"In vivo" leukocyte chemotaxis in experimental mice Schistosoma mansoni infection

The "in vivo" chemotaxis was studied in C57B1/10 mice 10, 30, 50 and 60 days after a Schistosoma mansoni infection in comparison to a control group (uninfected mice). Staphylococcal protein A was injected into a connective tissue air pouch of control and experimental mice and the leukocyte chemotaxis was counted. A decrease in polymorphonuclear (PMN) leukocyte response was found in infected mice in comparison to the control group (p<0.05). The 10 day infected mice showed a decreased PMN leukocyte response respecting the control group (p<0.05) and this finding became more evident 30 and 50 days post-infection. Although the PMN leukocyte response of 60 day infected mice increased in comparison to 50 day infected animals, it was still significantly lower the control response. The mononuclear leukocyte response was not significantly different between infected or uninfected mice.

Behaviour of albino and melanic variants of Biomphalaria glabrata Say, 1818 (Mollusca: Planorbidae) following infection by Schistosoma mansoni Sambon, 1907

The behaviour of the albino and melanic variants of Biomphalaria glabrata of Belo Horizonte (MG. Brazil) was studied comparatively, in terms of their respective susceptibilities to infection by Schistosoma mansoni of the same origin, through observation of the elimination of cercariae for a three-month period and the calculation of mortality and infection rates, in control and in infected snails. The number of amoebocytes, granulocytes and hyalinocytes in the circulating hemolymph during different periods of infection was analyzed. The evolution of the infection in the tissues was observed by means of histological cross-sections. The melanic variant showed greater susceptibility to infection and a higher mortality rate. The albino variant showed a higher number of circulating amoebocytes, both granulocytes and hyalinocytes. A higher number of degenerated sporocysts were seen in the histological cross-sections of the albino variant. The results suggest that the melanic variant of B. glabrata was more susceptible to infection by S. mansoni than was the albino variant.

The effects of the experimental infection from a focus of transmission of Schistosoma mansoni in a population of Biomphalaria tenagophila (D'Orbigny, 1835) in the region of "Vale do Ribeira de Iguape", Brazil

To observe the effects of the parasitic infection on the biology of B. tenagophila, field and laboratory populations of this mollusk from Itariri, in Vale do Ribeira, Brazil, were experimentally infected. Each mollusk received 10 miracidia of Schistosoma mansoni (SJ lineage) and was observed throughout the parasite's development. The biological variables were compared according to the criteria "group" and "infectious phase". The main damage caused by the parasitic infection manifested itself in reproduction, longevity and lesions on the shell of the mollusks in the patent phase. An infection rate of 58.8% was observed. Microanatomical study of the mollusk's digestive gland and ovotestis revealed the presence of evolving larval forms and cercariae. It was concluded that the effects of the parasitic infection on both populations were moderate, despite the low survival rate of the infected mollusks, the damage did not prevent either reproduction or the elimination of cercariae, which continued for a long time.

Lack of Galectin-3 Disturbs Mesenteric Lymph Node Homeostasis and B Cell Niches in the Course of Schistosoma mansoni Infection

Galectin-3 is a beta-galactoside-binding protein that has been shown to regulate pathophysiological processes, including cellular activation, differentiation and apoptosis. Recently, we showed that galectin-3 acts as a potent inhibitor of B cell differentiation into plasma cells. Here, we have investigated whether galectin-3 interferes with the lymphoid organization of B cell compartments in mesenteric lymph nodes (MLNs) during chronic schistosomiasis, using WT and galectin-3(-/-) mice. Schistosoma mansoni synthesizes GalNAc beta 1-4(Fuc alpha 1-3) GlcNAc(Lac-DiNAc) structures (N-acetylgalactosamine beta 1-4 N-acetylglucosamine), which are known to interact with galectin-3 and elicit an intense humoral response. Antigens derived from the eggs and adult worms are continuously drained to MLNs and induce a polyclonal B cell activation. In the present work, we observed that chronically-infected galectin-3(-/-) mice exhibited a significant reduced amount of macrophages and B lymphocytes followed by drastic histological changes in B lymphocyte and plasma cell niches in the MLNs. The lack of galectin-3 favored an increase in the lymphoid follicle number, but made follicular cells more susceptible to apoptotic stimuli. There were an excessive quantity of apoptotic bodies, higher number of annexin V(+)/PI(-) cells, and reduced clearance of follicular apoptotic cells in the course of schistosomiasis. Here, we observed that galectin-3 was expressed in nonlymphoid follicular cells and its absence was associated with severe damage to tissue architecture. Thus, we convey new information on the role of galectin-3 in regulation of histological events associated with B lymphocyte and plasma cell niches, apoptosis, phagocytosis and cell cycle properties in the MLNs of mice challenged with S. mansoni.

Schistosoma mansoni Tegument Protein Sm29 Is Able to Induce a Th1-Type of Immune Response and Protection against Parasite Infection

Background: Schistosomiasis continues to be a significant public health problem. This disease affects 200 million people worldwide and almost 800 million people are at risk of acquiring the infection. Although vaccine development against this disease has experienced more failures than successes, encouraging results have recently been obtained using membrane-spanning protein antigens from the tegument of Schistosoma mansoni. Our group recently identified Sm29, another antigen that is present at the adult worm tegument surface. In this study, we investigated murine cellular immune responses to recombinant (r) Sm29 and tested this protein as a vaccine candidate. Methods and Findings: We first show that Sm29 is located on the surface of adult worms and lung-stage schistosomula through confocal microscopy. Next, immunization of mice with rSm29 engendered 51%, 60% and 50% reduction in adult worm burdens, in intestinal eggs and in liver granuloma counts, respectively (p<0.05). Protective immunity in mice was associated with high titers of specific anti-Sm29 IgG1 and IgG2a and elevated production of IFN-gamma, TNF-alpha and IL-12, a typical Th1 response. Gene expression analysis of worms recovered from rSm29 vaccinated mice relative to worms from control mice revealed a significant (q<0.01) down-regulation of 495 genes and up-regulation of only 22 genes. Among down-regulated genes, many of them encode surface antigens and proteins associated with immune signals, suggesting that under immune attack schistosomes reduce the expression of critical surface proteins. Conclusion: This study demonstrates that Sm29 surface protein is a new vaccine candidate against schistosomiasis and suggests that Sm29 vaccination associated with other protective critical surface antigens is the next logical strategy for improving protection.

Peptides containing T cell epitopes, derived from Sm14, but not from paramyosin, induce a Th1 type of immune response, reduction in liver pathology and partial protection against Schistosoma mansoni infection in mice

Sm14 and paramyosin are two major Schistosoma mansoni vaccine candidate antigens. Recently, we have identified Sm14 and paramyosin epitopes that are recognized by T cells of resistant individuals living in endemic areas for schistosomiasis. Herein, mice were immunized with these peptides separately or in association in order to evaluate their vaccine potential. Immunization of mice with Sm14 peptides alone or mixed with paramyosin peptides was able to induce 26%-36.7% or 28%-29.2% of worm burden reduction, 67% or 46% of intestinal eggs reduction and also 54%-61% or 43%-52% of liver pathology reduction, respectively. Protection was associated with a Th1 type of immune response induced by Sm14 peptide immunization. In contrast, paramyosin peptide vaccination did not engender protective immunity or liver pathology reduction and immunization was associated with a Th2 type of immune response. (C) 2008 Elsevier B.V. All rights reserved.

Schistosoma mansoni: comparative evaluation of different routes of experimental infection

Experiments were carried out with Sw albino mice and it was concluded that the percutaneous route via abdominal skin was significatively more efficient than tail immersion method and subcutaneous infection; the subcutaneous injection was significatively more efficient than the percutaneous infection through the tail; this latter and the intraperitoneal injection, resulted in similar infections, but were significatively less efficient than the others. Significative difference was also observed in the comparison between the subcutaneous route and percutaneous infection through ear pinna. The influence of the site of skin infection by percutaneous route was also discussed.

The effect of Schistosoma mansoni infection on child morbidity in the State in Bahia, Brazil: II — Analysis at the individual level

The present investigation was carried out on a sample of 840 children (5 to 16 years old) from ten small towns of the State of Bahia in northeastern Brazil. The objetive was to study, by using a cross sectional methodology, the evolution of schistosomiasis morbidity (hepatic and splenic enlargement) in children, and the role of the intensity of S. mansoni infection in this process. The children were analised in three age groups (5 to 8, to 12 and 13 to 16 years old) and classified as uninfected, mildly infected, moderately infected and heavily infected according to the number of eggs in the stool. In children aged 5 to 8 years, increasing egg counts were not associated with increasing frequencies of hepatic or splenic enlargement. In the 9 to 12 years old group and association was observed with the prevalence of hepatic enlargement, but not with the prevalence of spleen enlargement. In the oldest group, 13 to 16 years old, an association was observed with the prevalence of enlargement of both organs. It was evident that in this population schistosomiasis morbidity develops in the early period of life as a gradual process starting with liver enlargement and followed by spleen enlargement some years later. It was found that the intensity of infection has a fundamental role in this process, although there is a latent period of some years before clinical splenomegaly appears in moderate-heavily infected children. The Authors suggest that the prevalence of splenomegaly in the 13 to 16 years old group is a good measure of the community level of schistosomiasis morbidity and could be used to measure the impact of control programs.

Experimental infection with Schistosoma mansoni of Biomphalaria straminea from different parts of the Northeast of Brazil

In the western part of the State of Bahia Biomphalaria straminea and B. glabrata both occur, but in the majority of cases they do not share the same habitat. In the State of Ceará, however, B. straminea is the sole snail host of Schistosoma mansoni. In this survey, no naturally infected B. straminea was found among snails collected from Bahia and Ceará, evidently because of the very low infection rates. The susceptibility of laboratory-reared specimens to infection with a Puerto Rican strain of S. mansoni was then tested experimentally. In general, the snails showed very low susceptibility. The infection rates were 1.1% among snails from Redenção (Ceará); 2.3% in those from Pentecoste (Ceará); 2.9% in snails from São Desidério (Bahia), while they were very high among an albino strain (NIH) of B. glabrata used as control. Another group of B. straminea from São Desidério was exposed to a Bahian strain of S. mansoni and the infection rate was still very low (3.6%) Apparently, the very low susceptibility of B. straminea, despite high snail density, is correlated with moderate infection rates with S. mansoni among humans, as shown by the results of stool examinations conducted by SUCAM in the municipalities of Redenção and Pentecoste, in Ceará.

Water-contact patterns and schistosoma mansoni infection in a rural community in northeast Brazil

We evaluated the influence of water-related human activities, contaminative behaviour, house location, education and socio-economic status on endemic Schistosoma mansoni infection. The study was conducted in a hilry non-irrigated area of rural northeast Brazil amongst a defined population of subsistence farmers, of whom 93% were infected by age 20. The area was mapped, water bodies were surveyed, and a detailed questionnaire was performed on each household. Infection was assessed by duplicate stool examinations using the sensitive Bell technique to quantify egg excretion. For each household, and index of intensity of infection was computed by grouping individual log-transformed egg counts as an age-sex adjusted Z score. Few households had a sanitary installation or a domestic water supply. However, neither water-contact nor contaminative behavior were indiscriminate. The people made considerable effort to defaecate far from a water source, to obtain household drinking water from the cleanest source, and to bathe only at certain sites where privacy is assured. Land ownership and literacy correlated poorly with the household index of intensity of infection. The key influence on infection status was the relative location of the house and snail-free or snail colonized water sources. In this area, a safe domestic water supply is the critical input needed to achieve definitive control of endemic Schistosomiasis.

Interrelation between parasitic infections and hypovitaminosis A: Schistosoma mansoni infection and the retinol blood levels in an endemic area of State of Minas Gerais (Brazil)

The interrelation between schistosomiasis and the retinol blood levels was studied in a double blind method, by comparing the serum vitamin A of the infected and non-infected group of an endemic area of Schistosoma mansoni infection. The infected group was characterized by 106 parasitized persons in the intestinal and hepatointestinal forms, who eliminated less than 500 eggs/gram of feces (Modified Kato's method); the non-infected group was characterized by 112 inhabitants of this endemic area without eggs in the stools and presenting negative intradermal reactions, and absence of previous specific treatment. The blood levels of retinol was determinated using trifluoracetic acid method, regarding the normal levels > 20,0mg/100ml. The results of this study point out the absence of correlation between S. mansoni infection and blood levels of vitamin A.

Validity of selected clinical signs and symptoms in diagnosis of Schistosoma mansoni infection

Sensitivity, specificity and positive predictive values of selected clinical signs and symptoms in the diagnosis of Schistosoma mansoni infection were evaluated in 403 individuals (69% of inhabitants over 1 year of age) in an endemic area in Brazil (Divino). Highest sensitivity (13%) was found for blood in stools. Specificity over 90% was found for blood in stools, palpable liver with normal consistency and palpable hardened liver at middle clavicular (MCL) or middle sternal lines (MSL). Hardened liver at MSL (83%) or MCL (75%), and blood in stools (78%) presented higher positive predictive values for S. mansoni infection, while palpable liver with normal consistency at MCL (45%) or MSL (48%) presented smaller values. Enlarged liver without specification of its consistency has been traditionally used as an indicator of the infection in areas where malaria or Kalazar are not endemic. Our results demonstrate that the probability that a person with blood in stools or hardened palpable liver is infected is higher than among those with palpable liver with normal consistency.

A multivariate analysis of socio-demographic factors, water contact patterns and Schistosoma mansoni infection in an endemic area in Brazil

Associations between socio-demographic factors, water contact patterns and Schistosoma mansoni infection were investigated in 506 individuals (87% of inhabitants over 1 year of age) in an endemic area in Brazil (Divino), aiming at determining priorities for public health measures to prevent the infection. Those who eliminated S. mansoni eggs (n = 198) were compared to those without eggs in the stools (n = 308). The following explanatory variables were considered: age, sex, color, previous treatment with schistosomicide, place of birth, quality of the houses, water supply for the household, distance from houses to stream, and frequency and reasons for water contact. Factors found to be independently associated with the infection were age (10-19 and > 20 yrs old), and water contact for agricultural activities, fishing, and swimming or bathing (Adjusted relative odds = 5.0, 2.4, 3.2, 2.1 and 2.0, respectively). This suggests the need for public health measures to prevent the infection, emphasizing water contact for leisure and agricultural activities in this endemic area.

Epidemiology of Schistosoma mansoni infection in a low-endemic area in Brazil: clinical and nutritional characteristics

A cross-sectional case-control study designed to evaluate the role of malnutrition in the association between the intensity of Schistosoma mansoni infection and clinical schistosomiasis, was conducted in an area with both low frequency of infection and low morbidity of schistosomiasis in Brazil. Cases (256) were patients with a positive stool examination for S. mansoni; their geometrical mean number of eggs/gram of feces was 90. Controls (256) were a random sample of the negative participants paired to the cases by age, sex and length of residence in the area. The clinical signs and symptoms found to be associated with S. mansoni infection, comparing cases and controls, were blood in stools and presence of a palpable liver. A linear trend in the relative odds of these signs and symptoms with increasing levels of infection was detected. Adjusting by the level of egg excretion, the existence of an interaction between palpable liver and ethnic group (white) was suggested. No differences in the nutritional status of infected and non-infected participants were found.

Schistosoma mansoni: evaluation of the activity of oxamniquine on schistosomules, at 24 hours after infection

Mice transcutaneously infected with about 400 cercariae were submitted to treatment with oxamniquine (400 mg/kg), 24 hours after infection. The recovery of schistosomules, at 4, 24, 48 and 72 hours and 35 days after treatment, showed the activity of the drug on the parasites, thus practically preventing their migration from the skin to the lungs. Worm recovery performed in the lungs (96 hours after treatment) showed recovery means of 0.6 worms/mouse in the treated group and 53.8 in the control group (untreated). The perfusion of the portal system carried out at 35 days after treatment clearly showed the elimination of all the parasites in the treated group, whereas a recovery mean of 144.7 worms/mouse was detected in the control group (untreated). These findings confirm the efficacy of oxamniquine at the skin phase of infection, and also show similarity with the immunization method that uses irradiated cercariae. The practical application of these findings in the medical clinic is discussed too