Functional morphology of the neuropeptidergic light-yellow-cell system in pulmonate snails

The light yellow neuropeptidergic cell system of the basommatophoran snail Lymnaea stagnalis is homologous to the R3-R14 system of the opisthobranch Aplysia californica, and produces three different neuropeptides. Systems homologous to the light yellow cells of Lymnaea stagnalis have been investigated morphologically in two Basommatophora (Lymnaea ovata, Bulinus truncatus) and three Stylommatophora (Helix aspersa, Cepaea nemoralis, Deroceras reticulatum). To this end, an antibody to synthetic light-yellow-cell peptide-II and oligonucleotides to mRNAs encoding parts of peptide-I and peptide-III, were used. The in situ hybridization probes gave negative results. On the other hand, neuronal cell clusters were observed in the central nervous system of all specias studied by immunocytochemistry. These clusters were located in the ganglia of the visceral complex. The neurons project axons into all nerves of these ganglia, especially into the pallial nerves, into the connective tissue of the central nervous system, and into the neuropile of various ganglia. The morphology of the systems is similar to that of the light-yellow-cell system of Lymnaea stagnalis. In all species, the wall of the aorta was innervated by immunoreactive axons. Peripheral innervation by the light-yellow-cell system was investigated in Helix aspersa and Deroceras reticulatum. Serial and alternate sections of whole snails were studied. Reconstructions were made of the heart-kidney-lung complex of these animals. In both species, the muscular vessels of the pulmonary system at the right side of the body were strongly innervated by immunoreactive axons. Furthermore, immunopositive innervation was observed to muscles in the secondary ureter-pneumostome area. The light-yellow-cell system of pulmonates is thus probably involved in the regulation of blood pressure and urine release.     

The distribution and variation of three species of land snail near Rickmansworth, Hertfordshire

The distribution of Arianta arbustorum, Cepaea nemoralis and C. hortensis , and the variations in the shell polymorphism of the Cepaea species have been studied in an area of low chalk downland and alluvial river valleys near Rickmansworth, Hertfordshire. The distribution of A. arbustorum is restricted, the species being largely confined to the bottoms of river valleys, especially in hedgerows. Variations in dampness seem the most likely limiting factor. C. hortensis is widespread and found in many habitats, but C. nemoralis is largely confined to woods, and is especially common in beechwoods. The morph-frequencies of C. nemoralis and C. hortensis vary in such a way as to suggest the operation of climatic and visual selection. In C. hortensis there are area effects for the presence or absence of pink five-banded shells, and there is an overall linkage disequilibrium resulting in an excess of unbandeds in non-yellow shells. Reasonable evidence for the operation of random factors in determining the morph-frequencies of populations of Cepaea in the area has been found only for three very disturbed populations. It is suggested that the large number of selective forces known to act on Cepaea populations in this area will account for the variation in morph-frequencies in populations from the same area or habitat.     

Structural diversity of NADPH diaphorase-reactive enteral networks in Stylommatophora (Gastropoda, Pulmonata)

Abstract. In this work we investigated the involvement of putative nitric oxide (NO)-forming neurons in enteric plexuses of stylommatophoran gastropods. The nitric oxide synthase (NOS)-containing cells were detected by NADPH diaphorase (NADPHd) histochemistry in the entreral nervous systems of several stylommatophoran species (Achatinacea: Achatina fulica , Helicacea: Cepaea hortensis, Cepaea nemoralis, Discus rotundatus, Helicella obvia, Helix lucorum, Helix lutescens, Monachoides umbrosa, Trichia hispida, Zebrina detrita , Succineacea: Succinea putris , Vertiliginacea: Clausilia dubia , Zonitacea: Arion ater, Arion subfuscus, Limax maximus ). We detected the NO synthesis of isolated midintestinal segments by Griess's quantification of nitrite, one end product of NO. Effects of the NOS substrate L-arginine and the NOS inhibitor Nω-nitro-L-arginine (NOARG) were also tested on nitrite production. We found NADPHd-reactive neurons and extrinsic nerves with NADPHd-stained fibers within the myenteric and submucosal networks of the midintestine of investigated members of Helicacea, Succineacea, and Vertiliginacea families. These networks innervated the midintestinal musculature and several nerve cells of the myenteric and submucosal plexi. In investigated members of Achatinacea and Zonitacea, NADPHd-stained networks were not detectable within the digestive tract. Administration of 1 mM L-arginine elevated, whereas 2 mM of NOARG diminished, the nitrite levels of the NADPHd-stained networks containing midintestine in C. nemoralis and H. lucorum . Enteral NADPHd staining was not detected in A. ater and L. maximus , and the nitrite production was not affected by L-arginine. Our results indicate a possible, but evolutionarily not conserved, NO-mediated enteral transmission in stylommatophoran gastropods.     

Differentiation of the prostate in the snail Helix aspersa Müll]

The differentiation of the snail Helix aspersa prostate gland is studied in animals from one to six months old. After the presentation of organogenesis the primary stage of secretion (3 months) and a stage where the prostate gland shows abundant secretions (6 months) were described; but the oviduct is still indifferentiated. The ultrastructural analysis shows the formation of secretory and ciliated cells in epithelioid prostatic tubes.     

Gene frequency changes in Cepaea snails on the Marlborough Downs over 25 years

In 1985 we resurveyed the sites on the Marlborough Downs in southern England at which Cain and Currey in 1960/61 sampled Cepaea snails and thence introduced the term 'area effects' to describe large areas of uniform morph frequency. Some sites no longer harboured Cepaea and at others the species composition had changed, with a general spread of Cepaea hortensis at the expense of Cepaea nemoralis. The majority, however, permitted comparison of morph frequencies between the two surveys. In C. nemoralis, we detected a significant overall decrease in the frequency of the brown morph and estimate selection as 5–9% per generation. There was no apparent change in frequencies of banded morphs. In C. hortensis we detected a significant overall increase in the frequency of unbanded shells (1–3% selection per generation) and an almost significant decrease in the frequency of fusions within the banded class. There was insufficient colour polymorphism in C. hortensis to allow analysis of colour morph frequencies. These changes—all in the direction of reduced absorption of solar energy—resemble others detected in both species at other localities in southern England. Possible explanations include large-scale climatic effects and changes in vegetation.     

The effect of temperature on the activity of three species of Helicid snail (Mollusca: Gastropoda)

The activity of the land snails Cepaea nemoralis (L.), C. hortensis (Müll.) and Arianta arbustorum (L.) was measured over 24 hours at the constant temperatures of 0, 3, 8, 17, 22 and 26°C at 100% relative humidity and with 16 hours light and 8 hours dark. The activity of all three species varies significantly with temperature and light, and there are differences in the activities of the species at each temperature. A. arbustorum is relatively more active than Cepaea at low rather than high temperatures, as is C. hortensis with respect to C. nemoralis , though to a lesser extent. There is little diurnal rhythm in activity at 0°C, but all species become more nocturnal as temperature increases; C. nemoralis is most, and A. arbustorum least nocturnal. The results obtained assist in explaining the observed geographical and local distributions of the species concerned.     

Poaceae in the natural diet of the snail Helix aspersa Müller (Gastropoda, Pulmonata).

The natural diet of Helix aspersa was studied in two populations by analysing faeces. Picris echioides, Carduus tenuifloris, Urtica dioica, Galium molugo (Dicotyledons) and Poaceae (Monocotyledons) were the principal resources of the studied populations. The species of Poaceae ingested by Helix aspersa in July were determined by analysing the phytoliths present in the faeces. Festuca rubra, Dactylis glomerata and Bromus hordeaceus seemed to be attractive for the snails whereas Elytrigia repens was rejected. These results were convergent with the ingestion rates of Poaceae in the laboratory but assimilation efficiencies did not explain these choices. Poaceae with the highest energetic values were not preferred to others. The texture, the mineral and organic contents of the Poaceae may be responsible for the snails feeding choices. The importance of Poaceae for this species is discussed.     

The reactivity of canine red cells with heterophile agglutinins having anti-A activity*

Heterophile A-reactive agglutinins, obtained from the seeds of Dolichos biflorus and from the snails Helix aspersa, Helix pomatia and Cepaea nemoralis, were tested for reactivity with the saliva and red cells of a random series of dogs. Saliva from dogs with the A-like Tr red cell antigen inhibited the agglutination of human A red cells by each of the reagents. However, none of the heterophile agglutinins distinguished Tr+ from Tr— red cells: the Dolichos agglutinin was non-reactive, while all of the snail reagents agglutinated both Tr+ and Tr— cells. It is concluded that the canine red cell bears a series of heterophile receptors which mask reactivity with the canine homologue of the red cell A-antigen, and that the restricted size of the binding sites of the agglutinins limits the informational significance of cross-reactivity obtained with these reagents.     

Food overlap in co-existing populations of the land snails Cepaea nemoralis (L.) and Cepaeahortensis (Müll)

Faecal analysis of adult Cepaea nemoralis and Cepaea hortensis from a mixed population on chalk grassland shows that the two snail species select the same plant material as food. Herbs are selected in preference to grasses and Urtica dioica is particularly favoured. C. hortensis has the more pronounced preference for senescent material. These results are discussed in relation to competition between the two species.     

Interacting effects of herbivory and fertility on a synthesized plant community

1 Outdoor microcosms were used to investigate the effects of invertebrate herbivory on plant community composition, and thereby infer possible effects on the rate of secondary succession, at differing levels of soil fertility.
2 A mixture containing 24 grassland plant species of widely different functional types was established, with 12 microcosms at each of three fertility levels. Four generalist herbivores ( Helix aspersa , Cepaea hortensis , Arianta arbustorum and Sitobion avenae ) were added to half of the microcosms. Above-ground biomass of each species was harvested after 2 years. Reproductive variables were also measured for one species, Poa annua .
3 At both moderate and high soil fertility generalist invertebrate herbivores fed selectively on early successional, fast-growing species, thus increasing the relative abundance of later successional, slow-growing species. This supports the hypothesis that herbivory increases the rate of secondary succession. Flowering and viable seed production of early successional ephemerals was also reduced by the invertebrate herbivores across a wide range of soil fertility. This would seriously reduce the ability of a species to persist in the community, thereby further hastening the rate of succession.     

The survival, weight-loss and behaviour of three species of land snail in conditions of low humidity

Cepaea nemoralis L., C. hortensis Mull, and Arianta arbustorum L. show differences in survival, rate of water-loss and behaviour under conditions of low humidity in laboratory studies. C. nemoralis survives better, ceases activity more rapidly, and loses proportionally less weight in low humidity than C. hortensis , which in turn shows the same properties with respect to A. arbustorum . The differences between these three species are reflected in differences in distribution in parts of Britain.     

The habitats of the land snail Cepaea nemoralis (L.) on downland and their ecogenetie significance

Population densities of adult Cepaea nemoralis (L.) were estimated for nine chalk grassland sites on the South Downs. Estimates of cover, species composition and height of vegetation were also obtained. The density of C. nemoralis was positively associated with Bromus erectus cover, and negatively with Festuca spp. cover. This finding, confirmed by an earlier survey, suggests that the density of C. nemoralis in chalk grassland is directly or indirectly governed by mammalian grazing pressure, and not by the availability of suitable food. Intensely grazed downland is an unsuitable, or at best a marginal habitat for C. nemoralis. This conclusion is discussed with particular reference to 'area effects' in the shell character polymorphism, often found in downland populations of C. nemoralis.     

Epithelial cell types of the primary ureter of Helix aspersa: Ultrastructural and cytochemical characteristics

The present study describes the morphological characteristics which determine the structural polarity of the principal and ciliated cells in the primary ureter epithelium of Helix aspersa. These characteristics are analysed on the basis of the function performed by both cell types. The presence of paniculate glycogen and the location of glycoconjugates associated with cell membranes of the epithelial cells is revealed by the method of Thiéry.     

The chromosomes of Cepaea vindobonensis (Pulmonata: Helicidae) and the relationship between this species and the rest of the genus

The karyotype of Cepaea vindobonensis (Pfeiffer) (2 n = 50) is defined and compared with those of C. sylvatica (Draparnaud), C. nemoralis (L.) and C. hortensis (Müller). Cepaea vindobonesis shares its chromosome number with C. sylvatica. The genus Cepaea thus consists of two pairs of species, one with 2 n = 50 the other with 2 n = 44. The karyotype of C. vindobonensis is distinct from that of C. sylvatica as it lacks any discontinuity in its range of chromosome length and has all three of its longest chromosome pairs nearly identical. The two 2 n = 50 species both have an unusual meiotic bivalent showing a 'satellite-type' structure. It is apparent that the 2 n = 50 species are more closely related to each other than to the 2 n = 44 species. No precise definition of the evolutionary history can be given but a working hypothesis is suggested.     

Evolutionary history of the land snail Helix aspersa in the Western Mediterranean: preliminary results inferred from mitochondrial DNA sequences

Intraspecific phylogeographic methods provide a means of examining the history of genetic exchange among populations. As part of a study of the history of Helix aspersa in the Western Mediterranean, we performed a phylogenetic analysis based on partial sequences of the mitochondrial large ribosomal subunit (16S) gene. Our samples include 31 H. a. aspersa populations from North Africa previously investigated for anatomical and biochemical characters. To clarify subspecific relationships, three individuals of the subspecies H. a. maxima were also studied. The molecular phylogeny inferred agrees largely with previous results, in splitting H. a. aspersa haplotypes into an eastern and a western group. H. a. maxima haplotypes form a third lineage arising before the H. a. aspersa groups. Divergence times estimated between the lineages suggest that dispersal during Pleistocene glaciation and vicariance events due to Pliocene geological changes in the western Mediterranean may both have played a significant part in the establishment of the present range of H. aspersa.     

A Neurosecretory Phenomenon in Some Cerebral Nervous Cells of the Land Snail Helix aspersa maxima (Mollusca,Gastropoda), and Its Relationship to Circadian Locomotor and Food-Intake Activities

Some cerebral neurosecretory cells of the land snail Helix aspersa maxima have been studied using light microscopy combined with image analysis. The amount of neurosecretion varies during the course of the day, displaying a peak at 22:00 GMT. This peak is in phasis with the maxima of the locomotor and food-intake activities recorded by means of a video apparatus.     

Field prevalence and laboratory susceptibility of southern Australian land snails to Brachylaima cribbi sporocyst infection

Brachylaima cribbi is a terrestrial trematode of birds and mammals with helicid and hygromiid land snails reported as first and second intermediate hosts. However, reports describing the first intermediate host range of B. cribbi have been limited to those snail species present in a small number of geographical locations in South Australia. The natural first intermediate host range, distribution and prevalence of B. cribbi in land snails in southern Australia were determined. A total of 6,432 introduced and native land snails were collected from eight geographical districts across 3,000 km of southern Australia and examined microscopically for B. cribbi sporocysts. Four introduced European snails, Theba pisana, Cernuella virgata, Cochlicella acuta and Cochlicella barbara were natural first intermediate hosts. Sporocyst-infected snails were detected in all districts from Victoria to the west coast of South Australia, a distance of over 1,300 km. Natural sporocyst infection was not observed in introduced European snails Microxeromagna armillata and Helix aspersa or in native Australian land snails Succinea australis and Strangesta gawleri. Egg feeding experiments in the laboratory with B. cribbi confirmed the susceptibility of those species of snails found to be natural first intermediate hosts. Of those species not found to be infected in nature, only M. armillata could be infected in the laboratory. Although this study has shown that five different species of European land snails are suitable first intermediate hosts for B. cribbi there are as yet no reports of B. cribbi from these snails in Europe or from other countries where they have been introduced. Further investigations are needed in Europe to clarify the origins of this parasite.     

Patterns of morphological and molecular polymorphism in the land snail Cepaea nemoralis

In the land snail Cepaea nemoralis , allele frequencies at loci controlling shell polymorphisms often show large areas of remarkable constancy which are separated by steep clines from neighbouring areas with strikingly different allele frequencies. It has recently been claimed that these 'area effects' exemplify a general tendency for population differentiation without geographic isolation in a variety of organisms of relatively low mobility. As such they could represent an early phase in the process of speciation. If this is true, population differentiation of shell polymorphisms in Cepaea would be expected to be accompanied by parallel differentiation at other gene loci, such as those detected by gel electrophoresis.
We have studied populations of C. nemoralis in North Wales and in the Valle de Aran of the Pyrenees. Levels of molecular heterogeneity are comparable to those found in related animals which show much less visible polymorphism. In spite of some statistical problems inherent in the analysis ol overlapping geographic patterns, there is no clear association between the patterns of geographic variation at the visible and molecular levels. Claims that genetic differentiation in the visible polymorphisms between C. nemoralis populations are a special case of the formation of geographic races are therefore probably not justified.     

Cepaea on Salisbury Plain: patterns of variation, landscape history and habitat stability

Patterns of variation in Cepaea nemoralis and C. hortensis on Salisbury Plain are related to its landscape history. Areas of arable or mixed farming, with hedgerows and woods, show patterns of variation with habitat and topography. Areas of downland, until recently heavily grazed by sheep, show area effects. Cepaea populations in these downland areas would have been much rarer, and more isolated when grazing was intense. This pattern is repeated in other parts of southern England, showing that area effects in Cepaea are strongly associated with areas of habitat instability, where Cepaea survived as small and isolated populations until recently. This evidence enhances the plausibility of explanations of area effects involving isolation, founder effect or local selection, and co-adaptation.     

The karyotype of Cepaea sylvatica (Pulmonata: Helicidae) and its relationship to those of C. hortensis and C. nemoralis

The karyotypes of Cepaea nemoralis (L.) and C. hortensis (Müller), with 2n=44 and a conspicuously large pair of chromosomes, are described and compared with that of C. sylvatica (Draparnaud) which has 2n=50. The karyotype of C. sylvatica also has a conspicuously large pair of chromosomes but the comparison suggests that these have an independent origin from those in the 2n = 44 species. There is no evidence that the large chromosomes in C. nemoralis and C. hortensis have originated from simple fusion of chromosomes from a 2n=50 karyotype with chromosomes all sub-equal such as is reported for C. vindobonensis. It may be that such a karyotype with little size differentiation amongst the chromosomes is not a primitive feature in the Helicinae. The relationship of shell colour and banding polymorphism to the chromosome architecture is discussed.