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PubMed1998

Recombinant human interleukin-1-mediated killing of Schistosoma mansoni primary sporocysts in Biomphalaria glabrata.

Previous work has indicated that injection of recombinant-human interleukin (rhIL)-1beta in Schistosoma mansoni-infected M-line Biomphalaria glabrata resulted in a significant reduction in the number of cercariae shed. The purpose of the present work was to determine if primary sporocysts were killed following rhIL-1beta injection in susceptible snails and, if so, to determine if killing was the direct result of hemocyte activity. Counting of primary sporocysts indicated a 50% reduction in the number surviving at 3 days PE in snails from 2 susceptible strains following injection. Histological analysis indicated that killing occurred with little-to-no observable hemocyte/parasite contact, whereas short-term culture of primary sporocysts with cell-free plasma (hemolymph) from injected snails rapidly initiated killing in vitro. Because levels of a snail IL-1-like molecule (SnaIL-1) drop significantly following schistosome exposure in M-line snails, because resistant snails maintain higher SnaIL-1 levels following infection, and because rhIL-1beta upregulates hemocyte cytotoxic mechanisms, these data support the contention that SnaIL-1 plays a role in determining resistance in B. glabrata. These data also indicate that schistosome death may be separated from parasite encapsulation by hemocytes and that an as yet unidentified humoral killing mechanism/factor may exist in B. glabrata. Lastly, these data further support the hypothesis that cytokine-like molecules are important, functionally conserved immunodefense mediators in both vertebrates and invertebrates.

Analysis of Variance↗
PubMed1998

Effects of Plagiorchis elegans (Digenea: Plagiorchiidae) infection on the reproduction of Biomphalaria glabrata (Pulmonata: Planorbidae).

Infection with the digenean parasite Plagiorchis elegans dramatically reduced the reproductive output of Biomphalaria glabrata exposed to the parasite as juveniles or adults. The total number of eggs produced by infected snails was reduced to approximately 7 and 13% of control values, respectively. Parasitic castration was attributed to the presence of mother sporocysts that readily established in the tissues of this incompatible host. Infection did not result in the production of cercariae but significantly shortened the life span of juvenile and adult B. glabrata by approximately 23 and 10%, respectively. Plagiorchis elegans also castrated its compatible host, Stagnicola elodes.

Animals↗
PubMed2001

Effects of the serotonin receptor ligand methiothepin on reproductive behavior of the freshwater snail Biomphalaria glabrata: reduction of egg laying and induction of penile erection.

The biogenic monoamine serotonin (5-HT) has been reported to enhance egg laying in the freshwater gastropod Biomphalaria glabrata, an intermediate host for human blood flukes. Methiothepin, a vertebrate 5-HT(1/2) receptor ligand which binds with high affinity to a 5-HT receptor (5-HTlym) in Lymnaea stagnalis was tested for its ability to block egg laying in B. glabrata as a possible target for snail control. A single 30-min application of methiothepin (1 microM) was sufficient to prevent egg laying for over 1 week and did so in a dose-dependent fashion. Furthermore, single applications of methiothepin (1 microM and 10 microM) induced penile erection in a high percentage of snails tested. Latency to erection was long (at least 8 hr), but the duration of erection was long-lasting (up to 48 hr). Despite the erections, methiothepin-treated snails failed to achieve copulations. The pharmacological effect of methiothepin on both male and female reproductive processes is similar to that produced in other molluscs, and points to the gene for the 5-HT receptor mediating or modulating both or either processes as a potential target of snail control strategies. J. Exp. Zool. 289:202-207, 2001.

Animals↗
PubMed2001

Parasite-responsive IgSF members in the snail Biomphalaria glabrata: characterization of novel genes with tandemly arranged IgSF domains and a fibrinogen domain.

Two novel genes of the immunoglobulin superfamily (IgSF), FREP3 and FREP7, are reported from the snail Biomphalaria glabrata, a prominent intermediate host of the human parasite Schistosoma mansoni. They resemble other B. glabrata genes that encode fibrinogen-related proteins (FREPs), but differ in that they encode proteins with two tandemly arranged IgSF domains followed by a C-terminal fibrinogen domain. FREPs are hemolymph proteins that increase in abundance following exposure to a digenetic trematode, Echinostoma paraensei, and that bind to and precipitate parasite antigens. Within each gene, the two IgSF-coding regions are dissimilar from one another: the N-terminal IgSF1 domain is encoded by a single exon whereas the downstream IgSF2 domain is encoded by three exons. For both FREPs 3 and 7, the IgSF2 domain belongs to the variable (V) type, whereas the IgSF1 domain is not easily classified with respect to IgSF type. The fibrinogen-encoding region in both genes is relatively conserved and lacks introns. FREP3 exhibits extensive variation in the IgSF1 region. A ratio of nonsynonymous versus synonymous substitutions of 2.56 suggests that this region is under positive selection. A genomic fragment identifiable as FREP7 but lacking an exon was also found, further suggestive of variability within FREP IgSF-encoding regions. Insofar as FREPs are hypothesized to function in nonself recognition, the identification of additional novel FREP genes as part of a growing gene family in B. glabrata is of interest. Such genes, particularly given their variable nature, serve as a model to study the complexity of invertebrate defense responses.

Amino Acid Sequence↗
PubMedDate not supplied

Characterization of fucosyllactose determinant-bearing glycoproteins probed by a Biomphalaria alexandrina lectin in Schistosoma mansoni cercariae.

Utilizing a Biomphalaria alexandrina-derived lectin (BaSII) of proven specificity to a Schistosoma mansoni-associated fucosyllactose [(Fuc alpha 1-2) Gal beta 1-4 Glc] determinant, two determinant-bearing glycoproteins of 40 and 37 kDa were found to be synthesized by the cercarial stage of the parasite. The two glycoproteins were isolated by BaSII affinity column chromatography from extracts of cercariae metabolically radiolabelled with 35S-methionine. Treatments with endoglycosidases, alkaline borohydride, as well as concanavalin A column chromatography and analysis by two-dimensional gels indicated that the two glycoproteins are synthesized as a single 33 kDa polypeptide backbone that is differentially glycosylated with one and/or two determinant-bearing N-linked complex-type glycan units of either the biantennary, or, to a lesser extent, the tri- or tetra-antennary types. The two glycoproteins lack other conventional high mannose-type or O-linked glycans, and the distinct structures of the complex-type oligosaccharides accounted solely for the expression of three isomorphs for each determinant-bearing glycoprotein. Based on the structural relatedness of the fucosyllactose determinant to the antigenic mammalian blood group H trisaccharide, our observations may have implications in mechanisms of host-parasite interactions as well as immunoprophylaxis.

Animals↗
PubMed1992

Genetics of Biomphalaria glabrata and its effect on the outcome of Schistosoma mansoni infection.

The genetics of the snail Biomphalaria glabrata is better characterized than that of any other intermediate host of schistosomes of humans. Using techniques of selective breeding, several snail stocks have been developed that consistently display resistant or susceptible phenotypes. Investigators using these stocks have learned that several snail and parasite genes influence the course of parasite development. Here, Charles Richards, Matty Knight and Fred Lewis discuss the importance of the snail's genetics in categorizing resistance in this complex invertebrate, some recent molecular evidence that may help us understand several of the problems that still remain, and some challenges lying ahead for investigators in this field.

Journal Article↗
PubMed1977

Glycogen metabolism in the snail, Biomphalaria glabrata.

1. The distribution of glycogen in different tissues of the snail, Biomphalaria glabrata was determined; the cephalopedal region contained 15-17 mg, the mantle region 23-29 mg, the hepatopancreas 31-45 mg, and the ovotestis 50 mg of glycogen/g wet wt. 2. There was a significant decrease in the glycogen content in the cephalopedal region after incubating the snails with 5-hydroxytryptamine (4 x 10(-5) M) in vivo. 3. Enzymatic degradation of glycogen from different tissues revealed that the degree of branching was 9% of the total glucosyl residues, and the length of outer branches was about 40% of the total glucosyl residues. 4. There was an active form of glycogen phosphorylase in the cephalopedal region and the mantle region. Phosphorylase in the hepatopancreas was generally inactive, but could be activated by an endogenous phosphorylase kinase. 5. Glycogen synthetase of the snail tissues required glucose-6-phosphate to be active.

Animals↗
PubMed2004

Dominant lethal effect of 60Co gamma radiation in Biomphalaria glabrata (SAY, 1818).

The dominant lethal effects of gamma radiation of 60Co in the snail Biomphalaria glabrata were studied. Three groups of 13 wild-type snails were irradiated with single doses of 2.5; 10 and 20 Gy. Crossings were carried out at intervals of 7, 17, 23, 30 and 36 days after irradiation. The dominant lethal effect was observed only at the first crossing occurring 7 days after irradiation with 2.5 Gy. With 10 and 20 Gy, the induction of lethal mutations was detected at 7, 17 and 23 days after irradiation; a dose-response effect was observed. The effect was stronger 7 days after irradiation, decreasing in the succeeding crossings up to 30 days. Cell-killing effects on germ cells were detected in the crossings at 23 days and 30 days after irradiation with 20 Gy. After 36 days, frequencies of malformations resumed background levels; crossing rates partially recovered. These results show that gamma radiation affected all the stages of spermatogenesis. Germ cells at later phases were more sensitive to the mutagenic effect of radiation and the cell killing effects were observed on the youngest cells. This response was similar to the highly homogeneous pattern observed in widely different species and allowed us to estimate some parameters of spermatogenesis in B. glabrata.

Age Factors↗
PubMed2006

Inhibition of cholinesterase activity by azinphos-methyl in two freshwater invertebrates: Biomphalaria glabrata and Lumbriculus variegatus.

In this study, some biochemical features and the extent of inhibition induced by the organophosphorous pesticide azinphos-methyl on the cholinesterase (ChE) activity present in whole soft tissue of two freshwater invertebrate species, the gastropod Biomphalaria glabrata and the oligochaete Lumbriculus variegatus were investigated. Both invertebrate organisms presented marked differences in ChE activity, type of enzymes and subcellular location. Acetylthiocholine was the substrate preferred by B. glabrata ChE. The enzyme activity was located preferentially in the supernatant of 11,000 x g centrifugation and was inhibited by increasing concentrations of substrate but not by iso-OMPA. Results showed that there were progressive inhibitions of the enzyme activity, with values 21%, 59%, 72%, 76%, and 82% lower than the control at levels of 1, 10, 50, 100 and 1000 microM of eserine, respectively. In contrast, L. variegatus ChE activity was distributed both in the supernatant and pellet fractions, with values approximately 6 and 20 times higher than B. glabrata, respectively. Studies with butyrylthiocholine and iso-OMPA suggested that about 72% of the activity corresponded to butyrylcholinesterase. A strong enzyme inhibition (88-94%) was found at low eserine concentrations (1-10 microM). ChE activity from L. variegatus and B. glabrata was inhibited by in vivo exposure to azinphos-methyl suggesting that both species can form the oxon derivative of this pesticide. However, both invertebrate species showed a very different susceptibility to the insecticide. The NOEC and EIC50 values were 500 and 1000 times lower for L. variegatus than for B. glabrata, reflecting that the oligochaetes were much more sensitive organisms. A different pattern was also observed for the recovery of the enzymatic activity when the organisms were transferred to clean water. The recuperation process was faster for the oligochaetes than for the gastropods. Mortality was not observed in either of the experimental conditions assayed, not even at concentrations that induced 90% of ChE inhibition. The differences in substrate specificity, sensitivity to inhibitors, and subcellular location between the ChEs of B. glabrata and L. variegatus could be the main factors contributing to the differential susceptibility to azinphos-methyl ChE inhibition found in the present study.

Animals↗
PubMed2001

Differential gene expression in haemocytes of the snail Biomphalaria glabrata: effects of Schistosoma mansoni infection.

Parasite encapsulation and destruction in Biomphalaria glabrata has been shown to involve the cellular component of the snail's internal defence system, the haemocytes. To identify genes involved in the immunobiology of these cells, we used the method of differential display reverse transcriptase polymerase chain reaction (DDRT-PCR) to investigate differential gene regulation in haemocytes isolated from Schistosoma mansoni exposed and unexposed snails. RNA isolated from circulating haemocytes from resistant snails (BS-90 stock), previously exposed to S. mansoni, was analysed using 12 different arbitrary primers in conjunction with an anchored Oligo d(T(11)CG) primer. Transcription profiles between haemocytes of parasite exposed and unexposed snails were compared and a total of 87 differentially regulated bands were identified and isolated. Of these, 65 bands were cloned and used as probes in Southern blots to show the presence of corresponding sequences in the snail genome. RT-PCR was performed to verify the regulation of these transcripts. DNA sequence analysis showed that the majority of the cloned sequences were novel, although a few showed a high degree of sequence similarity to other sequences in the DNA and protein databases. One of these included a differentially expressed transcript that showed a significant degree of sequence identity to E. coli transposase Tn5, an enzyme whose activity is normally associated with generating mobility and instability in the genome.

Amino Acid Sequence↗
PubMed2000

Calcium binding constituents of the organic shell matrix from the freshwater snail Biomphalaria glabrata.

The Ca2+ binding of an EDTA-free water-soluble (SM) and -insoluble (IM) organic matrix of the freshwater snail Biomphalaria glabrata was investigated, using a 45Ca2+ autoradiography after SDS-electrophoretical separation and a calcium binding assay. Electrophoresis of the SM showed a considerable amount of Alcian blue and Stains all positive material, regarded as glycosaminoglycans (GAGs) or proteoglycans (PGs). This part of the SM was slightly positive after 45Ca2+ autoradiography at pH 6.8. The Ca2+ binding increased, raising the pH to 7.4 and 8.0 and was especially strong when simulating the real conditions of the extrapallial space with a carbonate buffer of pH 7.4. The Ca2+ binding assay of the IM showed the same pH-dependency that was observed in the SM. The titration of the IM with Ca2+ at pH 8.0 lead to a dissociation constant of 7.5 x 10(-5) M. While Mg2+ displaced 45Ca2+ in the same way as nonradioactive Ca2+, an approximately 400-fold amount of Na+ was necessary to reduce the binding of 45Ca2+ to 50%. The Ca2+ binding of the organic matrix from the B. glabrata shell appears to be a process of low specificity, medium affinity and high pH-dependency. Apparently, acidic carbohydrate-rich PGs are the only calcium binding constituents of the organic shell matrix.

Animals↗
PubMed2000

Monoamines and their metabolites in the freshwater snail Biomphalaria glabrata.

Metabolism of the major monoamines and their functions were studied in the freshwater snail Biomphalaria glabrata. In both juvenile and adult snails, the plasma (cell-free hemolymph) appears to act as a reservoir for most of these monoamines and their metabolites including among others, L-dopa and dopamine as major constituents. Significant quantities of L-tryptophan, precursor of indoleamines, also was found in the plasma. L-dopa, serotonin, homovanillic acid and dopamine were prominently represented in the central nervous system of the snail, while serotonin and its metabolites, 5-hydroxyindole acetic acid and 5-hydroxytryptophol were found in the ovotestis. Catecholamines such as L-dopa, dopamine and homovanillic acid were identified in the albumen gland. Functional aspects of both dopamine and serotonin were studied using in vitro cultures of albumen glands, the site of perivitelline fluid and galactogen synthesis in B. glabrata. Dopamine was found to stimulate the release of secretory proteins when exogenously added to gland cultures and this process was inhibited by chlorpromazine, a dopamine receptor antagonist. Similarly, exogenous serotonin stimulated in vitro protein secretion by albumen glands. Thus, these results suggest that monoamines may play important roles in regulating reproductive activity of this snail and provides an excellent model for studying neurotransmitter function and metabolism in molluscs.

Animals↗
PubMed2002

Monoamines in the albumen gland, plasma, and central nervous system of the snail Biomphalaria glabrata during egg-laying.

The potential role of selected biogenic monoamines and related compounds in the reproductive physiology of the freshwater snail Biomphalaria glabrata was investigated. Extracts of the albumen gland (AG), plasma, and central nervous system (CNS) were subjected to high pressure liquid chromatography with electrochemical detection (HPLC-ED), and under the extraction and separation conditions employed the following amines were detected: tyrosine, dihydroxyphenylalanine (DOPA), dopamine, and tryptophan in the AG; DOPA, tyrosine, and tryptophan in the plasma; DOPA, tyrosine, dopamine and 5-hydroxytryptamine in the CNS. These compounds were then quantified in individual samples taken from snails known to be in a particular stage of the egg-laying process. AG dopamine levels were highest in snails in the first stage of the reproductive process, when the AG is secreting perivitelline fluid around each fertilized ovum. Significant decreases in AG protein content during the later stages of the egg-laying process were also evident. Plasma tyrosine and DOPA levels were lowest in snails that contained a fully packaged egg mass, while no changes in monoamine content were observed in the CNS. These data provide insights into the role(s) that monoamines, especially catecholamine-related compounds, may play in B. glabrata reproductive physiology.

Animals↗
PubMed2000

Compatibility of Schistosoma mansoni Cameroon and Biomphalaria pfeifferi Senegal.

The vectorial capacity of Biomphalaria pfeifferi from Ndiangue, Senegal, was investigated with an allopatric isolate of Schistosoma mansoni from Nkolbisson, Cameroon. The snail infection rate after exposure to a single miracidium per snail (MD1) was 56. 3 %, and 91.6%, for snails exposed to 5 miracidia per snail (MD5). The minimum pre-patent period was 21 days. The mean total cercarial production for the MDI group was 18,511 cercariae per snail, and 9757 cercariae for the MD5 group. The maximum production of cercariae for 1 day was 4892 observed in a snail from the MDI group at day 43 post-infection. The mean longevity of snails was higher in group MD1 (88 days p.i.) than in group MD5 (65 days p.i.). The chronobiological emergence pattern revealed a circadian rhythm with one shedding peak at mid-day. Comparisons are made with the vectorial capacity of the sympatric combination of B. pfeifferi Senegal/S. mansoni Senegal.

Animals↗
PubMed2003

Phylogeography of Biomphalaria glabrata and B. pfeifferi, important intermediate hosts of Schistosoma mansoni in the New and Old World tropics.

The historical phylogeography of the two most important intermediate host species of the human blood fluke Schistosoma mansoni, B. glabrata in the New World, and B. pfeifferi in the Old World, was investigated using partial 16S and ND1 sequences from the mitochondrial genome. Nuclear sequences of an actin intron and internal transcribed spacer (ITS)-1 were also obtained, but they were uninformative for the relationships among populations. Phylogenetic analyses based on mtDNA revealed six well-differentiated clades within B. glabrata: the Greater Antilles, Venezuela and the Lesser Antilles, and four geographically overlapping Brazilian clades. Application of a Biomphalaria-specific mutation rate gives an estimate of the early Pleistocene for their divergence. The Brazilian clades were inferred to be the result of fragmentation, due possibly to climate oscillations, with subsequent range expansion producing the overlapping ranges. Within the Venezuela and Lesser Antilles clade, lineages from each of these areas were estimated to have separated approximately 740 000 years ago. Compared to B. glabrata, mitochondrial sequences of B. pfeifferi are about 4x lower in diversity, reflecting a much younger age for the species, with the most recent common ancestor of all haplotypes estimated to have existed 880 000 years ago. The oldest B. pfeifferi haplotypes occurred in southern Africa, suggesting it may have been a refugium during dry periods. A recent range expansion was inferred for eastern Africa less than 100 000 years ago. Several putative species and subspecies, B. arabica, B. gaudi, B. rhodesiensis and B. stanleyi, are shown to be undifferentiated from other B. pfeifferi populations.

Africa↗
PubMed2004

Long-term controlled field experiment on the competition between two species of Biomphalaria (Mollusca, Basommatophora), the snail vectors of Schistosoma mansoni in Northeastern Brazil.

A long-term controlled field experiment on the interactions of the populations of Biomphalaria glabrata (target population) and B. straminea (competitor) was carried out in the county of Alhandra, state of Paraíba, Brazil, during the period 1980 through 1989. Results obtained in the current paper show that the snail B. straminea has strong competitive advantages over B. glabrata. In six out of nine streams the native population of B. glabrata were totally excluded and replaced by B. straminea. There is evidence showing that seasonal dryness has marked influence on the phenomenon studied in this paper. In all the streams were B. straminea already predominated, return of B. glabrata was never observed.

Journal Article↗
PubMed1996

[Schistosomiasis: new occurrence of Biomphalaria straminea in Belo Horizonte, Minas Gerais]

In Belo Horizonte, Minas Gerais, Brazil, the snail Biomphalaria straminea was found in the Pampulha region. Recently the snail was found in ditches in the old Santa Lúcia Reservoir, a now-extinct focus of schistosomiasis transmission by B. glabrata. The snails were collected and examined to verify whether they were naturally infected with Schistosoma mansoni. Negative specimens were used for breeding or infection with the LE strain of S. mansoni from the laboratory and another strain obtained from eggs found in the feces of a schoolchild (VGS) from Belo Horizonte. Among the 1890 snails collected from 1994 to 1995, none were infected with S. mansoni. Among 87 snails collected and exposed to the LE strain, 9 (10.3%) shed cercariae; among 83 snails from F1, and exposed to LE, 10 (12.0%) shed cercariae. Among 88 snails exposed to VGS, 10 (11.3%) shed cercariae. In Belo Horizonte, schistosomiasis is transmitted by B. glabrata and B. tenagophila. However, currently there exists the risk of appearance of a focus in which B. straminea could be the host if prophylactic measures are not taken by the authorities responsible for the construction of a park and lake at that site.

Journal Article↗
PubMed1966

The ecology of Bulinus truncatus and Biomphalaria alexandrina and its implications for the control of bilharziasis in the Egypt-49 project area.

The respective vectors of the two forms of bilharziasis in Egypt do not have the same ecological distribution. Bulinus truncatus is most abundant in large canals, and decreases in density as the water approaches and flows into drains. Biomphalaria alexandrina is most abundant in drains, and decreases in density upstream from these habitats. Both species are most abundant in the presence of aquatic vegetation, but they differ in their respective associations with the water hyacinth, Eichhornia crassipes. Biomph. alexandrina reaches maximum abundance in the presence of this plant, but Bul. truncatus is as uncommon in the absence of plants as in the presence of E. crassipes.Calculation of life-table parameters from field data shows that, under optimum field conditions, both species can double their populations in 14-16 days. The reproductive rates of both species are greatest in March and the death rates in midsummer. The observed peak densities in May and June give a false impression of optima because of undercollection of young snails, which are most abundant in March and April.Control operations should take advantage of the findings on population parameters. A single area-wide treatment with molluscicide in April is recommended. During the remainder of the year, search for isolated foci of snail breeding and individual treatment of these will effect large savings of chemical and will be effective in controlling the transmission of the parasites.

Animals↗