Schistosoma japonicum: disc electrophoretic protein patterns of the Japanese, Philippine, and Formosan strains

Soluble proteins of the Japanese, Philippine, and Formosan strains of Schistosoma japonicum were separated by disc electrophoresis using polyacrylamide gel columns. Differences between strains and sexes were investigated on the basis of R_f values of the bands, location of prominent peaks, quantitative comparisons of major bands, and overall densitometric profiles. With male extract, 29, 28, and 29, distinct bands were resolved for the Japanese, Philippine, and Formosan strains, respectively. Female extracts gave 31, 26, and 25 distinct bands for the Japanese, Philippine, and Formosan strains, respectively. Both qualitative and quantitative differences were found among the three strains and between sexes. The closest relationship of densitometric profile was between the Japanese and Formosan strains.     

Induction of resistance in Oncomelania hupensis nosophora against Schistosoma japonicum, but not against Paragonimus ohirai, using irradiated miracidia

Oncomelania hupensis nosophora snails sensitized with X-irradiated Schistosoma japonicum miracidia demonstrated resistance against a following challenge infection with non-irradiated homologous miracidia. The resistance in O. h. nosophora against S. japonicum was acquired within 1 day of sensitization, and it was strongest in a group challenged at an interval of 3 days. The resistance persisted for at least 4 weeks. Histological examinations revealed amoebocyte accumulation around the challenged S. japonicum sporocysts. On the other hand, when O. h. nosophora sensitized by exposure to X-irradiated P. ohirai or S. japonicum miracidia were subsequently challenged with normal P. ohirai miracidia, no resistance was observed, although they expressed the resistance against heterologous S. japonicum infection.     

Schistosoma malayensis n. sp.: a Schistosoma japonicum-complex schistosome from Peninsular Malaysia

Schistosoma malayensis n. sp., a member of the Schistosoma japonicum complex is described from Rattus muelleri in Peninsular Malaysia and 2 strains are characterized. The only morphological differences noted among adults from natural hosts were that S. malayensis are in general smaller than S. mekongi and S. japonicum. But these differences may be the result of host-induced variations and therefore are of little taxonomic value. To minimize the effects of host-induced variations, adult worms recovered from laboratory mice with similar worm burdens at 50-56 days postinfection were compared. These comparisons revealed only minor morphometric differences among these 3 species. Schistosoma malayensis eggs from naturally and experimentally infected hosts are most similar to those of S. mekongi, with eggs of both species being, in general, smaller than those of S. japonicum. The egg index for S. malayensis is usually higher than for S. japonicum and lower than for S. mekongi. Differences were noted in the developmental rates in mice for 2 isolates of S. malayensis, S. mekongi, and S. japonicum (Philippine strain), but relatively large differences observed between isolates of S. malayensis indicate that, in this case, the developmental rate is not a useful taxonomic character. Schistosoma malayensis is erected principally on the basis of differences, reported elsewhere, in the life histories and in the electrophoretic migration patterns of isoenzymes of adult worms as compared to S. mekongi and S. japonicum. These comparisons indicate that S. malayensis is more closely related to S. mekongi than to S. japonicum.     

Selection of genotypes of Schistosoma mansoni and their maintenance by sporocyst transplantation

Schistosoma mansoni was isolated by hatching eggs obtained from a naturally infected Rat in Grand Etang, Guadeloupe; fifty Biomphalaria glabrata were exposed to five miracidia each. The resulting cercariae were used to infect laboratory mice which were later sacrificed to provide worms for enzyme analyses and eggs for further infections. Seven enzymes in extracts of individual worms were examined by isoelectric focusing in polyacrylamide gels: AcP, G6PDH, PGM, GPI and HK showed no variation, whereas MDH and LDH proved to be polymorphic. Two MDH loci were recognised, MDH-2 was invariant whereas two alleles were assumed at the MDH-1 locus. It was not possible to make a genetic interpretation of the complex banding pattern of LDH, although 4 types (LDH-A, -B, -C, -D) were observed. Of the snail infections, one batch of snails was exposed to 5 miracidia per snail in the normal way whereas other snails were each exposed to a single miracidium. The latter were sacrificed to provide sporocysts to transplant into further groups of recipient snails. Cercariae from the recipient snails were used to infect mice and the adult worms were analysed and compared with the normally passaged material. In this way, three lines, defined by the possession of particular MDH and LDH types, were selected from the originally polymorphic population; two were identical. The combination of single miracidium infections and enzyme typing has illustrated the possibility of selecting parasite lines of known genotype; transplantation of sporocysts from snail to snail has demonstrated that such lines can be maintained exclusively in the intermediate host.(ABSTRACT TRUNCATED AT 250 WORDS)     

Comparative studies of the defense mechanism against Schistosoma japonicum of schistosome-susceptible and -resistant Oncomelania nosophora

The amphibious snail Oncomelania nosophora is an intermediate host of Schistosoma japonicum. Previously we reported that there are two strains of the snail, one resistant and one susceptible to a Mindoro, the Philippines, strain of S. japonicum. The resistant snails were collected from Nirasaki and susceptible snails from Kisarazu, Japan. To determine early cellular responses in the two snail strains, we examined histologic alterations in the snails for up to 18 h after the initial exposure to miracidia. In both strains, the penetrating miracidia were distributed in the foot, mantle, gills, heart, stomach, and kidney, and the mean number of penetrating miracidia was similar in both strains. After penetration, snail hemocytes migrated toward the larvae, and by 12 h after exposure, substantial numbers of penetrated larvae were surrounded and encapsulated by hemocytes. The percentage of larvae encapsulated by hemocytes during 12-18 h after the exposure was significantly higher in the resistant Nirasaki strain (60.9+/-19.8%) than in the susceptible Kisarazu strain (42.3+/-15.0%). In a few snails of the Nirasaki strain, all the larvae found were encapsulated by hemocytes. The differences in hemocyte responses between the two strains may explain the susceptibility of the snails to schistosome larvae.     

Parasitological studies at a new breeding area of Oncomelania hupensis chiui in Taipei county, Taiwan.

Oncomelania hupensis chiui was found at the Jui-Ping area, Linkow District, Taipei County, Taiwan, which is located on the north coast of the island approximately 10 km of the mouth of the Tamasui River. The habitat of the snails at this area is similar to the type locality at Alilao, on the slope of the hills along of coast. The snails from this new breeding area, are similar to the Alilao snails, and are highly susceptible to the infection with geographic strains of Schistosoma japonicum including the Changhua, Ilan, Japanese, Philippine and Chinese strains. Examinations of 7,758 snails and intradermal tests on 736 inhabitants along with stool examinations of 622 people indicated that there was no evidence of the presence of Schistosoma japonicum at this area. Results obtained from stool examination demonstrated high prevalence of hookworm infection (53.4%) and relatively low infection rates of Asearis lumbricoides (37.0%) and Trichuris trichiura (14.1%). The protozoan infections were low in general. A single Scotch-tape technique for Enterobius vermicularis among 499 school children yielded a 55.7% infection rate.     

Schistosoma japonicum and Paragonimus ohirai: antagonism between S. japonicum and P. ohirai in Oncomelania nosophora

Antagonistic interactions between Schistosoma japonicum and Paragonimus ohirai were examined in the snail host, Oncomelania nosophora. When P. ohirai-infected snails were exposed to S. japonicum miracidia at intervals of 4 to 18 weeks post-first exposure, only a few snails (0-7%) were found to be superinfected with S. japonicum sporocysts. Sporocysts were fewer in number than those of single infected controls. Mature S. japonicum cercariae were not observed. Furthermore, when the snails were examined at intervals of 14 to 18 weeks post-second exposure, neither sporocysts nor cercariae of S. japonicum were found. On the other hand, when the snails were exposed to miracidia of S. japonicum and P. ohirai simultaneously, they were easily infected with both parasites. At 26 weeks after simultaneous exposure, however, the infection rate of S. japonicum was significantly lower than that of controls. In contrast, when S. japonicum-infected snails were exposed to P. ohirai miracidia, they were superinfected with P. ohirai, although the infection rate was somewhat lower than that of controls. These results indicate the existence of antagonism between S. japonicum and P. ohirai in O. nosophora. Furthermore, P. ohirai was dominant over S. japonicum in the antagonistic interactions in this snail host.     

Expression of uptake hydrogenase and hydrogen oxidation during heterotrophic growth of Bradyrhizobium japonicum.

Strains I-110 ARS, SR, USDA 136, USDA 137, and AK13 1c of Bradyrhizobium japonicum induced Hup activity when growing heterotrophically in medium with carbon substrate and NH4Cl in the presence of 2% H2 and 2% O2. Hup activity was induced during heterotrophic growth in the presence of carbon substrates, which were assimilated during the time of H2 oxidation. Strains I-110 ARS and SR grown heterotrophically or chemoautotrophically for 3 days had similar rates of H2 oxidation. Similar rates of Hup activity were also observed when cell suspensions were induced for 24 h in heterotrophic or chemoautotrophic growth medium with 1% O2, 10% H2, and 5% CO2 in N2. These results are contrary to the reported repression of Hup activity by carbon substrates in B. japonicum. Bradyrhizobial Hup activity during heterotrophic growth was limited by H2 and O2 and repressed by aerobic conditions, and CO2 addition had no effect. Nitrogenase and ribulosebisphosphate carboxylase activities were not detected in H2-oxidizing cultures of B. japonicum during heterotrophic growth. Immunoblot analysis of cell extracts with antibodies prepared against the 65-kilodalton subunit of uptake hydrogenase indicated that Hup protein synthesis was induced by H2 and repressed under aerobic conditions.     

Susceptibility of Oncomelania hupensis formosana recombinants and hybrids with Oncomelania hupensis nosophora to infection with Schistosoma japonicum

Chiu J.-K., Ong S.-J., Yu J.-C., Kao C.-Y. and Iuima T. 1981. Susceptibility of Oncomelania hupensis formosana recombinants and hybrids with Oncomelania hupensis nosophora to infection with Schistosoma japonicum. International Journal for Parasitology11: 391–397. Three generations of Oncomelania hupensis formosana recombinants were produced by mating the Kaohsiung race with the Ilan and Changhua races of the snails. F₂ and F₃ recombinants were produced by back-crossing F₁ and F₂ with Kaohsiung O. h. formosana. Subsequently, susceptibility of recombinants to infection with the original strain of Schistosoma japonicum, Ilan or Changhua strain, was investigated. Results indicated that susceptibility of recombinants declined steadily generation by generation. Marked decline of infectivity was observed for Kaohsiung-Ilan recombinants as compared with Kaohsiung-Changhua recombinants. For example, the overall infection rate of Kaohsiung-Ilan F₁ recombinants was 7.3 % with a 51.4 % of control snails. The same figures for F₂ were 4.2 and 52.6%, and 1 and 40.3 % for F₃. On the other hand, the overall infection rate of Kaohsiung-Changhua F₁ recombinants was 21.9% with a 46.9% of control snails; and 11.9 and 50.3 % for F₂; and 7.6 and 33.2% for F₃. The F₃ hybrids of Oncomelania hupensis nosophora from Japan and O. h. formosana from Kaohsiung and Ilan were also produced, and susceptibility with the Japanese strain of S. japonicum was studied. A highly significant decline of susceptibility was observed among hybrids (4.4%) in contrast with control snails (85.6 %).Feasibility of applying O. h. formosana in biological control of S. japonicum was discussed. One must determine in the laboratory, prior to application, which race of O. h. formosana should be used depending on the Oncomelania snails of an endemic area. For S. japonicum prevalent in Yamanashi, Japan, the Ilan race of O. h. formosana was found to be better choice than the Kaohsiung race of the snails.     

The occurrence of two opecoeliid digeneans in Mullus barbatus and M. surmuletus from the Spanish south-eastern Mediterranean

The infection by Opecoeloides furcatus and Poracanthium furcatum (Opecoeliidae) was studied in 121 Mullus barbatus and 113 M. surmuletus collected from the Spanish south-eastern Mediterranean. The prevalence of infection was most frequent in M. surmuletus with values of 81.42% for O. furcatus and 38.05% for P. furcatum. In M. barbatus the prevalences of O. furcatus and P. furcatum were 54.54% and 14.88% respectively. Statistically significant differences were found between the infection of the two hosts with P. furcatum. No significant differences in worm burdens could be attributable to host size or to seasonal changes, although a lower infection of M. barbatus by O. furcatus occurred in the autumn. Furthermore, the electrophoretic mobility of the enzyme malic dehydrogenase (MDH) was also studied and both digeneans presented different patterns, corresponding in both cases to homozygotic genotypes.     

Allozyme Variability and Phylogenetic Relationships in Honey Bee (Hymenoptera: Apidae: Apis mellifera) Populations From Greece and Cyprus

Ten gene enzymic systems (alpha-GPDH, AO, MDH, ADH, LAP, SOD, ALP, ACPH, ME, and EST), corresponding to 12 genetic loci, were assayed from five Greek populations representing three subspecies of Apis mellifera, A. m. cecropia (Pthiotida, Kythira), A. m. macedonica (Macedonia), and the "Aegean race" of A. mellifera, which is supposed to be very similar to A. m. adami (Ikaria, Kasos), as well as a population from Cypus (A. m. cypria). ADH( *)-1, ADH( *)-2, and LAP( *) electrophoretic patterns discriminate the Cyprus population from the Greek populations. MDH( *)-1, EST( *)-3, SOD( *), ALP( *), and ME( *) loci were found to be polymorphic in almost all populations. The observed heterozygosity was found to range from 0.066 to 0.251. Allele frequencies of all loci were used to estimate Nei's genetic distance, which was found to range between 0.011 and 0.413 among the populations studied. UPGMA and neighbor-joining phylogenetic trees obtained by genetic distance matrix methods, as well as a Wagner tree based on the discrete character parsimony method, support the hypothesis that the most distant population is that from Cyprus. Our allozymic data support A. m. cypria as a distinct subspecies, but there was no allozymic support for the distinction of the other subspecies existing in Greece.     

Detection of inducible nitric oxide synthase in Schistosoma japonicum and S. mansoni

Schistosoma japonicum and S. mansoni were tested for reactivity with an anti-inducible nitric oxide (iNOS) antibody and the distribution of iNOS was studied by immunofluorescent tests in different stages of the parasites. Reactivity was associated with the tegument in both larval schistosomes (sporocysts and cercariae) and eggs. With adult worms, the majority of the immunofluorescence was predominantly subtegumental in S. japonicum and parenchymal in S. mansoni. Fluorescence was also observed in host tissues (snails and mouse liver). In Western blots, the enzyme of S. japonicum had an apparent molecular weight of about 210 kDa. The possible role of worm and host iNOS in the parasite-host interrelation remains to be clarified.     

Molecular detection of Schistosoma japonicum in infected snails and mouse faeces using a real-time PCR assay with FRET hybridisation probes

A real-time polymerase chain reaction (PCR) assay with fluorescence resonance energy transfer (FRET) hybridisation probes combined with melting curve analysis was developed to detect Schistosoma japonicum in experimentally infected snails and in faecal samples of infected mice. This procedure is based on melting curve analysis of a hybrid between an amplicon from the S. japonicum internal transcribed spacer region 2 sequence, which is a 192-bp S. japonicum-specific sequence, and fluorophore-labelled specific probes. Real-time FRET PCR could detect as little as a single cercaria artificially introduced into a pool of 10 non-infected snails and a single egg inoculated in 100 mg of non-infected mouse faeces. All S. japonicum-infected snails and all faecal samples from infected mice were positive. Non-infected snails, non-infected mouse faeces and genomic DNA from other parasites were negative. This assay is rapid and has potential for epidemiological S. japonicum surveys in snails, intermediate hosts and faecal samples of final hosts.     

日本血吸虫新抗原基因的筛选、克隆、 表达和免疫效果研究

为了寻找日本血吸虫 (Schistosoma japonicum, Sj) 新的疫苗候选基因并进行免疫效果研究,用 Sj 雌虫抗原免疫家兔制备血清,对Sj成虫 cDNA 文库进行免疫筛选,将获得的新基因 ( 命名为Sj-F1, GenBank 登录号为 AY261995) 克隆入原核表达载体 pTWIN1 和真核表达载体 pcDNA3 ,经 PCR 、限制性酶切筛选和鉴定阳性重组子. 将 pTWIN1/Sj-F1 质粒转化大肠杆菌 ER2566,在低温和低 IPTG 浓度下诱导表达可溶性重组融合蛋白 (rSj-F1/intein2),并经 SDS- 聚丙烯酰胺凝胶电泳 (SDS-PAGE) 和蛋白质印迹 (Western blot) 分析鉴定. 将 pcDNA3/Sj-F1 质粒转化大肠杆菌 ER2502 ,大量制备 DNA 疫苗. 用重组融合蛋白和 DNA 疫苗免疫小鼠,末次免疫后 2 周用Sj尾蚴进行攻击感染. 感染后 42 天剖杀冲虫,计算减虫率和减卵率. 感染前采血用 ELISA 法检测抗体. 免疫保护效果测定显示：重组蛋白疫苗以 FCA 作佐剂经皮下免疫和以壳聚糖作佐剂经粘膜免疫分别获得了 28.07%、 24.69% 的减虫率和 48.30% 、 46.38% 的减卵率; DNA 疫苗 (pcDNA3/Sj-F1) 单独免疫获得了 18.47% 的减虫率和 35.06% 的减卵率;用 DNA 疫苗启动免疫后用重组蛋白疫苗经皮下加强免疫,减虫率和减卵率分别提高到了 40.42% 和 56.17%;用 DNA 疫苗启动免疫后用重组蛋白疫苗经黏膜加强免疫,减虫率和减卵率增高更明显,分别提高到了 42.38% 和 62.87%. 结果表明,Sj-F1 重组蛋白疫苗及 DNA 疫苗均可诱导小鼠产生部分抗血吸虫感染的保护力,两者联合免疫保护效果优于单一疫苗.     

Immunization of mice with cells from juvenile worms of Schistosoma japonicum provides immunoprotection against schistosomiasis

To validate the protective efficacy against schistosomiasis by immunization with cells from juvenile Schistosoma japonicum in a murine model and to analyze possible factors related to protection, in this study, two independent repeated vaccination trials were performed. After three subcutaneous vaccina- tions, in trial one, in the absence of adjuvant, primary juvenile worm cells (pJCs) from S. japonicum induced remarkable average reductions in worm burden (54.3%), liver eggs per gram (LEPG) load (59.8%) as well as egg granulomas size (66.5%) compared to PBS control group (P<0.01), which were significantly higher than those elicited by fractions of juvenile worm cells (JCFs) or fractions of juvenile worms (JWFs) (P<0.05). Non-cell components of worms (WNCs) showed no significant protection. In trial two, compared to PBS control group, significant protective effect was also observed for cultured juvenile worm cells (cJCs) from S. japonicum with 58.4% worm reduction and 68.1% LEPG reduction (P<0.01). However, cultured adult worms cells (cACs) showed significantly higher worm burden (P<0.05) and egg burden (P<0.01) when compared to cJCs. Immunological analysis of trial two revealed that cJCs engendered a Th1-biased mixed Th1/Th2 type of immune response while cACs elicited a Th2-type response. Our data indicated that immunization with both primary and cultured cells from S. japonicum juvenile worms provided high immunoprotection, for which the physical character of immunogens, stage-specific parasite and the type of immune response induced might be responsible, suggesting that vaccination with whole cells from S. japonicum larvae is a promising approach to produce protec- tive immunity against schistosomiasis.     

Genetic control and developmental expression of malate dehydrogenase in Apis mellifera

Starch gel electrophoresis of extracts of Apis mellifera indicates that genetic variability exists for the enzyme cytoplasmic malate dehydrogenase (E.C. 1.1.1.37). Analysis of individuals throughout development indicates that the isozyme patterns are identical for larvae and adults and suggests a dimeric structure for the molecule. The isozyme pattern observed in pupae is more complex than that of larvae and adults may be due to an additional pupal-specific MDH gene being expressed or to an epigenetic modification of the isozymes. Forty-three colonies with artificially inseminated queens were used to study the Mendelian pattern of inheritance. The data revealed that the MDH isozymes are encoded by three alleles, Mdh-1A, Mdh-1B, and Mdh-1C. The frequency of the Mdh-1 alleles is different in two analyzed subspecies, A. m. adansonii (African bees) and A. m. ligustica (Italian bees), with Mdh-1A and Mdh-1B in the African bees being 0-768 and 0.202, respectively. For the Italian bees, these frequencies are 0.136 and 0:154, respectively.     

Miracidia of Schistosoma japonicum: approach and attachment to the snail host

Schistosoma japonicum miracidia swim directed along a chemical gradient toward the snails Oncomelania hupensis and Biomphalaria glabrata, and they turn back when the concentration of attractive chemicals decreases. The host signal for this chemotactic response has a molecular weight of more than 30,000. When swimming miracidia encounter the surface of O. hupensis or agar containing O. hupensis snail-conditioned water (SCW) they perform the host-specific responses "contact with return," "repeated investigation," and "attachment," but they do not exhibit such behavior when encountering B. glabrata surface or agar containing B. glabrata SCW. Thus S. japonicum miracidia respond to different host signals when they approach snails than when they attach to snails.     

Malate dehydrogenase and glucose-6-phosphate dehydrogenase, key markers for studying the genetic diversity of Actinobacillus actinomycetemcomitans

Abstract Cell-free extracts of strains belonging to the 5 serotypes of A. actinomycetemcomitans were screened for several enzymes. Enzymes representative of the pentose phosphate pathway/hexose monophosphate shunt and the TCA cycle were present. Of these glucose-6-phosphate dehydrogenase (G6PDH) and malate dehydrogenase (MDH) were the most readily detected and stable. MDH and G6PDH retained more than 50% of their activities at alkaline pHs (10–11) for up to 6 h and 3 h at 25°C, respectively, while at pH 6.5, 50% of their activities were lost within 2–3 h. The K _m for malate oxidation catalysed by MDH was 5.8×10⁻⁴ M while that for glucose-6-phosphate oxidation was 2.0×10⁻⁴ M. The pH optima for MDH and G6PDH oxidation activities were 10 and 9.5, respectively. Among the 5 designated serotypes of A. actinomycetemcomitans three groups were delineated by multilocus enzyme electrophoresis using MDH and G6PDH.     

Novel functions of the alpha-ketoglutarate dehydrogenase complex may mediate diverse oxidant-induced changes in mitochondrial enzymes associated with Alzheimer's disease

Measures in autopsied brains from Alzheimer's Disease (AD) patients reveal a decrease in the activity of alpha-ketoglutarate dehydrogenase complex (KGDHC) and an increase in malate dehydrogenase (MDH) activity. The present experiments tested whether both changes could be caused by the common oxidant H(2)O(2) and to probe the mechanism underlying these changes. Since the response to H(2)O(2) is modified by the level of the E2k subunit of KGDHC, the interaction of MDH and KGDHC was studied in cells with varying levels of E2k. In cells with only 23% of normal E2k protein levels, one-hour treatment with H(2)O(2) decreased KGDHC and increased MDH activity as well as the mRNA level for both cytosolic and mitochondrial MDH. The increase in MDH did not occur in cells with 100% or 46% of normal E2k. Longer treatments with H(2)O(2) inhibited the activity of both enzymes. Glutathione is a major regulator of cellular redox state and can modify enzyme activities. H(2)O(2) converts reduced glutathione (GSH) to oxidized glutathione (GSSG), which reacts with protein thiols. Treatment of purified KGDHC with GSSG leads to glutathionylation of all three KGDHC subunits. Thus, cellular glutathione level was manipulated by two means to determine the effect on KGDHC and MDH activities. Both buthionine sulfoximine (BSO), which inhibits glutathione synthesis without altering redox state, and H(2)O(2) diminished glutathione to a similar level after 24 h. However, H(2)O(2), but not BSO, reduced KGDHC and MDH activities, and the reduction was greater in the E2k-23 line. These findings suggest that the E2k may mediate diverse responses of KGDHC and MDH to oxidants. In addition, the differential response of activities to BSO and H(2)O(2) together with the in vitro interaction of KGDHC with GSSG suggests that glutathionylation is one possible mechanism underlying oxidative stress-induced inhibition of the TCA cycle enzymes.     

Isozyme patterns of Schistosoma japonicum and S. mansoni

Isozyme patterns of six enzymes, glucose-6-phosphate dehydrogenase, glucosephosphate isomerase, hexokinase, malate dehydrogenase, 6-phosphate dehydrogenase and phosphoglucomutase were examined in electrophoresed homogenates of adult male worms of Schistosoma japonicum and S. mansoni. In general, enzyme patterns obtained from the parasite homogenates differed from that of host (mouse) blood and muscle, indicating that electrophoretic patterns from parasite extracts are most probably of parasite origin. Adult male and female S. mansoni worms yielded identical patterns. However, all six enzyme patterns showed distinct differences between S. japonicum and S. mansoni. These results suggest that S. japonicum is clearly distinguishable from S. mansoni at the molecular level.