Search

Podocotyle is a genus of marine opecoelid digeneans, widespread in fishes and invertebrates (gastropods and crustaceans) in the neritic and littoral zones of mainly the Northern Hemisphere. The phylogeny, taxonomy and evolution of digeneans of this genus are quite mysterious due to the wide distribution of these parasites and their species richness. For a number of representatives of Podocotyle spp. a wide range of phenotypic variability has been described, which prevents species identification based solely on morphological characteristics. In this study, we apply molecular and morphological techniques to identify Podocotyle spp. from vertebrate and invertebrate hosts inhabiting the White Sea. Specimens of Podocotyle spp. were collected from pleuronectid and zoarcid fishes, as well as from periwinkles of the intertidal zone, and were studied using light microscopy, scanning electron microscopy, and molecular phylogenetic methods. A combination of molecular and morphological approaches revealed Podocotyle angulata and three species-level taxa awaiting nomenclatural proposals, Podocotyle sp. of Denisova et al. 2023, P. atomon sensu lato and Podocotyle cf. angulata of Sokolov et al. 2023. Conspecificity of one type of cercariae released from infected Littorina obtusata and L. saxatilis with adult digeneans identified as P. atomon sensu lato was established. We provide detailed descriptions of the cotylomicrocercous larvae of P. atomon sensu lato and Podocotyle sp. of Denisova et al. 2023, focusing on the features of the surface morphology of their tegument and associated structures in order to determine the general and diagnostic characteristics of these species. [ABSTRACT FROM AUTHOR]

Chromosomal rearrangements can lead to the coupling of reproductive barriers, but whether and how they contribute to the completion of speciation remains unclear. Marine snails of the genus Littorina repeatedly form hybrid zones between populations segregating for multiple inversion arrangements, providing opportunities to study their barrier effects. Here, we analyzed 2 adjacent transects across hybrid zones between 2 ecotypes of Littorina fabalis ("large" and "dwarf") adapted to different wave exposure conditions on a Swedish island. Applying whole-genome sequencing, we found 12 putative inversions on 9 of 17 chromosomes. Nine of the putative inversions reached near differential fixation between the 2 ecotypes, and all were in strong linkage disequilibrium. These inversions cover 20% of the genome and carry 93% of divergent single nucleotide polymorphisms (SNPs). Bimodal hybrid zones in both transects indicated that the 2 ecotypes of Littorina fabalis maintain their genetic and phenotypic integrity following contact. The bimodality reflects the strong coupling between inversion clines and the extension of the barrier effect across the whole genome. Demographic inference suggests that coupling arose during a period of allopatry and has been maintained for > 1,000 generations after secondary contact. Overall, this study shows that the coupling of multiple chromosomal inversions contributes to strong reproductive isolation. Notably, 2 of the putative inversions overlap with inverted genomic regions associated with ecotype differences in a closely related species (Littorina saxatilis), suggesting the same regions, with similar structural variants, repeatedly contribute to ecotype evolution in distinct species. [ABSTRACT FROM AUTHOR]

Marine snails (genus Littorina) have recently emerged as a model for speciation research, with repeated formation of different types of barriers to gene flow in contact zones between ecotypes and taxa. Inversion polymorphisms are found in an increasing number of species, including Littorina snails, and have been suggested to promote both local adaptation, ecotype formation and speciation, but their roles can vary. In Littorina , their role in later stages of speciation remains particularly unclear. A one-dimensional speciation continuum is frequently used to frame our understanding of the speciation process. However, a number of recent studies have argued that no single measure adequately summarises reproductive isolation and, therefore, the speciation process is best viewed in multiple dimensions. The many Littorina contact zones provide empirical data to illustrate how this principle can be applied. Speciation is a key evolutionary process that is not yet fully understood. Combining population genomic and ecological data from multiple diverging pairs of marine snails (Littorina) supports the search for speciation mechanisms. Placing pairs on a one-dimensional speciation continuum, from undifferentiated populations to species, obscured the complexity of speciation. Adding multiple axes helped to describe either speciation routes or reproductive isolation in the snails. Divergent ecological selection repeatedly generated barriers between ecotypes, but appeared less important in completing speciation while genetic incompatibilities played a key role. Chromosomal inversions contributed to genomic barriers, but with variable impact. A multidimensional (hypercube) approach supported framing of questions and identification of knowledge gaps and can be useful to understand speciation in many other systems. [ABSTRACT FROM AUTHOR]

The intertidal gastropod Littorina saxatilis is a model system to study speciation and local adaptation. The repeated occurrence of distinct ecotypes showing different levels of genetic divergence makes L. saxatilis particularly suited to study different stages of the speciation continuum in the same lineage. A major finding is the presence of several large chromosomal inversions associated with the divergence of ecotypes and, specifically, the species offers a system to study the role of inversions in this divergence. The genome of L. saxatilis is 1.35 Gb and composed of 17 chromosomes. The first reference genome of the species was assembled using Illumina data, was highly fragmented (N50 of 44 kb), and was quite incomplete, with a BUSCO completeness of 80.1% on the Metazoan dataset. A linkage map of one full-sibling family enabled the placement of 587 Mbp of the genome into 17 linkage groups corresponding to the haploid number of chromosomes, but the fragmented nature of this reference genome limited the understanding of the interplay between divergent selection and gene flow during ecotype formation. Here, we present a newly generated reference genome that is highly contiguous, with a N50 of 67 Mb and 90.4% of the total assembly length placed in 17 super-scaffolds. It is also highly complete with a BUSCO completeness of 94.1% of the Metazoa dataset. This new reference will allow for investigations into the genomic regions implicated in ecotype formation as well as better characterization of the inversions and their role in speciation. [ABSTRACT FROM AUTHOR]

Extreme weather events (e.g., droughts, excessive precipitation) are expected to increase in frequency and severity in the coming decades due to climate change, causing significant impacts on society and ecosystems. Because these events are rare and complex, they have been studied with manipulative experiments. Littorinidae snails inhabit a complex and variable environment in which they must deal with periodic extreme events and are thus considered excellent ecological models for these studies. Therefore, this study aimed to understand the effects of extreme climatic events on the survival and weight of the species Littorina fabalis and Littorina littorea in Spain and Littoraria angulifera and Littoraria flava in Brazil. Higher mortality rates and greater weight loss were observed in the desiccation resistance treatment compared to the control treatment in both countries. The results showed dependence on the species' body size. The submergence tolerance treatment indicated that the species from Spain are more susceptible to mortality in response to excessive rainfall and/or coastal flooding. Each species tested for the effect of extreme climatic events using an integrated response strategy with clear latitudinal differences. Understanding the organisms' responses at different latitudes is essential for conservation biology on a global scale. [ABSTRACT FROM AUTHOR]

Simple Summary: Nowadays, it is well known that the gut bacterial microbiome is crucially important for the adaptation of multicellular organisms to their environment. In this study, we aimed to identify the differences between the bacteriomes of two closely related marine snail species living in sympatry but feeding in different micro-niches. Although there were significant interspecies differences detected during the summer season, we did not observe this dissimilarity during the cold season. Moreover, the diversity of bacterial communities associated with snails decreased in autumn. We suggest that bacteria predominantly associated with one species degrade its toxic feeding substrate. These results help to understand the role of bacteriomes in the adaptation and divergence of closely related species. Symbiotic microorganisms may provide their hosts with abilities critical to their occupation of microhabitats. Gut (intestinal) bacterial communities aid animals to digest substrates that are either innutritious or toxic, as well as support their development and physiology. The role of microbial communities associated with sibling species in the hosts' adaptation remains largely unexplored. In this study, we examined the composition and plasticity of the bacteriomes in two sibling intertidal gastropod species, Littorina fabalis and L. obtusata, which are sympatric but differ in microhabitats. We applied 16S rRNA gene metabarcoding and shotgun sequencing to describe associated microbial communities and their spatial and temporal variation. A significant drop in the intestinal bacteriome diversity was revealed during the cold season, which may reflect temperature-related metabolic shifts and changes in snail behavior. Importantly, there were significant interspecies differences in the gut bacteriome composition in summer but not in autumn. The genera Vibrio, Aliivibrio, Moritella and Planktotalea were found to be predominantly associated with L. fabalis, while Granulosicoccus, Octadecabacter, Colwellia, Pseudomonas, Pseudoalteromonas and Maribacter were found to be mostly associated with L. obtusata. Based on these preferential associations, we analyzed the metabolic pathways' enrichment. We hypothesized that the L. obtusata gut bacteriome contributes to decomposing algae and detoxifying polyphenols produced by fucoids. Thus, differences in the sets of associated bacteria may equip their closely phylogenetically related hosts with a unique ability to occupy specific micro-niches. [ABSTRACT FROM AUTHOR]

Abstract Microbes can play a prominent role in the evolution of their hosts, facilitating adaptation to various environments and promoting ecological divergence. The Wave and Crab ecotypes of the intertidal snail Littorina saxatilis is an evolutionary model of rapid and repeated adaptation to environmental gradients. While patterns of genomic divergence of the Littorina ecotypes along the shore gradients have been extensively studied, their microbiomes have been so far overlooked. The aim of the present study is to start filling this gap by comparing gut microbiome composition of the Wave and Crab ecotypes using metabarcoding approach. Since Littorina snails are micro‐grazers feeding on the intertidal biofilm, we also compare biofilm composition (i.e. typical snail diet) in the crab and wave habitats. In the results, we found that bacterial and eukaryotic biofilm composition varies between the typical habitats of the ecotypes. Further, the snail gut bacteriome was different from outer environments, being dominated by Gammaproteobacteria, Fusobacteria, Bacteroidia and Alphaproteobacteria. There were clear differences in the gut bacterial communities between the Crab and the Wave ecotypes as well as between the Wave ecotype snails from the low and high shores. These differences were both observed in the abundances and in the presence of different bacteria, as well as at different taxonomic level, from bacterial OTU's to families. Altogether, our first insights show that Littorina snails and their associated bacteria are a promising marine system to study co‐evolution of the microbes and their hosts, which can help us to predict the future for wild species in the face of rapidly changing marine environments.

We studied the reproductive behaviour associated with mate choice by size in three species of White Sea molluscs of the genus Littorina—L. saxatilis, L. obtusata and L. fabalis. Three behavioural patterns were revealed after a comparative analysis of the behaviour of the male snails. The males of L. obtusata, regardless of their size, could mate with all females but preferred to mate with larger ones. The males of L. fabalis chose females of their own size or larger. The males of L. saxatilis mated with females of approximately their own size. In the first case (L. obtusata), there is a free exchange of genes between all the size groups in the population. In the second case (L. fabalis), the gene transfer from small males to females of any size is free but the gene transfer from large males to small females is limited. In the third case (L. saxatilis), the gene flow is limited between groups of individuals of dramatically different size. Thus, in the case of L. saxatilis and L. fabalis, to varying degrees restriction of random mating associated with size assortativity in choosing a mating partner was expressed, which under certain circumstances (divergence of individuals in different econiches, the presence of ecotypes differing in size, lack of stronger natural selection in the other direction) can favour sympatric speciation. The males of all the three periwinkles' species preferred larger females but sexual selection in favour of larger females was found only in L. obtusata. In the other two species, there were no statistically significant size differences between copulating and non-copulating females. This appears to be due to the occurrence of size-assortative mating in these species, which limits the ability of males to choose larger female partners. [ABSTRACT FROM AUTHOR]

Microbes can play a prominent role in the evolution of their hosts, facilitating adaptation to various environments and promoting ecological divergence. The Wave and Crab ecotypes of the intertidal snail Littorina saxatilis is an evolutionary model of rapid and repeated adaptation to environmental gradients. While patterns of genomic divergence of the Littorina ecotypes along the shore gradients have been extensively studied, their microbiomes have been so far overlooked. The aim of the present study is to start filling this gap by comparing gut microbiome composition of the Wave and Crab ecotypes using metabarcoding approach. Since Littorina snails are micro‐grazers feeding on the intertidal biofilm, we also compare biofilm composition (i.e. typical snail diet) in the crab and wave habitats. In the results, we found that bacterial and eukaryotic biofilm composition varies between the typical habitats of the ecotypes. Further, the snail gut bacteriome was different from outer environments, being dominated by Gammaproteobacteria, Fusobacteria, Bacteroidia and Alphaproteobacteria. There were clear differences in the gut bacterial communities between the Crab and the Wave ecotypes as well as between the Wave ecotype snails from the low and high shores. These differences were both observed in the abundances and in the presence of different bacteria, as well as at different taxonomic level, from bacterial OTU's to families. Altogether, our first insights show that Littorina snails and their associated bacteria are a promising marine system to study co‐evolution of the microbes and their hosts, which can help us to predict the future for wild species in the face of rapidly changing marine environments. [ABSTRACT FROM AUTHOR]

Renicolid digeneans parasitize aquatic birds. Their intramolluscan stages develop in marine and brackish-water gastropods, while metacercariae develop in molluscs and fishes. The systematics of renicolids is poorly developed, their life cycles are mostly unknown, and the statuses of many species require revision. Here, we establish based on integrated morphological and molecular data that adult renicolids from gulls Larus argentatus and Larus schistisagus and sporocysts and cercariae of Cercaria parvicaudata from marine snails Littorina spp. are life-cycle stages of the same species. We name it Renicola parvicaudatus and synonymized with it Renicola roscovitus. An analysis of the cox1 gene of R. parvicaudatus from Europe, North America and North Asia demonstrates a low genetic divergence, suggesting that this species has formed quite recently (perhaps during last glacial maximum) and that interregional gene flow is high. In Littorina saxatilis and L. obtusata from the Barents Sea, molecular analysis has revealed intramolluscan stages of Cercaria littorinae saxatilis VIII, a cryptic species relative to R. parvicaudatus. In the molecular trees, Renicola keimahuri from L. schistisagus belongs to another clade than R. parvicaudatus. We show that the species of this clade have cercariae of Rhodometopa group and outline morphological and behavioural transformations leading from xiphidiocercariae to these larvae. Molecular analysis has revealed 3 main phylogenetic branches of renicolids, differing in structure of adults, type of cercariae and host range. Our results elucidate the patterns of host colonization and geographical expansion of renicolids and pave the way to the solution of some long-standing problems of their classification. [ABSTRACT FROM AUTHOR]

Abstract Divergence of ecological niches in phylogenetically closely related species indicates the importance of ecology in speciation, especially for sympatric species are considered. Such ecological diversification provides an advantage of alleviating interspecies competition and promotes more efficient exploitation of environmental resources, thus being a basis for ecological speciation. We analyzed a group of closely related species from the subgenus Neritrema (genus Littorina, Caenogastropoda) from the gravel‐bouldery shores. In two distant sites at the Barents and Norwegian Sea, we examined the patterns of snail distribution during low tide (quantitative sampling stratified by intertidal level, presence of macrophytes, macrophyte species, and position on them), shell shape and its variability (geometric morphometrics), and metabolic characteristics (metabolomic profiling). The studied species diversified microbiotopes, which imply an important role of ecological specification in the recent evolution of this group. The only exception to this trend was the species pair L. arcana / L. saxatilis, which is specifically discussed. The ecological divergence was accompanied by differences in shell shape and metabolomic characteristics. Significant differences were found between L. obtusata versus L. fabalis and L. saxatilis / L. arcana versus L. compressa both in shell morphology and in metabolomes. L. saxatilis demonstrated a clear variability depending on intertidal level which corresponds to a shift in conditions within the occupied microhabitat. Interestingly, the differences between L. arcana (inhabiting the upper intertidal level) and L. compressa (inhabiting the lower one) were analogous to those between the upper and lower fractions of L. saxatilis. No significant level‐dependent changes were found between the upper and lower fractions of L. obtusata, most probably due to habitat amelioration by fucoid macroalgae. All these results are discussed in the contexts of the role of ecology in speciation, ecological niche dynamics and conservatism, and evolutionary history of the Neritrema species.

Abstract The presence of shell bands is common in gastropods. Both the marine snails Littorina fabalis and Lttorina saxatilis are polymorphic for this trait. Such polymorphism would be expected to be lost by the action of genetic drift or directional selection, but it appears to be widespread at relatively constant frequencies. This suggests it is maintained by balancing selection on the trait or on a genetically linked trait. Using long time series of empirical data, we compared potential effects of genetic drift and negative frequency‐dependent selection (NFDS) in the two species. The contribution of genetic drift to changes in the frequency of bands in L. fabalis was estimated using the effective population size estimated from microsatellite data, while the effect of genetic drift in L. saxatilis was derived from previously published study. Frequency‐dependent selection was assessed by comparing the cross‐product estimator of fitness with the frequency of the polymorphism across years using a regression analysis. Both studied species showed patterns of NFDS. In addition, in L. fabalis, contributions from genetic drift could explain some of the changes in banding frequency. Overdominance and heterogeneous selection did not fit well to our data. The possible biological explanations resulting in the maintenance of the banding polymorphism are discussed.

In the evolution of invertebrates, the transition from egg-layers to brooders occurred many times. However, the molecular mechanisms underlying this transition are still not well understood. Recently diverged species genus Littorina (Mollusca, Gastropoda, Caenogastropoda, Littorinimorpha): Littorina saxatilis, L. arcana, L. compressa, L. obtusata and L. fabalis might be a fruitful model for elucidation of these mechanisms. All five species sympatrically inhabit an intertidal zone. Only L. saxatilis is ovoviviparous while the other four species form clutches. Although in L. saxatilis jelly gland of the pallial oviduct function as a brood pouch, it is not deeply modified at the morphological level in comparison to egg-laying relatives. Comparative analysis of transcriptomic profiles of the pallial oviducts of these closely related species might help to uncover the molecular mechanisms of the egg-laying to brooding transition. Unraveling of the mechanisms underlying this transition in L. saxatilis is important not only in aspects of reproduction biology and strategy, but also in a broader view as an example of relatively fast evolutionary transformations. We generated an RNA-seq dataset (224 104 446 clean reads) for oviducts of five species genus Littorina. Libraries of all five species were sequenced using Illumina HiSeq 2500; additional reads for L. arcana were obtained using Illumina NovaSeq 6000. Transcriptomic profiles were analyzed in pooled samples (of three individuals) with two biological replicates for each species (each biological replicate was prepared and sequenced as a separate library). The transcriptome was assembled de novo and annotated with five assembles corresponding to each species. The raw data were uploaded to the SRA database, the BioProject IDs are PRJNA662103 (“obtusata” group) and PRJNA707549 (“saxatilis” group).

Divergence of ecological niches in phylogenetically closely related species indicates the importance of ecology in speciation, especially for sympatric species are considered. Such ecological diversification provides an advantage of alleviating interspecies competition and promotes more efficient exploitation of environmental resources, thus being a basis for ecological speciation. We analyzed a group of closely related species from the subgenus Neritrema (genus Littorina, Caenogastropoda) from the gravel‐bouldery shores. In two distant sites at the Barents and Norwegian Sea, we examined the patterns of snail distribution during low tide (quantitative sampling stratified by intertidal level, presence of macrophytes, macrophyte species, and position on them), shell shape and its variability (geometric morphometrics), and metabolic characteristics (metabolomic profiling). The studied species diversified microbiotopes, which imply an important role of ecological specification in the recent evolution of this group. The only exception to this trend was the species pair L. arcana / L. saxatilis, which is specifically discussed. The ecological divergence was accompanied by differences in shell shape and metabolomic characteristics. Significant differences were found between L. obtusata versus L. fabalis and L. saxatilis / L. arcana versus L. compressa both in shell morphology and in metabolomes. L. saxatilis demonstrated a clear variability depending on intertidal level which corresponds to a shift in conditions within the occupied microhabitat. Interestingly, the differences between L. arcana (inhabiting the upper intertidal level) and L. compressa (inhabiting the lower one) were analogous to those between the upper and lower fractions of L. saxatilis. No significant level‐dependent changes were found between the upper and lower fractions of L. obtusata, most probably due to habitat amelioration by fucoid macroalgae. All these results are discussed in the contexts of the role of ecology in speciation, ecological niche dynamics and conservatism, and evolutionary history of the Neritrema species. [ABSTRACT FROM AUTHOR]

The presence of shell bands is common in gastropods. Both the marine snails Littorina fabalis and Lttorina saxatilis are polymorphic for this trait. Such polymorphism would be expected to be lost by the action of genetic drift or directional selection, but it appears to be widespread at relatively constant frequencies. This suggests it is maintained by balancing selection on the trait or on a genetically linked trait. Using long time series of empirical data, we compared potential effects of genetic drift and negative frequency‐dependent selection (NFDS) in the two species. The contribution of genetic drift to changes in the frequency of bands in L. fabalis was estimated using the effective population size estimated from microsatellite data, while the effect of genetic drift in L. saxatilis was derived from previously published study. Frequency‐dependent selection was assessed by comparing the cross‐product estimator of fitness with the frequency of the polymorphism across years using a regression analysis. Both studied species showed patterns of NFDS. In addition, in L. fabalis, contributions from genetic drift could explain some of the changes in banding frequency. Overdominance and heterogeneous selection did not fit well to our data. The possible biological explanations resulting in the maintenance of the banding polymorphism are discussed. [ABSTRACT FROM AUTHOR]

Trematodes of the genus Himasthla are usual parasites of coastal birds in nearshore ecosystems of northern European seas and the Atlantic coast of North America. Their first intermediate hosts are marine and brackish-water gastropods, while second intermediate hosts are various invertebrates. We analysed sequences of partial 28S rRNA and nad1 genes and the morphology of intramolluscan stages, particularly cercariae of Himasthla spp. parasitizing intertidal molluscs Littorina spp. in the White Sea, the Barents Sea and coasts of North Norway and Iceland. We showed that only three Himasthla spp. are associated with periwinkles in these regions. Intramolluscan stages of H. elongata were found in Littorina littorea, of H. littorinae, in both L. saxatilis and L. obtusata, and of Cercaria littorinae obtusatae, predominantly, in L. obtusata. Other Himasthla spp. previously reported from Littorina spp. in North Atlantic are either synonymous with one of these species or described erroneously. Based on a comparison of newly generated 28S rDNA sequences with GenBank data, rediae and cercariae of C. littorinae obtusatae were identified as belonging to H. leptosoma. Some previously unknown morphological features of young and mature rediae and cercariae of the three Himasthla spp. are described. We provide a key to the rediae and highlight characters important for identification of cercariae. Genetic diversity within the studied species was only partially determined by their specificity to the molluscan host. The nad1 network constructed for H. leptosoma lacked geographical structure, which is explained by a high gene flow owing to highly vagile definitive hosts, shorebirds. [ABSTRACT FROM AUTHOR]

Littorina Férussac, 1822 is an abundant genus of small gastropods found in the upper littoral zone of rocky seashores worldwide. Although ecologically important, shell-based species identification in this genus is challenging due to phenotypic variation in shell morphology and lack of diagnostic characters among morphologically similar species. In this study, we revised the taxonomy of Korean Littorina species using morphological characters (shell and radula) and cox1 mitochondrial DNA sequences for three Korean species: L. brevicula, L. sitkana, and L. horikawai. Results suggest that L. sitkana was erroneously reported as L. kasatka in a previous study. A new record for Littorina horikawai (Matsubayashi & Habe in Habe, 1979), previously unknown from Korea, is described, which can be distinguished from L. sitkana by the presence of alternating white and brown spiral ribs on each whorl. Comparison of the mtDNA cox1 gene sequences shows very low intraspecific variation even between geographically distant populations. A phylogenetic tree supports a close relationship between L. horikawai and L. sitkana, consistent with earlier phylogenetic studies.

Sympatric coexistence of recently diverged species raises the question of barriers restricting the gene flow between them. Reproductive isolation may be implemented at several levels, and the weakening of some, e.g. premating, barriers may require the strengthening of the others, e.g. postcopulatory ones. We analysed mating patterns and shell size of mates in recently diverged closely related species of the subgenus Littorina Neritrema (Littorinidae, Caenogastropoda) in order to assess the role of premating reproductive barriers between them. We compared mating frequencies observed in the wild with those expected based on relative densities using partial canonical correspondence analysis. We introduced the fidelity index (FI) to estimate the relative accuracy of mating with conspecific females and precopulatory isolation index (IPC) to characterize the strength of premating barriers. The species under study, with the exception of L. arcana, clearly demonstrated preferential mating with conspecifics. According to FI and IPC, L. fabalis and L. compressa appeared reliably isolated from their closest relatives within Neritrema. Individuals of these two species tend to be smaller than those of the others, highlighting the importance of shell size changes in gastropod species divergence. L. arcana males were often found in pairs with L. saxatilis females, and no interspecific size differences were revealed in this sibling species pair. We discuss the lack of discriminative mate choice in the sympatric populations of L. arcana and L. saxatilis, and possible additional mechanisms restricting gene flow between them. [ABSTRACT FROM AUTHOR]

Genus Littorina subgenus Neritrema (Mollusca, Caenogastropoda) includes the “obtusata” group of closely related species (Littorina obtusata and L. fabalis). The anatomy of the adult reproductive system (pallial oviduct) is the only reliable feature used for species identification in females of these species. Reproductive system anatomy and reproduction-associated proteins often diverge between sibling species. Despite being of high evolutionary interest, the molecular basis of this divergence remains poorly understood. We performed proteotranscriptomic comparison of oviducts of L. obtusata and L. fabalis by RNA-seq on Illumina HiSeq 2500 and two-dimensional protein electrophoresis (2D DIGE) with MS/MS identification of the species-specific proteins. The interspecies differences in the oviduct were associated with (1) metabolic proteins reflecting overall physiological differences between L. obtusata and L. fabalis, (2) receptor proteins, and (3) transcripts related to transposable elements (TEs). Various receptors identified may recognize a wide variety of ligands from pathogen-associated molecular patterns to specific carbohydrates on the sperm surface. Therefore, these may participate in immune defense as well as in sperm storage and regulation. Species-specificity of multiple TE sequences (coding for reverse transcriptase and ribonuclease H) may indicate the important role of these genomic elements in the Littorina species divergence, which has not been reported previously.

Abstract The rough periwinkle Littorina saxatilis is characterised for showing shell phenotypic variation associated to environmental clines and this is found in parallel across shores. In order to disentangle the additive genetic effects and the environmental factors involved in this variation, we analysed natural populations subjected to a gradient in wave exposure and crab predation in 3 different Rias (primary estuaries) from Galicia (NW Spain). In addition to previously described "Crab" and "Wave" ecotypes, we studied a new phenotype ("Crab+") associated to the most sheltered habitats of the gradient. We obtained estimates of heritability and Q st (between population genetic differentiation) in embryos and P st (between population phenotypic differentiation) in adults for shell morphology traits. Heritability estimates were close to one for shell size and relative shell aperture, suggesting a low impact of environmental factors in embryos. Estimates of Q st between ecotypes were significant and typically much higher than published F st estimates from neutral markers, indicating a role for selection on shaping additive genetic variation. Estimates of P st between ecotypes for adults were substantially larger than embryos´ Q st , suggesting also a role of phenotypic plasticity in ecotype differentiation. Our results suggest that P st vs F st comparisons should be taken with caution because they might not reflect real additive genetic effects, but P st vs Q st comparisons could shed light into the role of phenotypic plasticity. Highlights • Morphometric analysis of Littorina saxatilis is carried out in adults and embryos. • We compared phenotypic (P ST) and genetic divergence (Q ST , F ST) between ecotypes. • High heritabilities are shown for shell traits (size and shell aperture) in embryos. • Q ST vs F ST comparisons show a role for natural selection in ecotype differentiation. [ABSTRACT FROM AUTHOR]

Abstract Dislodgment, in particular by breaking waves, is a widely acknowledged source of stress, if not mortality, in rocky shores intertidal gastropods. This raises the question of understanding if snails dislodged from a specific habitat have the ability to navigate back to their original substrate, and how this is achieved. In this context, we investigated the consequences of dislodgement on a population of Littorina littorea inhabiting two proximate habitats (a seawall ranging from the low to the high intertidal zone and the related low intertidal rocky platform) located on an isolated offshore reef over a three-month period. Using tagged undisturbed, disturbed, translocated and transplanted L. littorea , we assessed a potential tidal height preference and found that L. littorea showed a clear tendency to remain on their original tidal height, with recapture rates ranging between 100% and 88%, respectively 1 and 78 days after dislodgment. We subsequently measured the recapture rates of individuals dislodged from three distinct tidal levels on the seawall and from a horizontal platform. Dislodged L. littorea showed a clear tendency to return to their original tidal level, with return rates ranging between 32 and 48% 78 days after dislodgement; snails from the horizontal platform were never found on the proximate seawall. In addition, high frequency observations conducted on individually tagged dislodged L. littorea during the first 8 successive low tides of our survey indicate that individuals originating from the seawall consistently showed higher movement rates and directionality towards the seawall, while individuals originating from the platform showed no directionality and reduced migration rate. These results show that dislodged L. littorea navigate back to their original substrate and tidal height. Putative processes include a combination of geotaxis, chemotaxis and rheotaxis. At relatively short time scales (i.e. days to months), L. littorea show both tidal and habitat partitioning that is distinct from the movement patterns previously reported in littorinids over a range of temporal scales. [ABSTRACT FROM AUTHOR]

Littorina Férussac, 1822 is an abundant genus of small gastropods found in the upper littoral zone of rocky seashores worldwide. Although ecologically important, shell-based species identification in this genus is challenging due to phenotypic variation in shell morphology and lack of diagnostic characters among morphologically similar species. In this study, we revised the taxonomy of Korean Littorina species using morphological characters (shell and radula) and cox1 mitochondrial DNA sequences for three Korean species: L. brevicula, L. sitkana, and L. horikawai. Results suggest that L. sitkana was erroneously reported as L. kasatka in a previous study. A new record for Littorina horikawai (Matsubayashi & Habe in Habe, 1979), previously unknown from Korea, is described, which can be distinguished from L. sitkana by the presence of alternating white and brown spiral ribs on each whorl. Comparison of the mtDNA cox1 gene sequences shows very low intraspecific variation even between geographically distant populations. A phylogenetic tree supports a close relationship between L. horikawai and L. sitkana, consistent with earlier phylogenetic studies. [ABSTRACT FROM AUTHOR]

The evolution of complement system in invertebrates is poorly investigated. While the repertoire of complement genes in several Ecdysozoa lineages is found substantially different from that of Deuterostomia, the composition and function of the complement in the second protostome lineage, Lophotrochozoa, remains unclear. Here we report the general description of new transcriptomic data on the common periwinkle, Littorina littorea , and trace the evolutionary trajectories of the ancestral proto-complement repertoire. The repertoire is defined as immune cascade providing the minimum set of C3-associated molecules required for C3b amplification, opsonization of the targets and their phagocytosis: thioester protein (TEP) C3, serine protease C2/factor B (Bf) and complement receptors (CR). The reference transcriptome of L. littorea was built from the dual-species RNA-seq experiment with the periwinkle and its tissue digenean parasite Himasthla elongata . Five TEPs, including the ortholog of the C3, are found expressed in the in the mollusk's inflamed tissues. The homolog of the complement receptors CR1/CR2 is also expressed, however the ortholog of Bf is not. The extensive phylogenetic analysis showed that the C3 ortholog and the complement receptors are retained in all key lophotrochozoan taxa: Mollusca, Annelida and Brachiopoda. However, the Bf ortholog was lost at least three times independently in different lineages: i) Cephalopoda, ii) a common ancestor of all Gastropoda and iii) one of the Annelida lineage, Clitellata. Both C3 and Bf molecules were retained in bivalve species, brachiopods and annelid worms from the Polychaeta lineage. Hypothetically, the function of the lost Bf in these animals can be compensated by Factor L (Lf) – the serine protease first found in L. littorea and homologous to both, the Bf and the arthropod factor C (Cf). The contrast differences in proto-complement repertoire between the sister mollusk’ taxa, Bivalvia and Gastropoda (the conserved and modified sets, respectively), can underlie differences in their susceptibility to digenean infection. [ABSTRACT FROM AUTHOR]

Abstract: Reproductive isolation is the key attribute of biological species and establishment of the reproductive barriers is an essential event for speciation. Among the mechanisms of reproductive isolation, gamete incompatibility due to the variability of gamete interaction proteins may drive fast divergence even in sympatry. However, the number of available models to study this phenomenon is limited. In case of internally fertilized invertebrates, models to study gamete incompatibility and sperm competition mechanisms are restricted to a single taxon: insects. Here, we propose a group of closely related Littorina species as a new model for such studies. Particularly since periwinkles are already thoroughly studied in terms of morphology, physiology, ecology, phylogeny, and ecological speciation. Earlier, we have identified the first species‐specific Littorina sperm protein (LOSP) with no known conservative domains or homologies. LOSP is relatively abundant component of sperm extracts and might be involved in gamete incompatibility. Here, we characterize its definitive localization and mRNA expression pattern in the male reproductive system by immunocytochemistry and RNA in situ hybridization. We demonstrate that LOSP distribution is limited to the parasperm cells. Losp gene expression occurs only at the early stages of parasperm development. The protein is stored within granules of mature parasperm and, most likely, is released after ejaculation inside female reproductive system. Thus, LOSP is the only described molluscan paraspermal protein to date, and there is a possibility for LOSP to be involved in gamete incompatibility since heterospermy is a common phenomenon among Littorina. [ABSTRACT FROM AUTHOR]

With both global surface temperatures and the incidence and intensity of extreme temperature events projected to increase, the assessment of species' sensitivity to chronic and acute changes in temperature has become crucial. Sensitivity predictions are based predominantly on adult responses, despite the fact that early life stages may be more vulnerable to thermal challenge. Here, we compared the sensitivity of different life history stages of the intertidal gastropod Littorina obtusata using thermal death time curves, which incorporate the intensity and duration of heat stress, and used these to calculate upper critical thermal limits (CTmax) and sensitivity to temperature change (z). Early (larval) life stages had both a lower CTmax and a lower z than adults, suggesting they are less good at withstanding short-term extreme thermal challenges but better able to survive moderate temperatures in the long term. This result supports the predicted trade-off between acute and chronic tolerance to thermal stress, and is consistent with the different thermal challenges that these stages encounter in the intertidal zone. We conclude that different life history stages employ different thermal strategies that may be adaptive. Our findings caution against the use of predictions of the impact of global warming that are based on only adult responses and, hence, which may underestimate vulnerability. [ABSTRACT FROM AUTHOR]

Aim: The present study aims to focus on the role of common marine snails (Littorina littorea) as a vector for some trematode parasites. Materials and Methods: A total of 327 marine water L. littorea snails were collected during the summer of 2016 from a Qarun lake in the EL-Fayoum Governorate, Egypt. The snails were investigated for infection by trematode parthenitae through induction of cercarial shedding by exposure to light and crushing the snails. The species were stored in Search Laboratory of Zoology Department, Faculty of Science, Fayoum University. Results: Three species of Strigeid littorina cercaria were identified from the infected snails. They are described here and they identified in relation to close-up morphological features and linked to its snail hosts. They give the following names: Cercaria strigeid littorina type 1, C. strigeid littorina type 2, and C. strigeid littorina type 3. The incidence of infection by these cercariae was 33%, 25.7%, and 2.4%, respectively. Conclusion: This study is clarifying the importance of this marine snail as intermediate hosts for new trematode species. [ABSTRACT FROM AUTHOR]

There has been a significant increase in the literature regarding the effects of warming and acidification on the marine ecosystem. To our knowledge, there is very little information on the potential effects of both combined stressors on marine grazer-seaweed interactions. Here, we evaluated, for the first time several phenotypic responses (e.g periwinkle survival, condition index, consumption rates, seaweed photosynthetic activity and oxidative stress) of the temperate periwinkle Littorina obtusata (grazer) and the brown seaweed Ascophyllum nodosum (prey) to such climate change-related variables, for 15 days. Increased temperature (22 °C, pH 8.0) elicited a significant lethal effect on the periwinkle within a short-term period (mortality rate > 90%). Acidification condition (18 °C, pH 7.6) was the one that showed lower mortality rates (≈20%), reflected by lower impact on periwinkle fitness and consumption rates. Under a scenario of increased temperature and lowered pH the antioxidant defences of L. obtusata seemed to be supressed increasing the risk of peroxidative damage. The seaweed evidenced signs of cellular damage under such conditions. These results suggest that: i) lower pH per se seems to benefit the interaction between grazer and seaweed while, ii) a combined scenario of increased temperature and lowered pH may be negative for the interaction, due to the unbalance between periwinkle mortality rates and consumption rates. But most importantly, since grazing often plays an important role on structuring natural communities, such predator-prey disturbances can elicit cascading effects on the remaining community structure and functioning of the temperate rocky-shore ecosystems. [ABSTRACT FROM AUTHOR]

Cadmium (Cd) is one of the most harmful metals, being toxic to most animal species, including marine invertebrates. Among marine gastropods, the periwinkle (Littorina littorea) in particular can accumulate high amounts of Cd in its midgut gland. In this organ, the metal can elicit extensive cytological and tissue-specific alterations that may reach, depending on the intensity of Cd exposure, from reversible lesions to pathological cellular disruptions. At the same time, Littorina littorea expresses a Cd-specific metallothionein (MT) that, due to its molecular features, expectedly exerts a protective function against the adverse intracellular effects of this metal. The aim of the present study was, therefore, to assess the time course of MT induction in the periwinkle's midgut gland on the one hand, and cellular and tissue-specific alterations in the digestive organ complex (midgut gland and digestive tract) on the other, upon exposure to sub-lethal Cd concentrations (0.25 and 1 mg Cd/L) over 21 days. Depending on the Cd concentrations applied, the beginning of alterations of the assessed parameters followed distinct concentration-dependent and time-dependent patterns, where the timeframe for the onset of the different response reactions became narrower at higher Cd concentrations compared to lower exposure concentrations. [ABSTRACT FROM AUTHOR]

The paper describes the conditions of formation of large accumulative forms on the southeastern coast of the Baltic Sea and presents the results of a comparative analysis of these structures. It has been established that Baltic barrier-lagoon systems have a complex geological and geomorphologic structure. Sandy barriers, which are, in general, marine accumulative formations, often comprise various fragments of different genesis (moraine remnants, glaciolacustrine and deltaic plains, etc.). The formation and development of large accumulative forms in the southeastern Baltic occurred against the background of sea level fluctuations in the Littorina time following a unified scenario for the entire coast. [ABSTRACT FROM AUTHOR]

After the resolution of the 3D structure of the Cd9-aggregate of the Littorina littorea metallothionein (MT), we report here a detailed analysis of the metal binding capabilities of the wild type MT, LlwtMT, and of two truncated mutants lacking either the N-terminal domain, Lltr2MT, or both the N-terminal domain, plus four extra flanking residues (SSVF), Lltr1MT. The recombinant synthesis and in vitro studies of these three proteins revealed that LlwtMT forms unique M9-LlwtMT complexes with Zn(II) and Cd(II), while yielding a complex mixture of heteronuclear Zn,Cu-LlwtMT species with Cu(I). As expected, the truncated mutants gave rise to unique M6-LltrMT complexes and Zn,Cu-LltrMT mixtures of lower stoichiometry with respect to LlwtMT, with the SSVF fragment having an influence on their metal binding performance. Our results also revealed a major specificity, and therefore a better metal-coordinating performance of the three proteins for Cd(II) than for Zn(II), although the analysis of the Zn(II)/Cd(II) displacement reaction clearly demonstrates a lack of any type of cooperativity in Cd(II) binding. Contrarily, the analysis of their Cu(I) binding abilities revealed that every LlMT domain is prone to build Cu4-aggregates, the whole MT working by modules analogously to, as previously described, certain fungal MTs, like those of C. neoformans and T. mesenterica. It is concluded that the Littorina littorea MT is a Cd-specific protein that (beyond its extended binding capacity through an additional Cd-binding domain) confers to Littorina littorea a particular adaptive advantage in its changeable marine habitat. [ABSTRACT FROM AUTHOR]

The Littorina Sea stage (past c. 7000 years) development of the northern, and its interactions with the central Baltic Sea have been influenced by spatially different but in the north very strong glacio-isostatic uplift. Here we investigate the impact of the isostatic readjustment on the northern Baltic Sea environment and on the water exchange with the central Baltic Sea using geochemical and microfossil records from well-dated sediment cores of the Bothnian Sea/Bay that are compared to sedimentary records of the Gotland Basin and Landsort Deep. Benthic foraminifera and manganese carbonate layers in the sediments from northern Baltic Sea sub-basins imply oxic marine intrusions from c. 7.0 cal. ka BP. Related water-column stratification together with enhanced primary productivity caused oxygen-depleted bottom waters and deposition of organic-rich laminated sediments during the Holocene Thermal Maximum. Comparable sedimentary/geochemical records in early Littorina Sea sediments of e.g. the eastern Gotland Basin therefore imply a low impact of isostatic uplift on the northern Baltic Sea environment. A climate-controlled decline in hypoxia induced a gradual deposition of rather homogenous sediments in the central and northern Baltic Sea during the mid-Littorina Sea stage. Progressive isostatic uplift caused a diverging environmental development in both basins during the mid- to late Littorina Sea stage. The Åland sill shoaled markedly allowing only strong inflows to enter the northern Baltic Sea as late as 2.3 cal. ka BP. The restricted deep-water exchange with the central Baltic Sea probably resulted in a pronounced freshening and oxic bottom waters. This, together with an uplift-related sediment re-deposition induced accumulation of rather homogenous sediments throughout the remaining Littorina Sea stage. By contrast, a climate-controlled depositional environment in the central Baltic Sea during this time caused the formation of homogeneous sediments during cold phases and laminated sediments during warm phases. [ABSTRACT FROM AUTHOR]

The idea of parasitic systems, formulated by V.N. Beklemishev 70 years ago, represents a conceptual tool for the analysis of populational and biocoenotic roles of parasites. The questions concerning longterm stable persistence of host-parasite systems in communities can be discussed meaningfully only within this concept. Importantly, the set of terms elaborated within the parasitic system concept is applicable not only to parasitology, but also contributes to the general knowledge of life cycles of organisms and differences in the environment. This concept provides an opportunity for comprehensive analysis of systems, based on any type of stable biocoenotic interactions in the community (predation, commensalism, competition, etc.). Trematode-based parasitic systems, involving populations of intertidal mollusks of the genus Littorina, allow demonstrating how the strong 'negative' effect of parasites on hosts at the individual level (complete parasitic castration) can be compensated at the population level. Such compensation functions as a prerequisite for maintaining long-term stable interactions between populations of parasites and their hosts within parasitic system (the ecosystem, biocoenotic level). [ABSTRACT FROM AUTHOR]

The hypothesis on monogenic inheritance of white longitudinal shell bands in the snail Littorina obtusata was tested. The data showed that a single gene with two alleles was responsible for the presence of the bands on the snail shell. The presence of bands was the dominant character. [ABSTRACT FROM AUTHOR]

L. obtusata and Trivia sp. shells were systematically used for personal ornamentation by groups who occupied northern Iberia during the Mesolithic. The shells from El Mazo and El Toral III (Asturias, Spain) offer a unique opportunity for investigating raw material procurement, selection strategies, and manufacture processes developed by Asturian Mesolithic societies for beads production. By combining taphonomic, morphometric, and microscopic analyses, our results show that the shells were introduced and transformed in the caves. Mollusk consumption at the sites and bead manufacture indicate that the sites were occupied for both economic and symbolic purposes. The use of similar shell beads by contemporaneous societies located in different environments (coastal and interior) and relying on drastically different subsistence strategies mirrors the complex circulation network developed by Mesolithic foraging societies. [ABSTRACT FROM AUTHOR]

Littorina brevicula is one of the most common gastropods in the supralittoral zone around Japan. The northernmost population of this species is around Hokkaido and the determinant of this northern limit is likely seawater and air temperature. To reconstruct an evolutionary history of this species, we investigated genetic differentiation among 12 populations (three from Hokkaido, six from Honshu and three from Kyushu) using a mitochondrial DNA marker (partial sequence of the NADH dehydrogenase 6 gene). The haplotype network showed shallow genetic divergence within the species, suggesting a bottleneck followed by population expansion. One major haplotype that occurred in 70.5% of all individuals examined was the most frequent in every population sampled. A second major haplotype was abundant around Kyushu but not found in Hokkaido. This skewed haplotype distribution resulted in significant genetic differentiation along the north-south axis of Japan. The importance of the southern clade, which included the second major haplotype, was supported by population genetic analyses of datasets that excluded either the southern clade or the northern clades. The north-south differentiation remained when datasets that excluded the northern clades were used, but disappeared when datasets that excluded the southern clade were used. The combined evidence of shallow divergence and the north-south population structure suggests that the L. brevicula population around Japan once declined and then expanded and colonized northward. Although the time of population reduction and recolonization could not be precisely estimated, the observation that this species is absent further north in Japan suggests that it would have been unable to survive in northern Japan during the last glacial maximum (LGM) and therefore recolonization likely occurred after the LGM, probably from south to north. [ABSTRACT FROM AUTHOR]