Different behavioral profiles between invasive and native nudibranchs: means for invasion success?

Behavior is predicted to be a primary determinant of the success of the invasion process during the early phases of colonization. Comparing invaders with sympatric native species may provide a good approach to unravel behavioral traits involved in an invasion process. In this study, we carried out an experimental simulation of the introduction and the acclimatization phase into a new environment and assessed the expression of activity, alertness, and habituation in an invasive Mediterranean population of the South African nudibranch Godiva quadricolor comparing its profiles with those of the sympatric Mediterranean native nudibranchs Cratena peregrina and Caloria quatrefagesi. Individuals of these 3 species were subjected to 3 behavioral tests: spontaneous activity, carried out in the introduction phase (immediately after sampling) and after a week of acclimatization; alert test, in which a potential threat was simulated by means of a tactile stimulus, and habituation test, in which the same alert test stimulus was repeated 5 times at 30-min intervals. The invasive G. quadricolor showed higher levels of exploration activity, thigmotaxis, alertness, and sensitization than the native species. These behavioral traits may represent pivotal drivers of the ongoing invasion process.

See-Star: a versatile hydrogel-based protocol for clearing large, opaque and calcified marine invertebrates.

Studies of morphology and developmental patterning in adult stages of many invertebrates are hindered by opaque structures, such as shells, skeletal elements, and pigment granules that block or refract light and necessitate sectioning for observation of internal features. An inherent challenge in studies relying on surgical approaches is that cutting tissue is semi-destructive, and delicate structures, such as axonal processes within neural networks, are computationally challenging to reconstruct once disrupted. To address this problem, we developed See-Star, a hydrogel-based tissue clearing protocol to render the bodies of opaque and calcified invertebrates optically transparent while preserving their anatomy in an unperturbed state, facilitating molecular labeling and observation of intact organ systems. The resulting protocol can clear large (> 1 cm3) specimens to enable deep-tissue imaging, and is compatible with molecular techniques, such as immunohistochemistry and in situ hybridization to visualize protein and mRNA localization. To test the utility of this method, we performed a whole-mount imaging study of intact nervous systems in juvenile echinoderms and molluscs and demonstrate that See-Star allows for comparative studies to be extended far into development, facilitating insights into the anatomy of juveniles and adults that are usually not amenable to whole-mount imaging.

Effects of glyphosate-based herbicide on gut microbes and hepatopancreatic metabolism in Pomacea canaliculata.

Roundup®, a prominent glyphosate-based herbicide (GBH), holds a significant position in the global market. However, studies of its effects on aquatic invertebrates, including molluscs are limited. Pomacea canaliculata, a large freshwater snail naturally thrives in agricultural environments where GBH is extensively employed. Our investigation involved assessing the impact of two concentrations of GBH (at concentrations of 19.98 mg/L and 59.94 mg/L, corresponding to 6 mg/L and 18 mg/L glyphosate) during a 96 h exposure experiment on the intestinal bacterial composition and metabolites of P. canaliculata. Analysis of the 16 S rRNA gene demonstrated a notable reduction in the alpha diversity of intestinal bacteria due to GBH exposure. Higher GBH concentration caused a significant shift in the relative abundance of dominant bacteria, such as Bacteroides and Paludibacter. We employed widely-targeted metabolomics analysis to analyze alterations in the hepatopancreatic metabolic profile as a consequence of GBH exposure. The shifts in metabolites primarily affected lipid, amino acid, and glucose metabolism, resulting in compromised immune and adaptive capacities in P. canaliculata. These results suggested that exposure to varying GBH concentrations perpetuates adverse effects on intestinal and hepatopancreatic health of P. canaliculata. This study provides an understanding of the negative effects of GBH on P. canaliculata and may sheds light on its potential implications for other molluscs.

Aplacophoran traits in the late Ordovician septemchitonid polyplacophorans.

A sample of phosphatized, originally calcareous, mollusk shells from the Katian age uppermost Mójcza Limestone at its type locality yielded a few hundred polyplacophoran plates. The chelodids are very rare among them. Three septemchitonid species dominate. They represent a gradation from underived steep roof-like plates to almost cylindrical ones, leaving only a narrow ventral slit for the foot. Apparently, this represents the first step toward the extremely derived 'segmented clam' Bauplan of the Silurian Carnicoleus, with plates completely closed at the venter except for the mouth and anal openings. To enable growth, the plates became thinner and more flexible (or perhaps resorbed) along the dorsum. The tendency toward reduction of the ventral gap of the plates in the early Paleozoic septemchitonid polyplacophorans implies their lack of ability to cling to the substrate with a muscular foot. In compensation, their plates changed toward a more efficient protective function, covering the animal body sides more and more completely. This may explain the origin of the ventral furrow of extant solenogasters hiding the rudimentary foot. An opposite route was chosen by the coeval Acaenoplax lineage, in which the plates did not contact each other, exposing much of the soft body on the dorsum. In both cases the animals appeared to be worm-like, perhaps representing different ways of evolution from the Paleozoic chitons to the extant aplacophorans.

Characterization of natural radioactivity and relative dose due to consumption of edible bivalves inhabiting the Cochin Backwater Lagoon, Kerala, southwestern coast of India.

The study on natural radionuclides in edible bivalves from the Cochin backwater lagoon, Kerala, employing alpha spectrometry, revealed higher concentrations of 210Po and 210Pb compared to 238U and 232Th. The annual committed effective dose (ACED) for the adult coastal population was calculated at a range of 1494.9 to 5783.4 µSv y-1, with 210Po being the predominant contributor, responsible for about 85 % of the dose. This highlighted significant health risks, underscored by a calculated cumulative mortality and morbidity risk range between 5.23 × 10-3 and 2.02 × 10-2. These findings emphasized the need for further research and the development of regulatory measures to mitigate exposure risks.

Per- and polyfluoroalkyl substances (PFASs) in bivalve molluscs from Shandong Province, China: Occurrence, distribution, and implications for human consumption.

We examined the occurrence and levels of 19 legacy and emerging per- and polyfluoroalkyl substances (PFASs) in 7 species of marine bivalve molluscs collected from four coastal cities of Shandong Province, China. Perfluorooctanoic acid (PFOA) was the most prevalent component, accounting for 68.1 % of total PFASs. The total PFASs in bivalve molluscs ranged from 0.86 to 6.55 ng/g wet weight, with the highest concentration found in Meretrix meretrix L. The concentration of total PFASs in bivalve molluscs showed the following trend: clams > scallops > oysters > mussels. Estimation on the human intake of PFASs from consumption of bivalve molluscs resulted in hazard ratios (HR) ranging from 0.12 to 6.40. Five of the seven species had HR >1, indicating high exposure risks associated with PFASs. Therefore, the occurrence of PFASs in marine biota is particularly concerning and further investigations on the sources of PFASs in Shandong are warranted.

Transcriptional landscape of small non-coding RNAs reveals diversity of categories and functions in molluscs.

Small non-coding RNAs (sncRNAs) are non-coding RNA molecules that play various roles in metazoans. Among the sncRNAs, microRNAs (miRNAs) guide post-translational gene regulation during cellular development, proliferation, apoptosis, and differentiation, while PIWI-interacting RNAs (piRNAs) suppress transposon activity to safeguard the genome from detrimental insertion mutagenesis. While an increasing number of piRNAs are being identified in the soma and germlines of various organisms, they are scarcely reported in molluscs. To unravel the small RNA (sRNA) expression patterns and genomic function in molluscs, we generated a comprehensive sRNA dataset by sRNA sequencing (sRNA-seq) of eight mollusc species. Abundant miRNAs were identified and characterized in all investigated molluscs, and ubiquitous piRNAs were discovered in both somatic and gonadal tissues in six of the investigated molluscs, which are more closely associated with transposon silencing. Tens of piRNA clusters were also identified based on the genomic mapping results, which varied among different tissues and species. Our dataset serves as important reference data for future genomic and genetic studies on sRNAs in these molluscs and related species, especially in elucidating the ancestral state of piRNAs in bilaterians.

Do Antarctic bivalves present microdebris? The case of Livingston Island.

Marine microdebris (MD) seem to be widespread in benthic invertebrates, even in the most remote areas of the planet such as Antarctica, although the information available is still very scarce. Here we provide a detailed quantification and characterization of the MD found on three common bivalve species (Aequiyoldia eightsii, Thracia cf. meridionalis, and Cyclocardia astartoides) inhabiting shallow areas in Johnsons' Bay, Livingston Island (South Shetland Islands, Antarctica) as a snapshot of the MD present. On average, these bivalves contained 0.71 ± 0.89 items per individual and 1.49 ± 2.35 items per gram, being comparable to the few previous existing studies in other Antarctic areas. Nearly half of the organisms analysed here (45.6 %), contained at least one item. No significant differences were found in the three bivalve species. As far as we know, this is the first study to analyse and compare MD in three bivalve species in the Antarctic Peninsula. Although our results indicate bivalves are as not as polluted as in other areas of the planet, this is remarkable since this is considered one of the last pristine areas of the world. Our results point to local activities as the main source of MD pollution in Livingston Island, although global pollution cannot be discarded. We believe this research provides a useful baseline for future studies and will contribute to develop policies and strategies to preserve Antarctic marine ecosystems from MD pollution.

The Pacific oyster reproduction is affected by early-life exposure to environmental pesticide mixture: A multigenerational study.

Pesticides threat marine organisms worldwide. Among them, the Pacific oyster is a bivalve mollusc model in marine ecotoxicology. A large body of literature already stated on the multiple-scale effects pesticides can trigger in the Pacific oyster, throughout its life cycle and in a delayed manner. In particular, reproductive toxicity is of major concern because of its influence on population dynamics. However, past studies mostly investigated pesticide reprotoxicity as a direct effect of exposure during gametogenesis or directly on gametes and little is known about the influence of an early embryo exposure on the breed capacity. Therefore, we studied delayed and multigenerational consequences through gametogenesis features (i.e. sex ratio, glycogen content, gene expression) and reproductive success in two consecutive oyster generations (F0 and F1) exposed to an environmentally-relevant pesticide mixture (sum nominal concentration: 2.85 µg.L-1) during embryo-larval development (0-48 h post fertilization, hpf). In the first generation, glycogen content increased in exposed individuals and the expression of some gametogenesis target genes was modified. The reproductive success measured 48 hpf was higher in exposed individuals. A multigenerational influence was observed in the second generation, with feminisation, acceleration of gametogenesis processes and the sex-specific modification of glycogen metabolism in individuals from exposed parents. This study is the first to highlight the delayed effects on reproduction induced by an early exposure to pesticides, and its multigenerational implications in the Pacific oyster. It suggests that environmental pesticide contamination can have impacts on the recruitment and the dynamics of natural oyster populations exposed during their embryo-larval phase.

Genetic sex determination in three closely related hydrothermal vent gastropods, including one species with intersex individuals.

Molluscs have undergone many transitions between separate sexes and hermaphroditism, which is of interest in studying the evolution of sex determination and differentiation. Here, we combined multi-locus genotypes obtained from restriction site-associated DNA (RAD) sequencing with anatomical observations of the gonads of three deep-sea hydrothermal vent gastropods of the genus Alviniconcha living in the southwest Pacific. We found that all three species (Alviniconcha boucheti, Alviniconcha strummeri, and Alviniconcha kojimai) share the same male-heterogametic XY sex-determination system but that the gonads of XX A. kojimai individuals are invaded by a variable proportion of male reproductive tissue. The identification of Y-specific RAD loci (found only in A. boucheti) and the phylogenetic analysis of three sex-linked loci shared by all species suggested that X-Y recombination has evolved differently within each species. This situation of three species showing variation in gonadal development around a common sex-determination system provides new insights into the reproductive mode of poorly known deep-sea species and opens up an opportunity to study the evolution of recombination suppression on sex chromosomes and its association with mixed or transitory sexual systems.

Nutraceutical and Medicinal Importance of Marine Molluscs.

Marine molluscs are of enormous scientific interest due to their astonishing diversity in terms of their size, shape, habitat, behaviour, and ecological roles. The phylum Mollusca is the second most common animal phylum, with 100,000 to 200,000 species, and marine molluscs are among the most notable class of marine organisms. This work aimed to show the importance of marine molluscs as a potential source of nutraceuticals as well as natural medicinal drugs. In this review, the main classes of marine molluscs, their chemical ecology, and the different techniques used for the extraction of bioactive compounds have been presented. We pointed out their nutraceutical importance such as their proteins, peptides, polysaccharides, lipids, polyphenolic compounds pigments, marine enzymes, minerals, and vitamins. Their pharmacological activities include antimicrobial, anticancer, antioxidant, anti-inflammatory, and analgesic activities. Moreover, certain molluscs like abalones and mussels contain unique compounds with potential medicinal applications, ranging from wound healing to anti-cancer effects. Understanding the nutritional and therapeutic value of marine molluscs highlights their significance in both pharmaceutical and dietary realms, paving the way for further research and utilization in human health.

First record of Arionvulgaris Moquin-Tandon, 1855 (Arionidae) from Armenia.

Background: Arionvulgaris Moquin-Tandon, 1855 is amongst the fastest-spreading terrestrial slugs Europe-wide. In recent years, it has been recorded in Canada, Mexico and continues to expand eastwards into Eurasia. Renowned for its high invasiveness, combatting its swift spread creates significant challenges in organising effective preventative measures. New information: This study presents the first record of Arionvulgaris from Armenia, which is the second record of this species' invasion of the Caucasus. In 2022, a substantial population of A.vulgaris was observed close to the City of Stepanavan, which is also the first record in Armenia of the family Arionidae. How the species was introduced to Armenia remains unknown. Identification of Arionvulgaris was conducted, based on external and genital morphology and mitochondrial CO1 (cytochrome c oxidase subunit 1) gene sequencing, revealing notable similarities with Central European clades. Our results confirm the introduction and distribution of A.vulgaris to Armenia. Invasion of such species into Armenia will require additional monitoring and would be aided by further research on Armenia's mollusc fauna in the future.

Revisiting the evolution of Family B1 GPCRs and ligands: Insights from mollusca.

Family B1 G protein-coupled receptors (GPCRs) are one of the most well studied neuropeptide receptor families since they play a central role in many biological processes including endocrine, gastrointestinal, cardiovascular and reproduction in animals. The genes for these receptors emerged from a common ancestral gene in bilaterian genomes and evolved via gene/genome duplications and deletions in vertebrate and invertebrate genomes. Their existence and function have mostly been characterized in vertebrates and few studies exist in invertebrate species. Recently, an increased interest in molluscs, means a series of genomes have become available, and since they are less modified than insect and nematode genomes, they are ideal to explore the origin and evolution of neuropeptide gene families. This review provides an overview of Family B1 GPCRs and their peptide ligands and incorporates new data obtained from Mollusca genomes and taking a comparative approach challenges existing models on their origin and evolution.

Molecular cloning, characterisation and molecular modelling of two novel T-synthases from mollusc origin.

The glycoprotein-N-acetylgalactosamine ß1,3-galactosyltransferase, known as T-synthase (EC 2.4.1.122), plays a crucial role in the synthesis of the T-antigen, which is the core 1 O-glycan structure. This enzyme transfers galactose from UDP-Gal to GalNAc-Ser/Thr. The T-antigen has significant functions in animal development, immune response, and recognition processes. Molluscs are a successful group of animals that inhabit various environments, such as freshwater, marine, and terrestrial habitats. They serve important roles in ecosystems as filter feeders and decomposers but can also be pests in agriculture and intermediate hosts for human and cattle parasites. The identification and characterization of novel carbohydrate active enzymes, such as T-synthase, can aid in the understanding of molluscan glycosylation abilities and their adaptation and survival abilities. Here, the T-synthase enzymes from the snail Pomacea canaliculata and the oyster Crassostrea gigas are identified, cloned, expressed, and characterized, with a focus on structural elucidation. The synthesized enzymes display core 1 ß1,3-galactosyltransferase activity using pNP-α-GalNAc as substrate and exhibit similar biochemical parameters as previously characterised T-synthases from other species. While the enzyme from C. gigas shares the same structural parameters with the other enzymes characterised so far, the T-synthase from P. canaliculata lacks the consensus sequence CCSD, which was previously considered indispensable.

The Occurrence of Microplastics in Donax trunculus (Mollusca: Bivalvia) Collected along the Tuscany Coast (Mediterranean Sea).

Microplastics (MPs) (0.1 µm-5 mm particles) have been documented in oceans and seas. Bivalve molluscs (BMs) can accumulate MPs and transfer to humans through the food chain. BMs (especially mussels) are used to assess MPs' contamination, but the genus Donax has not been thoroughly investigated. The aim of this study was to detect and characterize MPs in D. trunculus specimens collected along the Tuscan coast (Italy), and to assess the potential risk for consumers. The samples (~10 g of tissue and intervalval liquid from 35 specimens) were digested using a solution of 10% KOH, subjected to NaCl density separation, and filtered through 5 µm pore-size filters. All items were morphologically classified and measured, and their mean abundance (MA) was calculated. Furthermore, 20% of them were analyzed by Raman spectroscopy and, based on the obtained results, the MA was recalculated (corrected MA) and the annual human exposure was estimated. In the 39 samples analyzed, 85 items fibers (n = 45; 52.94%) and fragments (n = 40; 47.06%) were found. The MA was 0.23 ± 0.17 items/grww. Additionally, 83.33% of the items were confirmed as MPs (polyethylene and polyethylene terephthalate). Based on the correct MA (0.18 MPs/grww), D. trunculus consumers could be exposed to 19.2 MPs/per capita/year. The health risk level of MPs was classified as level III (moderate).

Distribution of microplastics in seafloor sediments and their differential assimilation in nearshore benthic molluscs along the south-west coast of India.

Spatial and temporal distribution of microplastics (MPs) in the nearshore seafloor sediments along the Southwest coast of India and their patterns of accumulation in selected infaunal and epibenthic molluscs with diverse feeding strategies were investigated. Along the 300-km coastal stretch, which is one of the most productive and biodiversity rich regions of the eastern Arabian Sea, notable levels of MP contamination in both sediment (617.7 items/kg dry weight) and molluscs (5.39 items/g) was recorded. The concentration of MPs in sediments also varied seasonally, with a higher prevalence during the post-monsoon season. Among the four molluscan groups studied, the highest MP abundance was recorded among scavenging gastropod Pseudominolia biangulosa (9.13 items/g), followed by microcarnivore scaphopod Tesseracme quadrapicalis (5.96 items/g). In comparison, the suspension feeding bivalve, Anadara hankeyana and deposit feeding clam Jitlada philippinarum had lesser accumulation of MPs (2.98 items/g and 3.50 items/g respectively). The majority of MPs in sediments and within molluscs were less than 250 µm in size (89.14%) and were predominantly fibres and fragments. Chemical characterisation of MPs revealed eleven types of polymers dominated by polyethylene (PE) and polypropylene (PP). Present study identified positive correlations between ingested MP polymers and the feeding strategies of molluscs. Higher values for the ecological risk assessment indices (PHI, PLI and PERI) in most of the stations indicated the severity of plastic pollution in the region. Molluscs being a major contributor to the benthic food web is also a connecting link to higher trophic levels. Hence understanding the specificity in the MPs accumulation pattern within this group has far reaching significance in utilizing them as potential bioindicators for pollution studies in marine ecosystems.

Ancestors' Gift: Parental Early Exposure to the Environmentally Realistic Pesticide Mixture Drives Offspring Phenotype in a Larger Extent Than Direct Exposure in the Pacific Oyster, Crassostrea gigas.

Marine organisms are threatened by the presence of pesticides in coastal waters. Among them, the Pacific oyster is one of the most studied invertebrates in marine ecotoxicology where numerous studies highlighted the multiscale impacts of pesticides. In the past few years, a growing body of literature has reported the epigenetic outcomes of xenobiotics. Because DNA methylation is an epigenetic mark implicated in organism development and is meiotically heritable, it raises the question of the multigenerational implications of xenobiotic-induced epigenetic alterations. Therefore, we performed a multigenerational exposure to an environmentally relevant mixture of 18 pesticides (nominal sum concentration: 2.85 µg·L-1) during embryo-larval stages (0-48 hpf) of a second generation (F1) for which parents where already exposed or not in F0. Gene expression, DNA methylation, and physiological end points were assessed throughout the life cycle of individuals. Overall, the multigenerational effect has a greater influence on the phenotype than the exposure itself. Thus, multigenerational phenotypic effects were observed: individuals descending from exposed parents exhibited lower epinephrine-induced metamorphosis and field survival rates. At the molecular level, RNA-seq and Methyl-seq data analyses performed in gastrula embryos and metamorphosis-competent pediveliger (MCP) larvae revealed a clear F0 treatment-dependent discrimination. Some genes implicated into shell secretion and immunity exhibited F1:F0 treatment interaction patterns (e.g., Calm and Myd88). Those results suggest that low chronic environmental pesticide contamination can alter organisms beyond the individual scale level and have long-term adaptive implications.

Monospecific mangrove reforestation changes relationship between benthic mollusc diversity and biomass: Implication for coastal wetland management.

Anthropogenic causes are overtaking natural factors to reshape patterns of biodiversity and ecosystem functioning. Mangrove reforestation aimed at reversing losses of mangroves has been conducted worldwide for several decades. However, how reforestation influences the link between ecological processes that shape community diversity and the consequent effects on ecosystem functions such as biomass production is less well known. Here we used data collected before and after mangrove planting to examine the effects of reforestation on molluscan species richness and biomass production by testing the changes in species richness, compositional similarities, distance-decay effects (community similarity decreases with increasing geographical distance) in metacommunity across a regional scale of 480 km (23-27 °N) in southeast Chinese coasts. Additionally, we further detected the impact of landscape configuration caused by different intensities of reforestation on the mollusc community. After the mangrove reforestation, mollusc species richness and biomass increased significantly. The increases in species richness and biomass of mollusc community were mediated by reducing distance-decay effect, indicating an increase in relationship strength between species richness and biomass might be associated with a decrease in distance-decay effect with rising mangrove habitat. We highlight the importance of considering the effects of anthropogenic changes on the relationship between biodiversity and ecosystem functioning. Quantifying the distance-decay effect of these influences enables management decisions about coastal restoration to be based upon ecological mechanisms rather than wishful thinking or superficial appearance.

Using a metabolomics approach to investigate the sensitivity of a potential Arctic-invader and its Arctic sister-species to marine heatwaves and traditional harvesting disturbances.

Coastal species are threatened by fishing practices and changing environmental conditions, such as marine heatwaves (MHW). The mechanisms that confer tolerance to such stressors in marine invertebrates are poorly understood. However, differences in tolerance among different species may be attributed to their geographical distribution. To test the tolerance of species occupying different thermal ranges, we used two closely related bivalves the softshell clam Mya arenaria (Linnaeus, 1758), a cold-temperate invader with demonstrated potential for establishment in the Arctic, and the blunt gaper Mya truncata (Linnaeus, 1758), a native polar species. Clams were subjected to a thermal stress, mimicking a MHW, and harvesting stress in a controlled environment. Seven acute temperature changes (2, 7, 12, 17, 22, 27, and 32 °C) were tested at two harvesting disturbance intensities (with, without). Survival was measured after 12 days and three tissues (gills, mantle, and posterior adductor muscle) collected from surviving individuals for targeted metabolomic profiling. MHW tolerance differed significantly between species: 26.9 °C for M. arenaria and 17.8 °C for M. truncata, with a negligeable effect of harvesting. At the upper thermal limit, M. arenaria displayed a more profound metabolomic remodelling when compared to M. truncata, and this varied greatly between tissue types. Network analysis revealed differences in pathway utilization at the upper MHW limit, with M. arenaria displaying a greater reliance on multiple DNA repair and expression and cell signalling pathways, while M. truncata was limited to fewer pathways. This suggests that M. truncata is ill equipped to cope with warming environments. MHW patterning in the Northwest Atlantic may be a strong predictor of population survival and future range shifts in these two clam species. As polar environments undergo faster rates of warming compared to the global average, M. truncata may be outcompeted by M. arenaria expanding into its native range.

Trace elements assessment in Cerastoderma glaucum from port areas in the Tunisian Mediterranean coast: The influence of parasites on bioaccumulation.

In the present study, the seasonal concentration of seven trace elements was investigated in sediment and the cockle Cerastoderma glaucum tissues from two port areas in the North-East and South of Tunisia comparing cockles non-parasitized (NP) and parasitized (P) with digenean parasites. Elements concentration in sediments analyzed in both sites revealed that Zinc (Zn), Chromium (Cr), and Lead (Pb) were the most abundant ones, while Cadmium (Cd) and Mercury (Hg) were less abundant. The bioaccumulation of trace metals and Arsenic (As) in the tissues of cockles seems to be modulated by both the infection state and the parasite species. The relationship between bioaccumulation of metals and As, trematode species and abiotic parameters showed that the availability of certain metals for uptake by P cockles of both sites was influenced by the salinity and temperature of the water. Our results corroborate the possibility of using digenean infecting bivalves in biomonitoring aquatic ecosystems.