Unravelling epigenetic mechanisms in Cerastoderma edule genome: a comparison of healthy and neoplastic cockles.

Cancer is a multifaceted genetic disease characterized by the acquisition of several essential hallmarks. Notably, certain cancers exhibit horizontal transmissibility, observed across mammalian species and diverse bivalves, the latter referred to as hemic neoplasia. Within this complex landscape, epigenetic mechanisms such as histone modifications and cytosine methylation emerge as fundamental contributors to the pathogenesis of these transmissible cancers. Our study delves into the epigenetic landscape of Cerastoderma edule, focusing on whole-genome methylation and hydroxymethylation profiles in heathy specimens and transmissible neoplasias by means of Nanopore long-read sequencing. Our results unveiled a global hypomethylation in the neoplastic specimens compared to their healthy counterparts, emphasizing the role of DNA methylation in these tumorigenic processes. Furthermore, we verified that intragenic CpG methylation positively correlated with gene expression, emphasizing its role in modulating transcription in healthy and neoplastic cockles, as also highlighted by some up-methylated oncogenic genes. Hydroxymethylation levels were significantly more elevated in the neoplastic samples, particularly within satellites and complex repeats, likely related to structural functions. Additionally, our analysis also revealed distinct methylation and activity patterns in retrotransposons, providing additional insights into bivalve neoplastic processes. Altogether, these findings contribute to understanding the epigenetic dynamics of bivalve neoplasias and shed light on the roles of DNA methylation and hydroxymethylation in tumorigenesis. Understanding these epigenetic alterations holds promise for advancing our broader understanding of cancer epigenetics.

Oxidative response to accumulation of trace metals in tissue of two bivalves, the Asian green mussel Perna viridis and the blood cockle Tegillarca granosa, living in Pattani Bay, Thailand.

OBJECTIVE: Using bivalves to indicate aquatic pollutants was favorable for discerning the negative effects of high levels of metal accumulation in tissue. We investigated the correlation between trace metal accumulation and the tissue oxidative response of two bivalves. METHODS: The Asian green mussel Perna viridis and the blood cockle Tegillarca granosa were sampled along with seawater and sediments from three locations around Pattani Bay, Thailand. Accumulation of nine trace metals (cadmium, cobalt, copper, chromium, nickel, manganese, iron, zinc, and lead) in seawater, sediments, and tissue and the oxidative tissue response were evaluated. Metal bioaccumulation factor, biota-sediment accumulation factor, and histopathology were also indicated. RESULT: The present study found that P. viridis and T. granosa were macroconcentrators and bioaccumulative of cadmium, and their tissue accumulation of cadmium was strongly related to lipid peroxidation activation. Perna viridis exhibited a higher oxidative response than T. granosa, as indicated by malondialdehyde, catalase, and reduced glutathione levels. CONCLUSION: The present study indicated that P. viridis and T. granosa were macroconcentrators and bioaccumulative of cadmium, and their tissue accumulation of cadmium was strongly related to lipid peroxidation activation. Research has shown discernible negative effects of a high level of metal accumulation in tissue, and deformed and damaged tissues were present in the gills, digestive glands, intestines, and feet of P. viridis and T. granosa.

Heterogeneous microgeographic genetic structure of the common cockle (Cerastoderma edule) in the Northeast Atlantic Ocean: biogeographic barriers and environmental factors.

Knowledge of genetic structure at the finest level is essential for the conservation of genetic resources. Despite no visible barriers limiting gene flow, significant genetic structure has been shown in marine species. The common cockle (Cerastoderma edule) is a bivalve of great commercial and ecological value inhabiting the Northeast Atlantic Ocean. Previous population genomics studies demonstrated significant structure both across the Northeast Atlantic, but also within small geographic areas, highlighting the need to investigate fine-scale structuring. Here, we analysed two geographic areas that could represent opposite models of structure for the species: (1) the SW British Isles region, highly fragmented due to biogeographic barriers, and (2) Galicia (NW Spain), a putative homogeneous region. A total of 9250 SNPs genotyped by 2b-RAD on 599 individuals from 22 natural beds were used for the analysis. The entire SNP dataset mostly confirmed previous observations related to genetic diversity and differentiation; however, neutral and divergent SNP outlier datasets enabled disentangling physical barriers from abiotic environmental factors structuring both regions. While Galicia showed a homogeneous structure, the SW British Isles region was split into four reliable genetic regions related to oceanographic features and abiotic factors, such as sea surface salinity and temperature. The information gathered supports specific management policies of cockle resources in SW British and Galician regions also considering their particular socio-economic characteristics; further, these new data will be added to those recently reported in the Northeast Atlantic to define sustainable management actions across the whole distribution range of the species.

Trematode genetic patterns at host individual and population scales provide insights about infection mechanisms.

Multiple parasites can infect a single host, creating a dynamic environment where each parasite must compete over host resources. Such interactions can cause greater harm to the host than single infections and can also have negative consequences for the parasites themselves. In their first intermediate hosts, trematodes multiply asexually and can eventually reach up to 20% of the host's biomass. In most species, it is unclear whether this biomass results from a single infection or co-infection by 2 or more infective stages (miracidia), the latter being more likely a priori in areas where prevalence of infection is high. Using as model system the trematode Bucephalus minimus and its first intermediate host cockles, we examined the genetic diversity of the cytochrome c oxidase subunit I region in B. minimus from 3 distinct geographical areas and performed a phylogeographic study of B. minimus populations along the Northeast Atlantic coast. Within localities, the high genetic variability found across trematodes infecting different individual cockles, compared to the absence of variability within the same host, suggests that infections could be generally originating from a single miracidium. On a large spatial scale, we uncovered significant population structure of B. minimus, specifically between the north and south of Bay of Biscay. Although other explanations are possible, we suggest this pattern may be driven by the population structure of the final host.

Biochemical responses in mice induced by saxitoxins extracted from the cockles Acanthocardia tuberculatum.

Harmful algae blooms have increased in frequency and geographic range in recent decades, and they produce toxins strains such as saxitoxins (STXs). they block voltage-gated sodium channels and can lead to several poisonings and the death of organisms that pose a significant risk to public and environmental health. The study of STXs toxicity has been carried out but little is known about the response of antioxidant enzymes activities to STXs in mice. The purpose of this study was to evaluate biochemical responses and oxidative stress induced by STXs extracted from Acanthocardia tuberculatum. To this end, daily, mice were treated orally for 7 days with sublethal concentrations (10 mg/100 g mouse). The animal's liver was assessed using biomarkers such as activities of catalase (CAT), thiobarbituric acid reactive substances (TBARS), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and succinate dehydrogenase (SDH). In the blood, plasmatic markers were analysed as glutamic oxalic transaminase (GOT), glutamic pyruvic transaminase (GPT), creatinine phosphokinase (CPK), lactate dehydrogenase (LDH), urea and creatinine. Globally, test toxicity test showed a significant decrease in the weight at 10 mg /100 g mouse, and the results showed an increase of GPT, GOT, CPK, LDH, CAT and TBARS activities and the inhibitory effect of GAPDH activities but creatinine, urea and SDH activities showed no significative difference from the control. We concluded that STXs induce oxidative stress breaking in mice the balance of the defence system and causing oxidations reactions. Moreover, STXs affect energy metabolism in mice, however, renal function in mice is not affected by exposure to STXs.

Utilization of cockle shell powder as an adsorbent to remove phosphorus-containing wastewater.

The paper demonstrates the capability of using cockle shells as an adsorbent for phosphorus removal from simulated petrochemical wastewater, focusing on the actual condition of the petrochemical facultative pond. In this study, the physicochemical properties of shell powder were determined, such as the functional groups, surface morphology, crystalline structure, and surface area using FTIR, SEM, EDX, XRD, and BET. It was observed that the optimum conditions for effective phosphorus removal are under the presence of rotational speed (125 rpm), higher dosage (7 g/L), and larger surface area (smaller particle size) of the shell powder. Fine powder achieved up to 52.27% of phosphorus removal after 40 min compared to coarse powder which could only give 16.67% removal. Additionally, calcined shell powder demonstrated a higher phosphorus removal rate, i.e., up to 62.37%, compared to raw shell powders. The adsorption isotherm was studied using Langmuir and Freundlich models, but the isothermal data fit better for the Freundlich model (R2 = 0.9836). Overall, this study has successfully generated a greener and low-cost adsorbent.

Exploring the feasibility of cryopreserving larvae of the common cockle (Cerastoderma edule) for hatchery production.

The decline of natural populations of the common cockle (Cerastoderma edule) through the European coast is posing a threat to local small-scale fisheries. These declines are primarily attributed to the prevalence of several pathogens and the disseminated neoplasia in cockle populations. The institution of a biobank of cryopreserved larvae could enhance hatchery production and help the restocking. The present work aimed at the development of a cryopreservation protocol for larvae of the common cockle using the mollusk cryopreservation protocols designed in our laboratory. Toxicity bioassays and short-term cryopreservation experiments were performed for protocol optimization according with cellular tolerance. Once settled, the viability of cryopreserved larvae was studied long term. Toxicity tests evidenced high tolerance of larvae against detrimental effects of Cryoprotecting Agents (CPAs). Cryopreservation of 48 h-old D-larva showed a 100% survival when increasing the equilibrium time from 15 to 60 min and using Propylene-Glycol (PG) + 0.4 M Trehalose (TRE) in Filtered Sea Water (FSW) and 60 min of exposure to CPA solution before slow-cooling. However, when cryopreserving the older larvae, the variation in equilibrium times hardly showed any effect but 10% Ethylene-Glycol (EG) + 0.4 M TRE and 60 min of exposure yielded the best relative survivorship (100%). Cryopreservation caused a significant delay on the growth rate of the latest larval stage. However, cryopreserved larvae survived to day 4-6, while 30 ± 12.17% of control larvae developed into pediveliger stage, of which 50% settled and transformed into juvenile cockles. These results demonstrated the role of the cell-type specificity in cryopreservation and highlight the importance of studying potential long-term effects of this tool to ensure the viability of the protocols.

Long-term survey discloses a shift in the dynamics pattern of an emerging disease of cockles Cerastoderma edule, marteiliosis, and raises hypotheses to explain it.

Drivers of marine disease outbreaks are poorly understood in spite of their growing impact. We present here results from a unique case study examining how cockles Cerastoderma edule have responded to the introduction of the novel protistan Marteilia cochillia, which led in 2012 to cockle fishery collapse in Galician rias. Based on intensive survey for eight years (2011-2019) of two affected shellfish beds, inner and outer in the Ría de Arousa, involving monthly evaluation of cockle health status and estimation of mortality, detailed information is provided of the declining impact of marteiliosis over a wild cockle population with evidence suggesting its increasing resistance. Disease dynamics involved an annual "breaking wave" of prevalence and subsequent cockle mass mortality, causing the near extinction of every recruited cohort. A shift in this pattern, from a severe epidemic towards an endemic profile, was observed in the inner shellfish bed since the cohort that was recruited in 2016, suggesting the hypothesis of increasing marteiliosis resistance through natural selection. Risk factors that may contribute to trigger marteiliosis outbreaks were analysed. Host age and sex did not influence susceptibility to marteiliosis. No clear relationships between environmental conditions (temperature, salinity and upwelling index) or cockle density and disease dynamics were found. Spatial differences in disease dynamics could be due to differences in the abundance of infective stages hypothetically linked to spatial differences in the population dynamics of a putative planktonic intermediate host. All these findings have potential implications for the management of diseased populations.

Decline of Marteilia cochillia in Ría de Arousa may be due to increased resistance in host Cerastoderma edule.

A huge, unprecedented mortality of cockle Cerastoderma edule caused by the protist Marteilia cochillia, which had never before been detected in Galicia (NW Spain), brought on a cockle fishery collapse in the Ría de Arousa (Galicia) in 2012. Since then, the disease dynamic pattern in the shellfish bed of Lombos do Ulla (at the inner area of that ria) involved an overwhelming annual wave of infections and subsequent cockle mass mortality that caused the near extinction of every cohort recruited to that bed. However, a pattern shift was detected among wild cohorts recruiting since 2016, with progressive declines of marteiliosis prevalence and increments in cockle survival. This suggested 2 non-exclusive hypotheses: increasing marteiliosis resistance through natural selection, and reduced abundance and/or virulence of the parasite. A field experiment was performed to assess these hypotheses by comparing marteiliosis prevalence and severity, as well as mortality, in cockles that naturally recruited to this bed in 2017 and 2018 with those of naïve cockles collected from a marteiliosis-free area and transplanted into Lombos do Ulla in 2017 and 2018. Marteiliosis prevalence and cumulative cockle mortality quickly reached very high values among the transplanted cockles, demonstrating that the parasite remained present and virulent in the area. Conversely, marteiliosis prevalence and cockle mortality were much lower in the cockles that recruited to Lombos do Ulla, suggesting increased resistance that may have been driven by natural selection. The young age at which cockles start reproduction and the very high mortality caused by marteiliosis may have enhanced natural selection.

First report of Perkinsus olseni infections in blood cockles Tegillarca granosa on the south coast of Korea.

The protozoan parasite Perkinsus olseni has become a focus of attention since it has been responsible for mass mortalities and economic losses in a wide range of bivalve hosts globally. The P. olseni host range along the south coast of Korea may extend beyond what was previously understood, and blood cockles in the Family Arcidae are also suggested to be potential hosts of P. olseni. In the present study, we applied histology and molecular techniques to identify Perkinsus sp. infections in the blood cockles Tegillarca granosa, which have been commercially exploited on the south coast of Korea for several decades. Histology and molecular techniques, including genus-specific immunofluorescence assay, species-specific fluorescence in situ hybridization, and phylogeny based on the ribosomal DNA internal transcribed spacer region revealed that T. granosa is infected by P. olseni, although the prevalence was low (0.5%). Histology revealed massive hemocyte infiltrations in the mantle, gill, and digestive gland connective tissues, indicating that the infection exerts negative impacts on the host cockles.

Metal Contamination and Biomarkers in Cerastoderma glaucum: A Multi-level Approach.

In this study, we focused on evaluating the responses of the cockle, Cerastoderma glaucum to in situ exposures to metals at three sites in the Gulf of Gabes in the coastal zone of Tunisia differing in levels of metal contamination. Firstly, we examined the general physiological state of the organisms. Secondly, we evaluated the bioaccumulation of several metals (Cd, Cu, Zn, Ni) in the cockles. Thirdly, we focused on evaluating histologically changes in gametogenesis and sexual maturity of the organisms. Finally, we determined the expression of seven key genes encoding enzymes or proteins involved in responses to different types of environmental stressors. Results showed a decrease in the general physiological status of the cockles, including a reduced condition index, sex ratios skewed to females (70% and 80% females in the intermediate and the contaminated site, respectively) and greater mortalities in tests under anoxic conditions (i.e., stress on stress test) in cockles collected from the most contaminated site (LT50 = 2.88 days) compared to the cockles from the intermediate site (LT50 = 5 days) and the less contaminated site (LT50 = 6 days). Results for metal bioaccumulation showed that the levels of Cd, Cu, Zn and Ni in cockles were consistent with the contaminant gradient, with the highest levels in cockles from the most contaminated site (1.04; 4.92; 52.76 and 13.81 µg/g dw, respectively), followed by those from the intermediate site (0.34; 2.94; 36.94; 17.40 µg/g dw, respectively) and then the less contaminated site (0.065; 1.27; 21.62 and 5.40 µg/g dw, respectively). Results from the gametogenesis and maturity index showed few differences in the reproductive cycle of cockles collected from the three study sites. There were different patterns of gene expression that were divided into three groups in terms of responses: (1) expression of genes involved in metal detoxification, ATP Binding Cassette Subfamily B Member 1 (ABCB1) and metallothionein MT) and genes for superoxide dismutases (i.e., Mn SOD and CuZn SOD), which did not show any difference in their levels of expression; (2) heat shock protein 70 (HSP70) gene expression, which decreased in cockles according to the pollution gradient, and (3) expression of catalase (CAT) and cytochrome oxidase subunit 1 (COI) genes was threefold and 1000-fold higher in cockles from intermediate and most contaminated sites compared to the less contaminated site. Therefore, changes in overall physiological condition, sex ratios and expression of HSP70, CAT and COI genes may be appropriate biomarkers for in situ studies of the impacts of metals in cockles. However, these biomarkers should be coupled to proteomics studies.

Evaluation of the chemical, physical, and biological properties of a newly developed bioceramic cement derived from cockle shells: an in vitro study.

BACKGROUND: Tricalcium silicate is the main component of commercial bioceramic cements that are widely used in endodontic treatment. Calcium carbonate, which is manufactured from limestone, is one of the substrates of tricalcium silicate. To avoid the environmental impact of mining, calcium carbonate can be obtained from biological sources, such as shelled mollusks, one of which is cockle shell. The aim of this study was to evaluate and compare the chemical, physical, and biological properties of a newly developed bioceramic cement derived from cockle shell (BioCement) with those of a commercial tricalcium silicate cement (Biodentine). METHODS: BioCement was prepared from cockle shells and rice husk ash and its chemical composition was determined by X-ray diffraction and X-ray fluorescence spectroscopy. The physical properties were evaluated following the International Organization for Standardization (ISO) 9917-1;2007 and 6876;2012. The pH was tested after 3 h to 8 weeks. The biological properties were assessed using extraction medium from BioCement and Biodentine on human dental pulp cells (hDPCs) in vitro. The 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5[(phenylamino)carbonyl]-2 H-tetrazolium hydroxide assay was used to evaluate cell cytotoxicity following ISO 10993-5;2009. Cell migration was examined using a wound healing assay. Alizarin red staining was performed to detect osteogenic differentiation. The data were tested for a normal distribution. Once confirmed, the physical properties and pH data were analyzed using the independent t-test, and the biological property data were analyzed using one way ANOVA and Tukey's multiple comparisons test at a 5% significance level. RESULTS: The main components of BioCement and Biodentine were calcium and silicon. BioCement's and Biodentine's setting time and compressive strength were not different. The radiopacity of BioCement and Biodentine was 5.00 and 3.92 mmAl, respectively (p < 0.05). BioCement's solubility was significantly higher than Biodentine. Both materials exhibited alkalinity (pH ranged from 9 to 12) and demonstrated > 90% cell viability with cell proliferation. The highest mineralization was found in the BioCement group at 7 days (p < 0.05). CONCLUSIONS: BioCement exhibited acceptable chemical and physical properties and was biocompatible to human dental pulp cells. BioCement promotes pulp cell migration and osteogenic differentiation.

Effects of short-term hyposalinity stress on four commercially important bivalves: A proteomic perspective.

Increased heavy rainfall can reduce salinity to values close to 0 in estuaries. Lethal and sublethal physiological and behavioural effects of decreases in salinity below ten have already been found to occur in the commercially important clam species Venerupis corrugata, Ruditapes decussatus and R. philippinarum and the cockle Cerastoderma edule, which generate an income of ∼74 million euros annually in Galicia (NW Spain). However, studies of the molecular response to hyposaline stress in bivalves are scarce. This 'shotgun' proteomics study evaluates changes in mantle-edge proteins subjected to short-term hyposaline episodes in two different months (March and May) during the gametogenic cycle. We found evidence that the mantle-edge proteome was more responsive to sampling time than to hyposalinity, strongly suggesting that reproductive stages condition the stress response. However, hyposalinity modulated proteome profiles in V. corrugata and C. edule in both months and R. philippinarum in May, involving proteins implicated in protein folding, redox homeostasis, detoxification, cytoskeleton modulation and the regulation of apoptotic, autophagic and lipid degradation pathways. However, proteins that are essential for an optimal osmotic stress response but which are highly energy demanding, such as chaperones, osmoprotectants and DNA repair factors, were found in small relative abundances. In both months in R. decussatus and in March in R. philippinarum, almost no differences between treatments were detected. Concordant trends in the relative abundance of stress response candidate proteins were also obtained in V. corrugata and C. edule in the different months, but not in Ruditapes spp., strongly suggesting that the osmotic stress response in bivalves is complex and possibly influenced by a combination of controlled (sampling time) and uncontrolled variables. In this paper, we report potential molecular targets for studying the response to osmotic stress, especially in the most osmosensitive native species C. edule and V. corrugata, and suggest factors to consider when searching for biomarkers of hyposaline stress in bivalves.

Vortex-assisted dispersive solid-phase microextraction of ondansetron and domperidone using carbonized cockle shell modified with nitrogen and sulfur-doped carbon dots as a bio-based sorbent.

Herein, for the first time, the wasted cockle shells were modified with nitrogen and sulfur-doped carbon dots after carbonization and used as a suitable bio-based sorbent for the extraction of two important antiemetic drugs named ondansetron and domperidone. The extraction and separation were performed based on vortex-assisted dispersive solid-phase microextraction and high-performance liquid chromatography, respectively. Various techniques such as Fourier-transform infrared, fluorescence, field emission scanning electron microscopy, thermogravimetric analysis, nitrogen adsorption-desorption analysis, X-ray powder diffraction, and energy-dispersive spectroscopy were used to characterize the chemical composition of prepared sorbent. After examining the factors affecting the extraction efficiency and access to the optimal points using the response surface methodology, the linearity was in the range of 5-350 µg/L with acceptable coefficients of determination (R2 > 0.993). The limits of detection and quantification were in the range of 1.5-2.3  and 4.9-7.1 µg/L, respectively. In the end, the proposed method was applied for the quantitative determination of trace levels of target analytes from pharmaceutical tablets, serum, and urine samples. The recoveries were more than 95.2%, indicating the proposed method's excellent accuracy.

Discovery of the parasite Marteilia cocosarum sp. nov. In common cockle (Cerastoderma edule) fisheries in Wales, UK and its comparison with Marteilia cochillia.

Diseases of bivalve molluscs caused by paramyxid parasites of the genus Marteilia have been linked to mass mortalities and the collapse of commercially important shellfish populations. Until recently, no Marteilia spp. have been detected in common cockle (Cerastoderma edule) populations in the British Isles. Molecular screening of cockles from ten sites on the Welsh coast indicates that a Marteilia parasite is widespread in Welsh C. edule populations, including major fisheries. Phylogenetic analysis of ribosomal DNA (rDNA) gene sequences from this parasite indicates that it is a closely related but different species to Marteilia cochillia, a parasite linked to mass mortality of C. edule fisheries in Spain, and that both are related to Marteilia octospora, for which we provide new rDNA sequence data. Preliminary light and transmission electron microscope (TEM) observations support this conclusion, indicating that the parasite from Wales is located primarily within areas of inflammation in the gills and the connective tissue of the digestive gland, whereas M. cochillia is found mainly within the epithelium of the digestive gland. The impact of infection by the new species, here described as Marteilia cocosarum n. sp., upon Welsh fisheries is currently unknown.

Vibrio aestuarianus subsp. cardii subsp. nov., pathogenic to the edible cockles Cerastoderma edule in France, and establishment of Vibrio aestuarianus subsp. aestuarianus subsp. nov. and Vibrio aestuarianus subsp. francensis subsp. nov.

Cockle mortality events have been reported in northern France since 2012. In the present study, we describe and investigate the implication of a potential bacterial causative agent in cockle mortality. Bacteria isolated from five different cockle mortality events were characterized and studied. Using phenotypic analysis combined with DNA-DNA hybridization (DDH) and whole genome sequencing, the isolates were shown to belong to Vibrio aestuarianus, a species regularly detected in France during oyster mortality events. Comparison of the strains from cockles with strains from French oysters and the type strain showed that the strains from cockles were genetically different to those from oysters and also different to the V. aestuarianus type strain. Moreover, the cockle and oyster strains were classified into two different, but close, groups both separated from the type strain by: (1) analyses of the ldh gene sequences; (2) DDH assays between 12/122 3T3T (LMG 31436T=DSM 109723T), a representative cockle strain, 02/041T (CIP 109791T=LMG 24517T) representative oyster strain and V. aestuarianus type strain LMG 7909T; (3) average nucleotide identity values calculated on the genomes; and (4) phenotypic traits. Finally, results of MALDI-TOF analyses also revealed specific peaks discriminating the three representative strains. The toxicity of representative strains of these cockle isolates was demonstrated by experimental infection of hatchery-produced cockles. The data therefore allow us to propose two novel subspecies of Vibrio aestuarianus: Vibrio aestuarianus subsp. cardii subsp. nov. for the cockle strains and Vibrio aestuarianus subsp. francensis subsp. nov. for the Pacific oyster strains, in addition to an emended description of the species Vibrio aestuarianus.

Co-occurrence of pathogen assemblages in a keystone species the common cockle Cerastoderma edule on the Irish coast.

Despite coinfections being recognized as the rule in animal populations, most studies focus on single pathogen systems. Pathogen interaction networks and the drivers of such associations are lacking in disease ecology studies. Common cockle Cerastoderma edule populations are exposed to a great diversity of pathogens, thus making them a good model system to investigate. This study examined the diversity and prevalence of pathogens from different taxonomic levels in wild and fished C. edule on the Irish coast. Potential interactions were tested focussing on abiotic (seawater temperature and salinity) and biotic (cockle size and age, and epiflora on shells) factors. No Microsporidia nor OsHV-1µVar were detected. Single infections with Haplosporidia (37.7%) or Vibrio (25.3%) were more common than two-pathogen coinfected individuals (9.5%), which may more easily succumb to infection. Fished C. edule populations with high cockle densities were more exposed to infections. Higher temperature and presence of epiflora on cockle shells promoted coinfection in warmer months. Low seawater salinity, host condition and proximity to other infected host species influenced coinfection distribution. A positive association between two Minchinia spp. was observed, most likely due to their different pathogenic effect. Findings highlight the major influence that ecological factors have on pathogen interactions and host­pathogen interplay.

Effect of light on the trematode Himasthla elongata: from cercarial behaviour to infection success.

The cockle Cerastoderma edule, a socioeconomically important bivalve of the northeast Atlantic, is host to several trematodes, including Himasthla elongata. In the life cycle of this trematode, cercariae (free-living stages) emerge from the first intermediate host, a snail, to infect cockles as second intermediate hosts. During their lifespan (less than 2 d), cercariae must ensure successful host-to-host transmission via the surrounding water and therefore are exposed to and impacted by different environmental conditions, including abiotic factors. Given that the light:dark cycle is one of the major drivers of behaviour in aquatic habitats, we aimed to determine the influence of light on cercariae and host behaviour based on 3 hypotheses. First, by having a benthic second intermediate host, these cercariae will display a photonegative orientation; second, and conversely, host behaviour will not be influenced by light; and third, cercariae infection success will be light-dependent. Results showed that cercariae display a photopositive orientation (first hypothesis rejected), displaying movements towards light. Host activity (evaluated by oxygen consumption) was similar among conditions, i.e. dark vs. light (second hypothesis accepted), but hosts acquired more parasites when experimentally infected in the dark (third hypothesis accepted). This light-dependent infection of the host is explained by a change of cercarial behaviour when exposed to light, decreasing their infection success. This study highlights that trematode responses to external conditions may be linked to successful life cycle completion rather than being altered by the host habitat. Light influence on cercarial behaviour resulted in increased infection success that may affect trematode population dynamics and their distributional range.

Monorchis parvus and Gymnophallus choledochus: two trematode species infecting cockles as first and second intermediate host.

The most deleterious stage of a trematode life cycle occurs in the first intermediate host where the parasite penetrates as a miracidium and asexually multiplicates in sporocysts or rediae. When infection advances, other organs can be occupied with severe effects on host individual health and population dynamics. Existing studies focused on these host/parasite systems are still scarce due to the usual low prevalence in ecosystems. Using cockles (Cerastoderma spp.) and two trematode species (Monorchis parvus and Gymnophallus choledochus) infecting these bivalves as first and second intermediate host, the present work aimed to (1) summarize the most relevant literature and (2) provide new information regarding this host/parasite system, taking advantage of a 21-year monthly database from Banc d'Arguin (France). This long-term monitoring showed that different trematode species display varying host size range preference (6-38 and 31-36 mm for M. parvus and G. choledochus, respectively). The occurrence of coinfection was lower than expected, raising some questions related to parasite interspecific competition. This review improved our understanding of the processes shaping the prevalence and distribution of parasitism. This study highlighted that beyond constant trematode assemblage monitoring, there is a need to identify the main predictors of rediae/sporocysts infection, such as the definitive host dynamics and miracidium infection processes, for future better management of host severe disease and mortality episodes.

Detection of haplosporidian protistan parasites supports an increase to their known diversity, geographic range and bivalve host specificity.

Haplosporidian protist parasites are a major concern for aquatic animal health, as they have been responsible for some of the most significant marine epizootics on record. Despite their impact on food security, aquaculture and ecosystem health, characterizing haplosporidian diversity, distributions and host range remains challenging. In this study, water filtering bivalve species, cockles Cerastoderma edule, mussels Mytilus spp. and Pacific oysters Crassostrea gigas, were screened using molecular genetic assays using deoxyribonucleic acid (DNA) markers for the Haplosporidia small subunit ribosomal deoxyribonucleic acid region. Two Haplosporidia species, both belonging to the Minchinia clade, were detected in C. edule and in the blue mussel Mytilus edulis in a new geographic range for the first time. No haplosporidians were detected in the C. gigas, Mediterranean mussel Mytilus galloprovincialis or Mytilus hybrids. These findings indicate that host selection and partitioning are occurring amongst cohabiting bivalve species. The detection of these Haplosporidia spp. raises questions as to whether they were always present, were introduced unintentionally via aquaculture and or shipping or were naturally introduced via water currents. These findings support an increase in the known diversity of a significant parasite group and highlight that parasite species may be present in marine environments but remain undetected, even in well-studied host species.