Schistosoma mansoni and Schistosoma japonicum: Utilization of amino acids

, , and 1972. Schistosoma mansoni and Schistosoma japonicum: utilization of amino acids. International Journal for Parasitology 2: 425–430. The production of ¹⁴CO₂ from 12 labeled amino acids by S. mansoni and S. japonicum was studied. No ¹⁴CO₂ was detected from incubations with glycine, isoleucine, leucine, lysine or phenylalanine. Differences were found between sexes and/or species for the other amino acids studied. Species related differences included a greater rate of metabolism of glutamic and aspartic acid by S. mansoni than by S. japonicum. Proline and histidine were utilized by S. mansoni males and females, respectively. S. japonicum male worms did not utilize proline, while histidine was not utilized by the female of this species. Major sex related differences included greater ¹⁴CO₂ production from glutamic acid, aspartic acid and arginine by S. mansoni males than by females, and the utilization of histidine by male S. japonicum but not by females. Incubation in tyrosine resulted in the release of only small amounts of ¹⁴CO₂ by female worms of both species but no ¹⁴CO₂ production by male worms.     

A study of the death of Schistosoma mansoni cercariae during penetration of mammalian host skin: The influence of the ages of the cercariae and of the host

A study of the death of Schistosoma mansoni cercariae during penetration of mammalian host skin: the influence of the ages of the cercariae and of the host. International Journal for Parasitology 3: 789–794. The number of cercariae of S. mansoni which die during penetration of mouse abdominal skin steadily increases with age following emergence from the snail. An initial mortality level of about 30 per cent is observed for 2-h-old cercariae which rises to 50 per cent at 8 h and 85 per cent at 24 h. These increased losses in the skin are shown to account substantially for the known decrease in infectivity which accompanies ageing of cercariae. The number of cercariae which die in the skin of very young mice (2 days old) is less than one-third of the level in adult mice. Losses in the skin increase with age of the host up to about 28–35 days. This increased mortality in the skin is shown to account for the observed age resistance of mice, where fewer cercariae mature into adult worms in mice of 1 month or more, than in very young mice.     

Diplostomum spathaceum cercariae respond to a unique profile of cues during recognition of their fish host

During its normal life cycle, Diplostomum spathaceum cercariae attach to and invade fish intermediate hosts. They are also known to attach to various other aquatic animals in response to water currents, touch and carbon dioxide. The purpose of this study was to identify the specific stimuli used by D. spathaceum cercariae to recognise the appropriate fish host. We characterised the host cues which stimulate them to remain on the host (enduring contact) and to penetrate the skin. Cercariae were exposed to animal skin tissues and fish skin surface mucus, their extracts and chemical modifications integrated into agar or offered via membrane filters. Enduring contact was stimulated by hydrophilic extracts Mr<3kDa, which were sensitive to oxidation of carbohydrates. The stimulating cues are probably small molecular carbohydrates, as monosaccharides stimulated enduring contacts, but amino acids, urea, electrolytes and peptides did not. Penetration was stimulated by hydrophilic macromolecules, Mr>30kDa, and by lipids. The hydrophilic stimuli were protease resistant and precipitable with Alcian blue and they were sensitive to alkaline cleavage, to digestion with lysozyme and neuraminidase as well as to oxidation of sialic acids. They were considered to be glycoproteins with O-glycosidically linked carbohydrate chains and bound sialic acids as signal structures. The lipophilic penetration stimuli were contained exclusively in the fatty acid fractions, and the stimulating characteristics of these fatty acids resembled the stimulating penetrations in other cercarial species. Diplostomum spathaceum cercariae respond to a unique profile of cues in their sequence of host-recognition phases. These cues differ from those used in other fish parasites studied to date and underline the diversity of fish recognition strategies.     

Specificity of natural resistance to trematode infections in Biomphalaria glabrata

The 10-R2 strain of Biomphalaria glabrata was strongly resistant to various strains of Schistosoma mansoni in its laboratory of origin (NIH) and to three strains of S. mansoni we tested against it. However, subsequent development of three inbred lines of B. glabrata 10-R2 snails, separately maintained in our San Francisco laboratory, showed slight loss of resistance in one colony, very much less resistance (or partial susceptibility) in another, and retention of the original resistance in a third to the Puerto Rico (PR-1) strain of S. mansoni. No selection for resistance to infection was involved in the breeding protocol for these 10-R2 lines, so the changes were apparently random ones that became established in the separate inbred substrains. In spite of their changed response to the PR-1 strain of S. mansoni, all three 10-R2 substrains retained only slightly diminished resistance to S. mansoni Lc-1 strain and an essentially undiminished resistance to irradiated Echinostoma lindoense, E. paraensei and E. liei sporocysts. This suggests that natural resistance to S. mansoni PR-1 in B. glabrata is specific, a response that differs from the host response to either S. mansoni Lc-1 or to the echinostomes.     

Capacity of irradiated echinostome sporocysts to protect Schistosoma mansoni in resistant Biomphalaria glabrata

Three closely related species of Echinostoma flukes each has a distinctive pattern of protection of Schistosoma mansoni in schistosome-resistant Biomphalaria glabrata host snails. Protection of developing S. mansoni by irradiated E. paraensei sporocysts in the schistosome-resistant snail host was strong; protection induced by irradiated E. lindoense and E. liei sporocysts was weak or not measurable. The capacity of irradiated E. paraensei sporocysts to interfere with the host's innate anti-schistosome response also differed between strains of B. glabrata. Protection of S. mansoni strain Lc-1 was greater in B. glabrata strain 10-R2 than it was in strain M-RLc snails. Irradiated E. paraensei sporocysts also induced a different response to the two schistosome strains in a single host strain. Irradiated E. paraensei sporocysts induced in B. glabrata 10-R2 snails a stronger protection of S. mansoni strain PR-1 than of strain Lc-1. Exposure of each snail to the irradiated E. paraensei miracidia usually protected the following challenge schistosome infection better when 30 rather than 10 irradiated echinostome miracidia were used.     

Cloning and partial nucleotide sequence of Schistosoma japonicum paramyosin: a potential vaccine candidate against schistosomiasis.

, , , and 1992. Cloning and partial nucleotide sequence of Schistosoma japonicum paramyosin: a potential vaccine candidate against schistosomiasis. International Journal for Parasitology 22: 1187–1191. Paramyosin from the blood fluke, Schistosoma mansoni, has shown promise as a vaccine candidate for schistosomiasis mansoni. Here we report the cloning and partial nucleotide sequence of a cDNA encoding paramyosin from the related human parasite, Schistosoma japonicum. Affinity purified antibodies to this clone recognized a S. japonicum antigen of molecular weight 97 kDa, equivalent to the reported size of S. mansoni paramyosin. Alignment of the cDNA sequence with that of S. mansoni paramyosin revealed 90% identity. Comparison of the predicted amino acid sequences revealed 95% identity. Although these two parasites differ in many characteristics, the substantial homology demonstrated here between S. mansoni and S. japonicum paramyosin could have important implications for the development of a S. japonicum vaccine.     

Cross-reactivity between Necator americanus and Schistosoma mansoni in mice.

, , and 1992. Cross-reactivity between Necator americanns and Schistosoma mansoni in mice. International Journal for Parasitology 22: 1143–1149. Poly-parasitism is common in endemic communities and reactivity of sera from hookworm-infected patients against schistosomular antigens has been reported. Protective cross-immunity between N. americanus and S. mansoni was investigated in NIH and BALB/c mice. Protective resistance to homologous challenge with both parasites was confirmed in this model, however, functional immunity to heterologous challenge was not demonstrated. Sera from animals which had received homologous challenge with N americanus and from hookworm-infected mice, which had previously been exposed to radiation-attenuated S. mansoni, exhibited an enhanced IgGAM response to infective stage N. americanus somatic antigens. The implications of these results with respect to serodiagnosis are discussed.     

Schistosoma mansoni encodes SMT3B and SMT3C molecules responsible for post-translational modification of cellular proteins

The sumoylation pathway is a post-translational modification of nuclear proteins widespread among several organisms. SMT3C is the main protein involved in this process and it is covalently conjugated to a diverse assortment of nuclear protein targets. To date, 3 SUMO paralogues (SMT3C, A/B) have been characterized in mammals and plants. In this work we characterized two SUMO related genes, named SMT3B and SMT3C throughout Schistosoma mansoni life cycle. The SmSMTB/C encodes for proteins sharing significant amino acid homology with SMT3. Phylogenetical analyses revealed that both SmSMT3B/C are distinct proteins. Additionally, SmSMT3B and C are expressed in cercariae, adult worms, eggs and schistosomula however SmSMT3C gene showed an expression level 7 to 9 fold higher than SmSMT3B in eggs, schistosomula and adult worms. The comparison between the SmSMT3C genomic and cDNA sequences established that the encoding sequence is interrupted by 3 introns of 70, 37 and 36 bp. Western Blot has shown SMT3 conjugates are present in nuclear and total protein fractions of adults and cercariae. Therefore our results suggest a functional sumoylation pathway, and the presence of two paralogues also suggests the specificity of substrates for SMT3 in S. mansoni.     

Production of monoclonal antibodies against target schistosomal antigen secreted in the urine of Schistosoma mansoni-infected patients

This study was undertaken to develop an immunodiagnostic test of active human schistosomiasis mansoni using a monoclonal antibody which targets urinary schistosomal antigen. Polyclonal antisera raised in rabbits against the processed urine of Schistosoma mansoni-infected patients showed very high and significant reactivity with ES product of ova compared with other different S. mansoni antigens. The monoclonal antibody (4.23) was reactive with repetitive epitopes of S. mansoni soluble egg antigen and ES product of ova with molecular mass range of 65–23 kDa and 80–23 kDa, respectively. It recognised different stages of the parasite life-cycle, with no cross reaction with Fasciola or hydatid antigen. MAbs were characterised by isotyping, immunoelectrophoresis, SDS–PAGE and the enzyme-linked immunoelectrotransfer blot technique, ELISA, and their recognition of carbohydrate or protein antigenic epitopes by periodate oxidation and trichloroacetic acid treatment of the antigen. It was used for detection of circulating schistosomal antigen in an antigen capture antibody sandwich ELISA on sera and urines of 58 S. mansoni-infected patients, 17 S. haematobium-infected patients, 15 parasite-free negative healthy controls and sera from 13 schistosomiasis-free patients harbouring Fasciola or hydatid infections. The percentage sensitivity of the assay in the serum of S. mansoni-infected patients was 98.4% and in urine 94.8%. A positive correlation was found between the number of faecal S. mansoni eggs and the circulating antigen, both in serum and in urine. Antigen circulating in urine correlated with that in the sera of S. mansoni patients. These data provide a sensitive and non-invasive method almost comparable with the use of sera for immunodiagnosis of schistosomiasis and an indirect way to reflect the intensity of infection. Australian Society for Parasitology.     

Circadian Pattern of Cercarial Emergence in Schistosoma Mansoni(Platyhelminthes: Digenea) from Isolated Biomphalaria Glabrata

The present work aimed to compare the acrophases (peak hours) of emergence of Schistosoma mansoni cercariae among isolated individuals of the snail Biomphalaria glabrata. Laboratory stocks of melanic B. glabrata from the same biotope as the S. mansoni strain (Belo Horizonte, Minas Ger-ais) were used. Twenty-two snails individually exposed to five miracidia were tested. Chronobiological trials were performed outdoors after an acclimation period of at least a week. Three groups of snails were tested between November 1989 and April 1991. Cercarial emergence from individual isolated snails was quantified every 3 h for 3 consecutive days. In all trials, most cercariae were found to emerge during daytime (94.9%). Time series and chronograms showed recurrent peaks during the daytime. The periodogram suggested that 24 h was the period that best fitted cercarial emergence data in 90.9% of the snails. The single cosinor analysis confirmed 24-h rhythms in 95.5% of the snails. Acrophases of cercarial emergence among individual snails occurred between 14:15 and 17:02. They did not differ significantly. The population cosinor analysis indicated greater homogeneity in the 24-h rhythms of cercarial emergence than in the snail groups of each chronobiological trial. Acrophases of cercarial emergence occurred between 14:53 and 15:27 and did not differ significantly among all trials. Data from the three trials were pooled and analyzed using the population cosinor. This statistical method indicated a homogeneity in the 24-h rhythms of cercarial emergence from all snails, with acrophase occurring around 15:00. Results showed that the acrophases of cercarial emergence of S. mansoni are similar among isolated B. glubrura specimens. Data support the hypothesis of a “gate” rhythm in the dynamics of cercarial production and emergence. It is suggested that the adaptive importance of the “gate” mechanism is associated with the concentration of cercariae in the water at times when the vertebrate is present, optimizing the contact between the parasite and the host. The emergence of some cercariae at night (5.1% of the total number of emerged cercariae) suggests a flexible “gate” that could be associated with a residual light effect or with experimental procedures in the laboratory.     

Host reactions in Biomphalaria glabrata to Schistosoma mansoni miracidia, involving variations in parasite strains, numbers and sequence of exposures

M-line Biomphalaria glabrata snails are susceptible to Puerto Rican (PR-1) strain of Schistosoma mansoni, but are resistant to a St. Lucian (LC-1) strain. 10-R2 B. glabrata snails are resistant to both strains of S. mansoni. When 10-R2 snails were exposed repeatedly to PR-1 S. mansoni miracidia for 5 consecutive days, all of the sporocysts were encapsulated and destroyed by the snails. Thirty-four per cent of sporocysts examined in M-line snails with similar exposures were also degraded. In double concurrent infections of M-line B. glabrata with [³H]leucine-labeled and unlabeled PR-1 and Lc-1 S. mansoni, the incompatible Lc-1 miracidia were selectively attacked and destroyed. This destruction occurred irrespective of the sequence of exposure of the 2 strains of miracidia, and whether or not the miracidia were labeled. Successful superinfection of M-line B. glabrata with homologous S. mansoni miracidia was obtained at least 4 days after the primary exposure to the miracidia.     

Implication of Papio anubis in the transmission of intestinal schistosomiasis in three new foci in Kime area, Ethiopia.

Epidemiological studies were conducted in the Lake Langano area in the Rift Valley of Ethiopia to determine the occurrence of schistosomiasis and assess factors involved in its transmission. Microscopic examination of faecal specimens from free ranging Papio anubis (anubis baboon) troops from Bishan Gari and Burka Dita forest reserves revealed Schistosoma mansoni eggs with a prevalence of 12.1% (11/91) and 26.2%(34/130), respectively. The eggs were viable as confirmed by miracidial hatching and infectivity tests. Out of the total 12 communities (three schools, five villages and one herdsmen community) surveyed for schistosomiasis around Lake Langano, individuals excreting S. mansoni eggs were found in nine communities with prevalence of infection ranging from 1.4 to 43%. The intensity of infection ranged from 24 EPG (eggs per gram of faeces) to 243 EPG. Excretion of viable eggs by the baboons indicate that they play a role in maintenance of S. mansoni infection in the locality. The detection of S. mansoni eggs in young children, collection of cercarial-infected Biomphalaria pfeifferi in water bodies, and establishment of S. mansoni infection in lab-bred mice have confirmed establishment of transmission foci in Kime area, south-east of Lake Langano. However, the lake itself does not seem to support transmission of schistosomiasis since no snails were found along the shore of the Lake. Further investigations are indicated to fully elucidate the role baboons play in the epidemiology of schistosomiasis in the Rift Valley of Ethiopia. The risk of introduction of water-based development projects in these new endemic foci in relation to S. mansoni infection in the baboons is discussed.     

Serological differentiation of acute and chronic schistosomiasis using Schistosoma mansoni recombinant protein RP26

We obtained a recombinant protein encoded by Schistosoma mansoni gene which was able to differentiate acute from chronic schistosomiasis when applied as antigen in enzyme-linked immunosorbent assay (ELISA). A cDNA clone encoding a 26 kDa recombinant protein (RP26) was selected by screening of an adult worm S. mansoni λZAP expression library with rabbit sera produced against PIII, an adult worm protein fraction already known to possess protective and immunomodulating effects. The clone cDNA presented 99% identity with S. mansoni Sm22.3 gene. We assayed IgG reactivity of sera from 18 patients with acute, 25 patients with chronic S. mansoni infection and 20 uninfected donors with RP26 in ELISA. Our results showed that 89% of sera were positive in acute schistosomiasis group, and only 26% in chronic group, without false-positive reactions in uninfected group. In mice the immune response to RP26 increased up to week 9 after infection and then diminished. We proposed that production of antibodies binding to RP26 stopped at the chronic stage of disease. The testing of sera from eight other parasitic infections with RP26 revealed no positive reactions in majority of sera. However, we observed low positive reaction in sera from 20% of leishmaniasis patients. Our results indicate that a recombinant protein RP26 can be used as immunodiagnostic reagent for detection of acute phase of schistosomiasis mansoni.     

Responses of Littoraria irrorata say (Mollusca: Gastropoda) to water-borne chemicals: a comparison of chemical sources and orientation mechanisms

The salt marsh periwinkle, Littoraria irrorata, exhibits orientation behaviors in response to a number of chemical stimuli. In this study, we quantified the response of L. irrorata to chemical stimuli derived from three sources: Spartina alterniflora (a food source), the blue crab Callinectes sapidus (a predator), and crushed conspecifics. Animals consistently moved toward sources of S. alterniflora extract and crushed conspecifics relative to controls, but did not approach or avoid extracts from C. sapidus relative to controls. Animals had lower response rates to stimuli from S. alterniflora than to controls, but not to putatively aversive chemical stimuli (C. sapidus or crushed conspecifics). However, periwinkles that moved in experimental groups and control groups did not differ in how far they crawled. In addition, we measured turning angles of pathways of periwinkles to determine the orientation mechanism used. Littoraria irrorata displayed a taxis in response to crushed conspecifics, but a kinesis in response to C. sapidus and S. alterniflora. Our results suggest that the ecological context of chemical cues influence the evolution of orientation mechanisms. Cues that require immediate response (crushed conspecifics) elicit directed movement using taxis, whereas those that represent less alarming stimuli (a blue crab in the vicinity) or that represent a ubiquitous resource (Spartina) elicit movement by kinesis.     

Mechanisms of resistance to S. mansoni infection: the rat model

Human schistosomiasis is associated with IgE and eosinophilia, feature of a type 2 response. In experimental investigations, murine model has been widely used in order to dissect the immune responses involved in the expression of protective immunity or disease in Schistosoma mansoni infection. Collectively, observations made in this model and in humans demonstrated a strong contrast since a Th2 response in infected mice is involved in the expression of pathology, however, in infected humans the same type of response is rather beneficial for the host. This review will consider the relevance of extrapolating studies of immune responses from experimentally infected rats a semi-permissive host, to studies on S. mansoni infected humans.     

A comparison of the glycolipid compositions of cercarial and adult Schistosoma mansoni and their associated hosts

Glycolipid patterns of cercarial and adult Schistosoma mansoni were determined and compared with those of Biomphalaria glabrata snails and mouse (BALB/c strain) red blood cells by high-performance thin-layer chromatography. Differences in glycolipid content were found between cercariae and adult worms and between these stages and their respective host tissues.     

ITS2 ribosomal RNA indicates Schistosoma hippopotami is a distinct species

The internal transcribed spacer region of the ribosomal RNA, ITS2, was sequenced from a singlé specimen of S. hippopotami collected from a pulmonary artery of the hippopotamus, Hippopotamus amphibius in South Africa. The nucleotide sequence was aligned with those of S. mansoni, S. rodhaini, S. haematobium, S. intercalatum, S. curassoni. S bovis and S. japonicum. Both maximum parsimony and genetic distance analyses were performed on these data sets. Using S. japonicum as outgroup to the African schistosomes, a single most-parsimonious tree was obtained of length 64 steps with a consistency index of 1. S. hippopotami was the sister-group to the remaining African species. This species has lateral-spined eggs and its basal position in the tree suggests that this condition is primitive and that terminal-spined eggs developed secondarily. Molecular data clearly show that S. hippopotami cannot be considered synonymous with S. mansoni. Assuming the hippopotamus is the normal host of S. hippopotami, phylogenetic analysis is consistent with an ancient association between schistosomes and ungulates.     

Molecular cloning and enzymatic expression of the 28-kDa glutathione S-transferase of Schistosoma japonicum: evidence for sequence variation but lack of consistent vaccine efficacy in the murine host

Glutathione S-transferases (GSTs) have long been regarded as attractive vaccine (and drug) targets in schistosomes due to their suspected role in detoxification processes. Indeed, the 28-kDa GST of Schistosoma mansoni (SmGST28) has proven efficacy as an antigen for protective immunity reducing worm burden, female fecundity and egg viability. In contrast, the vaccinating effects of the bacterial expressed homologue of Philippine S. japonicum (SjpGST28) have proved disappointing, possibly because this recombinant form was an incomplete sequence, lacking five N-terminal amino acids which may have affected its vaccination efficacy. Here we describe the cloning and functional enzymatic expression of a complete cDNA encoding SjpGST28. We report also on the immunogenicity and vaccine efficacy of this molecule as a purified recombinant protein and as a DNA plasmid vaccine in the murine model. We further describe the cloning of several complete cDNAs encoding the Chinese homologue of SjpGST28 and the identification of 3 SjcGST28 sequence variants which are probably encoded by distinct alleles.     

外来种无瓣海桑化感作用研究初报*

种子萌发率和胚根长度作为测量指标,以化感效应敏感指数RI为评价指标,研究了无瓣海桑(Sonneratia apetala)各器官的水浸液对海桑(Sonneratia caseolaris)和无瓣海桑种子发芽的影响。结果表明:1)无瓣海桑的各器官水浸液对海桑种子萌发均起抑制作用,高浓度下抑制效应明显;2)不同浓度无瓣海桑根的水浸液对海桑的胚根长均起抑制作用,高浓度下显著抑制,随着浓度降低抑制作用减弱;无瓣海桑的枝、叶和果的水浸液对海桑的胚根长表现为"高抑低促",即高浓度下显著抑制,随着浓度降低抑制作用减弱甚至转为促进;3)无瓣海桑各器官的水浸液对本身种子的发芽表现为"高抑低促";4)无瓣海桑果的水浸液对本身的胚根长表现为高浓度下显著抑制,随着浓度降低抑制作用逐渐减弱;无瓣海桑枝、叶和根的水浸液对本身的胚根长表现为"高抑低促";5)无瓣海桑各器官的水浸液中果的水浸液对海桑或无瓣海桑种子发芽和胚根长的抑制作用最强。