Scanning electron microscope studies of miracidia suggest introgressive hybridization between Schistosoma haematobium and S. haematobium x S. mattheei in the Eastern Transvaal

Schistosoma haematobium miracidia were collected from a locality with a high prevalence of human infection with the animal parasite, S. mattheei, which hybridizes with S. haematobium, and from 2 localities with negligible infection rates. The terebratoria of the miracidia from these localities were compared with each other, with laboratory maintained S. haematobium and with four populations of S. mattheei by means of scanning electron microscopy. It was found that the terebratorial membrane of certain of the S. haematobium miracidia from the locality with a high S. mattheei prevalence in humans, resembled the more intricate membrane of S. mattheei. This suggests introgressive hybridization between S. haematobium and S. haematobium x S. mattheei.     

The influence of physical factors on the survival and infectivity of miracidia of Schistosoma manosni and S. haematobium I. Effect of temperature and ultra-violet light.

The influence of temperature and ultra-violet radiation on the degree of activity, survival and infectivity of schistosome miracidia is profound. Miracidia of Schistosoma mansoni and S. haematobium were affect eaually. Only miracidia classified as "active" or "slow" were capable of penetration, a capacity they retained for about 17 hours at 19 degrees C. Miracidia that were "lethargic" as a result of low temperature, old age or ultra-violet radiation lost their infective capacity. The conclusion, however, is that neither the temperatures encountered in the field nor the solar ultra-violet radiation penetrating turbid waters are likely to be harmful to miracidia and thus have no effect on the level of transmission.     

Efficacy of praziquantel treatment of schistosomiasis in a non-endemic country: a follow-up of parasitological, clinical and immunological parameters.

Schistosomiasis patients were immigrants to Czechoslovakia from Angola and Yemen. Most of them had light or moderate infections and felt subjectively healthy. They received treatment with praziquantel (two doses with a total of 40 mg/kg) and were followed up for several years. In nine of 13 patients, Schistosoma haematobium or S. mansoni eggs with undamaged miracidia were detected in biopsies from the bladder or the rectum one year or later after treatment. Granulomatous reactions in the rectum and bladder lesions of stage 1 including thickened bladder walls persisted in most of the patients. Antibody levels against adult S. mansoni worm antigen remained elevated for at least two years after therapy in some patients and declined in others. Among the nine patients, for whom pre- and post-treatment sera were available, the changes in relative levels of antibodies did not strictly correlate with the continued presence of schistosome eggs in, or their absence from, biopsies. We discuss the results obtained with sensitive diagnostic techniques in the absence of subjectively perceived disease.     

Epidemiology of mixed Schistosoma mansoni and Schistosoma haematobium infections in northern Senegal

Due to the large overlap of Schistosoma mansoni- and Schistosoma haematobium-endemic regions in Africa, many people are at risk of co-infection, with potential adverse effects on schistosomiasis morbidity and control. Nonetheless, studies on the distribution and determinants of mixed Schistosoma infections have to date been rare. We conducted a cross-sectional survey in two communities in northern Senegal (n=857) to obtain further insight into the epidemiology of mixed infections and ectopic egg elimination. Overall prevalences of S. mansoni and S. haematobium infection were 61% and 50%, respectively, in these communities. Among infected subjects, 53% had mixed infections and 8% demonstrated ectopic egg elimination. Risk factors for mixed infection - i.e. gender, community of residence and age - were not different from what is generally seen in Schistosoma-endemic areas. Similar to overall S. mansoni and S. haematobium infections, age-related patterns of mixed infections showed the characteristic convex-shaped curve for schistosomiasis, with a rapid increase in children, a peak in adolescents and a decline in adults. Looking at the data in more detail however, the decline in overall S. haematobium infection prevalences and intensities appeared to be steeper than for S. mansoni, resulting in a decrease in mixed infections and a relative increase in single S. mansoni infections with age. Moreover, individuals with mixed infections had higher infection intensities of both S. mansoni and S. haematobium than those with single infections, especially those with ectopic egg elimination (P<0.05). High infection intensities in mixed infections, as well as age-related differences in infection patterns between S. mansoni and S. haematobium, may influence disease epidemiology and control considerably, and merit further studies into the underlying mechanisms of Schistosoma infections in co-endemic areas.     

Responses of Echinostoma caproni miracidia to gravity, light, and chemicals.

In a four-tube vertical system, Echinostoma caproni miracidia exhibited a strong negative geotaxis which was dominated by a positive phototaxis. In horizontal chambers a positive phototactic response was also demonstrated. These miracidia showed a positive chemoresponse, as determined in phi-chambers, to glutamic and aspartic acids but not leucine. Positive responses were also elicited to snail-conditioned water and sulfuric and acetic acids. Ammonia, Mg2+, and HCl produced no significant reactions. Responses of E. caproni and Schistosoma mansoni miracidia, both of which develop in Biomphalaria glabrata snails, were similar providing further evidence that miracidia mimic the behavioral patterns of compatible snail species.     

Mating interactions between Schistosoma haematobium and S. mansoni

Schistosoma haematobium and S. mansoni are two medically important schistosomes, commonly occurring sympatrically in Africa and so potentially able to infect the same human host. Experiments were designed to study the mating behaviour of these two species in mixed infections in hamsters. Analysis of the data obtained showed that both heterospecific and homospecific pairs readily form. No significant difference was seen between the two species in their ability in forming pairs, however, S. mansoni showed a greater homospecific mate preference. Analysis of the data using the Mantel-Haenszel test suggests that mating competition does occur between S. haematobium and S. mansoni, the former being the more dominant species. Both species appeared to be able to change mate, with S. haematobium showing a greater ability in taking S. mansoni females away from S. mansoni males when introduced into a pre-established S. mansoni infection highlighting the competitiveness of S. haematobium. The significance of the results is discussed in relation to the epidemiological consequences occurring in Senegal, and other areas where both species are sympatric.     

Schistosoma mansoni,S. haematobium,and S. japonicum: early events associated with penetration and migration of schistosomula through human skin

Migratory pattern of schistosomula of Schistosoma mansoni, S. haematobium, and S. japonicum through human skin were analyzed in skin organ cultures. These studies showed that the schistosomula of S. mansoni and S. haematobium has similar migratory patterns through human skin. During the first 24h after infection nearly 90% of S. mansoni and S. haematobium schistosomula were present only in the epidermis. Majority of the schistosomula were found in the dermis only after 48h and they appear to reach the dermal vessels around 72h after infection. Migratory pattern of S. japonicum on the other hand was significantly different from the other two species in that over 90% of the parasites had already reached the dermis within the first 24h and schistosomula were present in the dermal vessels within 2h after infection. Analysis of the cytokine pattern at 8h after infection by a macro gene array and RT-PCR analysis showed that out of 24 different cytokines analyzed only IL-1ra, IL-10, and TNF-alpha were increased in the human skin following infections with S. mansoni and S. haematobium, whereas, after infection with S. japonicum there was significant increases in IL-1beta, IL-1ra, IL-2, IL-6, IL-8, IL-10, IL-15, IL-18, and TNF-alpha. Immunohistochemical analysis of epidermal sheets showed focal accumulation of HLA-DR(+) cells in areas where schistosomula of S. mansoni had entered the human skin.     

Schistosoma mansoni, S. haematobium, and S. japonicum: identification of genus- and species-specific antigenic egg glycoproteins

Immunoreactive egg glycoproteins of Schistosoma mansoni, S. haematobium, and S. japonicum which are genus- and species-specific, or react with sera of patients infected with other parasites, have been identified. Egg proteins were labeled with Iodine-125, and the concanavalin A-binding glycoproteins were immunoprecipitated with sera of patients infected with one of four species of Schistosoma or Trichinella spiralis, Taenia solium, Echinococcus granulosus, Entamoeba histolytica, or Wuchereria bancrofti. These immunoprecipitates were analyzed by two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Despite the strikingly different patterns of glycoproteins of the African species, the antibody immune responses of patients infected with S. mansoni and S. haematobium were found to be so similar that differentiation could not be established. In contrast, sera of patients infected with S. japonicum, S. mekongi, or parasites not of the genus Schistosoma, immunoprecipitated fewer of the major S. mansoni or S. haematobium glycoproteins. Likewise, antibody immune responses of patients infected with the Oriental schistosomes (S. japonicum and S. mekongi) could not be differentiated. Only a few quantitative differences were noted between our S. mansoni egg glycoprotein extract and a standardized soluble egg antigen extract. This study provides an explanation for the extensive cross-reactivity observed in diagnostic assays which utilize various fractions of schistosomal egg extracts as the antigen.     

IgM antibodies recognizing carbohydrate epitopes shared between schistosomula and miracidia of Schistosoma mansoni that block in vitro killing

A series of monoclonal antibodies (mAb) was raised in mice against Schistosoma mansoni, which recognized a carbohydrate determinant on a major Mr greater than 200,000 schistosomulum surface antigen. These mAb cross-reacted with the surface of cercariae and miracidia and with schistosomula of S. haematobium and S. bovis. Other mAb were generated that only recognized a Mr 20,000 schistosomulum surface antigen; they did not cross-react with eggs or miracidia and were species specific. The anti-Mr 20,000 mAb of the IgG1 isotype exhibited high levels of complement-dependent cytotoxicity to schistosomula in vitro. IgM mAb that recognized carbohydrate epitopes of the Mr greater than 200,000 surface antigen blocked the lethal activity of the anti-Mr 20,000 mAb. The IgM anti-Mr greater than 200,000 mAb also reduced complement-dependent cytotoxicity of serum from mice vaccinated with irradiated cercariae.     

Schistosoma mansoni and Schistosoma haematobium: identification and characterization of glycoconjugate antigens in the hemolymph of infected vector snails

Two carbohydrate epitopes were identified by monoclonal antibodies (KCS and E2) and characterized with respect to their immunoreactivity, monosaccharide structure, and location. Immunofluorescence demonstrated the presence of both epitopes on the surfaces of sporocysts, cercariae, and miracidia of Schistosoma mansoni, Schistosoma haematobium, and Schistosoma japonicum. However, spatial distribution and density of expression varied among species and developmental stages, and neither epitope was detectable on adult worm surfaces. Both glycans were found in the hemolymph of infected, but not uninfected, intermediate snail hosts. The presence of epitopes in hemolymph, as well as in schistosome eggs, is species-specific for KCS, recognizing only S. mansoni, and partly specific for E2, which reacted predominantly with S. haematobium. Immunoaffinity purification of target antigens for KCS and E2 from hemolymph of infected Biomphalaria and Bulinus, respectively, followed by carbohydrate composition analysis revealed a high content of fucose in both glycans. Methylation analysis demonstrated exclusively terminal fucose for the target antigen of KCS and terminal as well as internal fucose for the one of E2. Removal of terminal fucose abolished reactivity with both monoclonal antibodies. Both glycans are different from previously characterized schistosome carbohydrates. Their biological function(s) remain to be defined.     

Host parasite relationship among 3 Schistosoma mansoni and 6 Biomphalaria glabrata strains from epidemic and no epidemic Venezuelan areas is studied

Six lots of 18 B. glabrata from: La Victoria, Turmero, Cagua in Aragua state; Caserío El 25 in Carabobo state, Chabasquén in Portuguesa state and Humocaro Bajo in Lara state, were experimentally infected with miracidia of SM, C5 and C6 strains of Schistosoma mansoni (18 snails/Schistosoma mansoni strain). The averages of the intramolluscal period (IMP) obtained for the S. mansoni strains were very similar and comprised between 35.4 and 36.1 days. No significative statistical differences in the IMP were found according to the S. mansoni strain and the size of snails: < 7 mm and > 7 mm. However, significative statistical differences in the IMP were found, in relation to the B. glabrata strain and between the snails classified in two groups according to the S. mansoni dose (5 miracidia/snail and 10 miracidia/snail). The higher percentages of infection (PI) were found for the following parasite-snail combinations: C6-Cas. El 25 (80.7%), SM-La Victoria (73.1%) and C5-Cagua (62%). No significative statistical differences were found for the PI a) between the snail classified in two groups according to the size (< 7 mm and > 7 mm), b) in relation to the miracidium dosification (5 and 10 miracidia/snail and c) in accord to the S. mansoni strain. However, significative statistical differences were found for the PI obtained with different strains of the snail.     

Schistosoma haematobium and S. japonicum: analysis of the ribosomal RNA genes and determination of the "gap" boundaries and sequences

We have determined the intragenic organization of the rRNA genes of Schistosoma haematobium and S. japonicum and found them to be similar to that of S. mansoni and other eukaryotes. An entire ribosomal repeat approximately 10 kbp in size from each species was isolated as a SalI fragment from a genomic library constructed in bacteriophage lambda. The segments encoding both the small and large rRNAs have been identified using three cloned EcoRI fragments of S. mansoni as probes. There were three EcoRI fragments (4.2, 3.0, 1.6 kbp) from S. haematobium and four EcoRI fragments (4.6, 2.3, 1.7, 1.0 kbp) from S. japonicum. As in a wide variety of organisms within the protostome phyla, the 28S rRNA in schistosomes contains a "gap" which separates it into two fragments. The length of the gap sequence in S. haematobium is 54 bases and it is identical to that in S. mansoni in both length and sequence. However, in S. japonicum the sequence is between 64-67 bases long. In each case, irrespective of the species, the gap is located at the same position within the 28S rRNA. Secondary structures of the gap sequence derived by computer analysis predict a conformation with the minimum free energy that has an UAAU tract in a hairpin loop for S. haematobium and an UAUU tract for S. japonicum.     

Evidence for the presence of active cytochrome P450 systems in Schistosoma mansoni and Schistosoma haematobium adult worms

Extracts of the adult worms of both Schistosoma mansoni and Schistosoma haematobium can metabolise some typical P450 substrates but to differing degrees. S. mansoni worm extracts displayed a approximately 12-fold higher specific activity for an aminopyrine substrate than rat liver microsomes. At 4 mM substrate concentration the demethylation reaction with N-nitrosodimethylamine (NDMA) (5 nmol HCHO/mg protein/min) was only half that of rat liver microsomes, whereas in extracts of S. haematobium, no detectable activity was found towards NDMA. Using ethylmorphine as substrate the demethylation activity of S. mansoni extracts (1.82 nmol HCHO/mg protein/min) was 5.5-fold lower than that of rat liver microsomes. Benzphetamine demethylase activity was also readily detectable in S. mansoni worm extracts at 6.79 nmol HCHO/mg protein/min compared with 10.20 nmol HCHO/mg protein/min in the case of rat liver microsomes. When aniline was used as substrate, surprisingly, no activity was found in worm extracts of either S. mansoni or S. haematobium, whereas rat liver microsomes showed high activity towards this amine. The anti-P450 2E1 and 2B1/2 cross-reacted with both worm homogenates and gave a specific band corresponding to a protein of molecular weight of approximately 50.0 kDa. A study with anti-P450 IVA antibody revealed that while this protein was strongly expressed in S. haematobium worm extracts, no immunoreactivity was observed with extracts of S. mansoni. Immunoblotting analyses with anti-P450 IIIA and P450 1A1 did not detect immunoreactive protein in either S. mansoni or S. haematobium.     

Isolation and characterization of surface antigens from Schistosoma mansoni. II. Antigenicity of radiolabeled proteins from adult worms.

Adult Schistosoma mansoni were radiolabeled in vitro with 125I Bolton-Hunter reagent. Surface membrane antigens were solubilized with non-ionic detergent, then reacted with infection or normal serum. The antigen-antibody complexes were then precipitated with staphylococcal protein A immunoadsorbent, eluted with urea and SDS, and fractionated by SDS-PAGE. The results indicated the presence of 6 to 8 tegument antigens, depending on the type of antisera used. Human antisera to S. japonicum and S. haematobium reacted with some but not all of the antigens identified with human S. mansoni infection serum; this implies the presence of species-specific tegument antigens. The molecular weights of the radiolabeled antigens ranged from 10,000 to 100,000. A large (greater than 100,000) molecular weight glycoprotein and an uncharacterized lipid fraction appeared to be precipitated nonspecifically. Immunoprecipitation methods with anti-mouse IgG and anti-mouse whole serum failed to detect the presence of hostlike antigens in the labeled extracts. Several of the labeled proteins from S. mansoni were found to react with serum from patients infected with either S. haematobium or with S. japonicum.     

Schistosoma mansoni: comparison of a Caribbean and African strain and experimental crossing based on compatibility with intermediate hosts and Rattus rattus

To elucidate changes relative to compatibility with intermediate and definitive hosts affecting Schistosoma mansoni since it was introduced to the New World, the compatibility of S. mansoni from Africa (the Cameroons), from the Caribbean (Guadeloupe), and those resulting from experimental crosses with the gastropods Biomphalaria glabrata and B. pfeifferi has been studied. Results show that S. mansoni, regardless of its origin or its usual snail host, always infects B. pfeifferi more successfully than B. glabrata. The success rate with B. pfeifferi is 100% with 5 miracidia of S. mansoni from Guadeloupe and 97% with 5 miracidia from the Cameroons. On the other hand, in B. glabrata infection rate was 54% with 5 miracidia from Guadeloupe and 0% with 5 miracidia from the Cameroons (a rate of 19% is reached when using 10 miracidia). Hybrid miracidia infect B. pfeifferi and B. glabrata with a success rate of 60 and 86%, respectively, which are intermediate between those of the parent strains. Studies of S. mansoni development in Rattus rattus show that there is better infectivity and survival for the Caribbean strain than the Cameroon strain: the percentage worm recovery 4 weeks after exposure in 34% for S. mansoni from Guadeloupe, 14% for S. mansoni from the Cameroons, and 31% for the hybrids. The mortality rate between 4 and 12 weeks after exposure is 51% for S. mansoni from Guadeloupe, 87% for S. mansoni from the Cameroons, and 31% for the hybrids.(ABSTRACT TRUNCATED AT 250 WORDS)     

Methyl-beta-Cyclodextrin treatment and filipin staining reveal the role of cholesterol in surface membrane antigen sequestration of Schistosoma mansoni and S. haematobium lung-stage larvae

Ex vivo lung-stage larvae of Schistosoma mansoni and S. haematobium do not bind specific antibodies in the indirect membrane immunofluorescence test (IF), probably as a result of confinement of the surface membrane antigens in immobile, lipid-rich sites. Treatment with the membrane-impermeable, cholesterol-extracting drug methyl-beta-cyclodextrin (MBCD) and staining with filipin III (filipin), a fluorescent polyene antibiotic widely used for the detection and quantitation of cholesterol in biomembranes, allowed us to examine the role of cholesterol in surface membrane antigen sequestration of S. mansoni and S. haematobium ex vivo lung-stage larvae. Treatment of S. mansoni larvae with MBCD elicited appreciable cholesterol depletion as judged by filipin-cholesterol fluorescence diminution, which was accompanied by a considerable increase in specific antibody binding in IF, thus suggesting that cholesterol plays a predominant role in sequestration of the surface membrane antigens of S. mansoni lung-stage schistosomula. Despite that, MBCD induced an almost complete depletion of cholesterol from the outer membrane of S. haematobium larvae; no increase in specific antibody binding in IF was evident, implying that cholesterol is not responsible for masking surface membrane antigens of S. haematobium lung-stage larvae.     

Reaction of Philophthalmus rhionica (Trematoda, Philophthalmidae miracidia to light

The light response of miracidia was studied by means of several original methodics. The heterogeneity in the character of light response between various larvae of the same age population was determined in the course of experiments. It was shown that a part of miracidia possesses the strict positive phototaxis. Moreover, in their movement to the light the larvae seem to orientate themselves on the light intensity gradient. Not all the miracidia possess the positive phototaxis ("+" miracidia). Some of them ("-" miracidia) leave for the dark side of camera under the influence of direct ray of light. There are also larvae indifferent to the light conditions in thepopulation. They can move in different directions in spite of the light ray direction ("0" miracidia). The number of "+", "-" and "0" miracidia in the population is inconstant. The number of the negative phototactic larvae grows with age and respectively the number of "+" miracidia decreases. It is obvious the "+" miracidia can transform into "0" or "-" forms, while there is no opposite transformation. The reasons of differences in the movement character of "+" and "-" miracidia are under discussion.     

Frequency and possible consequences of hybridization between Schistosoma haematobium and S. mattheei in the Eastern Transvaal Lowveld

A survey was conducted at a locality in the Eastern Transvaal Lowveld where the prevalence of human infection with the bovine parasite, Schistosoma mattheei, is relatively high. It was found that, when compared to the number of S. haematobium eggs released into the environment, the number of S. mattheei eggs, with enclosed hybrid miracidia, is small. The consequences of backcrossing between the hybrids and S. haematobium was considered; a mathematical model indicated that a high percentage of the S. haematobium population should contain a small proportion of S. mattheei genes. The results indicate that it is highly unlikely that the two species will evolve into a single species, neither does it seem that the virulence of the parental species will be influenced.     

Sublethal effects of ultraviolet b radiation on miracidia and sporocysts of Schistosoma mansoni: intramolluscan development, infectivity, and photoreactivation

Schistosoma mansoni occurs in tropical regions where levels of ultraviolet B (UVB; 290-320 nm) light are elevated. However, the effects of UVB on parasite transmission are unknown. This study examines effects of UVB on the miracidia and sporocysts of S. mansoni, focusing specifically on intramolluscan development, infectivity, and the ability to photoreactivate (repair DNA damage using visible light). Histology revealed that miracidia irradiated with 861 J x m(-2) underwent abnormal development after penetrating Biomphalaria glabrata snails. Total number of sporocysts in snail tissues decreased as a function of time postinfection (PI), among both nonirradiated and irradiated parasites; however, this decrease was greater in the latter. Moreover, whereas the proportion alive of nonirradiated sporocysts increased PI, that of irradiated sporocysts, i.e., derived from irradiated miracidia, decreased. Irradiation of miracidia with UVB resulted in decreased prevalence of patent infection (defined by presence of daughter sporocysts) in a dose-dependent manner, and no infections occurred at a dose of 861 J x m(-2). Like many aquatic organisms, including the snail host, parasites were able to photoreactivate if exposed to visible light following UVB irradiation, even subsequent to penetrating snails. These photoreactivation results suggest cyclobutane-pyrimidine dimers in DNA as the primary mechanism of UVB damage, and implicate photoreactivation, rather than nucleotide excision, as the main repair process in S. mansoni.     

Studies on experimental mixed infections of Schistosoma mansoni and S. haematobium in hamsters

Golden hamsters were superinfected simultaneously with 100 Schistosoma haematobium cercariae, 1 and 3 weeks after initial infection with 100 S. mansoni cercariae. Results indicate that there was a higher degree of resistance to superinfection with S. haematobium at 1 week following initial infection with S. mansoni than that produced in the other two superinfections. This resistance was evidenced by a reduction in the number and size of worms of both species, decrease in S. haematobium egg extrusion per female and by a striking deviation in the egg distribution pattern of both species. Such an early host resistance was not recorded in previous works. Cross-mating was observed but no hybridization took place and the eggs produced were hatchable and typical of their species.