Guild structure of larval trematodes in the snail Helisoma anceps: patterns and processes at the individual host level

Factors that influenced the infracommunity structure of trematodes parasitizing the pulmonate snail Helisoma anceps were studied over a 15-mo period; the guild included 8 species of parasites. Infracommunities were depauperate, with double patent infections observed in only 7 of 1,485 infected snails; a total of 4,899 was examined. Halipegus occidualis-Haematoloechus longiplexus was the most common dual infection. Both species share the same definitive host and, in both cases, eggs are the infective stage for the snail. Switches and losses of infections in individual snails were observed, suggesting the occurrence of dynamic interactions within the guild. A dominance hierarchy was constructed based on field observations and experimental infections. Echinostomatids were dominant; species without rediae in their life cycles were subordinates. Halipegus occidualis (which has rediae) was intermediate in dominance. Spatial and temporal heterogeneity in the distribution and abundance of trematode infective stages indicate that not all the snails have the same probability of becoming infected. Habitat structure, behavior of the definitive host, the nature of the infective stages, and snail population dynamics (mortality, recruitment, and size structure) generated spatial and temporal heterogeneity in this system. As a consequence, predictions of the probabilities of interspecific interactions based on an analysis of observed and expected frequencies of multiple infections could be inappropriate unless the potential sources of heterogeneity are considered.     

Regulation and restoration of motoneuronal synaptic transmission during neuromuscular regeneration in the pulmonate snail Helisoma trivolvis

Regeneration of motor systems involves reestablishment of central control networks, reinnervation of muscle targets by motoneurons, and reconnection of neuromodulatory circuits. Still, how these processes are integrated as motor function is restored during regeneration remains ill defined. Here, we examined the mechanisms underlying motoneuronal regeneration of neuromuscular synapses related to feeding movements in the pulmonate snail Helisoma trivolvis. Neurons B19 and B110, although activated during different phases of the feeding pattern, innervate similar sets of muscles. However, the percentage of muscle fibers innervated, the efficacy of excitatory junction potentials, and the strength of muscle contractions were different for each cell's specific connections. After peripheral nerve crush, a sequence of transient electrical and chemical connections formed centrally within the buccal ganglia. Neuromuscular synapse regeneration involved a three-phase process: the emergence of spontaneous synaptic transmission (P1), the acquisition of evoked potentials of weak efficacy (P2), and the establishment of functional reinnervation (P3). Differential synaptic efficacy at muscle contacts was recapitulated in cell culture. Differences in motoneuronal presynaptic properties (i.e., quantal content) were the basis of disparate neuromuscular synapse function, suggesting a role for retrograde target influences. We propose a homeostatic model of molluscan motor system regeneration. This model has three restoration events: (1) transient central synaptogenesis during axonal outgrowth, (2) intermotoneuronal inhibitory synaptogenesis during initial neuromuscular synapse formation, and (3) target-dependent regulation of neuromuscular junction formation.     

Component community dynamics of larval trematodes in the freshwater snail Semisulcospira nakasekoae in the Uji River, central Japan

The component community of larval trematodes in the freshwater snail Semisulcospira nakasekoae (Caenogastropoda: Sorbeoconcha: Pleuroceridae) was surveyed over 13 months from April 1996 to April 1997 inclusive. Temporal and spatial fluctuation of trematode prevalence, the frequency of multiple infections, and the duration of cercarial shedding were examined as factors that might affect trematode community structure. The spatial prevalence of some species varied significantly, but the dynamics were too small to allow an explanation of the overall pattern. The prevalence of sanguinicolids fluctuated temporally, despite a stable size distribution in the host populations (> 6.0 mm shell width), suggesting the life-cycle phenology of this species. Some pairs of species had statistically positive associations, but no pairs had negative associations. This shows the importance of positive association possibly as a result of suppression of the host defensive response on trematode community structures in molluscan hosts. The length of the patent period, which is part of the persistent period, varied among trematode species, suggesting it to be one of the factors determining prevalence in the host population.     

Spatial heterogeneity in recruitment of larval trematodes to snail intermediate hosts

Spatial variation in parasitism is commonly observed in intermediate host populations. However, the factors that determine the causes of this variation remain unclear. Increasing evidence has suggested that spatial heterogeneity in parasitism among intermediate hosts may result from variation in recruitment processes initiated by definitive hosts. I studied the perching and habitat use patterns of wading birds, the definitive hosts in this system, and its consequences for the recruitment of parasites in snail intermediate hosts. Populations of the mangrove snail, Cerithidea scalariformis, collected from mangrove swamps on the east coast of central Florida are parasitized by a diverse community of trematode parasites. These parasites are transmitted from wading birds, which frequently perch on dead mangrove trees. I tested the hypothesis that mangrove perches act as transmission foci for trematode infections of C. scalariformis and that the spatial variation of parasitism frequently observed in this system is likely to emanate from the distribution of wading birds. On this fine spatial scale, definitive host behaviors, responding to a habitat variable, influenced the distribution, abundance and species composition of parasite recruitment to snails. This causal chain of events is supported by regressions between perch density, bird abundance, bird dropping density and ultimately parasite prevalence in snails. Variation between prevalence of parasites in free-ranging snails versus caged snails shows that while avian definitive hosts initiate spatial patterns of parasitism in snails through their perching behaviors, these patterns may be modified by the movement of snail hosts. Snail movement could disperse their associated parasite populations within the marsh, which may potentially homogenize or further increase parasite patchiness initiated by definitive hosts.     

Fine structure of photoreceptors in larval trematodes

Summary The fine structure of photoreceptors is described in miracidia of Fasciola hepatica, Heronimus chelydrae, Allocreadium lobatum, and Spirorchis sp., and in a spirorchiid cercaria. All have in common eyespots consisting of pigment cells with chambers occupied by rhabdomeres consisting of retinular cell dendrites with numerous microvilli. Photoreceptors of the miracidia show a bilateral asymmetry which is most pronounced in H. chelydrae with a pair of well separated eyespots unequal in size. The smaller right one consists of a pigment cell and two rhabdomeres; the larger left eyespot has an anterior pigment cell with two rhabdomeres and a posterior cell containing one rhabdomere. Photoreceptors in the other species of miracidia also have five rhabdomeres but contain only two pigment cells which are closely apposed. Each contains a pair of lateral rhabdomeres and a fifth one occupies a posteromedian extension of the left pigment cell. In the number of rhabdomeres, their relationship to pigment cells and the resulting asymmetry, photoreceptors are more alike in the distantly related species of miracidia studied than they are in ocellate cercariae or even in the miracidium and cercaria of the same species or two closely related ones. From the asymmetry of photoreceptors in larvae of certain flatworms other than digenetic trematodes, it seems that eyespots of miracidia have retained an ancestral pattern whereas the diversity of photoreceptors in cercariae reflects the varied phototactic behavior of those larvae which complete their life cycles by all the means known for cercariae with a free-swimming period. In both miracidia and cercariae, photoreceptors show an anterior-posterior organization that would seem to be concerned with orientation of the larvae with respect to light.Supported in part by a David Ross Fellowship of the Purdue Research Foundation and in part by U.S.P.H.S. Grants 1T1 GM 1392 01 and 2T1 Al 106 07. We express thanks to Dr. Keith Dixon for aid in obtaining and processing miracidia of Fasciola hepatica; to Prof. Clark P. Read for his valuable comments and suggestions; and to Profs. Charles W. Philpott and Richard H. White for advice concerning electron microscopy.     

Genotype frequency differences in Halipegus occidualis-infected and uninfected Helisoma anceps

Allozyme frequencies in Helisoma anceps infected with the hemiurid trematode, Halipegus occidualis, were compared with those of uninfected H. anceps from a small, North Carolina farm pond. Of 6 loci found to be polymorphic, the frequencies of esterase-1 and leucine aminopeptidase were different in infected and uninfected snails. Genetic heterozygosity, as determined by starch gel electrophoresis, was greater in uninfected H. anceps relative to infected individuals. These observations combined with the high prevalence (up to 60%), complete castration in patent infections, and the absence of an encapsulation response in infected snails, suggest that factors conferring incompatibility may have been selected for in the H. anceps population within the pond.     

Locomotion in the pulmonate snail Melampus--I. The motor pattern

1. The patterned neural activity that drives muscular locomotor movements in Melampus is generated within the central nervous system. 2. In the transition from quiescent state to crawling, the pattern recorded in nerves and connectives changes from short-duration bursts in many units to the 60-100 sec cycle of events recorded during tethered crawling in the semi-intact snail. 3. Extracellularly recorded bursts and individually recognizable spikes in pedal nerves are correlated with movements that occur at each stage of the cyclically repeated crawl-steps. 4. Intracellularly recorded pedal neurons involved in locomotion receive excitatory drive, inhibitory drive, or alternating excitatory and inhibitory drive during the step cycle.     

The unusual life cycle of Daubaylia potomaca, a nematode parasite of Helisoma anceps

Daubaylia potomaca is a parasitic nematode that exhibits a direct life cycle using planorbid snails as their only host. Within the snail host Helisoma anceps , all developmental stages of the parasite are present at any given time. The nematode has an unusual life cycle, with the adult female being the infective stage rather than the third-stage larvae (L(3)), as is commonly the case in many other parasitic nematode life cycles. In addition, length analysis showed that L(1) and L(2) were not present in tissues, suggesting that larvae hatch from eggs as the L(3). Previous studies by other investigators show that adult females abandon Biomphalaria glabrata at some point between 3 and 9 days of host death; in the present study, adult female D. potomaca leave H. anceps up to 59 days (and a mean of 14.8 days) before host death. This observation indicates a striking physiological difference between an experimental and a natural host for the parasite.     

Histology and ultrastructure of the olfactory organ of the freshwater pulmonate Helisoma trivolvis

Summary The olfactory organ of Helisoma trivolvis is located on the surface of the body at the base of the cephalic tentacles. An evagination of skin, the olfactory plica, at the base of the tentacle extends over the olfactory organ dorsally. The epithelium of the olfactory organs contains unspecialized epithelial cells, ciliated epithelial cells, basal cells, mucous secretory cells, and sensory dendrites. The surface of the epithelium has a complex brush border of thick plasmatic processes, which branch to form several terminal microvillar twigs. Long slender cytoplasmic processes form a dense spongy layer among the plasmatic processes beneath the level of the terminal twigs. Bipolar primary sensory neurons clustered beneath the epithelium of the olfactory organ send dendrites through the epithelium to the free surface. Some sensory endings have a few short cilia, but most bear only microvilli. Cilia of sensory endings and epithelial cells extend beyond the brush border of the epithelium. Small axons arise from the perikarya of the sensory neurons and enter a branch of the olfactory nerve. HRP tracing indicates that the axons pass to the cerebral ganglion without interruption. Histochemical tests indicate that the sensory neurons are neither aminergic nor cholinergic.     

The shell matrix of the pulmonate land snail Helix aspersa maxima

In mollusks, the shell mineralization process is controlled by an array of proteins, glycoproteins and polysaccharides that collectively constitute the shell matrix. In spite of numerous researches, the shell protein content of a limited number of model species has been investigated. This paper presents biochemical data on the common edible land snail Helix aspersa maxima, a model organism for ecotoxicological purposes, which has however been poorly investigated from a biomineralization viewpoint. The shell matrix of this species was extracted and analyzed biochemically for functional in vitro inhibition assay, for amino acid and monosaccharides compositions. The matrix was further analyzed on 1 and 2D gels and short partial protein sequences were obtained from 2D gel spots. Serological comparisons were established with a set of heterologous antibodies, two of which were subsequently used for subsequent immunogold localization of matrix components. Our data suggest that the shell matrix of Helix aspersa maxima may differ widely from the shell secretory repertoire of the marine mollusks studied so far, such as the gastropod Haliotis or the pearl oyster Pinctada. In particular, most of the biochemical properties generally attributed to soluble shell matrices, such as calcium-binding capability, or the capacity to interfere in vitro with the precipitation of calcium carbonate or to inhibit the precipitation of calcium carbonate, were not recorded with this matrix. This drastic change in the biochemical properties of the landsnail shell matrix puts into question the existence of a unique molecular model for molluscan shell formation, and may be related to terrestrialisation.     

Chemosensory stimulation of the oral cavity of the snail Helisoma

Food extracts, perfused through the oral cavity of the snailHelisoma trivolvis, lead to synaptic activation of identifiedbuccal ganglia motor neurons. Both retractor and protractormotor neurons displayed cyclic bursts of firing characteristicof that observed during expression of the central feeding motorprogram(CFM). The possibility that leakage of food extracts from theoral cavity had a pharmacological effect on buccal neurons wasconsidered. Direct application of the extracts to the exposedganglionic surface did not evoke similar neuronal activity.Oral perfusion with a behaviorally aversive compound inhibitedboth the activity evoked by acceptable taste solutions and "spontaneously"generated activity in some preparations. It is concluded thatoral chemosensory receptors in the snail exert both an excitatoryand inhibitory influence on buccal motor neurons. The significanceof these results for cellular neurophysiological investigationof the synaptic events underlying the central processing ofafferent chemosensory information is discussed.     

Long-term analysis of Charlie's Pond: fecundity and trematode communities of Helisoma anceps

Charlie's Pond (North Carolina) harbors a diverse community of trematodes that infect the planorbid snail Helisoma anceps. Research at the Pond began in 1984 and serves as a foundation on which to investigate long-term changes in trematode communities. In 2002, 2005, and 2006 average size and fecundity of H. anceps were calculated each month, and seasonal trends analyzed with randomization tests. Concomitantly, trematode infections were recorded, and the community composition compared to those from previous studies. Helisoma anceps in 2002, 2005, and 2006 were smaller and less fecund than snails in 1984. The trematode community was consistently diverse, with 11 species recovered in 2006 versus 7 in 1984. However, the prevalence of Halipegus occidualis was much lower than previously observed (60% in 1984) and never exceeded 20% during the latter years. The decline of emergent vegetation is likely contributing to these changes. Aquatic macrophytes increase the surface area for growth of periphyton, the food source of these snails. Limited food supplies result in lower snail growth rates and fecundity. Similarly, emergent vegetation creates foci of transmission for H. occidualis between the frog definitive host and the snail intermediate host. When these areas are lost from the Pond, probability of transmission is reduced, and prevalence in the snail declines.     

Life history cost of trematode infection in Helisoma anceps using mark-recapture in Charlie's Pond

Parasitism has the potential to affect key life history traits of an infected host. Perhaps the most studied interactions are in snail-trematode systems, where infection can result in altered growth rates, survival, and/or fecundity of the individual. Positive correlations between host size and parasite prevalence are often attributed to changes in growth rates or mortality, which have been observed in the laboratory. Extending lab-based conclusions to the natural setting is problematic, especially when environmental conditions differ between the laboratory and the field. The present study uses reproduction experiments and mark-recapture methods to directly measure key life history traits of the pulmonate snail Helisoma anceps in Charlie's Pond. Based on previous laboratory and field experiments on H. anceps, we predict a significant reduction in fecundity, but not growth rate or survival, of infected snails. Individual capture histories were analyzed with multistate models to obtain estimates of survival and infection probabilities throughout the year. Recaptured individuals were used to calculate specific growth rates. Trematode infection resulted in complete castration of the host. However, neither survival nor growth rates were found to differ between infected and uninfected individuals. The probability of infection exhibited seasonal variation, but it did not vary with size of the snail. These results suggest that the correlation between host size and trematode prevalence is not due to differential mortality or changes in growth rates. Instead, the infection accumulates in large snails via the growth of smaller, infected individuals.     

Infrapopulation sizes of co-occurring trematodes in the snail Ilyanassa obsoleta

This study addresses the infrapopulation sizes of 2 larval trematode species Himasthla quissetensis and Zoogonus rubellus as they co-occur within their estuarine snail host Ilyanassa obsoleta. Rediae of H. quissetensis and sporocysts of Z rubellus were counted in snails singly infected with each parasite and in snails infected with both. Comparisons of the counts indicate that infrapopulations of H. quissetensis were unaffected by co-occurrence with Z rubellus. However, Z. rubellus infrapopulations were reduced when co-occurring with H. quissetensis. It is proposed that this situation does not result from an interspecific interaction between parasite species. Although this double infection is relatively frequent in certain snail populations, it is contended that these trematode species do not co-occur often enough to evolve responses to one another. However, the host environment must be encountered in each life cycle, and both trematode species must be adapted to use it. On this basis, whatever happens when these 2 species occupy the same host is based on adaptations of the parasites to the host. It is proposed that these parasites are adapted to self-limit their infrapopulations in the snail host. They can, thus, preserve and use the host for many years and thereby enhance total cercarial transmission (fitness). Infrapopulation sizes would be determined by host resource levels, which, among other factors, would be influenced by the presence of multiple parasite species. In single infections, by far the most common situation, host resource levels would be set by the nutritional status or age (size) of the host (or both). The reduced infrapopulation sizes of Z rubellus on co-occurrence suggest that this trematode is more sensitive to host resource levels than is H. quissetensis.     

Impact of larval trematode infection on the nitrogen excretion and ornithine-urea cycle enzymes of the pulmonate snail host,Lymnaea luteola

The freshwater gastropod Lymnaea luteola infected with xiphidiocercariae of Prosthogonimus sp. showed striking changes in nitrogen excretion. Infected snails excreted significantly less total Kjeldahl nitrogen into the amibient medium. A significant drop in urea nitrogen alone accounted for this drop in total nitrogen excreted, as there was no change in ammonia excretion. While no significant change was seen in the activity of ornithine carbamyltransferase and arginosuccinate lyase in infected snails, arginase activity invariably dropped. The present study thus has revealed that it is not the urea production that occurs in the digestive gland that is affected upon infection, but it is arginolysis that occurs in other tissues, like foot and mantle, that is curtailed.     

Oral water ingestion in the pulmonate freshwater snail,Lymnaea stagnalis

InLymnaea stagnalis, oral uptake of ambient medium was studied using⁵¹Cr-ethylenediaminetetra-acetic acid. In normal snails Cr-ethylenediaminetetra-acetic acid uptake showed two components: a high uptake rate within the first hour followed by moderate uptake proportional with time. The tracer accumulated mainly in the digestive system. All animals showed initial, transient uptake. Moderate uptake proportional with time did not occur in snails in which the buccal ganglia had been extirpated, in which both the buccal ganglia had been extirpated and the oesophagus was sectioned, or in snails provided with an oesophageal fistula. These snails did not accumulate tracer in the intestinal system. This type of tracer accumulation clearly represented oral ingestion of surrounding water. The oral water ingestion rate ranged from 8 to 12 μl·h⁻¹·g⁻¹. Assuming complete absorption, this accounts for 20–30% of the urine production rate. At low external concentrations the contribution of oral water ingestion to Na⁺ balance is negligible. However, its importance will grow with increasing external concentrations and becomes a major factor at higher concentrations. The ingestion rate increased almost sixfold when starving snails were allowed to feed. It is suggested that oral water ingestion is a consequence of making bite cycles and swallowing.     

Genetic structure of the common terrestrial pulmonate snail,Cryptozona siamensis (Pfeiffer, 1856), in Thailand

Allozyme variation was examined in 432 specimens of the pulmonate snail, Cryptozona siamensis, from 24 localities across three regions of Thailand plus an additional population from Malaysia. Although this snail is a cosmopolitan species it was predicted that C. siamensis should exhibit a large degree of genetic subdivision across these regions due to a low level of inter-population gene flow. To test this hypothesis, 10 enzyme systems were screened using horizontal starch gel electrophoresis, from which 14 presumed allozyme loci were scored and 13 found to be polymorphic. For these 13 polymorphic loci the expected heterozygosity levels were moderate (H_exp = 0.036–0.183, mean = 0.101). Genetic differentiation among the samples was low, with a low F_ST value (0.254) and genetic distance (D = 0.000–0.124, mean = 0.034) and no significant isolation by distance (IBD) across the whole sampled range of Thailand or Thailand plus Malaysia. However, an IBD pattern in the population substructure in the central Thailand region was detected. The low F_ST and moderate H_exp values in various populations of C. siamensis suggests a high degree of gene flow/migration between populations across the three geographic regions of Thailand (northern, central and southern) as well as the sampled population in Malaysia, perhaps associated with the anthropochoric effect of snail dispersal. The apparent IBD in the central Thailand region could be the consequence of local dynamics though this region.