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Studies on resistance in snails. 5. Tissue reactions to Echinostoma lincloense in naturally resistant Biomphalaria glabrata.

Laboratory-raised juvenile albino Biomphalaria glabrata snails show a wide range of natural resistance to a single infection with 50 or 100 miracidia of Echinostoma lindoense. In the most resistant snails all sporocysts are destroyed in peripheral tissues soon after miracidial penetration. In less resistant snails some sporocysts reach the heart where they are encapsulated. In fully susceptible snails, all sporocysts rapidly migrate to the heart, where they mature and continue to develop. The greater part of our B. glabrata colony consists of snails in which sporocysts reaching the heart will survive, but in which a varying number of sporocysts will be destroyed in the tissues. These snails are usually considered susceptible, as they do become infected. Tissue reactions induced by sporocysts following a single infection in naturally resistant snails are similar to reactions in snails with an acquired resistance. In fully susceptible snails, the amebocyte-producing organ remains small and inactive. It is slightly to moderately stimulated in partially resistant snails in which destruction of sporocysts occurs in the tissues and surviving larvae are found in the ventricle. In snails in which amebocyte aggregates or capsules develop in the ventricle, the organ becomes markedly enlarged. Migration of sporocysts in the snail appears not to be continuous, as periodic rests seem to occur. Migration follows intrusion of the sporocyst through the tissues, induced by bodily distension and contraction, and then proceeds within the arteries against the blood flow, passing from one endothelial attachment site to another, possibly aided by negative pressure during ventricular diastole.

Animals↗

Intermediate hosts of schistosoma; African Biomphalaria and Bulinus. I.

This study is an attempt to classify all described species of African Biomphalaria and Bulinus. It is based upon the author's examination of a great number of specimens collected from many parts of Africa. The variations attributable to age, environment and genetic factors which may be noted in the taxonomic characters are discussed, and some new species and subspecies are established. For each recognized species and subspecies the author states the distinguishing characters, indicates the geographical distribution, and gives a list of synonyms.

Africa↗

On the pathogenicity of attenuated Schistosoma mansoni cercariae released from metabolically disturbed Biomphalaria alexandrina.

Biomphalaria alexandrina were treated with sublethal concentrations (LC10) of dry powdered leaves Solanum nigrum or whole dry Ambrosia maritima. The two plants affected the development of schistosome parasite within snails by disturbing the glycolytic flux, the most important metabolic pathway for schistosome-infected snails. Attenuated cercariae released from treated snails were used to infect male albino mice to evaluate their pathogenicity compared to control cercariae shed from untreated snails. The mean number of worms established declined from about 70 +/- 17.6 worms/mouse infected with control cercariae to 23.4 +/- 20.33 and 14.18 +/- 10.06 worms/mouse infected with S. nigrum and A. maritima-attenuated cercariae respectively. Most males and females detected in these animals measured 0.4-0.6 and 0.6-1.2 mm respectively compared to 1.2-1.4 and 1.4-1.7 mm in males and females released from mice infected with normal cercariae. Egg count in the liver and intestine of mice infected with attenuated cercariae was remarkably lower showing reduced fecundity of worms developed from attenuated cercariae. Number and size of granulomatous reactions showed remarkable reduction in attenuated cercariae-infected mice. Biochemical analyses for pathogenicity achieved with attenuated cercariae showed that while serum aspartate and alanine aminotransferases (AST &ALT) were more or less similar, depleted glycogen and elevated lipid peroxides were normalized when compared to those infected with normal cercariae.

Ambrosia↗

Host-parasite relationships between Schistosoma mansoni and Echinostoma liei and their intermediate host Biomphalaria alexandrina using RAPD-PCR analysis.

Biomphalaria alexandrina are known to be intermediate hosts for both Schistosoma mansoni and Echinostoma liei. RAPD-PCR assay offers a new approach to host-parasite relationships. This was performed by investigating the genetic variation and compatibility among S. mansoni, E. liei and their intermediate host B. alexandrina with special emphasis on variations occurring in snails infected with S. mansoni and/or E. liei. Six primers were screened for DNA analysis and gave total patterns from 28 to 37 reproducible bands for each species. All specimens analyzed by the RAPD-PCR gave interpretable electrophoretic banding patterns that were polymorphic and compatible in the amplified products of these primers within each species.

Animals↗

Molluscicidal and cercaricidal efficacy of Acanthus mollis and its binary and tertiary combinations with Solanum nigrum and Iris pseudacorus against Biomphalaria alexandrina.

Molluscicidial activity of leaves of Acanthus mollis against Biomphalaria alexandrina were evaluated. Its petroleum ether extract (LC50 values = 6.92 mg/L) was more potent than Solanum nigrum and Iris pseudacorus extracts. A binary combination (1:1) of A. mollis and S. nigrum, as well as, a binary combination (1:1) of A. mollis and I. pseudacorus extracts showed additive effect on snails (24 hr LC50: 5.09 mg/l and 3.76 mg/l respecttively). A tertiary combination (1:1:1) of A. mollis, S. nigrum and I. pseudacorus extracts (24 hr LC50: 4.01 mg/l) showed good result. Also, petroleum ether extract of A. mollis leaves killed Schistosoma mansoni cercariae at concentrations of 20, 10 and 5 mg/l within 30, 45 min. and an hour respectively. Mortality increased with increasing exposure time and concentration.

Acanthaceae↗

Schistosoma mansoni infections in neonatal Biomphalaria glabrata snails.

In areas endemic for schistosomiasis, the population dynamics of the snail intermediate hosts have a direct effect on parasite transmission. The present study focused on the potential for neonatal Biomphalaria glabrata snails to become infected with Schistosoma mansoni and to produce cercariae under various conditions. It was found that snails as small as 0.74 mm in shell diameter could survive miracidial penetration and could release cercariae when as small as 1.6 mm in diameter. Cercariae produced by small snails were equally infectious for mice when compared with those shed by larger snails. Likewise, histological examination of neonatally exposed snails revealed normally developing parasites at all stages of infection. It was found that in 2 snail populations expressing either high or low susceptibility to the parasite, peak susceptibility occurred at 25 days of age in both groups. Daily cercarial production for neonatally exposed snails was initially low but increased dramatically as the snails grew, eventually reaching values as high as 2,100 cercariae/snail/day. A moderate to high percentage of snails infected as neonates was eventually capable of simultaneously producing both eggs and cercariae. These studies emphasize the potential importance of neonatal and preadult snails in helping to maintain foci of S. mansoni infection in endemic areas.

Animals↗

Effects of excretory-secretory products of Echinostoma paraensei larvae on the hematopoietic organ of M-line Biomphalaria glabrata snails.

Responses of the hematopoietic organ (HO) in Biomphalaria glabrata snails to extracts and excretory-secretory (E-S) products of Echinostoma paraensei larvae were studied to understand the HO-activating mechanism. M-line B. glabrata snails were injected with materials from E. paraensei larvae, and the size of the HO was ascertained in histological sections. The size of HO in snails injected with extracts and E-S products from sporocysts and rediae was significantly larger than that in snails injected with culture medium. E-S products of sporocysts were fractionated using ultrafiltration membranes, polyacrylamide gel electrophoresis, and electrophoretic elution. Examination of fractionated E-S products of sporocysts revealed that specific components of E-S products were responsible for HO-stimulating activity.

Animals↗

Characterization of surface lectins binding and SDS-PAGE protein patterns of Biomphalaria alexandrina haemocytes infected with Schistosoma mansoni.

Seven biotinylated lectins were tested with fluorescein labeled Avidin D for binding to haemocyte surfaces of non-inflected and infected Biomphalaria alexandrina snails with Schistosoma mansoi at 24 to 72 hours post infection (PI). Five lectins (Con A, DBA, SBA, RCA120 and WGA) yielded positive results. Two lectins (PNA and UEA-I) were negative. For the five positive lectins, the use of specific inhibition sugar indicated presence of glucose/mannose, galactose, N-acetyl-galactosamine (GalNAc) and N-acetyl-glucosamine (GlcNAc). The UEA-I lectin was negative and this indicates the absence of L-Fucose. There were no changes in the carbohydrates of haemocyte surfaces during the short periods (0 hr to 72 hrs) PI with S. mansoni. Electrophoresis of haemocyte homogenates of both non-infected and infected snails with S. mansoni yielded a complex pattern of polypeptides. The separated profiles demonstrate the occasional appearance or absence of certain bands in infected snails through four weeks PI. Post S. mansoni infection, dominant proteins appeared only through four weeks PI which were of average molecular masses 36, 9.3 and 5.3 kDa. At fourth week PI a remarkable decrease in the number of haemocyte protein fractions as well as the protein densities were reduced in compared to control snails. The results showed that qualitative and quantitative differences were detectable in the haemocyte proteins between non-infected and infected snails during four weeks of infection.

Animals↗

Biological and biochemical studies on Biomphalaria alexandrina snails, treated with low concentrations of certain molluscicides (synthetic and of plant origin).

The effect of low concentrations of different synthetic and natural mollusciciding agents may introduce to fresh water environment on reproduction and biochemical aspects of Biomphalaria alexandrina was studied. Different mollusciciding agents (copper sulphate, Bayluscide, Uccmaluscide, Agave filifera & A. attenuate) inhibited egg production, induced marked increased the percent of abnormal laid eggs and induced marked reduction in their hatchability. The maximal reductions in egg hatchability resulted with Bayluscide (0.0%) and Uccmaluscide (18%), A. filifera (21%) and A. attenuata (15%). All the antimolluscal materials caused a successful killing effect against miracidia and cercariae of Schistosoma mansoni. CuSo4, Bayluscide and Uccmaluscide killed 40% of the exposed miracidia and 50% of cercariae after an hour exposure. The plants sublethal concentration killed 100% of cercariae and miracidia after 6 hours exposure. Water leaving behaviour among the exposed snails was noticed especially during the first three weeks, showing maximal percentage (60%) after one week of exposure to Bayluscide. A general decrease in the activity of alkaline phosphatase (ALP) especially with Bayluscide (48.4%) and in acetylcholine esterase activity in the haemolymph especially on applying plant molluscicide A. filifera (50.8%) was noticed. Transaminases showed marked elevations in activities during the 1st three weeks, then began to drop (ASAT: 61.5%, with Bayluscide & ALAT: 50.8% with Uccmaluscide). The results reflect the effect of the metabolic disorders on life, egg laying, egg hatchability, hepatic cells damages, lack of smooth transmission at nerve junction, loss of muscular coordination and convulsions, then snails' death.

Animals↗

Effect of Commiphora molmol on adults, egg masses and egg-deposition of Biomphalaria arabica under laboratory conditions.

Commiphora molmol (Myrrh) has molluscicidal effect on Biomphalaria arabica snails at low concentration (40 ppm) after 48 hours exposure. The number of dead-snails increased with increasing the time of exposure. One day-old egg masses were more susceptible (death 100% with 80ppm) to the ovicidal effect of C. molmol than the five-day old ones (Death 95% with 80ppm). However, the eggs were more resistant to the C. molmol effect than the adult snails, embryogenesis began to stop at 20 ppm and eggs were all killed at 60 & 80 ppm. B. arabica fecundity decreased at 1 ppm. Based on safety to man and animals, C. molmol is recommended as a safe molluscidide.

Animals↗

A comparative study of the effects of phebrol on the respiratory chains of rat liver, Biomphalaria glabrata and Oncomelania nosophora.

1. Phebrol (sodium 2,5-dichloro-4-bromophenol), a synthetic molluscicide against Oncomelania nosophora, showed a dual effect on rat liver submitochondria, acting as an uncoupler at low concentrations (approximately 10 microM) and an inhibitor of succinate-cytochrome c reductase at high concentrations. 2. Phebrol also inhibited the enzymes responsible for succinate-fumarate conversion, i.e. the succinate-cytochrome c reductase, fumarate reductase and NADH-cytochrome c reductase of the mitochondrial fraction from Biomphalaria glabrata. 3. Kinetic inhibition studies showed succinate-cytochrome c reductase of B. glabrata and O. nosophora to be more sensitive than that of rat liver toward phebrol. 4. Phebrol accumulated in whole tissues of B. glabrata and O. nosophora and had significant effects on the production of succinate, fumarate and malate by these snails. 5. On the basis of these results, the possible sites of inhibition by phebrol of snail respiratory chains are proposed.

Animals↗

Molluscicidal effect of fenitrothion and anilofos on Lymnaea natalensis and Biomphalaria alexandrina snails and on the free larval stages of Schistosoma mansoni.

Psticides; fenitrothion and anilofos (aniloguard) were testd as molluscicides against Lymnaea natalensis and Biomplhalaria alexandrina. The LC10 & LC90 of fenitrothion was 0.12 & 0.21 ppm for L. nalalensis and 0.17 & .26 ppm for B. alexandrina, respectively. The LC50 & LC90 anilofos was 2.61 & 6.47 ppm for Lymnaea and 3.07 & 8.6 ppm for Biomphalaria. The effect of sublethal concentrations (LC0, LC5 & C10) of Feni-rothion on B. alexandrina growth rate, eggs hatchability and on free larval stages of Schistosonma mansoni (miracidia & cerca-riae) were studied. The results obtained showed that sublethal concentrations of fenitrothion caused reduction in growth rate of B. alexandrina and reduction in the hatchibility of snails eggs. The mortality rates of miracidia and cercariae were elevated by increasing both the concentrations of fenitrothion and the time of exposure. The results showed that fenitrothion was more toxic to the free larval stages of S. mansoni than to their snails. The results showed a significant reduction in total protein of treated snails when compared with controls in haemolymph while there was an increase of protein contents of the tissue. The AlkP enzyme activity was slightly increased in the haemolymph of experimental groups than the control and in the tissues the values were significantly higher when compared with control. ALT enzyme activity in haemolymph of experimental groups was higher than controls while its activity in tissue was lower. AST enzyme activity was higher in haemolymph and tissue of experimental groups than in controls.

Animals↗

Surface morphology and characteristics of hemocytes of Biomphalaria glabrata (Pulmonata: Planorbidae) from two geographic sources.

Biomphalaria glabrata is a fresh water snail of medical importance since it is the intermediate host of the trematode Schistosoma mansoni. The internal defense system of mollusks is mostly represented by circulating elements of the hemolymph (hemocytes). The infectivity of S. mansoni to B. glabrata is determined by genetic factors and evolutionary adaptations. In the present work factors about the parasite/snail relationship were evaluated, especially those related to the morphology and characteristics of the surface of cells present in the hemolymph of two strains of B. glabrata: a strain with high susceptibility to S. mansoni (Puerto Rico, PR) and a strain with medium susceptibility (Caripe, Ca). Hemolymph was collected by cephalopodal puncture; a total and a differential count of hemocytes were done in dyed preparations and through scanning electron microscopy (SEM). Results from both strains show a high quantitative variability of the total hemocyte count. Hemocytes dyed with May-Grünwald Giemsa showed cells with a basophilic nucleus predominantly in PR (61.7%), dense and strongly basophilic in Ca (71.2%) with significant differences between them. Through SEM round cells with a corrugated surface were observed (6-10microm), hemocytes with an irregular spongy surface (12microm), others with many projections (16microm) and cells not reported in similar to erythrocytes (21microm). Hemocytic cells from both strains confirmed cytoadherence and encapsulation were confirmed in the hemocytic cells from both strains, while no differences associated to the susceptibility of the strains were observed after 2 h of parasite-hemocyte incubation.

Animals↗

Effect of the plant Cupressus macro-carpa (Cupressacea) on some haematolo-gical and biochemical parameters of Biomphalaria alexandrina snails.

The dry powder of the plant aereal part; Cupressus macro-carpa (Cupressacea) was tested against Biomphalaria alexandrina. LC50 & LC90 values were 59.5 & 98.8 ppm, respec-tively. Exposure of B. alexandrina to sublethal concentrations (LC0, LC10 & LC25) of C. macrocarpa for three weeks signi-ficantly decreased the number of circulating hemocytes. The magnitude of reduction was increased with increasing of the tested concentration. The main type of cell in the hemolymph of B. alexandrina was the granulocyte (71.8%), followed by large round cells or hyalinocytes (19.0%) and small round cells or undifferentiate cells (9.2%). The percentage of different hemocyte categories was changed in treated snails. In snails maintained at LC25, showed significantly higher percentages of small round cells than controls, 56.2% & 9.2% respectively. Maintainence of B. alexandrina in sublethal concentrations for three weeks significantly reduced protein & hemoglobin content in the hemolymph. Reduction in enzyme activities occurred in the hemolymph and tissues of treated snails. The enzymes were pyruvate kinase (PK), lactat dehydrogenase (LDH), hexokinase (HK) and phosphoenol pyruvate carboxy kinase (PEPCK) which are very important in metabolism of the protein and carbohydrate. The infectivity of Schistosoma mansoni miracidia was greatly reduced by exposure to the sublethal concentrations (LC0, LC10 & LC25) of Cupressus. Infection rate of B. alexandrina reached to 54.5%, 37.5% & 16.7%, respectively compared to control (81.25%). Duration of cercarial shedding and the total periodic cercarial production/snail showed significant reduction while the parasite incubation period was significantly longer (p<0.05).

Animals↗

Excretory-secretory products of Echinostoma paraensei sporocysts mediate interference with Biomphalaria glabrata hemocyte functions.

Miracidia of Echinostoma paraensei were cultured in medium containing 14C-labeled amino acids, allowed to transform into sporocysts, and their excretory/secretory products (E-S) were collected and characterized by sodium dodecyl sulfate polyacrylamide gel electrophoresis and autoradiography. Effects of E-S on hemocytes of Biomphalaria glabrata were also assessed. E-S collected during day 1 of culture (E-S1) contained several polypeptides, none of which were labeled, suggesting that E-S1 are largely preformed. E-S1 significantly depressed the ability of hemocytes to phagocytose sheep red blood cells (SRBC), but otherwise had little effect on hemocyte structure or behavior. E-S released by sporocysts in day-2 cultures (E-S2) and in older cultures generally were similar and also contained several polypeptides, many of which were labeled, indicating active synthesis of E-S in vitro. E-S2 strongly inhibited hemocyte uptake of SRBC. Also, hemocytes pretreated with E-S2 assumed a spherical shape and failed to spread normally. E-S obtained through 10 days of culture mediated this effect. Active components of E-S2 were greater than 100 kDa in their native configuration, were heat- and trypsin-labile, and were bound by anti-E-S antibodies. Both greater than 200- and 80-kDa bands were prominent in anti-E-S immunoprecipitates. Hemocytes derived from snails of the 13-16-R1 strain of B. glabrata (a strain resistant to infection with Schistosoma mansoni), when pretreated with E-S2, bound to sporocysts of S. mansoni but lost their ability to damage such sporocysts. E-S2 interfered with hemocyte functions in ways inferred from earlier classic in vivo studies of trematode-snail interactions.

Animals↗

Embryonic development of the shell in biomphalaria glabrata (Say).

During embryogenesis of the fresh water snail Biomphalaria glabrata (Say) (Pulmonata, Basommatophora) shell formation has been studied by light and electron microscopical techniques. The shell field invagination (SFI), the secretion of the first shell layers, the development of the shell-forming mantle edge gland and spindle formation have been investigated. During embryonic development at 28 degrees C environmental temperature, the shell field invaginates after 35 h. After 40 h the SFI is closed apically by cellular protrusions and scale-like precursors of the periostracum. The first electron translucent layer of the periostracum stems from electron dense vesicles of the cells which lie at the opening of the SFI. A second electron dense layer appears some hours afterwards. When the shell appears birefringent in the polarizing microscope (45 h of development) calcium can be detected in it using energy dispersive x-ray analysis. As calcification occurs the intercrystalline matrix appears under the periostracum and the SFI begins to open. In embryos of 60 h the mantle cavity appears at the left caudal side. When the mantle edge groove develops (65 h of development) lamellate units are added to the outer layer of the periostracum, but no distinct lamellar layer is formed in B. glabrata. In addition to the lamellar cell and the periostracum cell, a secretory cell can be observed in the developing groove. After 65 h of development, spindle formation starts and the shell begins to coil in a left hand spiral. After 5 days of development the embryos are ready to leave the egg capsules.

Animals↗

[Effects of eugenol and derivatives on Biomphalaria glabrata].

Biomphalaria glabrata snails and egg-masses were exposed, for six to twenty-four hours to concentrations of 1, 10, 100 and 1000 ppm of Eugenol, O-methyleugenol, O-benzyleugenol and dehydrodieugenol. Only at 10 ppm O-benzyleugenol enhanced mortality of snails and egg-masses. The other substances showed ovicidal and molluscicidal activity only at 100 and 1000 ppm concentrations, causing a significant cardiac frequency reduction in snails after 6 to 24 hours of exposure as well as perduring low cardiac rates until 24 hours afterwards. Two specimen exposed to 100 ppm O-methyleugenol presented anesthetic effect and extrusion of copulator and urethral organs. No schistosomicide or anesthetic effects were observed in mice experimentally infected with Schistosoma mansoni and treated during 5 days with oral doses of 150 mg/kg of Eugenol, O-methyleugenol and O-benzyleugenol.

Administration, Oral↗

[Repopulation of breeding habitats of Biomphalaria glabrata after treatment with niclosamide].

Experiments were undertaken both in the laboratory and in the field between 1980-1984 to evaluate the causes of repopulation of breeding places of Biomphalaria glabrata following treatment with Niclosamide. Laboratory bioassays showed that the susceptibility to emulsifiable Niclosamide of B. glabrata collected monthly from an irrigation ditch system varied during the year. Lethal concentrations (LC90) ranged between 0.15 mg/l-1 and 0.60 mg/l-1. Statistically significant differences (alpha = 0.01) were evident between the months of May/82 and January/83 and December/82 and January/83, and were related to snail nutrition. In the field two types of foci of B. glabrata were treated with 10 ppm of Niclosamide. The first one consisted of a reservoir of 12000 1 of water in which 14.5% of snails were infected with Schistosoma mansoni. One application of molluscicide followed by cleaning of the reservoir eliminated all the snails. The second one consisted of an irrigation system in which 5.6% of the snails were infected with S. mansoni. One application of molluscicide without cleaning the ditches reduced the density of snails by 98%. The causes of the survival of 2.0% of the snails in the ditches are discussed in relation to the substratum of the breeding places and the treatment technique.

Animals↗