Snail-conditioned Water Research Articles

Identification of Schistosoma mansoni miracidia attractant candidates in infected Biomphalaria glabrata using behaviour-guided comparative proteomics.

Schistosomiasis, caused by infection with Schistosoma digenetic trematodes, is one of the deadliest neglected tropical diseases in the world. The Schistosoma lifecycle involves the miracidial infection of an intermediate freshwater snail host, such as Biomphalaria glabrata. Dispersing snail host-derived Schistosoma miracidia attractants has been considered a method of minimising intermediate host infections and, by extension, human schistosomiasis. The attractiveness of B. glabrata to miracidia is known to be reduced following infection; however, the relationship between duration of infection and attractiveness is unclear. Excretory-secretory proteins (ESPs) most abundant in attractive snail conditioned water (SCW) are key candidates to function as miracidia attractants. This study analysed SCW from B. glabrata that were naïve (uninfected) and at different time-points post-miracidia exposure (PME; 16h, 1-week, 2-weeks and 3-weeks PME) to identify candidate ESPs mediating Schistosoma mansoni miracidia behaviour change, including aggregation and chemoklinokinesis behaviour (random motion, including slowdown and increased turning rate and magnitude). Miracidia behaviour change was only observed post-addition of naïve and 3W-PME SCW, with other treatments inducing significantly weaker behaviour changes. Therefore, ESPs were considered attractant candidates if they were shared between naïve and 3W-PME SCW (or exclusive to the former), contained a predicted N-terminal signal peptide and displayed low identity (<50%) to known proteins outside of the Biomphalaria genus. Using these criteria, a total of 6 ESP attractant candidates were identified, including acetylcholine binding protein-like proteins and uncharacterised proteins. Tissue-specific RNA-seq analysis of the genes encoding these 6 ESPs indicated relatively high gene expression within various B. glabrata tissues, including the foot, mantle and kidney. Acetylcholine binding protein-like proteins were highly promising due to their high abundance in naïve and 3W-PME SCW, high specificity to B. glabrata and high expression in the ovotestis, from which attractants have been previously identified. In summary, this study used proteomics, guided by behavioural assays, to identify miracidia attractant candidates that should be further investigated as potential biocontrols to disrupt miracidia infection and minimise schistosomiasis.

A new accumulation assay of Schistosoma mansoni miracidia using square capillary glass tubes

Current control measures for schistosomiasis have only been partially successful in endemic areas due to socioeconomic constraints. One possibility for controlling the disease is to aim at the miracidial stage of the trematode to avoid infecting intermediate snail hosts by introducing more attractive substances for miracidia in the environment. Here, we introduce an accumulation assay of Schistosoma mansoni miracidia using a square glass tube for analysis of the positive responses of miracidia toward several substances, including snail-conditioned water of Biomphalaria glabrata, Bulinus globosus and insusceptible snails collected in the Nagasaki area in Japan. The substances are not proteins because miracidia accumulated in boiled snail-conditioned water and the secretion or emission level of substances depended on the feeding conditions of Biomphalaria glabrata. The present study also showed that substances emitted from Biomphalaria glabrata with a molecular weight around 10 kDa accumulated Schistosoma mansoni miracidia. Further, we showed that Schistosoma mansoni miracidia did not accumulate in response to mono- or disaccharides tested in the study.

Changes in snail chemical profiles through host-parasite interactions

Host behavior may be modified by their parasites to increase the likelihood of transmission, but mechanisms underlying these interactions are not well understood. Hosts and parasites release chemical signaling molecules, like oxylipins, that may affect transmission. Oxylipins are oxygenated metabolites of fatty acids that function as signaling molecules and have essential physiological and functional roles. Yet, the limited taxonomic and contextual scope of these studies constrains our ability to understand their role in parasite‐modified behavior. We characterized oxylipins in field-collected File Ramshorn snails, Planorbella pilsbryi. We tested for differences in oxylipin profiles based on infection status (infected with the trematode Echinostoma trivolvis lineage a and uninfected) and parasite activity (high and low). Snail-conditioned water samples were produced by placing five snails into artificial spring water for four hours. Oxylipins were extracted from snail-conditioned water samples and quantified using high performance liquid chromatography-tandem mass spectrometry. Infected snails emitted 69 oxylipins in higher amounts, with 37 only released by this group. Within infected snails, 18 oxylipins were emitted in higher amounts in snails with increased parasite activity. Oxylipins emitted in higher amounts by infected snails with increased parasite activity were predominantly derived from the cytochrome P450 pathway. As infected snails emit different oxylipin profiles than uninfected snails, their production may play a role in altering transmission success. By characterizing the oxylipins produced by snails, and how they are altered by infection, we can test their physiological and ecological roles in freshwater systems.

Density-Dependent Prophylaxis in Freshwater Snails Driven by Oxylipin Chemical Cues.

While animal aggregations can benefit the fitness of group members, the behaviour may also lead to higher risks of parasite infection as group density increases. Some animals are known to moderate their investment in immunity relative to the risk of infection. These animals exhibit density-dependent prophylaxis (DDP) by increasing their immune investment as group density increases. Despite being documented in many taxa, the mechanisms of DDP remain largely unexplored. Snails are known to aggregate and experience large fluctuations in density and serve as required hosts for many parasites. Further, they are known to use chemical cues to aggregate. To test whether freshwater snails exhibit DDP and investigate the role that chemical signaling compounds may play in triggering this phenomenon, we performed four experiments on the freshwater snail Stagnicola elodes, which is a common host for many trematode parasite species. First, we tested if DDP occurred in snails in laboratory-controlled conditions (control vs snail-conditioned water) and whether differences in exposure to chemical cues affected immune function. Second, we used gas chromatography to characterize fatty acids expressed in snail-conditioned water to determine if precursors for particular signaling molecules, such as oxylipins, were being produced by snails. Third, we characterized the oxylipins released by infected and uninfected field-collected snails, to better understand how differences in oxylipin cocktails may play a role in inducing DDP. Finally, we tested the immune response of snails exposed to four oxylipins to test the ability of specific oxylipins to affect DDP. We found that snails exposed to water with higher densities of snails and raised in snail-conditioned water had higher counts of haemocytes. Additionally, lipid analysis demonstrated that fatty acid molecules that are also precursors for oxylipins were present in snail-conditioned water. Trematode-infected snails emitted 50 oxylipins in higher amounts, with 24 of these oxylipins only detected in this group. Finally, oxylipins that were higher in infected snails induced naïve snails to increase their immune responses compared to sham-exposed snails. Our results provide evidence that snails exhibit DDP, and the changes in oxylipins emitted by infected hosts may be one of the molecular mechanisms driving this phenomenon.

Alterations in Lipids and Minerals in Relation To Larval Trematode Infections of Nerita Polita Marine Snails.

SummaryMarine snails provide an important source of vitamins, proteins, minerals, and essential fatty acids, and their mucus has a therapeutic significance. Parasitic infection of larval trematodes in these snails affects their nutritional value. The present study aimed to screen Nerita polita marine snails for the prevalence of cercarial infections and to evaluate the changes in lipids and some minerals in the infected as well as non-infected ones. Snails were collected randomly from February 2018 to January 2019 from the Red Sea Obhor bay, Jeddah city, Saudi Arabia. The amount of triglycerides, cholesterol, and phospholipids in the Digestive Gland Gonad Complex (DGG), hemolymph, and Snail Conditioned Water (SCW) of non-infected and infected Nerita polita snail was estimated using a spectrophotometer. Minerals content such as Ca, Zn, Pb, Na, Mn, Mg, K, Fe, Cu, and Cd, in the DGG and shell in the infected and non-infected snails were analyzed using an Inductively Coupled Plasma Optical Emission Spectrometer (ICP-OES). Two types of cercariae were detected, ocellate furcocercus cercaria (Trichobilharzia regent) and xiphidiocercaria (Litorina saxatilis VII). The study showed that there is a fl uctuation in the concentration of lipids and minerals between increase and decrease in DGG, hemolymph, shell, and SCW in infected snails. The nutritive value of the snails is affected with infection through the decrease of some lipids and minerals in infected snails. Further qualitative studies are needed.

Comparative study of excretory–secretory proteins released by Schistosoma mansoni-resistant, susceptible and naïve Biomphalaria glabrata

BackgroundSchistosomiasis is a harmful neglected tropical disease caused by infection with Schistosoma spp., such as Schistosoma mansoni. Schistosoma must transition within a molluscan host to survive. Chemical analyses of schistosome-molluscan interactions indicate that host identification involves chemosensation, including naïve host preference. Proteomic technique advances enable sophisticated comparative analyses between infected and naïve snail host proteins. This study aimed to compare resistant, susceptible and naïve Biomphalaria glabrata snail-conditioned water (SCW) to identify potential attractants and deterrents.MethodsBehavioural bioassays were performed on S. mansoni miracidia to compare the effects of susceptible, F1 resistant and naïve B. glabrata SCW. The F1 resistant and susceptible B. glabrata SCW excretory–secretory proteins (ESPs) were fractionated using SDS-PAGE, identified with LC-MS/MS and compared to naïve snail ESPs. Protein-protein interaction (PPI) analyses based on published studies (including experiments, co-expression, text-mining and gene fusion) identified S. mansoni and B. glabrata protein interaction. Data are available via ProteomeXchange with identifier PXD015129.ResultsA total of 291, 410 and 597 ESPs were detected in the susceptible, F1 resistant and naïve SCW, respectively. Less overlap in ESPs was identified between susceptible and naïve snails than F1 resistant and naïve snails. F1 resistant B. glabrata ESPs were predominately associated with anti-pathogen activity and detoxification, such as leukocyte elastase and peroxiredoxin. Susceptible B. glabrata several proteins correlated with immunity and anti-inflammation, such as glutathione S-transferase and zinc metalloproteinase, and S. mansoni sporocyst presence. PPI analyses found that uncharacterised S. mansoni protein Smp_142140.1 potentially interacts with numerous B. glabrata proteins.ConclusionsThis study identified ESPs released by F1 resistant, susceptible and naïve B. glabrata to explain S. mansoni miracidia interplay. Susceptible B. glabrata ESPs shed light on potential S. mansoni miracidia deterrents. Further targeted research on specific ESPs identified in this study could help inhibit B. glabrata and S. mansoni interactions and stop human schistosomiasis.

A Biomphalaria glabrata peptide that stimulates significant behaviour modifications in aquatic free-living Schistosoma mansoni miracidia.

The human disease schistosomiasis (or bilharzia) is caused by the helminth blood fluke parasite Schistosoma mansoni, which requires an intermediate host, the freshwater gastropod snail Biomphalaria glabrata (the most common intermediate host). The free-swimming parasite miracidia utilise an excellent chemosensory sense to detect and locate an appropriate host. This study investigated the biomolecules released by the snail that stimulate changes in the behaviour of the aquatic S. mansoni miracidia. To achieve this, we have performed an integrated analysis of the snail-conditioned water, through chromatography and bioassay-guided behaviour observations, followed by mass spectrometry. A single fraction containing multiple putative peptides could stimulate extreme swimming behaviour modifications (e.g. velocity, angular variation) similar to those observed in response to crude snail mucus. One peptide (P12;—R-DITSGLDPEVADD-KR—) could replicate the stimulation of miracidia behaviour changes. P12 is derived from a larger precursor protein with a signal peptide and multiple dibasic cleavage sites, which is synthesised in various tissues of the snail, including the central nervous system and foot. P12 consists of an alpha helix secondary structure as indicated by circular dichroism spectroscopy. This information will be helpful for the development of approaches to manipulate this parasites life cycle, and opens up new avenues for exploring other parasitic diseases which have an aquatic phase using methods detailed in this investigation.

Host-parasite relationship between Biomphalaria amazonica (Paraense, 1966) and Schistosoma mansoni (Sambon, 1907).

Biomphalaria amazonica is a planorbid species considered a potential host of Schistosoma mansoni. It is widely distributed in the Neotropical zone, particularly in the North and Centre-West of Brazil and in the North of Bolivia. The aim of the present study was to determine the host-parasite relationship between B. amazonica and S. mansoni (BH and SJ strains). Specimens of B. amazonica and their snail-conditioned water were examined in terms of their ability to attract miracidia. The infectivity of the mollusks was determined by exposing them to 20 miracidia of both strains. Sporocyst development and amebocyte reactions were studied after each mollusk specimen was exposed to 100 miracidia. Although no cercariae were eliminated, specimens of B. amazonica proved capable of attracting 77% of the miracidia they were exposed to. Viable sporocysts with no amebocyte reaction were found 96 hours after the exposure to miracidia. These results indicate the susceptibility of B. amazonica to the BH and SJ strains of S. mansoni, and therefore demonstrate the importance of this planorbid species as a potential vector of the trematode in the areas where it occurs.

Quality attracts parasites: host condition‐dependent chemo‐orientation of trematode larvae

Summary Environmental factors impairing physiological condition of organisms are assumed to predispose them to parasite infections. This is because host immune function is typically condition‐dependent. However, poor physiological condition has been reported to reduce host susceptibility to parasites in various systems. We examined whether such an effect can be due to altered exposure of hosts to active parasite transmission stages by investigating chemo‐orientation of free‐swimming cercariae larvae of a parasite Echinoparyphium aconiatum towards its snail host Lymnaea stagnalis. We used both long‐term and short‐term feeding treatments to manipulate the body condition and physiological traits related to food processing in experimental (i.e. target) snails and measured the preference of cercariae towards snail‐conditioned water (SCW) over clean water. We found that chemo‐orientation of cercariae depended on the nutritional status of target snails. High physiological condition (long‐term feeding) attracted parasites, but cercariae did not show preference towards SCW from individuals in poor physiological condition (long‐term starvation). Food processing (short‐term feeding treatments) did not affect chemo‐orientation. Our results suggest that host condition‐dependent chemo‐orientation of parasite larvae is a likely mechanism explaining the reduced susceptibility of snails to infection due to food limitation in our study system. In general, the use of condition‐dependent host cues can be highly beneficial for parasites as it increases their transmission to high‐quality hosts. Furthermore, evolving counter adaptations to such a transmission strategy can be very difficult for hosts.

Protection motivation theory in predicting intention to engage in protective behaviors against schistosomiasis among middle school students in rural China.

BackgroundAmong millions of people who suffer from schistosomiasis in China, adolescents are at increased risk to be infected. However, there is a lack of theory-guided behavioral prevention intervention programs to protect these adolescents. This study attempted to apply the Protection Motivation Theory (PMT) in predicting intentions to engage in protective behaviors against schistosomiasis infection.MethodsThe participants were selected using the stratified cluster sampling method. Survey data were collected using anonymous self-reported questionnaire. The advanced structural equation modeling (SEM) method was utilized to assess the complex relationship among schistosomiasis knowledge, previous risk exposure and protective measures in predicting intentions to engage in protective behavior through the PMT constructs.Principal FindingsApproximately 70% of participants reported they were always aware of schistosomiasis before exposure to water with endemic schistosomiasis, 6% of the participants reported frequency of weekly or monthly prior exposure to snail-conditioned water. 74% of participants reported having always engaged in protective behaviors in the past three months. Approximately 7% were unlikely or very unlikely to avoid contact with snail-conditioned water, and to use protective behaviors before exposure. Results from SEM analysis indicated that both schistosomiasis knowledge and prior exposure to schistosomiasis were indirectly related to behavior intentions through intrinsic rewards and self-efficacy; prior protective behaviors were indirectly related to behavior intentions through severity, intrinsic rewards and self-efficacy, while awareness had an indirect relationship with behavior intentions through self-efficacy. Among the seven PMT constructs, severity, intrinsic rewards and self-efficacy were significantly associated with behavior intentions.ConclusionsThe PMT can be used to predict the intention to engage in protective behaviors against schistosomiasis. Schistosomiasis intervention programs should focus on the severity, intrinsic rewards and self-efficacy of protection motivation, and also increase the awareness of infection, and enrich the contents of schistosomiasis education.

Heredity of specific host-finding behaviour in Schistosoma mansoni miracidia.

Two strains of Schistosoma mansoni were used to investigate the hereditary basis of species-specific host recognition by analysing behavioural responses of miracidia to snail-conditioned water. An Egyptian strain of S. mansoni, capable of distinguishing its host snail Biomphalaria alexandrina from other snails was cycled repeatedly through Biomphalaria glabrata, the intermediate host of a Brazilian strain known to respond even to non-susceptible snails with high intensity. After 5 cycles in the non-natural host, miracidia of the Egyptian strain still retained their preference for the original host snail. In a second experiment, host-finding behaviour of hybrids between these two parasite strains was studied. In the F1 generation, hybrids of both parental combinations showed the same low degree of specificity as the pure-bred Brazilian strain. Approximately one quarter of F2 hybrids proved to be as discriminatory as the Egyptian strain, confirming dominant Mendelian inheritance of non-specificity in schistosome miracidial host-finding behaviour. Moreover, hybrids seem to have lost the ability to develop in B. alexandrina, possibly suggesting a link between host recognition and host compatibility. The heredity of this behavioural trait is of evolutionary and epidemiological significance, since a shift to low host-finding specificity might have been a prerequisite for S. mansoni to acquire new host snails after being introduced to South America by the slave trade.

Thin Layer Chromatographic Analysis of Neutral Lipids in Snail Conditioned Water and Feces of Biomphalaria glabrata Infected with Echinostoma caproni

Thin layer chromatography (TLC) was used to determine neutral lipids in snail conditioned water (SCW) and feces from Biomphalaria glabrata snails infected with Echinostoma caproni. Samples were extracted in chloroform–methanol (2:1), and the neutral lipids were separated on silica‐gel plates with a mobile phase consisting of petroleum ether–diethyl ether–glacial acetic acid (80:20:1). Zones were detected with phosphomolybdic acid reagent and quantified by densitometry. Qualitative analysis of SCW samples showed the presence of free sterols, free fatty acids, and triacylglycerols; qualitative analysis of fecal samples showed free sterols, free fatty acids, and unidentified pigments. Quantitative analysis of SCW with feces (SCW‐WF) from infected snails showed 7.35 ± 2.3 µg 2−1 mL−1 free fatty acids compared with uninfected controls, which had 8.05 ± 0.54 µg 2−1 mL−1. Quantitative analysis of SCW without feces (SCW‐WOF) showed that infected snails contained 4.76 ± 0.64 µg 2−1 mL−1 of free fatty acids compared with uninfected controls, which had 5.88 ± 0.34 µg 2−1 mL−1. Free fatty acids were the major fraction in snail feces. Samples from infected snails had 0.0224% ± 0.0022% compared with the uninfected controls, which had 0.0353% ± 0.0036%. The free fatty acids composition of the feces for infected snails was significantly lower than that of the uninfected samples (Student's t‐test, P < 0.05).

Miracidia of an Egyptian strain of Schistosoma mansoni differentiate between sympatric snail species.

The host-finding behavior of miracidia of 2 strains of Schistosoma mansoni from Egypt and Brazil was studied by recording their responses to snail-conditioned water (SCW) from the Egyptian sympatric snails, Biomphalaria alexandrina, Physa acuta, Lymnaea cailliudi, and Balinus truncatus, as well as from Biomphalaria arabica and Biomphalaria glabrata. Miracidia of the Egyptian strain significantly preferred SCW from their compatible hosts B. alexandrina and B. arabica and showed no or a weak response to SCW from the other sympatric species, whereas miracidia of the Brazilian strain did not differentiate between SCW from different snail species.

The effect of isolation on reproduction and growth of Pseudosuccinea columella (Pulmonata: Lymnaeidae): a snail-conditioned water experiment.

A snail-conditioned water experiment was conducted in Pseudosuccinea columella to test the possible role of a chemical interaction between snails on the diminished growth and fecundity rates found for snails raised in pairs compared to those raised in complete isolation. The results permit to discard the hypothesis of an inhibition of growth and reproduction between snails due to factors released into the water.

Snail Host Finding by Fasciola hepatica and Trichobilharzia ocellata: Compound Analysis of “Miracidia-Attracting Glycoproteins”

Kalbe, M., Haberl, B., and Haas, W. 2000. Snail host finding by Fasciola hepatica and Trichobilharzia ocellata: Compound analysis of “miracidia-attracting glycoproteins.” Experimental Parasitology96, 231–242. The glycoconjugates from snail-conditioned water of Lymnaea truncatula and L. stagnalis which elicit typical host finding behavior in miracidia of Fasciola hepatica and Trichobilharzia ocellata were separated by anion-exchange chromatography and a two-step size-exclusion chromatography. We obtained fractions attractive for the parasites with MW of about 106 Da in both snail species. These fractions still contained species-specific information since miracidia responded only to molecules from their respective host snail. Analysis of the amino acid composition from the protein backbone revealed a similar composition in the effective fractions of both snails. Amounts of serine and threonine were higher than 30 mol %, which is typical for mucin-type glycoproteins. The carbohydrate moieties consisted mainly of galactose and fucose, but nine different other monosaccharides also were identified in smaller amounts. The heterogeneity of the molecules was also confirmed by the binding of six different lectins. Because of these characteristics, the effective molecules were termed “miracidia-attracting glycoproteins” (MAGs). MAGs may play an important role for parasite transmission, as they may increase the chance of an encounter between parasite and host and enable the miracidia to discriminate between their specific intermediate host and other unsuitable snail species.

Host-finding in Echinostoma caproni: miracidia and cercariae use different signals to identify the same snail species.

The snail host signals releasing host-finding responses in miracidia and cercariae of Echinostoma caproni were analysed by fractionation of snail-conditioned water (SCW). Cercariae responded non-specifically to organic and hydrophilic, low molecular weight components of SCW showing their typical turning response. Hydrolysis of peptides in SCW had no effect on cercarial responses. An artificial mixture of amino acids in concentrations determined from SCW as well as glycine alone in a concentration corresponding to the total concentration of amino acids in SCW showed nearly the same efficacy as SCW itself. Miracidia responded to a high molecular weight glycoprotein fraction, which could be isolated from SCW by ion-exchange and size-exclusion chromatography. In contrast to an Egyptian Schistosoma mansoni strain, the echinostome miracidia were not able to differentiate between different snail species. The results show for the first time that miracidia and cercariae of the same species may use different signals to identify the same snail host species. This indicates an independent evolution of host-finding mechanisms in the two parasite stages.

Effects of snail-conditioned water from Biomphalaria glabrata on hatching of Echinostoma caproni miracidia.

In vitro studies were done on Echinostoma caproni eggs with fully developed miracidia to determine the effects of snail-conditioned water (SCW) from Biomphalaria glabrata on miracidial hatching in the light. Observations were made on miracidial hatching at 4, 8, 12, 16, 20, and 24 h in multiwell chambers in the presence of SCW (experimentals) versus controls in artificial spring water (ASW). The number of hatched eggs was significantly greater (Student's t-test, P<0.05) in SCW at all times as compared with those maintained in ASW. Significantly greater hatching was also obtained when snails were maintained in intact or perforated dialysis sacs in multiwell chambers as compared with sacs without snails. Agar plugs impregnated with SCW or the hydrophilic fraction of SCW that had been extracted in chloroform-methanol (2:1) did not influence significant hatching. However, the lipophilic fraction of the SCW extract caused significant hatching. Substances in SCW significantly increase hatching of E. caproni miracidia, but details on what these compounds are remain obscure.

High-Performance Thin-Layer Chromatographic Analysis of Sugars in Snail-Conditioned Water and Mucus from Biomphalaria glabrata, Helisoma trivolvis, and Lymnaea elodes

The presence of sugars, particularly glucose, maltose, and sucrose, in the SCW of Biomphalaria glabrata, the two strains of Helisoma trivolvis, and Lymnaea elodes was variable, although only maltose was quantifiable in two samples of snail conditioned waters from B. glabrata. The only sugar detected in mucus was glucose, but it was below the lowest quantifiable amount in the mucus samples of B. glabrata and H. trivolvis. The mean percentage of glucose in the mucus of L. elodes was 0.0040% ± 0.00067%.

Chemo-orientation of echinostome cercariae towards their snail hosts: the stimulating structure of amino acids and other attractants

The cercariae of Pseudechinoparyphium echinatum and Echinostoma revolutum locate their host snails by turning back when swimming in decreasing gradients of the small molecular weight fraction (< 500) of snail conditioned water. Fractionation and chemical modifications of snail conditioned water from Lymnaea stagnalis showed that amino acids are necessary for the stimulating activity of snail conditioned water. A complete mixture of amino acids in concentrations determined from snail conditioned water had a high attraction. However, differently composed mixtures of amino acids and even single amino acids also had the same attraction as this complete mixture when used in concentrations corresponding to the total concentration of amino acids in snail conditioned water. Experiments with analogues and derivatives of amino acids showed that the primary α-amino group and the α-carboxyl group are necessary for the full effectiveness of amino acids. The highest effect was elicited by l-amino acids with a primary α-amino group, whereas the amino acid type and the chain length seemed to be unimportant. However, the full attraction of snail conditioned water was not achieved by amino acids alone. Chemical modifications of snail conditioned water suggested that the additional stimuli were neither inorganic ions nor organic acids or lipids. As the full attraction of snail conditioned water was obtained when the amino acid mixture of snail conditioned water was combined with its content of urea and ammonia, we conclude that the cercariae use only these excretory products as additional signals for their chemo-orientation. Chemo-orientation to amino acids, urea and ammonia seems to reflect a strategy to locate a broad spectrum of aquatic hosts.

Chemo-orientation of echinostome cercariae towards their snail hosts: amino acids signal a low host-specificity

The cercariae of Pseudechinoparyphium echinatum and Echinostoma revolutum approach their intermediate host snails by turning back when swimming in decreasing concentration gradients of snail conditioned water. Host stimuli are small molecular amino compounds, probably amino acids. This response was inhibited reversibly when the cercariae were treated with 0.45 μM-silver nitrate. As silver nitrate binds to the ciliated papillae of the cercarial surface, papillar chemoreceptors may be involved. Snail conditioned water from different snail species stimulated different intensities of chemo-orientation in both species of digeneans. However, when the samples of water conditioned with different snail species or even fish, tadpoles and leeches were diluted to the same total amino acid concentrations, the intensities of cercarial responses were similar. Therefore, some specificity of the chemo-orientation seemed to be achieved only by the total concentration of amino acids and not by particular mixtures of amino acids. In fact, amino acid spectra released by freshwater snails varied not only among different snail species, but also within the same species.