Immunoblot analysis of membrane antigens of Schistosoma mansoni, Schistosoma intercalatum, and Schistosoma haematobium against Schistosoma-infected patient sera

Antigens present in aqueous n-butanolic extracts (BE) of Schistosoma mansoni (Venezuelan JL strain), Schistosoma intercalatum (Cameroon EDEA strain), and Schistosoma haematobium (Yemen strain) adult worm membranes were compared in immunoblot against sera of patients infected with S. mansoni, S. intercalatum, S. haematobium, Schistosoma japonicum, or Schistosoma mekongi looking for similarities (common antigens) and differences (species-specific antigens). About 17 S. mansoni BE polypeptides (M _r ∼8 to >80 kDa) were commonly recognized by S. mansoni-infected patient sera from Venezuela, Senegal, and Ethiopia. S. intercalatum-, S. haematobium-, or S. japonicum-infected sera were almost unreactive with S. mansoni BE. Nonetheless, S. mekongi-infected sera weakly cross-reacted with a ∼10–15-kDa subset of S. mansoni BE. About 72.7% of S. intercalatum-infected patient sera reacted with a ∼19–21-kDa complex in S. intercalatum BE and cross-reacted with a similar complex in S. haematobium BE. Conversely, all S. haematobium-infected patient sera reacted with a ∼19–21-kDa complex in S. haematobium BE and cross-reacted with the ∼19–21-kDa complex in S. intercalatum BE; S. mansoni- and S. japonicum-infected patient sera did not react with S. intercalatum or S. haematobium BE. Results showed the presence of a common membrane antigen between African schistosome species and species-specific antigens in S. mansoni BE that could be useful to discriminate between species and/or to detect Schistosoma infections.     

Circulating antigens in schistosomiasis: detection of 31/32-kDa proteins in sera from patients infected with Schistosoma japonicum,S. mansoni,S. haematobium,or S. intercalatum

A sandwich enzyme-linked immunosorbent assay (ELISA) was developed to detect 31/32-kDa schistosome proteins as circulating antigens in sera from schistosomiasis patients. A monoclonal antibody was used as a capture antibody and rabbit antiserum raised against purified 31/32-kDa proteins was the detecting antibody. Positive results were obtained with patients infected with Schistosoma japonicum (88%; n=69), S. mansoni (80%; n=56), S. haematobium (100%; n=40), or S. intercalatum (94%; n=65). Sera from uninfected Chinese and African individuals and from Chinese patients with trichinosis, cysticercosis, or paragonimiasis did not react in the assay. This ELISA appears to be valuable in diagnosing infections by all major human schistosome species.     

Molecular arguments for splitting of Schistosoma intercalatum, into two distinct species

The taxonomic status of the two known strains of Schistosoma intercalatum, the Lower Guinea strain (originating from Edea, Cameroon) and the Zaire strain (originating from Kinshasa, Democratic Republic of Congo, formerly Zaire) was examined using random amplified polymorphic DNA (RAPD) markers. Two additional species within the S. haematobium group, S. haematobium and S. mattheei, were included in the study. DNA was extracted from four male and four female worms of each species and strain under investigation. In all, 13 primers gave reproducible and informative marker patterns; the monomorphic bands in all the males and females of each sample were scored, and 138 bands were included in the final analysis. Overall, 14 RAPD fragments were shared by all the schistosomes studied, and 19 RAPD fragments were considered to be sex markers. Only 22% (20/91) of the RAPD fragments were shared between S. intercalatum Zaire and S. intercalatum Cameroon. The mean values recorded for the Nei and Li's genetic distances between S. haematobium and S. mattheei and between S. intercalatum Zaire and S. intercalatum Cameroon were 0.546 and 0.596, respectively. A principal component analysis and one-way analysis of variance (ANOVA/MANOVA) showed a significant separation between S. intercalatum Zaire and S. intercalatum Cameroon. The data support the hypothesis that S. intercalatum Zaire and S. intercalatum Cameroon are distinct species. Additional molecular-biology studies are in progress that involve the use of nuclear and mitochondrial markers to confirm the extent of the genetic divergence prior to the establishment of final decision on the taxonomic status of the two strains of S. intercalatum. Received: 6 June 2000 / Accepted: 11 July 2000     

Immunodiagnosis of Schistosomiasis mansoni with APIA (alkaline phosphatase immunoassay)

The previously shown antigenicity of Schistosoma mansoni (JL venezuelan strain) alkaline phosphatase (Mg2+, pH 9.5) allowed its use in an immunodiagnosis assay, that consisted in the immunoadsorption of the enzymatic activity. Protein-A coated polyvinyl plates were used as solid phase to capture IgG from sera of infected human patients. After buffered saline washings, the plates were incubated with an enzyme-rich fraction (a n-butanol aqueous extract of adult worm obtained from pairs). Immunoadsorbed alkaline phosphatase (AP) was revealed by adding rho-nitrophenyl phosphate. Anti-AP antibodies were detected in 93% of coproparasitologically proven S. mansoni-infected venezuelan patients but not in parasite-free control sera and sera from patients infected with parasitosis other than schistosomiasis. The APIA did not correlate with cure but the anti-AP antibody response was progressively reduced after treatment. The use of an AP substrate amplifying system allowed an improvement of the assay sensitivity without loss of specificity. The data suggest that the APIA could be used as a marker of infection by S. mansoni.     

A comparison of the susceptibility to praziquantel of Schistosoma haematobium,S. japonicum,S. mansoni,S. intercalatum and S. mattheei in hamsters

Summary Groups of six- to eight-week-old hamsters each experimentally infected with Sudanese and South African strains of S. haematobium, Puerto Rican and Liberian strains of S. mansoni, S. japonicum from mainland China, a Congo strain of S. intercalatum and a Nelspruit South African strain of S. mattheei, were treated with different dosage regimens of a new schistosomicide — praziquantel.The drug is more effective against S. haematobium when administered by the intramuscular route than per os, a complete parasitological cure being obtained following a single intramuscular injection of 200 mg/kg bwt. Per os the best results were obtained using the three highest regimens of 5×100 mg/kg, 5×50 mg/kg and 3×100 mg/kg bwt. It was observed that S. haematobium female worms are more susceptible to the compound than male worms.The results show that S. japonicum in the hamster is very susceptible to the compound, a dosage of 100 mg/kg administered orally for three days resulting in a complete parasitological cure. More than a 90% reduction in adult worms was obtained at all the dosage regimens used except 1×50 mg/kg (73.2%). Female worms were again found to be more susceptible to the drug than male worms. Hatching tests performed on ova in liver tissue of control and treated animals were positive up to four weeks after treatment, thus showing that praziquantel has no ovicidal properties.Immature S. japonicum worms were found to be markedly less susceptible to treatment than the mature schistosomes (5×100 mg/kg reduced mature adults by 99.7%, but immature forms were reduced by only 54.2%).A 100% cure rate was obtained in the hamster infected with the Liberian strain of S. mansoni following treatment with praziquantel at 3×100 mg/kg given on consecutive days, and at 3×50 mg/kg administered in one day. Treatment of the hamsters infected with the Puerto Rican strain of S. mansoni also resulted in substantial reductions of adult worms. Praziquantel was also found to be highly effective against S. intercalatum and S. mattheei.It is noted that the efficacy of the compound against S. haematobium, S. japonicum and S. mattheei in the hamster is significantly greater than that of metrifonate (Bilarcil). Following treatment with praziquantel most S. haematobium and S. japonicum worms undergo a classic hepatic shift and are trapped and killed in the liver tissue.It is considered that the present study shows that this new compound exhibits a high degree of activity against the three major schistosome species S. haematobium, S. japonicum and S. mansoni, against S. intercalatum, and against the cattle schistosome S. mattheei in the hamster, with no apparent significant differences in efficacy against the different geographical strains of the parasites used in the trials.

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Zusammenfassung Gruppen von 6–8 Wochen alten Goldhamstern wurden jeweils mit einem sudanesischen und südafrikanischen Stamm von S. haematobium, mit einem puertoricanischen und liberianischen Stamm von S. mansoni, mit S. japonicum aus Kontinentalchina, S. intercalatum vom Kongo und mit S. mattheei aus Nelspruit, Südafrika, infiziert und mit verschiedenen Dosierungen des neuen Schistosomenmittels Praziquantel behandelt.Praziquantel ist gegen S. haematobium bei intramuskulärer Gabe wirksamer als bei oraler Gabe. Eine parasitologische Heilung konnte durch eine einmalige intramuskuläre Injektion von 200 mg/kg erreicht werden. Bei oraler Gabe ergaben die drei höchsten Dosen 5×100, 5×50 und 3×100 mg/kg die besten Ergebnisse. Es wurde beobachtet, daß die Weibchen von S. haematobium empfindlicher gegen Praziquantel sind als die Männchen.Die Ergebnisse zeigen, daß S. japonicum im Goldhamster sehr empfindlich gegen Praziquantel ist. Eine parasitologische Heilung wird durch 3×100 mg/kg erreicht. Mit allen verabreichten Dosen wurde die Zahl der adulten Parasiten um mehr als 90% reduziert. Nur bei Gabe von 1×50 mg/kg betrug die Parasitenreduktion 73,2%. Wieder waren die Weibchen empfindlicher gegen Praziquantel als die Männchen. Der Miracidienschlüpfversuch wurde an Eiern aus der Leber behandelter und unbehandelter Kontrolltiere durchgeführt. Er war bis zu vier Wochen nach der Behandlung positiv; Praziquantel wirkt also nicht ovizid.Jugendliche S. japonicum waren gegen die Behandlung deutlich unempfindlicher als adulte Schistosomen (5×100 mg/kg bewirkten bei Adulten eine Reduktion um 99,7%, bei den Jugendlichen eine um 54,2%).Eine parasitologische Heilung wurde bei Goldhamstern, die mit einem liberianischen Stamm von S. mansoni infiziert waren, mit 100 mg/kg (verabreicht an drei aufeinanderfolgenden Tagen) und mit 3×50 mg/kg/die erreicht. Die Behandlung von Goldhamstern, die mit einem puertoricanischen S. mansoni-Stamm infiziert waren, ergab ebenfalls eine sehr starke Reduktion der Anzahl adulter Schistosomen. Gegen S. intercalatum und S. mattheei war Praziquantel ebenfalls sehr gut wirksam.Es wurde gefunden, daß die Wirksamkeit von Praziquantel gegen S. haematobium, S. japonicum und S. mattheei im Hamster deutlich besser ist als die von Metrifonat (Bilarcil). Nach Behandlung mit Praziquantel zeigen die meisten S. haematobium- und S. japonicum-Würmer eine klassische liver shift und werden im Lebergewebe festgehalten und abgetötet.Die Untersuchung zeigt, daß Praziquantel gegen alle drei wichtigen Schistosomenarten, S. haematobium, S. japonicum und S. mansoni, und auch gegen S. intercalatum sowie gegen den Rinderparasiten S. mattheei im Hamster hoch wirksam ist. Zwischen den in dieser Untersuchung verwendeten Parasitenstämmen unterschiedlicher geographischer Herkunft bestehen in der Wirksamkeit keine wesentlichen Unterschiede.

     

Enzymatic basis for the lack of oxamniquine activity in Schistosoma haematobium infections

The notion that oxamniquine is active against Schistosoma mansoni but inactive against S. haematobium was confirmed using in vitro cultures of adult worms. Since oxamniquine and hycanthone have been shown to become effective upon activation by a schistosome enzyme, enzymatic tests were carried out to detect possible differences between the enzyme of S. mansoni and that of S. haematobium . It was found that the S. mansoni enzyme could activate hycanthone and, to a lesser extent, oxamniquine. The S. haematobium enzyme, on the other hand, was capable of activating hycanthone but virtually incapable of activating oxamniquine. It is concluded that the different activity of oxamniquine in the two species is due to differences in the drug-activating enzyme.     

Schistosoma haematobium,S. intercalatum,S. japonicum,S. mansoni,and S. rodhaini in mice: relationship between patterns of lung migration by schistosomula and perfusion recovery of adult worms

The development of five schistosome species was compared in mice by the recovery of schistosomula from chopped lung tissue and of adult worms by portal perfusion. Three developmental patterns appeared. (1) Schistosoma japonicum was unique in showing an early establishment of schistosomula in and a rapid departure from the lungs together with the highest worm recovery; (2) S. haematobium contrasted by establishing later and persisting in the lungs for at least 2 weeks while yielding the lowest adult worm recovery; and (3) S. intercalatum, S. mansoni, and S. rodhaini had an intermediate pattern – they resided in the lungs for several days, then disappeared and produced intermediate numbers of adults. Lung petechiae, known to accompany the migration of S. japonicum, were never detected after infection with the other species. We speculate that the three migration patterns of schistosomes are related to the size of the relative spectra of naturally infected definitive hosts. Received: 31 August 1997 / Accepted: 15 October 1997     

Schistosomiasis at Loum,Cameroun; Schistosoma haematobium,S. intercalatum and their natural hybrid

Summary A survey of 500 schoolchildren in Loum in 1968 revealed an overall infection rate of 54.2% with Schistosoma intercalatum and this was the only species of schistosome encountered. In 1972 a number of children were found to be passing schistosome eggs in their urine and these eggs ranged in shape and size from the forms characteristic for S. haematobium to those of S. intercalatum. Preliminary laboratory studies demonstrated that hybridisation between the two species was occurring. Subsequent field surveys showed that the snail hosts for the two parasites (B. rohlfsi for S. haematobium and B. forskali for S. intercalatum) were both present in the river Mbette and its tributaries in Loum and the distribution of the two snail species coincided closely with the distribution of the schistosomes in the human population. Detailed study of a small group of children passing hybrid eggs in their urine revealed that few of them were passing eggs in their faeces and that those eggs which were found in faeces were not viable.Analysis of schistosome egg-shape by plotting cumulative size-frequency data on probability paper demonstrated that the graph obtained from a natural hybrid series was different from that given by a known mixture of the two separate species. The hybrid series included a number of exceptionally large eggs resembling those of S. bovis but isolation of these eggs and subsequent laboratory passage of the parasites showed that they were part of the series and were not evidence of the presence of a third species. Hybridisation experiments in the laboratory showed that the cross S. haematobium × S. intercalatum is fully viable but that the reverse mating is not successful, thus accounting for the failure of the faecal eggs recovered from children with hybrid infections. Histological results from laboratory passaged hybrids suggest that the Ziehl-positive staining reaction of the egg-shells of S. intercalatum may be a recessive character.The observations reported here indicate that S. haematobium has only recently become established in Loum and that it is, through introgressive hybridisation, replacing the indigenous S. intercalatum. A suggested explanation for the change in the parasite fauna is offered and this depends upon ecological changes resulting from forest clearance and agricultural development providing improved conditions for the spread of B. rohlfsi, the snail host for S. haematobium. It is suggested that, in contrast to recent reports on the spread of S. intercalatum, this species is in fact retreating and being replaced by S. haematobium in areas where forest clearance is taking place.In conclusion it is suggested that introgressive hybridisation of this kind may have been responsible for the evolution of certain characteristic local strains of African schistosomes.     

Schistosoma mansoni Infection Impairs Antimalaria Treatment and Immune Responses of Rhesus Macaques Infected with Mosquito-Borne Plasmodium coatneyi

Malaria and schistosomiasis are the world''s two most important parasitic infections in terms of distribution, morbidity, and mortality. In areas where Plasmodium and Schistosoma species are both endemic, coinfections are commonplace. Mouse models demonstrate that schistosomiasis worsens a malaria infection; however, just as mice and humans differ greatly, the murine-infecting Plasmodium species differ as much from those that infect humans. Research into human coinfections (Schistosoma haematobium-Plasmodium falciparum versus Schistosoma mansoni-P. falciparum) has produced conflicting results. The rhesus macaque model provides a helpful tool for understanding the role of S. mansoni on malaria parasitemia and antimalarial immune responses using Plasmodium coatneyi, a malaria species that closely resembles P. falciparum infection in humans. Eight rhesus macaques were exposed to S. mansoni cercariae. Eight weeks later, these animals plus 8 additional macaques were exposed to malaria either through bites of infected mosquitos or intravenous inoculation. When malaria infection was initiated from mosquito bites, coinfected animals displayed increased malaria parasitemia, decreased hematocrit levels, and suppressed malaria-specific antibody responses compared to those of malaria infection alone. However, macaques infected by intravenous inoculation with erythrocytic-stage parasites did not display these same differences in parasitemia, hematocrit, or antibody responses between the two groups. Use of the macaque model provides information that begins to unravel differences in pathological and immunological outcomes observed between humans with P. falciparum that are coinfected with S. mansoni or S. haematobium. Our results suggest that migration of malaria parasites through livers harboring schistosome eggs may alter host immune responses and infection outcomes.     

Rapid Detection of a Schistosoma mansoni Circulating Antigen Excreted in Urine of Infected Individuals by Using a Monoclonal Antibody

Schistosoma circulating antigens were used to indicate the infection intensity and to assess cure. An immunoglobulin G2a (IgG2a) mouse monoclonal antibody was used in a fast dot-enzyme-linked immunosorbent assay (ELISA; FDA) for rapid and simple diagnosis of schistosomiasis in the field. Seven hundred Egyptians were parasitologically examined for Schistosoma mansoni and other parasitic infections. A rectal biopsy was done as a “gold standard” for individuals showing no S. mansoni eggs in their feces. Egg counts were obtained by the Kato smear method for only 100 of 152 individuals with eggs in their feces. Specific anti-schistosome IgG antibodies were evaluated in sera by ELISA. Urine samples from the 700 individuals were tested by FDA for detection of the circulating antigen. The assay showed a sensitivity of 93% among 433 infected individuals and a specificity of 89% among 267 noninfected individuals. FDA showed the highest efficiency of antigen detection (91%) compared with the efficiency of antibody detection by ELISA (75%) and stool analysis (60%). In addition, FDA detected infected patients with 20 eggs/g of feces. Also, the sensitivity of FDA ranged from 90 to 94% among samples from patients with different clinical stages of schistosomiasis. All the assay steps can be completed within 30 min at room temperature for 96 urine samples. The monoclonal antibody identified a 74-kDa antigen in different antigenic extracts of S. mansoni and Schistosoma haematobium and in the urine of infected individuals. In addition, a 30-kDa degradation product was identified only in the urine samples. On the basis of these results, FDA should be used as a rapid tool for the sensitive and specific diagnosis of Schistosoma infection.     

Immunodiagnosis of Schistosomiasis Mansoni With Apia (Alkaline Phosphatase Immunoassay)

The previously shown antigenicity of Schistosoma mansoni (JL venezuelan strain) alkaline phosphatase (Mg²⁺, pH 9.5) allowed its use in an immunodiagnosis assay, that consisted in the immunoadsorption of the enzymatic activity. Protein-A coated polyvinyl plates were used as solid phase to capture IgG from sera of infected human patients. After buffered saline washings, the plates were incubated with an enzyme-rich fraction (a n-butanol aqueous extract of adult worm obtained from pairs). Immunoadsorbed alkaline phosphatase (AP) was revealed by adding p-nitrophenyl phosphate. Anti-AP antibodies were detected in 93% of coproparasitologically proven S. mansoni-infected venezuelan patients but not in parasite-free control sera and sera from patients infected with parasitosis other than schistosomiasis. The APIA did not correlate with cure but the anti-AP antibody response was progressively reduced after treatment. The use of an AP substrate amplifying system allowed, an improvement of the assay sensitivity without loss of specificity. The data suggest that the APIA could be used as a marker of infection by S. mansoni.     

On factors possibly restricting the distribution ofSchistosoma intercalatum Fisher, 1934

Summary Two hypotheses have been postulated explaining the limited distribution ofSchistosoma intercalatum.The first hypothesis is correlated with physical factors and behaviour of cercariae. Histochemical and ultrastructural studies have shown that in response to increased temperature change the cercariae ofS. intercalatum form aggregates, unlike other schistosome cercariae of man, which are noninfective to the definitive host. The aggregates are formed by the release of the adhesive post-acetabular gland secretion which causes the cercariae to stick together. It is suggested that ifS. intercalatum spread from streams within tropical rain forest to pools and laybys of streams in the savannah, cercariae would be subjected to greater daily temperature changes thus triggering the release of post-acetabular gland secretion, thereby impairing invasion of the definitive host.The second hypothesis is based on the natural occurrence of hybridisation betweenS. intercalatum andSchistosoma haematobium. With some strains of these two species there are no genetical isolating mechanisms. It is suggested that ifS. intercalatum extended into a savannah environment from tropical rain forest, hybridisation betweenS. intercalatum andS. haematobium would eventually occur. Experimental studies indicate that probably, as a result of introgressive hybridisation, a new strain ofS. haematobium would eventually supersede the originalS. intercalatum.     

Epidemiology,of bilharzias (schistosomiasis) in Uganda from 1902 until 2005

Background

Schistosoma mansoni was observed and reported in Kuluva hospital Arua District in north western Uganda as early as 1902. S. mansoni is widely distributed in Uganda along permanent water bodies.

Objective

To review the litreture on scistosomiasis in Uganda, since 1902.

Method

The core literature for this short review was searched from reports and publications by the British colonial Ministry of Health Districts Medical officers and Entomologists. Additional information was obtained from Makerere University Medical School library archives, London School of Hygiene and Tropical Medicine library archives, University of Antrwap, and post independence publications on schistosomiasis in Uganda in various journals.

Results

Since it was first detected in 1902 Schistosoma (S) mansoni is more widely distributed in Uganda than S. haematobium. However Schistosoma mansoni and S. haematobium are of public health importance in Uganda and the importance of migrants and fishermen in disseminating infections into non-infested areas and intensifying infection in areas already infested have been reported.

Conclusion

S. mansoni has been on the increase in Uganda whereas S. haematobium is localized in sporadic foci in the north of Uganda. Treatment with praziquantel the drug of choice in Uganda used in schistosomiasis control programme has reduced development of severe schistosomiasis.     

Anthelmintic activity of carvacryl acetate against Schistosoma mansoni

Blood flukes of the genus Schistosoma are the causative agents of human schistosomiasis, a debilitating disease that afflicts over 200 million people worldwide. Praziquantel is the drug of choice but concerns over praziquantel resistance have renewed interest in the search for alternative drug therapies. Carvacrol, a naturally occurring monoterpene phenol and food additive, has been shown high medicinal importance, including antimicrobials activities. The aim of this study was to evaluate in vitro effect of carvacryl acetate, a derivative of carvacrol, on Schistosoma mansoni adult worms. We demonstrated that carvacryl acetate at 6.25 μg/mL has antischistosomal activity, affecting parasite motility and viability. Additionally, confocal laser scanning microscopy pictures revealed morphological alterations on the tegumental surface of worms, where some tubercles appeared to be swollen with numerous small blebs emerging from the tegument around the tubercles. Furthermore, experiments performed using carvacryl acetate at sub-lethal concentrations (ranging from 1.562 to 6.25 μg/mL) showed an inhibitory effect on the daily egg output of paired adult worms. Thus, carvacryl acetate is toxic at high doses, while at sub-lethal doses, it significantly interferes with the reproductive fitness of S. mansoni adult worms. Due to its safety and wide use in the industry, carvacryl acetate is a promising natural product-derived compound and it may represent a step forward in the search for novel anthelmintic agents, at a time when there is an urgent need for novel drugs.     

Parasitological evaluation of Ro 15-9268, a 9-acridanone-hydrazone derivative against Schistosoma mansoni in mice, and observations on changes in serum enzyme levels

Schistosoma mansoni is one of the major causes of schistosomiasis prevalent in tropical and subtropical areas, especially in poor communities. It is estimated that at least 90 % of those requiring treatment for schistosomiasis live in Africa. The primary control strategy employed for schistosomiasis is mass drug administration (MDA).The aim is to reduce disease through treatments with a single lower dose of Ro 15-9268 as a new antischistosomal drug. In the present search, the efficacy of Ro 15-9268 was studied in mice using a dose of 12.5 mg/kg of body weight (b.wt.) against an Egyptian strain of S. mansoni. This was carried out at 2 days and 3, 4, and 6 weeks post–cercarial exposure of mice. The criteria used were the worm load, oogram pattern and number of ova in the liver and intestine, hepatic enzyme activity, and liver histopathology. The tested agent has led to a significant reduction in worm burden (89.80 %) in liver and portomesenteric veins concurrent with a hepatic shift at the second week posttreatment followed by a complete disappearance of worms, 4 weeks postmedication. The oogram of infected animals treated revealed an increased number of dead ova 2 days posttreatment and complete absence of immature and mature ova 2 weeks later. The hepatic and intestinal egg counts significantly declined by about 96 and 98 %, respectively, 6 weeks after treatment, and the fecal egg count completely disappeared from stool 4 weeks after medication. The hepatic histopathological changes were improved, ova were markedly degenerated, and worms showed fragmentation and degeneration after drug administration. In conclusion, when Ro 15-9268 was administered to mice infected with the Egyptian strain of S. mansoni, at a low dose level (12.5 mg/kg b.wt.), encouraging results were obtained. The drug showed high efficacy against schistosomal worms as well as histopathological inflammatory changes.     

A monoclonal antibody against Schistosoma haematobium soluble egg antigen: efficacy for diagnosis and monitoring of cure of S. haematobium infection

A monoclonal antibody (mAb), 2F/11F, raised against Schistosoma haematobium soluble egg antigen (SEA) was found to be nonreactive with S. mansoni SEA or other parasite antigens (Fasciola hepatica, Echinococcus granulosus). This IgG₁ mAb recognized a repetitive epitope on S. haematobium SEA in the molecular-weight regions of 70, 42, and 35 kDa. It was employed as both an antigen-capture and a biotinylated detection antibody in a sandwich enzyme-linked immunosorbent assay (ELISA) for the detection of circulating schistosome antigen (CSA) and had a detection limit of <1 ng S. haematobium SEA/ml. CSA levels were measured in serum and urine samples from 116 S. haematobium-infected rural students before therapy and at 4, 8, and 12 weeks after praziquantel treatment. Serum and urine samples from 50 S. mansoni -infected patients, 15 patients harboring other parasites, and 30 noninfected individuals were also assessed. CSA was detected in 90.5% of serum samples and 94% of urine samples from S. haematobium-infected patients. CSA was undetectable in serum from the 15 patients harboring other parasites and in 94% of serum samples and 84% of urine samples from S. mansoni-infected patients. In the S. haematobium-infected group a positive correlation was detected between CSA levels in serum and urine samples and the egg load per 10 ml urine. A significant reduction in CSA levels was detected in serum and urine samples after praziquantel therapy. CSA was undetectable in 87% of serum samples and 81.5% of urine samples from schistosomiasis haematobium patients at 12 weeks post-treatment. These data demonstrate that the use of mAb 2F/11F for detection of CSA provides a sensitive method for the immunodiagnosis and monitoring of cure of schistosomiasis haematobium. Received: 5 December 1998 / Accepted: 26 June 1999     

Detection of Schistosoma mansoni Membrane Antigens by Immunoblot Analysis of Sera of Patients from Low-Transmission Areas

Schistosoma mansoni surface membrane components play a relevant role in the host-parasite interaction, and some are released in vivo as circulating antigens. n-Butanol extraction favors the release of membrane antigens like alkaline phosphatase, which has been shown to be specifically recognized by antibodies from S. mansoni-infected humans and animals. In the present study, components in the n-butanol extract (BE) of the adult S. mansoni worm membrane fraction were separated by one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis (1D SDS-PAGE [15%]) and further analyzed by immunoblotting (immunoglobulin G) using defined sera. S. mansoni-infected patient sera, but not sera of uninfected patients or sera obtained from patients infected with other parasite species, specifically and variably recognized up to 20 polypeptides in the molecular mass range of ~8 to >80 kDa. There were some differences in the number, intensity, and frequency of recognition of the BE antigens among sera from Venezuelan sites of endemicity with a different status of schistosomiasis transmission. Antigens in the 28- to 24-kDa molecular mass range appeared as immunodominants and were recognized by S. mansoni-positive sera from all the sites, with recognition frequencies varying between 57.5 and 97.5%. Immunoblotting with BE membrane antigens resulted in a highly sensitive (98.1%), specific (96.1.0%), and confirmatory test for the immunodiagnosis of schistosomiasis in low-transmission areas.     

Salmonella typhimurium exacerbates injuries but resolves fibrosis in liver and spleen during Schistosoma mansoni infection

BackgroundIn most developing or undeveloped countries, patients are often co-infected with multiple pathogens rather than a single pathogen. While different pathogens have their impact on morbidity and mortality, co-infection of more than one pathogen usually made the disease outcome different. Many studies reported the co-infection of Schistosoma with Salmonella in pandemic areas. However, the link or the underlying mechanism in the pathogenesis caused by Schistosoma-Salmonella co-infection is still unknown.MethodsIn this study, Salmonella typhimurium (S. typhimurium) was challenged to Schistosoma mansoni (S. mansoni)-infected mice. Further experiments such as bacterial culture, histopathological examination, western blotting, and flow cytometry were performed to evaluate the outcomes of the infection. Cytokine responses of the mice were also determined by ELISA and real-time quantitative PCR.ResultsOur results demonstrated that co-infected mice resulted in higher bacterial excretion in the acute phase but higher bacterial colonization in the chronic phase. Lesser egg burden was also observed during chronic schistosomiasis. Infection with S. typhimurium during schistosomiasis induces activation of the inflammasome and apoptosis, thereby leading to more drastic tissue damage. Interestingly, co-infected mice showed a lower fibrotic response in the liver and spleen. Further, co-infection alters the immunological functioning of the mice, possibly the reason for the observed pathological outcomes.ConclusionCollectively, our findings here demonstrated that S. mansoni-infected mice challenged with S. typhimurium altered their immunological responses, thereby leading to different pathological outcomes.     

Effect of a novel benzimidazole derivative in experimental Schistosoma mansoni infection

Currently, praziquantel is the only drug of choice for treatment of schistosomiasis. Reports of praziquantel resistance raise concerns about future control of the disease. Therefore, the search for new schistosomicidal drugs is eminent. In this study, the effect of a novel benzimidazole-derived compound (compound BTP-Iso) was assessed in mice harboring adult Schistosoma mansoni (Egyptian strain). Mice were treated 42 days p.i. with compound BTP-Iso using two treatment regimens (200 or 300 mg/kg). In both regimens, there were significant reductions in the number of recovered S. mansoni worms especially females and in immature ova, in addition to a significant reduction in the number and size of hepatic granulomata. A dose of 300 mg/kg resulted in a significant decrease in intestinal and hepatic tissue egg loads. Effect on schistosomes was confirmed by scanning electron microscopy, where adult worms recovered from mice treated with 200 mg/kg of compound BTP-Iso revealed tegumental alternations, characterised by swelling of tegumental ridges, bleb formation, and mild erosion in male worms; however in females, there were extensive erosion and destruction of the tegumental surface. These promising results may encourage future use of compound BTP-Iso in the treatment of schistosomiasis. However, more research is needed to detect the effect of compound BTP-Iso on early developmental stages of S. mansoni and on other species of human schistosomes.