isoenzyme pattern similar to the standard T. cruzi Z1, showing that either xenodiagnosis or hemoculture can used without selecting parasite subpopulation from naturally infected opossums. Preveous isoenzyme patterns reported for human T.cruzi isolates from same region were completely different. This isoenzyme dissimilarity between sylvatic and domiciliar environments suggests the existence of two independent T. cruzi transmission cycles in Bambuí. The epidemiological implicatinos of these results are discussed.

Applications of the MuSIASEM approach to study changes in the metabolic pattern of Catalonia

This document presents an integrated analysis of the performance of Catalonia based on an analysis of how the energy consumption (measured at the societal level for the Catalan Society) is used within both the productive sectors of the economy and the household, to generate added value, jobs, and to guarantee a given level of material standard of living to the population. The trends found in Catalonia are compared to the trends of other European Countries to contextualize the performance of Catalonia with respect to other societies that have followed different paths of economic development. The first part of the document consists of the Multi-Scale Integrated Analysis of Societal and Ecosystem Metabolism (MuSIASEM) approach that has been used to provide this integrated analysis of Catalan Society across different scales (starting from an analysis of the specific sectors of the Catalan economy as an Autonomous Community and scaling up to an intra-regional (European Union 14) comparison) and across different dimensions of analyses of energy consumption coupled with added value generation. Within the scope of this study, we observe the various trajectories of changes in the metabolic pattern for Catalonia and the EU14 countries in the Paid Work Sectors composed of namely, the Agricultural Sector, the Productive Sector and the Services and Government Sector also in comparison with the changes in the household sector. The flow intensities of the exosomatic energy and the added value generated for each specific sector are defined per hour of human activity, thus characterized as exosomatic energy (MJ/hour) (or Exosomatic Metabolic Rate) and added value (€/hour) (Economic Labour Productivity) across multiple levels. Within the second part of the document, the possible usage of the MuSIASEM approach to land use analyses (using a multi-level matrix of categories of land use) has been conducted.

Reversibility of cardiac fibrosis in mice chronically infected with Trypanosoma cruzi, under specific chemotherapy

This investigation was performed to verify the effect of specific chemotherapy (Benznidazole or MK-346) on the inflammatory and fibrotic cardiac alterations in mice chronically infected with the strains 21 SF (Type II) and Colombian (Type III) of Trypanosoma cruzi. To obtain chronically infected mice, two groups of 100 Swiss mice each, were infected with either the 21 SF or the Colombian strain (2x 10 [raised to the power of] 4 and 5x 10 [raised to the power of] 4 blood forms respectively). The rate of morality in the acute phase was of 80% for both groups. Twenty surviving mice chronically infected with the 21 SF strain and 20 with the Colombian strain were then divided in treated and untreated groups. Excluding those that died during the course of treatment, 14 mice chronically infected with the 21 SF strain and 15 with the Colombian strain were evaluated in the present study. Chemotherapy was performed with Benznidazole (N-benzil-2-nitro-1-imidazolacetamide) in the dose of 100mg/k.b.w/day, for 60 days, or with the MK-436(3(1-methyl-5 nitroimidazol-2-yl) in two daily doses of 250 mg/k.b.w, for 20 days. Parasitological cure tests were performed (xenodiagnosis, haemoculture, subinovulation of the blood into newborn mice), and serological indirect immunofluorescence test. The treated and untreated mice as well as intact controls were killed at different periods after treatment and the heart were submitted to histopathological study with hematoxilineosin and picrosirius staining; ultrastructural study; collagen immunotyping, fibronectin and laminin identification by immunofluorescence tests. Results: the untreated controls either infected with 21 SF or Colombian strain, showed inflammatory and fibrotic alterations that were mild to moderate with the 21 SF strain and intense with the Colombian strain. Redpicrosirius staining showed bundles of collagen in the interstitial space and around cardiac fibers. Increased deposits of mitritial components and collagen fibers, macrophages and fibroblasts appeared at the ultra structural examination. Deposits of fibronectin, laminin, pro-III and IV collagens were seen, most intense in those infected with the Colombian strain. Treated nice, parasitologically cured, presented clear-cut regression of the inflammatory lesions and of the interstitial matrix thickening. Mice infected with the Colombian strain and treated with MK-436, was parasitologically cured in 5/6 cases and showed mild inflammatory infiltration and fibrosis. The mice treated with Benznidazole (Colombian strain) did not cure and showed moderate fibrosis and inflammation. Treatment of the nice infected with the 21 SF with Benznidazole determined parasitological cure of all animals, that showed mild inflammation and fibrosis of the myocardium. The cured mice of all groups and treated but uncured showed collagen degradation at electronmicroscopy and decrease of immunofluorescence pattern of the matrix.

Stability of isoenzyme and kinetoplast DNA (k-DNA) patterns in successively cloned Trypanosoma cruzi populations

Four Trypanosoma cruzi strains from zymodermes A, B, C and D were successively clonedon BHI-LIT-agar-blood BLAB). Twenty clones from the first generation (F1), 10 from The second (F2) and 4 from the third (F3) from the strains A138, B147 and C23 were isolated. The D150 strain provied 29 F1 and F2 clones. The strains and clones had their isoenzyme and K-DNA patterns determined. The clones from A138, Bl47 and C231 strains presented isoemzyme and K-DNA patterns identical between thewmselves and their respective parental strains. Therefore showing the homogenety and stability of isoenzyme and K-DNA patterns after successive cloning. The Dl50 strain from zymodeme D (ZD) showed heterogeneity. Twenty-eight out of 29 clones of the first generation were of zymodeme A and only one was of zymodeme C, confirming previous reports that ZD strains consisted of ZA and ZC parasite populations. The only D150 strain clone of zymodeme C showed a K-DNA pattern identical to its parental strain. The remining clones although similar among themselves were different from the parental strain. Thus the T. cruzi strains had either homonogeneus or heterogeneous populations. The clones produced by successive cloning provided genetically homonogeous populations. Their experimental use will make future results more reliable and reproducible.

Dynamics of fibrosis production and resorption in intestinal schistosomiasis of mice

A histological, morphometric and immunocytochemical study of schistosomal periovular granulomas in the liver and intestines of mice revealed that intestinal granulomas are smaller and contain less collagen than those in the liver. After curative treatment intestinal granulomas undergo a relatively more rapid resorption, although the general pattern of collagen degradation apparently does not differ from that observed in the liver. Tendency to form scattered, usually isolated granulomas that are only mildly fibrogenic, coupled with a well-balanced process of resorption appear as the explanation why intestinal fibrosis is not an outstanding feature of schistosomiasis as it is in the liver.

Kinetics of antigen specific and non-specific polyclonal B-cell responses during lethal Plasmodium yoelii malaria

In order to study the kinetics and composition of the polyclonal B-cell activation associated to malaria infection, antigen-specific and non-specific B-cell responses were evaluated in the spleens of mice infected with Plasmodium yoelii 17 XL or injected with lysed erythrocytes or plasma from P. yoelii infected mice or with P. falciparum culture supernatants. Spleen/body weigth ratio, numbers of nucleated spleen cells and Immunoglobulin-containing and Immunoglobulin-secreting cells increased progressively during the course of infection,in parallel to the parasitemia. A different pattern of kinetics was observed when anti-sheep red blood cell and anti-trinitrophenylated-sheep red blood cell plaque forming cells response were studied: maximum values were observed at early stages of infection, whereas the number of total Immunoglobulin-containing and Immunoglobulin-secreting cells were not yet altered. Conversely, at the end of infection, when these latter values reached their maximum, the anti-sheep red blood cell and anti-trinitrophenylated-sheep red blood cell specific responses were normal or even infranormal. In mice injected with Plasmodium-derived material, a higher increase in antigen-specific PFC was observed, as compared to the increase of Immunoglobulin-containing and Immunoglobulin-secreting cell numbers. This suggested a "preferential" (antigen-plus mitogen-induced) stimulation of antigen-specific cells rather than a generalized non-specific (mitogen-induced) triggering of B-lymphocytes. On the basis of these and previous results, it is suggested that polyclonal B-cell activation that takes place during the course of infection appears as a result of successive waves of antigen-specific B-cell activation.