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Description of the adult worms of a new Brazilian isolate of Echinostoma paraensei (Platyhelminthes:Digenea) from its natural vertebrate host Nectomys squamipes by light and scanning electron microscopy and molecular analysis.

Echinostoma paraensei Lie and Basch, 1967 (Echinostomatidae:Platyhelminthes), a 37 collar spine echinostome of the "revolutum group", has been used extensively as a model organism to study the interactions of digenetic trematodes with both their snail and vertebrate hosts. This worm was first isolated from the snail Biomphalaria glabrata from Belo Horizonte (BH isolate), Minas Gerais State, Brazil, by Lie and Basch [J Parasitol (1967) 53:1192-1199]. The natural definitive host for the BH isolate was never determined, and it has been maintained in the laboratory since 1967 in B. glabrata and hamsters. In this study, using light and scanning electron microscopy and molecular analysis, we describe an echinostome recently obtained from its natural vertebrate host, the wild rodent Nectomys squamipes (Rodentia: Sigmodontinae) from Sumidouro, Rio de Janeiro State, Brazil (RJ isolate). This echinostome was also compared to the laboratory-maintained BH isolate of E. paraensei. We observed that adult worms of both BH and RJ isolates could be differentiated from other echinostome species by the relatively small size of the dorsal collar spines relative to lateral and corner collar spines. SEM confirmed the similarity of this morphological character between the two isolates. As additional diagnostic features, the tegumentary spines are scale-like and the region between the genital pore and the acetabulum lacks scales. There is a folded protuberance with a pore just posterior to the genital pore. The tegument of the acetabulum is unspined and radially wrinkled, and there are numerous randomly distributed small, domed, ciliated papillae. The sequences of the internal transcribed spacers of the nuclear rDNA complex of the RJ and BH isolates are identical. Together these shared features provide strong evidence that both isolates are the same and can be referred to as E. paraensei. In conclusion, we have identified, for the first time, one of the natural definitive hosts for E. paraensei, the rodent N. squamipes, and have extended the known geographical distribution of this species to include Sumidouro in Rio de Janeiro State, Brazil.

Animals↗

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

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

Animals↗

Morphology of neurosecretory cells in basommatophoran snails homologous with egg-laying and growth hormone-producing cells of Lymnaea stagnalis.

In a light and electron microscope study, neurosecretory cells morphologically homologous with the egg-laying hormone-producing caudodorsal cells (CDC) and growth hormone-producing dorsal cells (DC, light green cells) of the freshwater basommatophoran snail Lymnaea stagnalis have been found in five genera (seven species) of Basommatophora, viz. in Lymnaea palustris and Lymnaea ovata, in Planorbis planorbis and Planorbis vortex, in Planorbarius corneus, in Bulinus truncatus, and in Biomphalaria glabrata. It is concluded that the functions of these cells are homologous as well. The homologies of the respective neuron types regard their locations in the cerebral ganglia, their clustering in groups, location of their neurohemal area (CDC: cerebral commissure; DC: median lip nerves), and ultrastructural characteristics (e.g., abundance of rough endoplasmic reticulum, well-developed Golgi apparatus, presence of two types of neuron-specific secretory granules, and release of granule contents by exocytosis into the hemolymph). In addition, CDC show large electron-dense granules and DC reveal infoldings of the plasma membrane at the abaxonal side of the soma as well as synaptic input. On the other hand, each neuron type shows species-specific characteristics, particularly with regard to the number of cells and the structure of the neurohemal area. Furthermore, the CDC show marked differences between genera in the morphology (especially the mean diameter) of type 2 and, particularly, type 1 secretory granules. The morphology of the two types of secretory granules in the DC differs strongly between species. The possible relation between the morphology and the chemical contents of secretory granules has been discussed.

Animals↗

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

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

Animals↗

A comparative study of hemocytes from six different snails: morphology and functional aspects.

Hemocytes taken from six different gastropod snails, Achatina achatina, A. fulica, Biomphalaria glabrata, Bulinus natalensis, Helix aspersa, and Lymnaea stagnalis, were compared for morphology, peroxidase activity, and, using methods developed for L. stagnalis, the ability to generate reactive oxygen inermediates upon phagocytic stimulation. Numbers of hemocytes per milliliter hemolymph and hemocytes' microscopical morphology showed some variation among the snail species. Peroxidase activity was demonstrated in all snail hemocytes except in those of B. glabrata and A. fulica. Hemocytes of all species generated superoxide upon phagocytic stimulation with zymosan (tested by superoxide dismutase-inhibitable reduction of nitroblue tetrazolium). When tested, hemocytes of A. achatina and of A. fulica displayed luminol-dependent chemiluminescence activity.

Animals↗

Molluscicidal saponins from Catunaregam nilotica.

Two new saponins were isolated from the fruits of Catunaregam nilotica Stapf, syn. Lachnosiphonium nilotica; Randia nilotica; Xeromphis nilotica. Their structures were determined mainly by spectroscopic methods as 3- O-[O-alpha-L-rhamnopyranosyl-(1-->3)-O-[O-beta-D-glucopyranosyl-(1 -->3)]- beta-D-glucopyranosyl]oleanolic acid and 28-O-beta-D- glucopyranosyl-3-O-[O-alpha-L-rhamnopyranosyl-(1-->3)-O[O-beta-D- glucopyranosyl]-beta-D-glycopyranosyl]oleanolate. The monodesmosidic saponin is a potent molluscicide against the schistosomiasis transmitting snail Biomphalaria glabrata with a LC50 value of 3 ppm. In addition two known saponins, 3-O-[2', 3'-di-O-(beta-D-glucopyranosyl)-beta-D- glucopyranosyl]oleanolic acid and 3-O-[O-beta-D-glucopyranosyl(1-->3)- beta-D-glucopyranosyl]oleanolic acid, were identified and their molluscicidal activity determined, the LC50 values being 26 and 3 ppm, respectively. Initial molluscicidal screening of the crude water and ethanol extracts revealed 100% snail mortality at concentrations of 100 and 50 ppm, respectively. The haemolytic activity of the molluscicidal saponins was determined as well and the HC50 values towards bovine erythrocytes found to be 3 ppm for the new saponin, and 16 and 2 ppm, respectively, for the two known saponins.

Animals↗

Use of praziquantel in patients with schistosomiasis mansoni previously treated with oxamniquine and/or hycanthone: resistance of Schistosoma mansoni to schistosomicidal agents.

Fourteen patients with active schistosomiasis mansoni in spite of previous treatment with oxamniquine and/or hycanthone were treated with praziquantel, single oral dose of 45 to 50 mg/kg body-weight. All underwent clinical, laboratory and electrocardiographic examination before and after treatment. Untoward effects (dizziness, drowziness, nausea and abdominal pain) were observed in ten. Laboratory findings disclosed no significant alteration and the electrocardiograms showed no abnormalities. Monthly follow-up examinations of 13 patients for six consecutive months showed parasitological cure in all. Before praziquantel treatment strains of Schistosoma mansoni were isolated from two patients, one treated three times with oxamniquine and the other with hycanthone once and oxamniquine twice. Progenies of these strains were maintained in Biomphalaria glabrata and mice. Groups of these infected mice were then treated with oxamniquine, hycanthone, niridazole and praziquantel and results compared with the BH strain maintained in our laboratory for many years. Schistosomicidal activity was assessed by the localization of worms in the portal vein system and oogram changes. Progenies from the strains isolated in this study were resistant to oxamniquine and hycanthone but sensitive to niridazole and praziquantel. The BH strain was sensitive to all four drugs. The serial runs of S. mansoni strains through intermediate and definitive hosts have not influenced their reactions to these schistosomicides.

Adolescent↗

Foci of Schistosoma mansoni in Assiut province in middle Egypt.

Following detection of Schistosoma mansoni in a 12 years old boy who had both schistosomal polyposis and periportal fibrosis with hepatosplenomegaly, epidemiological studies to confirm local transmission were performed in his and 30 other villages in Assiut Governorate, Egypt. The index case's brother and 6 of 380 inhabitants of his village who provided stool specimens were infected with S. mansoni and a farmer with dysentery and hepatosplenomegaly had a positive rectal snip biopsy. All had hepatic abnormalities on ultrasound examination. Two of 221 schoolchildren in another village had mixed infections with S. mansoni and S. haematobium; 17 others had only S. haematobium. None of 419 inhabitants living near the infected boys had S. mansoni infection. Snails from canals and drains near both villages were netted, identified, counted and checked for infection: in the second village one of 1039 Bulinus truncatus was infected with Schistosoma sp. but none of 859 Biomphalaria alexandrina was infected. Schistosomiasis mansoni is being focally transmitted in 2 villages in Assiut Governorate and appears to be spreading from Lower to Middle and Upper Egypt. We believe B. alexandrina, which has been present in some of the waterways for at least 15 years, were infected recently by local inhabitants returning from Iraq or by cattle traders or military recruits from the Delta.

Animals↗

The production of toxic oxygen metabolites by hemocytes of different snail species.

The phagocytic hemocytes of four snail species were investigated for their ability to generate reactive oxygen metabolites upon stimulation by foreign material. Hemocytes of the pond snail Lymnaea stagnalis and of the garden snail Helix aspersa showed a luminol-dependent chemiluminescence (CL) when they phagocytosed zymosan particles. This CL was inhibited by superoxide dismutase (SOD) and sodium azide, indicating the involvement of oxygen intermediates. Hemocytes of the planorbid snails Planorbarius corneus and Biomphalaria glabrata did not give a detectable CL response. This is probably due to the presence of hemoglobin in the hemolymph; after isolation of the cells and subsequent stimulation, however, still no CL could be measured. Hemocytes of all four snail species showed a SOD-sensitive nitroblue tetrazolium (NBT) reduction, indicating the generation of superoxide anions. Regarding the NBT reaction, no differences were observed between strains of B. glabrata that were susceptible or resistant to PR-1 Schistosoma mansoni; neither did exposure to the parasite have an effect on the ability of the hemocytes to reduce NBT. Also, hemocytes from Trichobilharzia ocellata-infected L. stagnalis did not react differently from hemocytes of uninfected snails. It is now clear that phagocytically stimulated hemocytes of several molluscan species can generate reactive forms of oxygen; the relevance of this fact for the phylogeny of killing systems operative in leukocytes is discussed.

Animals↗

Movements of molluscs by computer simulation.

The movements of two kinds of molluscs (Paludina viviparus and Heliosoma (taphius) Nigricans) (in Argentina Biomphalaria tenagophila) performing feeding activities have been simulated by a digital computer. Successive positions of the organisms were determined by a computer program which was based on Rashevsky's (1973) Principle of Adequate Design.

Animals↗

Search for antifungal, molluscicidal and larvicidal compounds from African medicinal plants.

African medicinal plants provide a rich source of biologically-active natural products. By the use of simple bioassays in conjunction with an approach involving bioactivity guided fractionation, it is possible to isolate novel compounds with interesting properties. Several hundred plant extracts have been submitted to different screens, including bioautography on TLC plates with the fungi Cladosporium cucumerinum and Candida albicans. Among the other test systems are a bioassay for molluscicidal activity with the schistosomiasis-transmitting snail Biomphalaria glabrata and a benchtop assay for larvicidal activity with larvae of the mosquito Aedes aegypti, the vector of yellow fever. Using a variety of modern separation techniques, a number of compounds with activity in these bioassays have been isolated. These include prenylated xanthones, tetracyclic phenols and saponins. The role of saponins and saponin-containing plants in the control of schistosomiasis is outlined.

Animals↗

Molluscicidal activity of synthetic lapachol amino and hydrogenated derivatives.

A series of new amino derivatives and a new partially hydrogenated derivative of the natural naphthoquinone lapachol were assayed for molluscicidal activity against Biomphalaria glabrata. These derivatives showed low to medium LC(50) values, and a 3.1 microg/mL value for the most potent derivative of the series. The toxicity is in agreement with the decrease of polar character of the tested compounds.

Amines↗

New 1,2,3,4-tetrahydro-1-aza-anthraquinones and 2-aminoalkyl compounds from norlapachol with molluscicidal activity.

New nitrogen derivatives from norlapachol, including four new diastereomeric 1,2,3,4-tetrahydro-1-aza-anthraquinones obtained from the Prins cyclization on suitable aminoacetaldehyde dimethylacetal derivatives with formic acid, were found to exhibit molluscicidal activity against Biomphalaria glabrata. These derivatives showed low to medium LC(50) values, similar to those reported previously for the homologous series of nitrogen derivatives of lapachol. The toxicity profile against Artemia salina was also determined for all compounds.

Alkylation↗

Molecular cloning and characterization of Mn-superoxide dismutase from disk abalone (Haliotis discus discus).

The mitochondrial enzyme manganese superoxide dismutase (mitMn-SOD) is one of the antioxidant enzymes involved in cellular defense against oxidative stress and catalyzes the conversion of O(2)(-) into the stabler H(2)O(2). In this study, a putative gene encoding Mn-SOD from disk abalone (Haliotis discus discus, aMn-SOD) was cloned, sequenced, expressed in Escherichia coli K12(TB1) and the protein was purified using pMAL protein purification system. Sequencing resulted ORF of 681 bp, which corresponded to 226 amino acids. The protein was expressed in soluble form with molecular weight of 68 kDa including maltose binding protein and pI value of 6.5. The fusion protein had 2781 U/mg activity. The optimum temperature of the enzyme was 37 degrees C and it was active in a range of acidic pH (from 3.5 to 6.5). The enzyme activity was reduced to 50% at 50 degrees C and completely heat inactivated at 80 degrees C. The alignment of aMn-SOD amino acid sequence with Mn-SODs available in NCBI revealed that the enzyme is conserved among animals with higher than 30% identity. In comparison with human mitMn-SOD, all manganese-binding sites are also conserved in aMn-SOD (H28, H100, D185 and H189). aMn-SOD amino acid sequence was closer to that of Biomphalaria glabrata in phylogenetic analysis.

Amino Acid Sequence↗

Effects of diet and larval trematode parasitism on lutein and beta-carotene concentrations in planorbid snails as determined by quantitative high performance reversed phase thin layer chromatography.

High performance thin layer chromatography (HPTLC) was used to quantify the concentrations of beta-carotene and lutein in Biomphalaria glabrata and Helisoma trivolvis (Colorado and Pennsylvania strains) snails under various conditions. These conditions were: snails fed a lettuce (L) vs. a yolk (Y) diet; B. glabrata infected with Echinostoma caproni vs. uninfected snails; and H. trivolvis (PA) infected with Echinostoma trivolvis vs. uninfected snails. The pigments were extracted from the snail whole bodies and digestive gland-gonad complexes, separated by reversed phase HPTLC, and quantified by densitometric scanning with standard calibration curves. Snails on the L-diet showed significant increases (Student's t-test, P<0.05) in the concentrations of beta-carotene and lutein compared to snails on the Y-diet. Snails infected with echinostomes showed no significant differences (Student's t-test, P>0.05) in the concentrations of lutein and beta-carotene compared to the uninfected cohorts. Our results were compared with previous studies that analyzed beta-carotene and lutein in snails infected with larval trematodes. Variations in the results of our study compared with others reflect intrinsic differences in the larval trematode-snail systems used.

Animals↗

Comparative study on the susceptibility of freshwater species to copper-based pesticides.

Copper compounds have been intentionally introduced into water bodies as aquatic plant herbicides, algicides and molluscicides. Copper-based fertilizers and fungicides have been widely used in agriculture as well. Despite the fact that copper is an essential element for all biota, elevated concentrations of this metal have been shown to affect a variety of aquatic organisms. Nonetheless, comparative studies on the susceptibility of different freshwater species to copper compounds have seldom been performed. This study was conducted to compare toxicity of copper-based pesticides (copper oxychloride, cuprous oxide and copper sulfate) to different freshwater target (Raphidocelis subcapitata, a planktonic alga and Biomphalaria glabrata, a snail) and non-target (Daphnia similis, a planktonic crustacean and Danio rerio, a fish) organisms. Test water parameters were as follows: pH = 7.4 +/- 0.1; hardness 44 +/- 1 mg/l as CaCO3; DO 8-9 mg/l at the beginning and > 4 mg/l at the end; temperature, fish and snails 25 +/- 1 degrees C, Daphnia 20 +/- 2 degrees C, algae 24 +/- 1 degrees C. D. similis (immobilization), 48-h EC50s (95% CLs) ranging from 0.013 (0.011-0.016) to 0.043 (0.033-0.057) mg Cu/l, and R. subcapitata (growth inhibition), 96-h IC50s from 0.071 (0.045-0.099) to 0.137 (0.090-0.174) mg Cu/l, were the most susceptible species. B. glabrata (lethality), 48-h LC50s from 0.179 (0.102-0.270) to 0.854 (0.553-1.457) mg Cu/l, and D. rerio (lethality), 48-h LC50s 0.063 (0.045-0.089), 0.192 (0.133-0.272) and 0.714 (0.494-1.016) mg Cu/l, were less susceptible than Daphnia to copper-based pesticides. Findings from the present study therefore suggest that increased levels of copper in water bodies is likely to adversely affect a variety of aquatic species.

Animals↗

Geographic information systems as a tool for control program management for schistosomiasis in Egypt.

During a 4-year study a geographic information system (GIS) risk model was constructed for predicting the relative risk of schistosomiasis in Kafr El-Sheikh governorate, Egypt. A 1-year 1990-1991 time series on diurnal temperature difference (dT) prepared from the advanced very high resolution radiometer (AVHRR) sensor on the NOAA-11 satellite was used to develop a regional risk model for the Nile delta based on thermal-hydrological domains. A May 15, 1990 Landsat TM scene (path 177, Row 38) was used to develop a local 'village-scale' environmental risk model based on higher resolution satellite sensor data (30 m picture element size at earth surface). Four of ten classes derived from a tasseled cap (Tcap) transformation of the Landsat TM scene were shown to be significantly related to a 5-year Schistosoma mansoni prevalence database from the Ministry of Health. A risk model was developed based on dT and the proportional area of the four Tcap classes in 5 km(2) buffer zones centered on rural health unit (RHU) reporting units. Available historical data on S. mansoni and its snail host Biomphalaria alexandrina, as well as recent field collected data were gathered and incorporated as separate themes. Model validation was done using data collected on snail population bionomics-infection rates, water quality, underground water table and cercariometry at 13 hydrologically representative sites. The role of soil type, water table and water quality was studied at 79 of 154 rural health unit sites. The model permitted retrieval of relevant data by RHU point location. For the first time in Egypt, the Kafr El-Sheikh GIS schistosoma prediction model can support MOH efforts to make more accurate control program decisions based on environmental predilection sites of endemic Schistosomiasis mansoni.

Animals↗

Satellite climatology and the environmental risk of Schistosoma mansoni in Ethiopia and east Africa.

Annual and seasonal composite maps prepared from the normalized difference vegetation index (NDVI) and earth surface maximum temperature (T(max)) satellite data from the archives of the Global land 1-km program of the United States Geological Survey (USGS) were studied for. their potential value, using geographic information system (GIS) methods, as surrogates of climate data in the development of environmental risk models for schistosomiasis in Ethiopia. Annual, wet season and dry season models were developed and iteratively analyzed for relationships with Schistosoma mansoni distribution and infection prevalence rates. Model-predicted endemic area overlays that best fit the distribution of sites with over 5% prevalence corresponded to values of NDVI 125-145 and T(max) 20-33 degrees C in the annual composite map, NDVI 125-145 and T(max) 18-29 degrees C for the wet season map, and NDVI 125-140 and T(max) 22-37 degrees C for the dry season map. The model-predicted endemic area was similar to that of a prior model developed using an independent agroecologic zone data set from the United Nations Food and Agriculture Organization (FAO). Results were consistent with field and laboratory data on the preferences and limits of tolerance of the S. mansoni-Biomphalaria pfeifferi system. Results suggest that Global 1-km NDVI and T(max), when used together, can be used as surrogate climate data for development of GIS risk assessment models for schistosomiasis. The model developed for Ethiopia based on global 1-km satellite data was extrapolated to a broader area of East Africa. When used with FAO agroecologic zone climate data limits of <27 degrees C for average annual mean temperature and annual moisture deficits (annual rain-annual potential evapotranspiration) of <-1300 mm, the model accurately represented the regional distribution of the S. mansoni-B. pfeifferi system in the East Africa extrapolation area.

Animals↗