Studies on granuloma formation in Syrian hamsters experimentally infected with Schistosoma mansoni

Juvenile rats are more susceptible to the acute toxicity of the phosphorothionate insecticides parathion and chlorpyrifos than are adult rats. Developmental changes in brain acetylcholinesterase and hepatic aliesterase (carboxylesterase), cytochrome P450, and the P450-mediated metabolism of these two phosphorothionate insecticides were investigated in male Sprague-Dawley rats. Specific activities of acetylcholinesterase in cerebral cortex, but not medulla oblongata, and of liver aliesterases increased with age, indicating the presence of both more target esterases and more protective esterases, respectively, in the adult compared to the juvenile animal. Sensitivity of the brain acetylcholinesterase to inhibition by paraoxon and chlorpyrifosoxon, as measured by IC50 values, did not change significantly with age, whereas the hepatic aliesterase sensitivity to inhibition decreased with age. Progressive increases in activities of P450-mediated activation (desulfuration) (6- to 14-fold) and detoxication (dearylation) (2- to 4-fold), as well as concentrations of P450 (7-fold) and protein (2-fold), were observed between neonate and adult hepatic microsomes. Microsomal pentoxyresorufin O-dealkylase activity followed a developmental pattern similar to desulfuration and dearylation, displaying a 16-fold increase between neonates and adults. However, microsomal ethoxyresorufin O-deethylase activity increased until 21 days of age, displaying a 16-fold increase, then decreased in adulthood to a level 10-fold higher than neonates. These results indicate that target enzyme sensitivity is not responsible for age-related toxicity differences, nor is the potential for hepatic bioactivation, whereas lower levels of hepatic aliesterase-mediated protection and P450-mediated dearylation probably contribute significantly to the greater sensitivity of juveniles to phosphorothionate toxicity.     

Pairing of Schistosoma mansoni males in infected Syrian hamsters

Pairing of male schistosomes in the liver of infected hamsters was recorded with Egyptians S. mansoni strain. The homospecific male pairs never carried each other in the gynaecophoric duct, but they being closed in either central or hepatic veins. Other perfused males and females en copula showed normal mating behaviour. The paired males were more or less in the same size. The random sexed miracidia used resulted in obtaining 1:2.1 female/male ratio. It is concluded that the random increase of male schistosomes may create the male pairing behaviour. Also, the migration of female against the blood stream to the mesenteric plexus of the host and the failure of male to catch them may lead to this homosexual pairing. The black haemozoin-like substance seen in mature females was also observed in the pairing males and this probably reflects the effect of scarcity or migration of females to the mesenteric plexus.     

Effect of wide spectrum anti-helminthic drugs upon Schistosoma mansoni experimentally infected mice

An important issue of concern for drug analysis in hair is the change in the drug concentration induced by the cosmetic treatment of hair. The products used for this treatment are strong bases and they are expected to cause hair damage. As a result drugs may be lost from the hair matrix or, under conditions of environmental contamination, be more easily incorporated into the hair matrix. We investigated the effects of cosmetic treatment in vivo by analysing hair samples selected from people who had treated their hair by bleaching or dyeing before sample collection. All of the subjects admitted a similar drug consumption during the time period for which the strands were analysed. Samples were viewed under a microscope to establish the degree of hair damage. Treated and untreated portions from each lock of hair were then selected, separated and analysed by standard detection procedures for cocaine, opiates, cannabinoids and nicotine. In all cases the drug content in hair that had undergone cosmetic treatment decreased in comparison to untreated hair. The majority of the mean differences were in the range of 40%-60% (cocaine, benzoylecgonine, codeine, 6-acetylmorphine and THC-COOH). For morphine the mean difference was higher than 60%, and two cases (THC and nicotine) differed by approx. 30%. These differences depended not only on the type of cosmetic treatment, as bleaching produced higher decreases than dyeing, but also on the degree of hair damage i.e. the more damaged the hair, the larger the differences in the concentration levels of drugs.     

Effect of Schistosoma mansoni infection on offsprings born from infected mothers

Offsprings C57BL/6 mice (4 weeks old) coming from either moderately infected (40 S. mansoni cercariae) or heavily infected (100 S. mansoni cerariae) mothers, were exposed to 40 S. mansoni cercariae each. Seven weeks post infection (P.I.), Offsprings were sacrificed. In both groups there was significant reduction in the worm load, both hepatic and intestinal tissue egg count. The oogram profile was not altered. Humoral immune response as regards the level of anti S. mansoni SEA Ab was elevated in both groups in comparison to their parallel controls at 2 weeks post delivery and 7 weeks P.I. The level of antibodies was significantly higher in heavily infected Offsprings than that present in offsprings coming from moderately infected mothers. Delayed footpad swelling and hepatic granuloma size were significantly reduced in both groups comparing with their corresponding controls.     

Vaccination of patas monkeys experimentally infected with Schistosoma haematobium using a recombinant glutathione S-transferase cloned from S. mansoni

The capacity of a recombinant glutathione S-transferase from Schistosoma mansoni (rSm28GST) to vaccinate primates (Erythrocebus patas) against a heterologous infection with Schistosoma haematobium has been tested. Two injections of the purified molecule with Muramyl-Di-Peptide (MDP) as adjuvant resulted in a high level antibody response in the five immunized animals and in a significant reduction in worm fecundity compared to the controls which received adjuvant alone. Mean levels of daily egg excretion in urine an faeces were reduced by respectively 55% and 74% although perfusion revealed that worm burdens were similar in both groups. The protective effect was long lasting since it was maintained up to the end of the experiment, 42 weeks after infection. Hatching rates and the numbers of intra-uterine eggs were also significantly affected by the vaccination. Tissue eggs were also drastically diminished in the urogenital system (-80%) but the reduction was not statistically significant. One animal was not protected by the immunization. There was a good correlation between parasitological data and the intensity of bladder lesions assessed by microscopic examination. Polypoid formations together with an intense exudation of the lamina propria were frequently seen in the controls but rarely in the vaccinated group where formation of scar tissue was predominant. These results underline the vaccine potential of the recombinant Sm28GST as a possible valuable prophylactic tool for the control of egg-induced pathology and transmission of African schistosomes.     

Detection of Schistosoma mansoni M antigen in circulating immune-complexes and in kidneys of infected hamsters

Circulating M antigen (CMA) of Schistosoma mansoni was found in the trichloroacetic acid (TCA) soluble fraction of a polyethylene glycol precipitate of serum from infected hamsters. It was also found as a TCA-soluble component in an immunoglobulin-containing fraction eluted from infected hamster kidneys. It was not found in similarly treated control sera nor in the products of acid dissociation of circulating immune complexes (CIC) from infected hamster sera. CMA was not detected in the kidneys of normal hamsters. Precipitating anti-M antigen antibodies were present in one of 10 sera from infected hamsters, but not in the eluates of hamster kidneys. These results indicate that CMA is present in circulating immune complexes in infected hamsters. The presence of CMA in kidneys from the same hamsters suggests a possible role for circulating antigens in immune-complexed form in the aetiology of glomerulonephritis in S. mansoni infection.     

An ultrasonographic study of liver fibrosis in patients infected with Schistosoma mansoni in north-east Brazil

Between August 1988 and July 1990, 176 patients with Schistosoma mansoni infection attending the University Hospital, Recife, Brazil received a complete clinical examination including stool examination for intestinal parasites, liver function tests, and ultrasonography. The majority were also examined by upper digestive tract endoscopy. The clinical distribution of their disease was as follows: 26.7% intestinal, 13.6% hepato-intestinal, 53.4% compensated hepatosplenic and 6.3% decompensated hepatosplenic. Infection intensity was high, with a median of 360 eggs/g of faeces. Ultrasonography showed a good correlation between the degree of hepatic periportal fibrosis and the clinical stage of disease (P < 0.0001). Of the patients with the intestinal form of schistosomiasis, 12.8% had grade I fibrosis and the others had no fibrosis; 33.3% of patients with hepatointestinal schistosomiasis had grade I fibrosis, 8.3% had grade II fibrosis, and 58.4% had no fibrosis; all the patients with hepatosplenic disease had grade II or grade III fibrosis. The degree of liver fibrosis detected by ultrasonography correlated with the degree of oesophageal varices detected by endoscopy (P = 0.0001). The degree of oesophageal varices also correlated with the presence of haemorrhage (P < 0.0001). Ultrasonography is considered superior to liver biopsy, permitting a dynamic approach to the study of schistosomiasis morbidity with precise diagnosis and simple sequential follow-up of post-treatment results.     

Studies on experimental mixed infection of Schistosoma haematobium and Schistosoma mansoni

Swiss Albino mice have been infected with S. haematobium and challenged 4 weeks later with S. mansoni. Parasitological, pathological and ultrastructural studies were done. The results revealed cross mating between the two species. A reduction in S. mansoni worm load, egg count, hepatic granuloma number and size was noticed. The presence of heterologous immunity was suggested.     

Protection of mice against Schistosoma mansoni infection by passive transfer of sera from infected rabbits

Twelve patients with bilateral comparable gingival recessions were treated using a split mouth design, to compare the relative success of root coverage by two regenerative procedures. The areas of recession treated were Class I or II according to Miller's classification and caused either an esthetic problem and/or root sensitivity. The symmetrical defects, on the maxillary canines, 4 mm or deeper, were randomly assigned in each patient to surgical procedures with either a bioresorbable matrix barrier (test) or a non-resorbable expanded polytetrafluoroethylene membrane (control). Gingival recession, clinical attachment level, probing depth, and extension of keratinized tissue were measured at baseline and at 6 months postsurgically. Both procedures resulted in significant root coverage (P < 0.0001) and attachment gain (P < 0.0001). The gingival recession decreased from 4.75 +/- 0.22 mm to 0.83 +/- 0.24 mm and from 4.75 +/- 0.22 mm to 0.75 +/- 0.22 mm, corresponding to a mean root coverage of 82.4% and 83.2%, at the test and control sites respectively. The average clinical attachment gain was 4.33 +/- 0.44 mm at the test sites compared to 4.42 +/- 0.48 mm for the non-resorbable barrier. No significant changes were found for probing depth and keratinized tissue. Data analysis did not demonstrate any significant difference between the two procedures for any of the variables included. However, a questionnaire given to each patient revealed the single-step surgery to be the patients' choice.     

Fc epsilonRI-deficient mice infected with Schistosoma mansoni mount normal Th2-type responses while displaying enhanced liver pathology

The IgE/Fc epsilonRI interaction is postulated to play an important role in resistance to helminths both at the level of anti-parasitic effector cell function and in the initiation of Th2 responses through IL-4 produced by Fc epsilonRI+ non-B, non-T (NBNT) cells. To formally evaluate the role of IgE/Fc epsilonRI signaling in the host response to helminths we studied Schistosoma mansoni infection in Fc epsilonRI knockout (KO) mice. Infected wild-type (wt) and KO animals showed comparable adult worm and tissue egg burdens, arguing against a role for Fc epsilonRI interactions in host resistance. Significantly, NBNT cells from infected KO, in contrast to wt animals, did not secrete IL-4 when stimulated with anti-IgE Ab or soluble parasite Ag. Nevertheless, serum IgE levels and Th2 cytokine production profiles were comparable in both strains of mice, demonstrating that the Ag-dependent stimulation of IL-4 secretion by NBNT cells is not essential for helminth-induced Th2 differentiation. However, when stimulated with low Ag doses, splenocytes from infected Fc epsilonRI-deficient mice produced less IL-4 in vitro than similar cultures from infected wt animals, an effect attributable to their defective NBNT cell function. Moreover, infected KO mice showed enhanced egg granuloma formation and hepatic fibrosis, revealing that the IgE/Fc epsilonRI interaction, while not essential for Th2 response development or resistance to primary infection, plays a significant role in down-regulating host pathology.     

An immunodiffusion assay for the detection of canine leishmaniasis due to infection with Leishmania infantum

An immunodiffusion assay (IDA) with polyethylene glycol (PEG) was tested for usefulness as diagnostic test for canine leishmaniasis (CL). A comparative analysis of dog sera was made using IDA with PEG, immunofluorescence assay (IFA) and enzyme immunosorbent assay (ELISA) techniques. Fourty-four dogs from Italy with CL (endemic dogs) and eight Dutch dogs with CL contracted in South Europe (expatriate dogs) were tested together with 40 endemic and 35 expatriate controls. Specificity did not differ substantially among the serotests, ELISA in endemic dogs being the least specific (mean specificity given in IFA, IDA and ELISA, 100%, 98% and 93.5%, respectively). Sensitivity in expatriate dogs was 100% for all serotests but was highly variable in endemic dogs. In parasite-negative dogs, IFA had the most sensitivity, i.e., 80.5% compared to 69% for both ELISA and IDA. In contrast, ELISA in parasite-positive endemic dogs had a sensitivity of 100% whereas both IFA and IDA gave a sensitivity of 93%. Despite its slightly lesser sensitivity than IFA or ELISA (2-6% and 5% respectively) in endemically infected dogs, IDA with PEG method may help to bring the diagnosis of CL within reach of the veterinary practitioner.     

Do intestinal parasites interfere with the seroepidemiologic surveillance of Schistosoma mansoni infection?

In view of the known cross-reactivity of sera from patients with intestinal parasites to some Schistosoma mansoni antigens, field work was conducted in an area of Venezuela non-endemic for schistosomiasis using the routine immunoenzymatic assay (ELISA) with soluble egg antigen (SEA). False positive reactions represented 15.3% of the total population as determined by SEA-ELISA. SEA-immunoblotting of the false positive sera indicated that protein fractions of 91 and 80 kDa appear to be responsible for cross-reactivity. Sera from hookworm infected individuals produced a higher frequency and intensity of cross-reaction than other sera. SEA-fractions of 105, 54, 46, 42, 32, 25 and 15 kDa were the most specific.     

The identification of markers segregating with resistance to Schistosoma mansoni infection in the snail Biomphalaria glabrata

Both snail and parasite genes determine the susceptibility of the snail Biomphalaria glabrata to infection with the trematode Schistosoma mansoni. To identify molecular markers associated with resistance to the parasite in the snail host, we performed genetic crosses between parasite-resistant and -susceptible isogenic snails. Because resistance to infection in adult snails is controlled by a single locus, DNA samples from individual F2 and F1 backcross progeny, segregating for either the resistant or susceptible phenotypes, were pooled (bulked segregant). Genotypes for both parents were determined with 205 arbitrary decamer primers by random amplified polymorphic DNA-PCR. Of the 205 primers, 144 were informative, and the relative allele frequencies between the pools for these primers were determined. Two primers, OPM-04 and OPZ-11, produced fragments in the resistant parent of one cross that were inherited in a dominant fashion in the resistant F2 and backcross-bulked segregant progeny. Subsequent typing of DNA samples of individual progeny snails showed that the 1.2-kb marker amplified by primer OPM-04 and the 1.0-kb marker produced by primer OPZ-11 segregated in the same dominant fashion with the resistant phenotype. Sequence analysis of the 1.2-kb marker showed that it corresponds to a repetitive sequence in the snail genome with no homology to existing DNA sequences in the public databases. Analysis of the 1. 0-kb marker showed that it also corresponds to a repetitive sequence in the B. glabrata genome that contains an imperfect ORF, with homology to retrovirus-related group-specific antigens (gag) polyprotein.     

Relation of intensity of infection to disease in hamsters with acute schistosomiasis mansoni

Groups of young hamsters were exposed to 3, 20, 40, 80, or 160 cercariae. A highly significant correlation was observed between the number of cercariae, worm burdens, and liver and fecal egg counts. The most heavily infected animals were the first to lose weight and die. Hamsters exposed to 20 or more cercariae and harboring a mean of 4.2 or more worm pairs developed significant hepatosplenic disease by 10 weeks after infection as assessed by hepatomegaly, splenomegaly and the development of portal hypertension. Lightly infected animals with single worm pairs did not develop significant disease.     

Large intestine obstruction complicated with perforation: a rare manifestation of Schistosoma mansoni infection

The authors report a case of a 25 year old Brazilian man with a history of crampy abdominal pain in the left iliac fossa for 2 weeks, abdominal distention, mucous diarrhea and anorexia. The patient presented signs of hemodynamic instability and a hard mass palpated in the left iliac fossa presented peritoneal irritation. At laparotomy, fecal peritonitis and a punched-out perforation of the midsigmoid colon were found. A left hemicolectomy was performed with terminal colostomy. Specimen examination revealed a thickened rectosigmoid wall, narrow lumen and multiple mucosal polyps. Microscopically, chronic granulomatous colitis with Schistosoma mansoni eggs confirmed the etiology. To the authors' knowledge, this is the first case of obstruction complicated with perforation due to mansoni schistosomiasis reported in the literature.     

Modification of the lung recovery assay for schistosomula and correlations with worm burdens in mice infected with Schistosoma mansoni

The kinetics of recovery of schistosomula from the lungs of previously-unexposed CF1 mice were studied following infection with Schistosoma mansoni cercariae. Lung tissue fragments were incubated from 3 to 48 hr and the completeness of recovery of viable schistosomula was determined. The recovery of schistosomula from the lungs was shown to correlate closely with 7-week worm recoveries. With the additional incubation described, the lung recovery assay may provide a more defined indicator of protective immunity. Based on the data generated by these analyses, it is predicted that under these conditions schistosomula arrive in the lungs from 3 to 7 days after infection, and that they sojourn there for a 3-day period.