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Molluscicidal activity of 2-hydroxy-3-alkyl-1,4-naphthoquinones and derivatives.

In the search for new molluscicidal agents we tested the activity of lapachol and other 2-hydroxy-3-alkylnaphthoquinones possessing nitrogenated alkyl chains, against the snail Biomphalaria glabrata. Lapachol, isolapachol and nor-lapachol showed strong molluscicidal activity against the adult snail (LD(90)<10 ppm) and significant toxicity against snail egg masses (LD(90)<0.2 ppm). As lapachol is easily extracted, and the derivatives can be synthesised without any difficulty, large-scale synthesis and field tests can be conducted, with a view to large-scale molluscicidal programs.

Animals↗

A newly-identified lineage of Schistosoma.

Because of their role in causing schistosomiasis, flukes of the genus Schistosoma are the best known of all digeneans. The genus has traditionally been divided into four familiar species groups. Here we report on three poorly known species of Schistosoma, one of which, Schistosoma hippopotami, is known from the hippopotamus, one of which is provisionally identified as Schistosoma edwardiense, another hippo parasite, and a third that has not previously been described. All were collected from freshwater snails obtained from Lake Edward, western Uganda, the type locality for both known hippo schistosomes. The three different kinds of schistosome cercariae differ from one another in size, and all are readily differentiated by their long tail stems from the cercariae of human-infecting species. Furthermore, each was recovered from a different genus of snail host, Biomphalaria sudanica, Bulinus truncatus or Ceratophallus natalensis. Molecular analysis, based on 8350 bases of combined nuclear and mitochondrial DNA, groups these three long tail-stem cercariae into a well supported clade that does not associate with any of the recognised species groups. The placement of this clade, basal to all African species plus several Asian species, suggests that there has been an ancient association between Schistosoma and hippos. This new African Schistosoma clade advocates the need for further modification of the traditional species group-based classification. Two of the four species groups are paraphyletic. It also suggests that Schistosoma has been remarkably plastic with respect to adapting to snail hosts-three distantly related genera of planorbid snails have been exploited by worms within a single clade. Finally, it adds a new layer of complexity to deciphering the origins of Schistosoma, often considered to be African but recently challenged as being Asian. In the late Cenozoic the distribution of hippo species straddled both Africa and Asia and they may have provided a means for the introduction of blood flukes to Africa.

Animals↗

Inland valley swamp rice development: malaria, schistosomiasis, onchocerciasis in south central Sierra Leone.

The prevalence of malaria, schistosomiasis and onchocerciasis was determined in 1,106 residents of five villages in the Moyamba District, Southern Sierra Leone, to determine whether inland valley swamp (IVS) development was associated with changes in the prevalence of malaria, schistosomiasis and onchocerciasis in these villages. These parasitic diseases were studied in four villages receiving IVS, Food and Agricultural Organization (FAO) assistance and in one village not receiving FAO assistance. Malaria was the most prevalent infection, detected in 42.6% of the persons examined, followed by O. volvulus (17.7%), S. haematobium (0.6%) and S. mansoni (0.3%). Plasmodium falciparum accounted for 90.4% of the malaria infections, followed by P. malaria (2.1%), P. ovale (0.5%), and mixed infections (7.0%). The trend of infection to O. volvulus increased significantly with an increase in age. S. haematobium (0.6%) and S. mansoni (0.3%) infections were low and no Biomphalaria pfeifferi and Bulinus globosus were found in 33 IVS development swamps examined. These data indicate that IVS development is associated with an increase in the prevalence of malaria infection, but not in the prevalence of O. volvulus, S. haematobium and S. mansoni.

Adult↗

Schistosomiasis control in Saudi Arabia with special reference to the period 1983-1988.

Schistosomiasis control operations carried out between 1983 and 1988 are described, together with information of the impact of schistosomiasis. The prevalence of schistosomiasis has declined from 9.5% in 1983 to 1.8% in 1988 (P less than 0.05). Bulinus truncatus, B. beccarii, and B. reticulatus wrighti are the intermediate hosts of S. haematobium in Saudi Arabia and are distributed unevenly in the different regions. Biomphalaria arabica is the intermediate host of S. mansoni and is found wherever the disease is endemic. The influx of expatriate workers from countries where schistosomiasis is endemic, inaccessible water sources, the constant movement of large numbers of bedouins in some regions, and manpower shortages in some regions are considered to be the main obstacles for schistosomiasis control programmes in Saudi Arabia.

Animals↗

Aquatic modules for bioregenerative life support systems based on the C.E.B.A.S. biotechnology [correction of biotechnilogy].

Most concepts for bioregenerative life support systems are based on edible higher land plants which create some problems with growth and seed generation under space conditions. Animal protein production is mostly neglected because of the tremendous waste management problems with tetrapods under reduced weightlessness. Therefore, the "Closed Equilibrated Biological Aquatic System" (C.E.B.A.S.) was developed which represents an artificial aquatic ecosystem containing aquatic organisms which are adapted at all to "near weightlessness conditions" (fishes Xiphophorus helleri, water snails Biomphalaria glabrata, ammonia oxidizing bacteria and the rootless non-gravitropic edible water plant Ceratophyllum demersum). Basically the C.E.B.A.S. consists of 4 subsystems: a ZOOLOGICAL (correction of ZOOLOGICASL) COMPONENT (animal aquarium), a BOTANICAL COMPONENT (aquatic plant bioreactor), a MICROBIAL COMPONENT (bacteria filter) and an ELECTRONICAL COMPONENT (data acquisition and control unit). Superficially, the function principle appears simple: the plants convert light energy into chemical energy via photosynthesis thus producing biomass and oxygen. The animals and microorganisms use the oxygen for respiration and produce the carbon dioxide which is essential for plant photosynthesis. The ammonia ions excreted by the animals are converted by the bacteria to nitrite and then to nitrate ions which serve as a nitrogen source for the plants. Other essential ions derive from biological degradation of animal waste products and dead organic matter. The C.E.B.A.S. exists in 2 basic versions: the original C.E.B.A.S. with a volume of 150 liters and a self-sustaining standing time of more than 13 month and the so-called C.E.B.A.S. MINI MODULE with a volume of about 8.5 liters. In the latter there is no closed food loop by reasons of available space so that animal food has to be provided via an automated feeder. This device was flown already successfully on the STS-89 and STS-90 spaceshuttle missions and the working hypothesis was verified that aquatic organisms are nearly not affected at all by space conditions, i.e. that the plants exhibited biomass production rates identical to the sound controls and that as well the reproductive, and the immune system as the embryonic and ontogenic development of the animals remained undisturbed. Currently the C.E.B.A.S. MINI MODLULE is prepared for a third spaceshuttle flight (STS-107) in spring 2001. Based on the results of the space experiments a series of prototypes of aquatic food production modules for the implementation into BLSS were developed. This paper describes the scientific disposition of the STS-107 experiment and of open and closed aquaculture systems based on another aquatic plant species, the Lemnacean Wolffia arrhiza which is cultured as a vegetable in Southeastern Asia. This plant can be grown in suspension culture and several special bioreactors were developed for this purpose. W. arrhiza reproduces mainly vegetatively by buds but also sexually from time to time and is therefore especially suitable for genetic engineering, too. Therefore it was used, in addition, to optimize the C.E.B.A.S. MINI MODULE to allow experiments with a duration of 4 month in the International Space Station the basic principle of which will be explained. In the context of aquaculture systems for BLSS the continuous replacement of removed fish biomass is an essential demand. Although fish reproduction seems not to be affected in the shortterm space experiments with the C.E.B.A.S. MINI MODULE a functional and reliable hatchery for the production of siblings under reduced weightlessness is connected with some serious problems. Therefore an automated "reproduction module" for the herbivorous fish Tilapia rendalli was developed as a laboratory prototype. It is concluded that aquatic modules of different degrees of complexity can optimize the productivity of BLSS based on higher land plants and that they offer an unique opportunity for the production of animal protein in lunar or planetary bases.

Ammonia↗

Identification and localization of a schistosome-associated fucosyllactose determinant expressed by Fasciola hepatica.

A Biomphalaria alexandrina-derived lectin (BaSII), of proven specificity to a Schistosoma mansoni-associated fucosyllactose [(Fuc alpha1-2) Gal beta1-4 Glc] determinant, was employed to investigate the putative antigenic cross-reactivity between Schistosoma mansoni and Fasciola hepatica in terms of this structurally-defined oligosaccharide sequence. BaSII affinity column chromatography of extracts of adult worms metabolically radiolabelled with 35S-methionine and analysis by two-dimensional gels established the expression of the fucosyllactose determinant in multiple copies among heterogeneous, acidic glycoproteins synthesized by adult Fasciola hepatica. Direct fluorescence microscopy revealed that determinant-bearing glycoproteins were localized to the external glycocalyx and perikarya of the tegument as well as the epithelial lining of the intestinal caeca and vitelline ducts and glands. Determinant expression was also evident in embryonated cells of eggs and miracidia as well as the intermediate cellular wall of encysted metacercariae, suggesting its conservation during the course of development of the parasite. Based on the structural relatedness of the cross-reactive fucosyllactose determinant to the antigenic mammalian blood group H trisaccharide, our observations may have implications in serodiagnosis and immunoprophylaxis of schistosomiasis/fascioliasis.

Animals↗

Isolation and characterization of an alpha-macroglobulin from the gastropod mollusc Helix pomatia with tetrameric structure and preserved activity after methylamine treatment.

A proteinase inhibitor with M(r) 697000 and 20.3% (w/w) carbohydrate was isolated from the haemolymph of the snail Helix pomatia and characterized. It was shown to have a tetrameric structure with subunits disulphide linked by two. It inhibited the activity of several types of proteinases against large substrates but not that of trypsin against N-alpha-benzoyl-DL-arginine-4-nitroanilide. This indicated a nonspecific and steric hindrance mode of inhibition. The ratio of trypsin molecules inactivated per inhibitor amounted to 1.5. This interaction led to a cleavage of the subunits into two equal fragments and to a slow to fast conformational change of the whole molecules. Experiments with 125I-labelled trypsin indicated that the proteinase had become covalently linked to one of the fragments. Heating of the inhibitor led to autolytic cleavage products but not when methylamine treated. Thiol titration after trypsin or methylamine treatment indicated the presence of one thiol ester bond per subunit. These facts are all indicative of an alpha-macroglobulin type of inhibitor. However, unlike for most of them the methylamine treatment did not induce a conformational change nor suppress its proteinase inhibitory activity. Moreover, invertebrate alpha-macroglobulins are mostly dimeric in structure but tetramers likewise do occur in Biomphalaria glabrata.

Animals↗

The molluscicidal activity of plants used in Brazilian folk medicine.

In a continuing search for new compounds for the control of the vectors of schistosomiasis, we have tested the activity of some Brazilian medicinal plants as sources of molluscicidal natural compounds, using two molluscicidal bioassays. Twenty-seven crude extracts, from twenty-six species belonging to nineteen families, were tested. Seven extracts showed significant molluscicidal activity against Biomphalaria glabrata adults with DL50 values of less than 50 ppm, and five of them were very active in the test using egg masses. The species most active against B. glabrata adults (LD50 value = 3.65 ppm) and their egg masses (LD50 value = 0.13 ppm) was Derris sp. Annona muricata [LD50 value (adult) = 11.86 ppm and LD50 value (egg) = 49.62 ppm], Jatropha elliptica (from Goiás state) [LD50 value (adult) = 24.80 ppm and LD50 value (egg) = 3.03 ppm] and Renealmia exaltata [LD50 value (adult) = 28.03 ppm and LD50 value (egg) = 21.67 ppm], were also considered promising molluscicidal plants.

Animals↗

Synthesis, electrochemistry, and molluscicidal activity of nitroaromatic compounds: effects of substituents and the role of redox potential.

Abstract-Molluscicidal bioassays and electrochemical studies (measurement of first wave reduction potential, Epcl) were performed on several synthetic nitroaromatics, in relation to possible correlation between biological activity, redox potential and structural effects. Five of them presented a significant molluscicidal activity on Biomphalaria glabrata (LD50 < 20 ppm). The Epc1 values ranged from -0.532 to -0.857 V versus Ag/AgCl (0.1 M) (-0.260 to -0.585 V versus NHE), all of them, in the favorable range for reduction in vivo. Data comparison between Epc1 and molluscicidal activity indicates that the presence of the electroactive nitro group is important for the biological activity. Correlation with redox potential, however, was not evident. Structural effects seem to be the most important parameter. Higher activity is noticeable for phenols, including the para-nitro azo or hydrazo-containing compounds. No activity was observed for compounds having the benzylic substituent in meta position to the nitro group. These results suggest that activity undoubtedly involves more than reduction characteristics and that the possible formation of electrophilic species, after nitro reduction, can play an important role in molluscicidal activity against B. glabrata.

Animals↗

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.

Adolescent↗

Interspecific stimulation of parthenogenesis in Schistosoma intercalatum and S. mansoni.

Experimental studies of mating behaviour of Schistosoma intercalatum and Schistosoma mansoni in mixed infections in mice showed that in simultaneous infections, without the possibility of choice of mate, heterologous pairing occurs readily. The paired females reach sexual maturity, are inseminated and lay parthenogenetic eggs. Miracidia originating from the S. mansoni male x S. intercalatum female are non infective to either Biomphalaria glabrata or Bulinus forskalii, whereas those from the reverse cross show a very low infectivity to only B. glabrata. The resulting haploid male cercariae also show a very low infectivity to mice (1.1%) and consequently only a very small number of adult worms develop. It appears from this study, on chromosomal and electrophoretic evidence, that generative (haploid) parthenogenesis occurs in S. mansoni females paired with S. intercalatum males.

Animals↗

Intergeneric reproductive stimulation and parthenogenesis in Schistosoma mansoni.

When mice were infected with female cercariae of Schistosoma mansoni and male cercariae of Schistosomatium douthitti, many mixed pairs formed. The paired females were approximately the same size as those in unisexual infections, far smaller than females paired with S. mansoni males. Although the Sch. douthitti males possessed well-developed testes, sperm were not found in their female partners, which developed scanty vitelline glands and produced laterally spined eggs typical of S. mansoni. Such eggs yielded swimming miracidia infective to the snail host of S. mansoni, Biomphalaria glabrata, but not to the lymnaeid snail host of Sch. douthitti. Sporocysts arising from these miracidia were haploid and produced cercariae infective to mice. Parthenogenetically derived female cercariae in mice co-infected with either parthenogenetically derived male or normal diploid male S. mansoni developed to large adults of normal appearance, whose eggs yielded diploid miracidia and subsequent generations of normal diploid schistosomes. Parthenogenetically derived females co-infected with Sch. douthitti males also paired and produced some eggs containing viable miracidia, which gave rise once again to haploid sporocysts. These observations confirm previous suggestions that the stimulus for maturation in female S. mansoni is distinct from that for growth, and is independent of insemination and fertilization. It is concluded that both diploid and haploid S. mansoni females are capable either of parthenogenesis or of bisexual reproduction when appropriately stimulated.

Animals↗

Schistosoma mansoni: quantification of skin penetration and early migration by differential external radioassay and autoradiography.

Eleven gamma-emitting radionuclides (49Sc, 54Mn, 59Fe (+2 and +3), 60Co, 65Zn, 75Se (as selenomethionine, selenocystine, selenite and selenate), 109Cd, 125Sb, 133Ba, 137Cs and 203Hg) were screened as labelling agents for Schistosoma mansoni cercariae by incubation of infected Biomphalaria glabrata snails in radioactive solution according to the technique of Christensen (1977). Only [75Se]methionine yielded satisfactorily labelled cercariae. Multiple regression analysis of volume, number of cercariae and radioactivity from a series of 10 aliquots of unwashed cercarial suspensions yielded estimates of unbound and cercarial bound radioactivity that were equivalent or superior to estimates based on assay of washed cercariae and eliminated loss of cercariae. Washing of cercarial suspensions over 8 micrometers pore diameter Millipore filters was found to result in entanglement of 60-90% of the cercariae on the filter disc. Differential external radioassay, a new technique employing partial body shielding within a total body counter, permitted separate estimation of tail and body radioactivity of conscious mice previously exposed by tail immersion to 75Se-labelled cercariae, with measurements repeated as often as desired. Approximately 39% of the 75Se present in emergent cercariae was retained by schistosomula transformed in vitro but this was subject to considerable variation, especially in schistosomula transformed in vivo. Secreted or catabolized label from penetrant cercariae and schistosomula was rapidly removed from the skin by the bloodstream. Numbers of schistosomula in tail skin were directly proportional to the number of reduced silver foci counted on tail autoradiograms; only a very small fraction of tail radioactivity represented unbound ('spurious') label. Migration of schistosomula away from skin was 50% complete at 3.8-4.3 days, as determined by probit analysis of autoradiographic data.

Animals↗

Schistosomicidal activities of Lymnaea stagnalis haemocytes: the role of oxygen radicals.

Macrophage-like defence cells (haemocytes) of the pond snail Lymnaea stagnalis mediate cytotoxicity through reactive oxygen intermediates (ROIs). This activity is NADPH-oxidase dependent, as in mammalian phagocytes during the respiratory burst. In this study, mother sporocysts of schistosomes, the compatible Trichobilharzia ocellata and the incompatible Schistosoma mansoni evoke in vitro ROI activities (detected by luminol dependent chemiluminescence, LDCL) from L. stagnalis haemocytes. S. mansoni is encapsulated by haemocytes and eliminated, whereas T. ocellata escapes encapsulation and survives. Both schistosomes were equally susceptible to in vitro oxidative damage from exposure to hydrogen peroxide and to ROIs generated by a xanthine/xanthine oxidase system. Protocatechuic acid, a specific antagonist of NADPH-oxidase, delayed the killing of T. ocellata and S. mansoni sporocysts by haemocytes of resistant snails (Biomphalaria glabrata and L. stagnalis, respectively). We conclude that ROIs take part in haemocyte-mediated cytotoxicity. However, neither a snail's capability to generate ROIs, nor a schistosome's susceptibility to ROIs, determine snail/schistosome incompatibility. Snail/schistosome compatibility is rather determined by the parasite's ability to modulate haemocyte behaviour such that effective encapsulation and the generation of lethal concentrations of ROIs are prevented.

Animals↗

Description of a tandem repeated DNA sequence of Echinostoma caproni and methods for its detection in snail and plankton samples.

Echinostome larval stages in the snail have a great potential as effective competitors for the control of schistosomes and adult worms can cause painful intestinal diseases in humans. Ecology and transmission of the larval stages of trematodes are poorly understood, especially because their identification in field-collected samples by microscopy is difficult. We cloned, sequenced and analysed a 192 bp tandem repreated DNA sequence of Echinostoma caproni (EcSau3A), an often discussed antagonist of Schistosoma mansoni in Biomphalaria snails. PCR primers against this sequence can detect less than 10 fg of E. caproni DNA, 2 miracidia in snails 1 day p.i., 1 metacercaria in 50 mg snail tissue and 1 cercaria in 50 mg plankton with high specificity. Methods described in this study can support the discovery of fundamental ecological principles on distribution, host specificity and epidemiology of E. caproni larvae under field conditions.

Animals↗

Evaluation of biological activities of triterpenoid saponins from Maesa lanceolata.

From a bioassay-guided fractionation procedure using Herpes simplex virus type 1 as the target model, a virucidal saponin mixture (maesasaponin mixture B) was isolated from the MeOH extract of leaves of Maesa lanceolata. The maesasaponin mixture B consisted of six homologous oleanane-type triterpenoid saponins 1-6, identified by 1H, 13C, and 2D NMR spectroscopy and mass spectrometry. The maesasaponin mixture B (1-6) showed several biological activities expected for saponins. It exhibited a moderate virucidal activity against enveloped viruses. The maesasaponin mixture B (1-6) was tested for further properties. The saponin mixture was found to be highly hemolytic and molluscicidal: it hemolyzed 50% of human erythrocytes (1% suspension) at a concentration estimated at 1.6 microg/mL and exerted against Biomphalaria glabrata snails a severe toxic effect with LD95 and LD50 values of 4.1 and 2.3 ppm, respectively. In addition, the maesasaponin mixture B (1-6) showed moderate fungistatic and antimutagenic properties. The evaluation of these diverse activities is described.

Animals↗

Daubaylia helicophilus n. sp. (Daubayliidae: Nematoda) a parasite of the snail, Gyraulus spirillus from Taiwan.

The nematode, Daubaylia helicophilus sp. n., is described from the snail, Gyraulus spirillus, originating from Taiwan. This species is distinguished from previously described members of the genus by the shape of the tail in both sexes, as well as the overall length and size and shape of the spicules and gubernaculum, respectively. Populations of infected snails have been maintained for a period of years and ingestion of the gravid female nematode seems to be at least one mode of infection. D. helicophilus will also infect other snails, experimentally, including the schistosome-bearing snail, Biomphalaria glabrata.

Animals↗

Susceptibility of rodents to infection with Schistosoma mansoni in Richard-Toll (Senegal).

The susceptibility of Arvicanthis niloticus, Mastomys huberti, Mastomys erythroleucus and Mus musculus was studied to assess the capacity of these rodents to transmit Schistosoma mansoni. The susceptibility was determined by the percentage of adult schistosomes recovered, the number of eggs per gramme of faeces, the viability of these eggs and the capacity of the rodents to maintain the life cycle of Schistosoma mansoni. The percentages of adult worms recovered were respectively 18%, 11.5%, 8.4% and 20.5% in A. niloticus, M. huberti, M. erythroleucus and M. musculus. After infection, they liberate in the environment viable eggs whose miracidia are infectious for the intermediate host (Biomphalaria pfeifferi). The mean egg load was 300 +/- 327.8 in A. niloticus; 664 +/- 673.5 in M. huberti; 240 +/- 304.8 in M. erythroleucus; 400 +/- 361.5 in M. musculus.

Animals↗