Pulmonate Land Research Articles

Update of available knowledge about the non-marine molluscs occurring in the “Republic of El Salvador”, Central America

With the invaluable participatory assistance and support of local researchers and collaborating informants in field, accompanied for the fundamental revision of the regional bibliography, once again through the AM Project (Brazil) it was possible to achieve the incorporation of new informations for the inventory in progress about the current non-marine land and freshwater molluscs present in the Central American territory of El Salvador Republic, including five (5) new gastropod species not previously reported, being four (4) pulmonate land/ terrestrial forest and one (1) operculate limnic/ freshwater, bringing the number of confirmed inventoried forms to 35. Additionally, the record of another two (2) terrestrial forest species previously registered for the territory is still presented, reinforcing the confirmation of its valid presence/ occurrence in the region.

Pulmonate slug evolution is reflected in the de novo genome of Arion vulgaris Moquin-Tandon, 1855

Stylommatophoran pulmonate land slugs and snails successfully completed the water-to-land transition from an aquatic ancestor and flourished on land. Of the 30,000 estimated species, very few genomes have so far been published. Here, we assembled and characterized a chromosome-level genome of the “Spanish” slug, Arion vulgaris Moquin-Tandon, 1855, a notorious pest land slug in Europe. Using this reference genome, we conclude that a whole-genome duplication event occurred approximately 93–109 Mya at the base of Stylommatophora and might have promoted land invasion and adaptive radiation. Comparative genomic analyses reveal that genes related to the development of kidney, blood vessels, muscle, and nervous systems had expanded in the last common ancestor of land pulmonates, likely an evolutionary response to the terrestrial challenges of gravity and water loss. Analyses of A. vulgaris gene families and positively selected genes show the slug has evolved a stronger ability to counteract the greater threats of external damage, radiation, and water loss lacking a protective shell. Furthermore, a recent burst of long interspersed elements in the genome of A. vulgaris might affect gene regulation and contribute to rapid phenotype changes in A. vulgaris, which might be conducive to its rapid adaptation and invasiveness.

Low genetic diversity and the phylogeny of Achatina fulica, an intermediate host of Angiostrongylus cantonensis in Thailand, inferred from 16S mitochondrial sequences.

The giant African land snail, Achatina fulica, is a pulmonate land snail of the Achatinidae family. The snail is native of East Africa and has spread to tropical and subtropical areas in different parts of the world. Achatina fulica is an important agricultural pests and the intermediate host for Angiostrongylus spp., especially Angiostrongylus cantonensis, which causes eosinophilic meningoencephalitis in humans. However, information on the genetic diversity and phylogenetic relationships among A. fulica populations in Thailand is limited. The goal of this study was to evaluate the genetic diversity and analyze haplotype network relationships between A. fulica populations in Thailand and other areas of the world. We collected A. fulica from 22 provinces in 6 regions of Thailand and analyzed a phylogenetic tree and haplotype network based on a fragment of 239 base pairs of the 16S rRNA sequence. The maximum likelihood, neighbor joining, and Bayesian inference trees for the 133 A. fulica 16S rRNA sequences revealed only one group, closely related only to A. fulica haplotype C, in all regions of Thailand. The haplotype network analysis showed that haplotype C is associated with F, P, E, Q, H and D. The genetic distance between the haplotype and A. fulica isolates in Thailand varies from 0 and 0.024, with a total of 21 variable sites. Haplotype C is predominant in Thailand and associated with other haplotypes from several areas. As inferred from 16S rDNA sequences, this study demonstrated low genetic diversity in A. fulica in Thailand. Low genetic diversity in the population can increase susceptibility to A. cantonensis infection, which may be associated with the current distribution of A. cantonensis.

Assessment of mitochondrial genomes for heterobranch gastropod phylogenetics

BackgroundHeterobranchia is a diverse clade of marine, freshwater, and terrestrial gastropod molluscs. It includes such disparate taxa as nudibranchs, sea hares, bubble snails, pulmonate land snails and slugs, and a number of (mostly small-bodied) poorly known snails and slugs collectively referred to as the “lower heterobranchs”. Evolutionary relationships within Heterobranchia have been challenging to resolve and the group has been subject to frequent and significant taxonomic revision. Mitochondrial (mt) genomes can be a useful molecular marker for phylogenetics but, to date, sequences have been available for only a relatively small subset of Heterobranchia.ResultsTo assess the utility of mitochondrial genomes for resolving evolutionary relationships within this clade, eleven new mt genomes were sequenced including representatives of several groups of “lower heterobranchs”. Maximum likelihood analyses of concatenated matrices of the thirteen protein coding genes found weak support for most higher-level relationships even after several taxa with extremely high rates of evolution were excluded. Bayesian inference with the CAT + GTR model resulted in a reconstruction that is much more consistent with the current understanding of heterobranch phylogeny. Notably, this analysis recovered Valvatoidea and Orbitestelloidea in a polytomy with a clade including all other heterobranchs, highlighting these taxa as important to understanding early heterobranch evolution. Also, dramatic gene rearrangements were detected within and between multiple clades. However, a single gene order is conserved across the majority of heterobranch clades.ConclusionsAnalysis of mitochondrial genomes in a Bayesian framework with the site heterogeneous CAT + GTR model resulted in a topology largely consistent with the current understanding of heterobranch phylogeny. However, mitochondrial genomes appear to be too variable to serve as good phylogenetic markers for robustly resolving a number of deeper splits within this clade.

Metagenomic Analysis of the Gut Microbiome of the Common Black Slug Arion ater in Search of Novel Lignocellulose Degrading Enzymes.

Some eukaryotes are able to gain access to well-protected carbon sources in plant biomass by exploiting microorganisms in the environment or harbored in their digestive system. One is the land pulmonate Arion ater, which takes advantage of a gut microbial consortium that can break down the widely available, but difficult to digest, carbohydrate polymers in lignocellulose, enabling them to digest a broad range of fresh and partially degraded plant material efficiently. This ability is considered one of the major factors that have enabled A. ater to become one of the most widespread plant pest species in Western Europe and North America. Using metagenomic techniques we have characterized the bacterial diversity and functional capability of the gut microbiome of this notorious agricultural pest. Analysis of gut metagenomic community sequences identified abundant populations of known lignocellulose-degrading bacteria, along with well-characterized bacterial plant pathogens. This also revealed a repertoire of more than 3,383 carbohydrate active enzymes (CAZymes) including multiple enzymes associated with lignin degradation, demonstrating a microbial consortium capable of degradation of all components of lignocellulose. This would allow A. ater to make extensive use of plant biomass as a source of nutrients through exploitation of the enzymatic capabilities of the gut microbial consortia. From this metagenome assembly we also demonstrate the successful amplification of multiple predicted gene sequences from metagenomic DNA subjected to whole genome amplification and expression of functional proteins, facilitating the low cost acquisition and biochemical testing of the many thousands of novel genes identified in metagenomics studies. These findings demonstrate the importance of studying Gastropod microbial communities. Firstly, with respect to understanding links between feeding and evolutionary success and, secondly, as sources of novel enzymes with biotechnological potential, such as, CAZYmes that could be used in the production of biofuel.

Radiocarbon Ecology of the Land Snail Helix Melanostoma in Northeastern Libya

AbstractTerrestrial gastropods are problematical for radiocarbon (14C) measurement because they tend to incorporate carbon from ancient sources as a result of their dietary behavior. The 14C ecology of the pulmonate land snail, Helix melanostoma in Cyrenaica, northeastern Libya, was investigated as part of a wider study on the potential of using terrestrial mollusk shell for 14C dating of archaeological deposits. H. melanostoma was selected out of the species available in the region as it has the most predictable 14C ecology and also had a ubiquitous presence within the local archaeology. The ecological observations indicate that H. melanostoma has a very homogenous 14C ecology with consistent variations in F14C across sample sites controlled by availability of dietary vegetation. The majority of dated specimens from non-urbanized sample locations have only a small old-carbon effect, weighted mean of 476±48 14C yr, with between ~1% and 9% of dietary F14C from non-organic carbonate sources. Observed instabilities in the 14C ecology can all be attributed to the results of intense human activity not present before the Roman Period. Therefore, H. melanostoma and species with similar ecological behavior are suitable for 14C dating of archaeological and geological deposits with the use of a suitable offset.

Hygromia cinctella (Draparnaud, 1801) (Mollusca: Gastropoda: Hygromiidae): a new adventive land snail for New Zealand

ABSTRACTHygromia cinctella is a medium-sized, pulmonate land snail native to the Mediterranean region and is undergoing rapid range expansion in Europe. Several living specimens were collected from an urban garden in the Wellington suburb of Brooklyn in 2015 and 2016, and constitute the first records of this species from New Zealand. Searches in surrounding suburbs have so far failed to find further snails. Eradication from Brooklyn may be viable but it seems likely that the species occurs elsewhere and the potential threat it poses to native species and agriculture, should it become established in New Zealand, appears low.

A new species of Leiostracus (Gastropoda, Pulmonata, Orthalicoidea) from Espírito Santo, Brazil

A remarkable new species of pulmonate land snail was found in the collection of the Senckenberg Forschungninstitut und Naturmuseum Frankfurt (Frankfurt am Main, Germany) and is described here as Leiostracus faerie sp. nov. It can be easily identified by its small and translucent shell with fine axial light brown bands and its protoconch sculpture. It was collected in the Rio Doce ("Doce River") region in Espírito Santo, Brazil, an area known for a high diversity and endemicity of land snails. This discovery shows how little this fauna is known and reinforces the importance of museum collections in the study of biodiversity and conservation.

Population dynamics and reproductive biology ofAchatina fulica Bowdich, 1822 (Mollusca, Gastropoda) in Salvador - Bahia

The risks following introduction of invasive species are enormous and incalculable. Achatina fulica is considered one of the 100 species of greatest invasive potential, stands out among the land pulmonate snails, mostly for its high reproductive potential that accelerates the process of dispersion, increasing the damages related to health (disease transmission), to economy (crops destruction) and to environment (biodiversity loss). In order to investigate the relationship of the African snail with the environment, trying to relate sexual activity with climatic variables (temperature, rainfall and humidity), a study was conducted on aspects of population dynamics and reproductive biology in Salvador city. The snails were collected by hand, monthly. Morphometric data were obtained in the field and dissections to analyze the reproductive system at laboratory. To test if there was a greater sexual activity in wet periods was performed first a Principal Component Analysis (PCA) with climatic variables, followed by a regression between precipitation (the most influential environmental variable) and the variable of interest (sexual activity). The results showed that there is an apparent annual cycle forA. fulica, with a recruitment period covering the end of rainy season and the dry season (August to December 2006 and February 2007). Increase of shell size (height) and of sexual activity were observed during the rainy season, although snails were found in full sexual activity during the whole period of study. The von Bertalanffy curve showed that the oldest specimen collected would have three years and eleven months. The regression between precipitation and sexual activity was significant (P = 0.002) showing that the higher rainfall increased sexual activity. In addition, it was observed that there is an increase in the thickness of peristome as the specimens reaches sexual maturity however this relationship isn’t precise (it were found individuals with sexual structures not consistent with the reproductive stage given by the thickness of peristome) and should be set for each region studied. As the eradication becomes impossible because of high level of invasion found in Salvador, it is recommended to control the population of African snails by continuously removing specimens, especially in the rainy periods.

Common effect of the mucus transferred during mating in two dart-shooting snail species from different families

Several taxa of pulmonate land snails exhibit a conspicuous mating behaviour, the shooting of so-called love darts. During mating, such land snail species stab a mating partner with a mucus-coated dart. It has previously been shown that the sperm donor physiologically influences the sperm recipient via the mucus covering the dart and thereby decreases the number of sperm digested by the recipient. However, the generality of this effect of the dart's mucus is unclear, because almost all the previous studies on the effect of the mucus used the brown garden snail Cornu aspersum from the family Helicidae. Therefore, the relationship between the acquisition of the mucus effect on the recipient and the evolution of the dart itself, and its mucus, is still open to debate. To test the commonality of the physiological effect of the dart mucus, we examined this in Euhadra peliomphala, a species from the Bradybaenidae family, and compared our findings with the results of previous work using C. aspersum. Our experiments showed that in E. peliomphala, the dart mucus had a physiological effect and lowered the accessibility of the gametolytic organ, as found in C. aspersum. This indicates that in various dart-bearing species the mucus from the dart glands targets the same organ and that the inhibition of sperm digestion has played a crucial role in the evolution of the dart and its mucus.

Phenotypic diversity, population structure and stress protein-based capacitoring in populations of Xeropicta derbentina, a heat-tolerant land snail species.

The shell colour of many pulmonate land snail species is highly diverse. Besides a genetic basis, environmentally triggered epigenetic mechanisms including stress proteins as evolutionary capacitors are thought to influence such phenotypic diversity. In this study, we investigated the relationship of stress protein (Hsp70) levels with temperature stress tolerance, population structure and phenotypic diversity within and among different populations of a xerophilic Mediterranean snail species (Xeropicta derbentina). Hsp70 levels varied considerably among populations, and were significantly associated with shell colour diversity: individuals in populations exhibiting low diversity expressed higher Hsp70 levels both constitutively and under heat stress than those of phenotypically diverse populations. In contrast, population structure (cytochrome c oxidase subunit I gene) did not correlate with phenotypic diversity. However, genetic parameters (both within and among population differences) were able to explain variation in Hsp70 induction at elevated but non-pathologic temperatures. Our observation that (1) population structure had a high explanatory potential for Hsp70 induction and that (2) Hsp70 levels, in turn, correlated with phenotypic diversity while (3) population structure and phenotypic diversity failed to correlate provides empirical evidence for Hsp70 to act as a mediator between genotypic variation and phenotype and thus for chaperone-driven evolutionary capacitance in natural populations.

Historic data analysis reveals ambient temperature as a source of phenotypic variation in populations of the land snailTheba pisana

Pulmonate land snails are often polymorphic in their shell coloration pattern. To quantify the contribution of environmental parameters to the nondirectional change in phenotypic variation, we used a historic dataset on Theba pisana morph frequencies and climate data for statistical modelling. We found significant correlations of the degree of phenotypic diversity between juveniles and corresponding adult individuals within the same and the subsequent generation. Among climate parameters, the phenotypic diversity of adults correlated significantly and positively with the mean and maximum ambient temperatures in the winter and spring only. There was no correlation between high or low temperatures and the frequency of distinct morphs. Akaike's information criterion-based model selection revealed the particular importance of only parental phenotypic diversity for next generation juvenile phenotypic diversity. By contrast, phenotypic diversity of the juveniles of the preceding year and the mean temperatures in winter and spring were important for the phenotypic diversity of adult snails. Approximately two-thirds of the explicable variation in phenotypic diversity of adults was explained by inheritance and approximately one-third was expained by ambient temperature. The present study shows that genetics and temperature interact to generate nondirectional changes in phenotypic variation within populations, which also can be reflected by changes in the phenotype of individuals

Limicolaria flammea (Müller, 1774), Another Potentially Invasive African Land Snail in Tropical Asia

The pulmonate land snail, Limicolaria flammea (Müller, 1774), is native to West Africa and its invasive potential is hitherto unknown. We report this species in Asia, specifically on the tropical island of Singapore. The presence of this alien species is of grave concern because its congener ( L. aurora) has invaded the French West Indies and the confamilial giant African land snail ( Achatina fulica) has colonized the globe to an extent where it is regarded as one of the world's worst 100 invasive alien species. In order to curb the spread of L. flammea into the native forests in Singapore, local authorities should start employing mechanical eradication methods (e.g., handpicking). We hypothesize that L. flammea arrived together with an exotic plant species, whose identity and import origin must be determined urgently to prevent the spread of this species into the rest of tropical Asia.

Slug(-gish) science, or an annotated catalogue of the types of tropical vaginulid and agriolimacid pulmonates (Mollusca, Gastropoda), described by Heinrich Simroth (1851-1917), in the Natural History Museum Berlin

At the beginning of the 20th century, the teacher of natural history and professor of zoology in Leipzig Heinrich Simroth (1851–1917) was one of the best known German malacologists. In addition to his encyclopaedic work and important review on molluscs and gastropods in particular, Simroth made his name as an expert of air-breathing terrestrial, hermaphrodite slugs, or pulmonate land snails without shell. Being a life-long passion for him Simroth described a plethora of new taxa, but never established his own reference collection. Contrary to recent assumptions, though, that there is no Simroth collection, a total of 43 of his slug types, in particular of tropical vaginulids from South America (10), Africa (10) and SE Asia (6), as well as agriolimacids from Ethiopia and the Caucasus (14 þ 1) as well as palearctic limacids (2), were found to be

Rapid upregulation of heart antioxidant enzymes during arousal from estivation in the Giant African snail ( Achatina fulica)

Estivation is an adaptive response to environments characterized by elevated temperatures and desiccative stress, as may occur during summer dry seasons. Similar to diapause and hibernation, it is characterized by low levels of activity, a drastically suppressed metabolic rate and enhanced stress resistance. We tested the hypothesis that Achatina fulica, a pulmonate land snail, enhances stress resistance during estivation and/or arousal by upregulating intracellular antioxidant defenses in the heart, kidney, hepatopancreas and foot tissues. No statistically significant changes in mitochondrial or cytosolic superoxide dismutase levels or activities, or glutathione peroxidase, glutathione reductase or catalase activities were associated with estivation in any tissue, however. In contrast, during arousal from estivation, activities of several antioxidant enzymes increased in heart, hepatopancreas and foot. In heart, a rapid increase in MnSOD protein levels was observed that peaked at 2 h post arousal, but no such change was observed in CuZnSOD protein levels. Glutathione peroxidase activity was upregulated at 1 h post arousal and remained elevated until 8 h post arousal in heart tissue. Glutathione peroxidase was also upregulated at 24 h post arousal in foot tissue. Glutathione reductase activity was upregulated at 4 h post arousal in heart and foot tissues whereas catalase activity showed no changes. Markers of lipid peroxidation and protein damage revealed no significant increases during estivation or arousal. Therefore, antioxidant enzymes may play a role in oxidative stress defense specifically during arousal from estivation in A. fulica.

Pulmonate land mollusks from three rainforest transects in northeastern Madagascar: part 1 of 2, with descriptions of 12 new species and one new subspecies: (Gastropoda: Pulmonata)

Pulmonate land mollusks from three rainforest transects in northeastern Madagascar: part 1 of 2, with descriptions of 12 new species and one new subspecies: (Gastropoda: Pulmonata)

Pulmonate land mollusks from three rainforest transects in northeastern Madagascar: part 2 of 2, with descriptions of 21 new species: (Gastropoda: Pulmonata: Stylommatophora)

Pulmonate land mollusks from three rainforest transects in northeastern Madagascar: part 2 of 2, with descriptions of 21 new species: (Gastropoda: Pulmonata: Stylommatophora)

Characterization of eight microsatellite loci in the Galápagos endemic land snail Bulimulus reibischi, and their cross‐species amplification

Variation in and amplification conditions for eight polymorphic microsatellite loci initially identified from Bulimulus akamatus, a pulmonate land snail from Galápagos, are described. Intraspecific polymorphism and heterozygosity of the eight markers were studied in 19 populations of Bulimulus reibischi, a closely related species of B. akamatus. Furthermore, the eight loci were also cross-amplified in six other closely related bulimulid species. The number of alleles across populations of B. reibischi at six loci is moderate (three to 10), but considerable for two other loci (19 and 20). There is no strong evidence for linkage among any of the loci examined.

Heat shock proteins (Hsp70) and water content in the estivating Mediterranean Grunt Snail ( Cantareus apertus)

Pulmonate land snails often are able to estivate to survive dry hot seasons were water and food are scarce. The aperture of the shell is closed with an epiphragm, and metabolism is depressed to approximately one fourth of basal metabolism. We investigated a molecular aspect of estivation focussing on the heat shock protein 70 (Hsp70) stress response during estivation in the Mediterranean Grunt Snail Cantareus apertus. Sequences of a new inducible hsp70 and of actin are presented and expression of the hsp70 gene as well as Hsp70 protein content was measured in estivating animals. Both Hsp70 protein and mRNA do not show a significant change from the control, although there is a trend that hsp70 mRNA is less abundant in estivating specimens. After heat shock, the expression of hsp70 increased and a higher Hsp70 protein content was detected. Water relations were also investigated. After a period of 6 months in the dormant state, the snails contained 14% less water than active ones, implying a constricted protection against desiccation, compared to the desert snail Sphincterochila zonata, and a Mediterranean-type water economy.

The first outdoor find of an American snail Zonitoides arboreus (Say, 1816) from the Czech Republic

The first outdoor record of the pulmonate land snail Zonitoides arboreus in the Czech Republic is presented. Our living population has survived the complete destruction of greenhouses and has been living in the open at the locality for several years.The Czech Republic is the fourth European mainland country with outdoor occurrence of this species.