Search

Identifying marine trematode parasites in host tissue can be complicated when there is limited morphological differentiation between species infecting the same host species. This poses a challenge for regular surveys of the parasite communities in species of socio-economic and ecological importance. Our study focused on identifying digenean trematode species infecting the marine bivalve Cerastoderma edule across Europe by comparing morphological and molecular species identification methods. Cockles were sampled from ten locations to observe the trematode parasites under a stereomicroscope (morphological identification) and to isolate individuals for phylogenetic analyses using two gene markers, the small sub-unit ribosomal (18S) RNA gene (SSU rDNA) and the mitochondrial cytochrome c oxidase subunit 1 (cox1). For the first time, we compared both morphological identification and phylogenetic analyses for each of the 13 originally identified species. First, we identified a group of five species for which morphological identification matched molecular results ( Bucephalus minimus , Monorchis parvus , Renicola parvicaudatus , Psilostomum brevicolle , Himasthla interrupta ). Second, we identified a group of six species for which molecular results revealed either misidentifications or cryptic diversity ( Gymnophallus choledochus , Diphterostomum brusinae , Curtuteria arguinae , Himasthla quissetensis , H. elongata , H . continua ). Third, our analyses showed that all sequences of two expected species, Gymnophallus minutus and G. fossarum , matched between the two, strongly suggesting that only G. minutus is present in the studied area. Our study clearly demonstrates that molecular tools are necessary to validate the trematode species composition. However, with 17 distinct genetic lineages detected, some of which are not fully identified, future studies are needed to clarify the identity and status (regular vs. accidental infection) of some of these cryptic trematode species., Competing Interests: None., (© 2024 The Authors.)

The parasitic species Perkinsus olseni (=  atlanticus ) (Perkinsea, Alveolata) infects a wide range of mollusc species and is responsible for mortality events and economic losses in the aquaculture industry and fisheries worldwide. Thus far, most studies conducted in this field have approached the problem from a "one parasite-one disease" perspective, notably with regards to commercially relevant clam species, while the impact of other Perkinsus species should also be considered as it could play a key role in the disease phenotype and dynamics. Co-infection of P. olseni and P. chesapeaki has already been sporadically described in Manila clam populations in Europe. Here, we describe for the first time the parasitic distribution of two Perkinsus species, P. olseni and P. chesapeaki , in individual clam organs and in five different locations across Arcachon Bay (France), using simultaneous in situ detection by quantitative PCR (qPCR) duplex methodology. We show that P. olseni single-infection largely dominated prevalence (46-84%) with high intensities of infection (7.2 to 8.5 log-nb of copies. g -1 of wet tissue of Manila clam) depending on location, suggesting that infection is driven by the abiotic characteristics of stations and physiological states of the host. Conversely, single P. chesapeaki infections were observed in only two sampling stations, Ile aux Oiseaux and Gujan, with low prevalences 2 and 14%, respectively. Interestingly, the co-infection by both Perkinsus spp., ranging in prevalence from 12 to 34%, was distributed across four stations of Arcachon Bay, and was detected in one or two organs maximum. Within these co-infected organs, P. olseni largely dominated the global parasitic load. Hence, the co-infection dynamics between P. olseni and P. chesapeaki may rely on a facilitating role of P. olseni in developing a primary infection which in turn may help P. chesapeaki infect R. philippinarum as a reservoir for a preferred host. This ecological study demonstrates that the detection and quantification of both parasitic species, P. olseni and P. chesapeaki , is essential and timely in resolving cryptic infections and their consequences on individual hosts and clam populations., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Itoïz, Mouronvalle, Perennou, Chailler, Smits, Derelle, Metz, Le Goïc, Bidault, de Montaudouin, Arzul, Soudant and Chambouvet.)

Transmissible cancers are malignant cell lineages that spread clonally between individuals. Several such cancers, termed bivalve transmissible neoplasia (BTN), induce leukemia-like disease in marine bivalves. This is the case of BTN lineages affecting the common cockle, Cerastoderma edule, which inhabits the Atlantic coasts of Europe and northwest Africa. To investigate the evolution of cockle BTN, we collected 6,854 cockles, diagnosed 390 BTN tumors, generated a reference genome and assessed genomic variation across 61 tumors. Our analyses confirmed the existence of two BTN lineages with hemocytic origins. Mitochondrial variation revealed mitochondrial capture and host co-infection events. Mutational analyses identified lineage-specific signatures, one of which likely reflects DNA alkylation. Cytogenetic and copy number analyses uncovered pervasive genomic instability, with whole-genome duplication, oncogene amplification and alkylation-repair suppression as likely drivers. Satellite DNA distributions suggested ancient clonal origins. Our study illuminates long-term cancer evolution under the sea and reveals tolerance of extreme instability in neoplastic genomes., (© 2023. The Author(s), under exclusive licence to Springer Nature America, Inc.)

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.

The effect of sediment dredging and disposal work on benthic communities was compared according to two sampling strategies, grab (1-mm mesh) and dredge (1-cm mesh). Nine subtidal, sandy sites were selected in Arcachon Bay (French Atlantic coast), where these operations were performed during winter. Fauna was sampled several months before and one year after the dredging or disposal work. The sediments were fine to medium sands and their characteristics were not modified. In dredging sites, abundance, diversity and community structure of grab sampled fauna were minimally affected by the activities, while abundance and community structure displayed significant changes for dredge sampled fauna. In disposal sites, there was no work effect on fauna, although environmental conditions changed, especially when initial sediments were covered by mussels or seagrass. This study suggests that dredge sampling can be an alternative to grab sampling for monitoring changes related to dredging in sandy shallow channels., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

In recent field studies, suspected gymnophallid metacercariae were histologically located in the mantle of mussels from the Norwegian Sea. Mussels from the sites in which that infection was detected also presented abnormally high pearl numbers. It has been previously described that gymnophallid metacercariae could cause pearl formation processes in mussels, as a host reaction to encapsulate these metacercariae. Given the pathological host reaction these parasites elicit, a study was performed to identify gymnophallid metacercariae found in mussels collected from Tromsø at morphological and molecular level and to assess, by the use of molecular tools, the relationship between the parasite and the biological material inside the pearls. As a result, Gymnophallus bursicola metacercariae infecting Norwegian Mytilus edulis were identified according to morphological characters, along with the first 18S rDNA and COI sequences for this trematode species. In addition, parasite DNA from the core of the pearls was extracted and amplified for the first time, confirming the parasitological origin of these pearls. This procedure could allow identifying different parasitic organisms responsible for the generation of pearls in bivalves.

Infection by parasites with complex life cycles such as trematodes depends on many environmental factors which may result in a time-lag between host biomass fluctuations and parasite density in hosts. A cockle (marine bivalve, second intermediate host) population and its associated parasite community were monitored over 15 years. A time-shift correlation analysis suggests that trematode abundance in cockles responds to cockle biomass after a long delay (8 year time-lag). Thus, these parasites can sustainably support a deficit of their intermediate host., (Copyright © 2023 Australian Society for Parasitology. Published by Elsevier Ltd. All rights reserved.)

Parasite species richness is influenced by a range of drivers including host related factors (e.g. host size) and environmental factors (e.g. seawater temperature). However, identification of modulators of parasite species richness remains one of the great unanswered questions in ecology. The common cockle Cerastoderma edule is renowned for its diversity and abundance of parasites, yet drivers of parasite species richness in cockles have not been examined to investigate the association of both macro and microparasite communities. Using cockles as a model species, some of the key drivers of parasite prevalence and parasite species richness were investigated. Objectives of this 19-month survey were to determine the influence of the environment, host-parasite dynamics and parasite associations on parasite species richness and prevalence at two different geographic latitudes, chosen based on environmental differences. The highest parasite species richness was recorded in the northern sites, and this was potentially influenced by a range of interactions between the host, the pathogens and the environment. Parasite prevalence increased with host size and age, and parasite species richness increased with reduced salinity. A number of interactions between parasites, and between parasites and pathologies may be influencing parasite infection dynamics. New and concerning information is also presented regarding interactions between parasites and their environment. A number of parasites and potential pathogens (bacteria, Trichodina ciliates, metacercariae, trematode sporocysts) may be advantaged under climate change conditions (warming seas, increased precipitation), increasing disease incidence, which may prove detrimental not just for cockles, but for other bivalve species in the future., Competing Interests: The authors have declared that no competing interests exist.

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.

The aetiological agent Perkinsus olseni is globally recognised as a major threat for shellfish production considering its wide geographical distribution across Asia, Europe, Australia and South America. Another species, Perkinsus chesapeaki, which has never been known to be associated with significant mortality events, was recently detected along French coasts infecting clam populations sporadically in association with P. olseni. Identifying potential cryptic infections affecting Ruditapes philippinarum is essential to develop appropriate host resource management strategies. Here, we developed a molecular method based on duplex real-time quantitative PCR for the simultaneous detection of these two parasites, P. olseni and P. chesapeaki, in the different clam tissues: gills, digestive gland, foot, mantle, adductor muscle and the rest of the soft body. We firstly checked the presence of possible PCR inhibitors in host tissue samples. The qPCR reactions were inhibited depending on the nature of the host organ. The mantle and the rest of the soft body have a high inhibitory effect from threshold of host gDNA concentration of 2 ng.µL -1 , the adductor muscle and the foot have an intermediate inhibition of 5 ng.µL -1 , and the gills and digestive gland do not show any inhibition of the qPCR reaction even at the highest host gDNA concentration of 20 ng.µL -1 . Then, using the gills as a template, the suitability of the molecular technique was checked in comparison with the Ray's Fluid Thioglycolate Medium methodology recommended by the World Organisation for Animal Health. The duplex qPCR method brought new insights and unveiled cryptic infections as the co-occurrence of P. olseni and P. chesapeaki from in situ tissue samples in contrast to the RFTM diagnosis. The development of this duplex qPCR method is a fundamental work to monitor in situ co-infections that will lead to optimised resource management and conservation strategies to deal with emerging diseases., (Copyright © 2021. Published by Elsevier Inc.)

Reproduction and parasites have significant impacts on marine animal populations globally. This study aimed to investigate the associative effects of host reproduction and a host-parasite interplay on a marine bivalve, along a geographic gradient of latitude. Cockles Cerastoderma edule were sampled from five European sites (54°N to 40°N), between April 2018 and October 2019. A histological survey provided data on trematode (metacercaria and sporocyst life stages), prevalence, and cockle stage of gametogenesis to assess the influence of a latitudinal gradient on both interplays. Sex ratios at the northernmost sites were skewed toward females, and spawning size was reduced at the lower latitudes. Trematode infection did not follow a latitudinal gradient. Localized site-related drivers, namely seawater temperature, varied spatially, having an impact on cockle-trematode interactions. Spawning was related to elevated temperatures at all sites. Prolonged spawning occurred at southern latitudes, where seawater temperatures were warmer. Trematode prevalence and the impact of trematodes on gametogenesis were found to be spatially variable, but not latitudinally. Therefore, it is not possible to determine the likelihood of boom and bust events in cockles, based on the latitudinal location of a population. In terms of sublethal impacts, it appeared that energy was allocated to reproduction rather than somatic growth in southern populations, with less energy allocated to reproduction in the larger, northern cockles. The demonstrated spatial trend of energy allocation indicates the potential of a temporal trend of reduced cockle growth at northern sites, as a result of warming sea temperatures. This awareness of the spatially varying drivers of populations is crucial considering the potential for these drivers/inhibitors to be exacerbated in a changing marine environment., Competing Interests: None declared., (© 2021 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd.)

Bivalve stocks have been decreasing in the last decades largely due to emergent diseases and consequent mass mortality episodes. Cerastoderma edule (the edible cockle) is one of the most exploited bivalves in Europe and is among the most common hosts for trematodes, the most prevalent macroparasites in coastal waters but yet poorly studied. Therefore, in the present study, this bivalve species was used as host model to determine if trematode infection exerts a negative effect on bivalve energy metabolism and balance and if the tissues targeted by different trematodes influence the metabolic cost, with physiological and biochemical consequences. Cockles were experimentally infected with two trematode species, Himasthla elongata and Renicola roscovitus, that infect the foot and palps, respectively. Trematode infection exerted a negative effect on the metabolism of C. edule, the second intermediate host, by reduction of oxygen consumption. A different host biochemical response was found depending on trematode species, especially in regard to the level of oxygen consumption decrease and the preferential accumulation of lipids and glycogen. This study represents a step towards the understanding of host-trematode relationships that can be used to better predict potential conservation threats to bivalve populations and to maximize the success of stock and disease management., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Knowledge mobilisation is required to "bridge the gap" between research, policy and practice. This activity is dependent on the amount, richness and quality of the data published. To understand the impact of a changing climate on commercial species, stakeholder communities require better knowledge of their past and current situations. The common cockle (Cerastoderma edule) is an excellent model species for this type of analysis, as it is well-studied due to its cultural, commercial and ecological significance in west Europe. Recently, C. edule harvests have decreased, coinciding with frequent mass mortalities, due to factors such as a changing climate and diseases. In this study, macro and micro level marine historical ecology techniques were used to create datasets on topics including: cockle abundance, spawning duration and harvest levels, as well as the ecological factors impacting those cockle populations. These data were correlated with changing climate and the Atlantic Multidecadal Oscillation (AMO) index to assess if they are drivers of cockle abundance and harvesting. The analyses identified the key stakeholder communities involved in cockle research and data acquisition. It highlighted that data collection was sporadic and lacking in cross-national/stakeholder community coordination. A major finding was that local variability in cockle populations is influenced by biotic (parasites) and abiotic (temperature, legislation and harvesting) factors, and at a global scale by climate (AMO Index). This comprehensive study provided an insight into the European cockle fishery but also highlights the need to identify the type of data required, the importance of standardised monitoring, and dissemination efforts, taking into account the knowledge, source, and audience. These factors are key elements that will be highly beneficial not only to the cockle stakeholder communities but to other commercial species., Competing Interests: The authors have declared that no competing interests exist.

Although parasites represent a substantial part of marine communities' biomass and diversity, their influence on ecosystem functioning, especially via the modification of host behaviour, remains largely unknown. Here, we explored the effects of the bopyrid ectoparasite Gyge branchialis on the engineering activities of the thalassinid crustacean Upogebia pusilla and the cascading effects on intertidal ecosystem processes (e.g. sediment bioturbation) and functions (e.g. nutrient regeneration). Laboratory experiments revealed that the overall activity level of parasitized mud shrimp is reduced by a factor 3.3 due to a decrease in time allocated to burrowing and ventilating activities (by factors 1.9 and 2.9, respectively). Decrease in activity level led to strong reductions of bioturbation rates and biogeochemical fluxes at the sediment-water interface. Given the world-wide distribution of mud shrimp and their key role in biogeochemical processes, parasite-mediated alteration of their engineering behaviour has undoubtedly broad ecological impacts on marine coastal systems functioning. Our results illustrate further the need to consider host-parasite interactions (including trait-mediated indirect effects) when assessing the contribution of species to ecosystem properties, functions and services., (© 2020 British Ecological Society.)

The invasive species Spartina anglica arose in Europe by a cross between the Afro-European species S. maritima (native, paternal ancestor) and the introduced North American S. alterniflora (invasive, maternal ancestor). Aqueous methanolic extracts were prepared from plant tissue for chemotaxonomical comparison between the three species and determination of the phenolic pattern inheritance in S. anglica. A total of 20 phenolic compounds were detected in the aerial tissues of S. anglica and S. alterniflora, but only seven in S. maritima. They were isolated from their respective crude extracts, and their structures were determined according to spectroscopic data analysis and chemical evidence. They all belong to the flavonoid class, with 13 of them identified as C-glycoflavonoid and seven as O-glycoflavonoid. All these products were detected for the first time from S. anglica, fourteen of them for the first time from S. alterniflora, and three of them for the first time from S. maritima. The individual concentrations in the three species were determined by quantitative HPLC. The two parental species were found to differ markedly in their foliar phenolic fingerprint, whereas that of S. anglica showed a clear maternal dominance. Eight of the fourteen major compounds identified were of maternal origin among which, six were over-expressed, only three were from paternal origin but under-regulated, while two originated from the two parents. As far as we know, this work represents the first exhaustive report of the phenolic fingerprints of S. alterniflora and S. anglica and of the phenolic pattern inheritance in S. anglica. The similarity in the phenolic chemistry of the introduced and invasive S. alterniflora to its progeny could play a role in the physiological vigour and invasion success of S. anglica. This work provide a foundation for further studies, considering the reported biological activities of C-glycosidic flavonoids and tricin derivatives, and the lack of knowledge of the ecological chemistry of the genus Spartina., Competing Interests: Declaration of competing interest The authors declare that there is no conflict of interest., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Coastal habitats provide many important ecosystem services. The substantial role of shellfish in delivering ecosystem services is increasingly recognised, usually with a focus on cultured species, but wild-harvested bivalve species have largely been ignored. This study aimed to collate evidence and data to demonstrate the substantial role played by Europe's main wild-harvested bivalve species, the common cockle Cerastoderma edule, and to assess the ecosystem services that cockles provide. Data and information are synthesised from five countries along the Atlantic European coast with a long history of cockle fisheries. The cockle helps to modify habitat and support biodiversity, and plays a key role in the supporting services on which many of the other services depend. As well as providing food for people, cockles remove nitrogen, phosphorus and carbon from the marine environment, and have a strong cultural influence in these countries along the Atlantic coast. Preliminary economic valuation of some of these services in a European context is provided, and key knowledge gaps identified. It is concluded that the cockle has the potential to become (i) an important focus of conservation and improved sustainable management practices in coastal areas and communities, and (ii) a suitable model species to study the integration of cultural ecosystem services within the broader application of 'ecosystem services'., Competing Interests: Declaration of competing interest None., (Copyright © 2020 The Authors. Published by Elsevier Ltd.. All rights reserved.)

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.

Very few studies have characterized the concentrations of pollutants in bioturbating species. These species are considered as ecosystem engineers and characterizing stressors, such as contaminants, that impact them could lead to a better understanding of the functioning of ecosystems. In addition to contaminants, bioturbators are affected by a wide range of stressors, which can influence their physiological status and their ability to accumulate pollutants. Among these stressors, parasitism is of particular concern due to the ubiquity of parasites in natural environments and their influence on the fitness of their host. This study aims to assess the relationship between parasitism and metal accumulation in the bioturbating mud shrimp Upogebia cf. pusilla. A one-year seasonal survey was conducted in Arcachon Bay, France, with the aims of (1) characterizing the levels of metals in the mud shrimp and (2) evaluating the influence of two macroparasites (a bopyrid isopod and a trematode) on the variation of the metal content in mud shrimp. The bopyrid parasite castrates its female host and a particular attention has therefore been paid to the reproductive cycle of female mud shrimp by quantifying the expression of the vitellogenin gene that encodes the major yolk protein in female crustaceans. The levels of contaminants in mud shrimp appeared low compared to those reported in other crustaceans in areas of higher pollution. Even at these low contamination levels, we observed a significant impact by the bopyrid parasite that depends on season: bopyrid-infested organisms are generally more contaminated than their uninfested conspecifics except in summer when the opposite trend was observed. We suggest that the bopyrid indirectly interferes with the metal accumulation process by altering the reproductive capabilities of the mud shrimp. On the opposite, very low influence of the trematode parasite on the metal content of the host was found., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Recently, we reported an emerging pathology named Brown Muscle Disease (BMD) affecting Asari clams inhabiting the most productive area for this species in France, the Arcachon Bay. The main macroscopic feature of the pathology relies on the atrophy of the posterior adductor muscle, affecting the ability of clams to burry. The research of the etiological agent of BMD privileged a viral infection. Contrary to healthy clams, infected animals are always found at the surface of the sediment and exhibit 30 nm virus-like particles in muscle, granulocytic and rectal cells. In order to get more insights on the etiology and impacts of the BMD on clams, we took advantage in the present study of next generation sequencing technologies. An RNA-Seq approach was used (i) to test whether viral RNA sequences can be specifically found in the transcriptome of diseased animals and (ii) to identify the genes that are differentially regulated between diseased and healthy clams. Contrary to healthy buried animals, in diseased clams one sequence showing extensive homologies with retroviridae-related genes was detected. Among the biological processes that were affected in diseased clams, the synaptic transmission process was the most represented. To deepen this result, a new sampling was carried out and the transcription level of genes involved in synaptic transmission was determined in healthy and diseased clams but also in clams with no visible sign of pathology but located at the surface of the sediment. Our findings suggest that muscle atrophy is a latter sign of the pathology and that nervous system could be instead a primary target of the BMD agent., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Parasites are widespread in natural environments, and their impacts on the fitness of their host and, at a broader scale, on ecosystem functioning are well recognized. Over the last two decades, there has been an increasing interest in the effects of parasites in conjunction with other stressors, especially pollutants, on the health of organisms. For instance, parasites can interfere with the bioaccumulation process of contaminants in their host leading to parasitized organisms exhibiting lower pollutants burdens than unparasitized individuals for example. However, the mechanisms underlying these patterns are not well understood. This study examined how the bopyrid parasite Gyge branchialis could lower the cadmium (Cd) uptake of its mud shrimp host Upogebia cf. pusilla. When exposed to water-borne Cd, parasites were able to bioaccumulate this trace metal. However, the uptake of Cd by the parasite was low and cannot entirely explain the deficit of Cd contamination of the host. The weight of gills of parasitized organisms was significantly reduced compared with unparasitized organisms. We suggest that by reducing the surface for metal uptake, parasites could lower the contaminant burden of their host.

Anthropogenic activities, especially those involving substances that pollute the environment can interfere with bivalve populations, as well as parasitism, a fundamental ecological interaction often neglected. In marine environments, organisms are concomitantly exposed to pollutants and parasites, a combination with synergistic, antagonistic or additive effects representing a potential threat to aquatic communities sustainability. In the present study, Cerastoderma edule (the edible cockle)-Himasthla elongata (trematode) was used as host-parasite model. Cockles are worldwide recognized as good sentinel and bioindicator species and can be infected by several trematodes, the most abundant macroparasites in coastal waters. Tested hypotheses were: 1) cockles exposed to increasing parasite pressure will present greater stress response; 2) cockles exposure to arsenic (single concentration test: 5.2 μg L -1 ) will change parasite infection success and cockles stress response to infection. Arsenic was used for being one of the most common pollutants in the world and stress response assessed using biochemical markers of glycogen content, metabolism, antioxidant activity and cellular damage. Results showed that intensity of parasite pressure was positively correlated to biochemical response, mainly represented by higher metabolic requirements. Contamination did not affect parasite infection success. Compared to arsenic, trematode infection alone exerted a stronger impact: higher glycogen storage, metabolism and cellular damage and antioxidant activity inhibition. In interaction, parasitism and arsenic reduced hosts metabolism and cellular damage. Therefore, to a certain extent and in a contamination scenario, cockles may benefit from trematode infection, working as a protection for the pollutant accumulation in the organisms, reducing overall ROS production, which can consequently led to less toxic effects. These findings highlighted the deleterious effects of trematode infection in their hosts and showed the importance of including parasitology in ecotoxicological studies., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Mud shrimp are considered as among the most influential ecosystem engineers in marine soft bottom environments because of their significant bioturbation activity and their high density. These organisms play a key role on the physical structure of sediments through intense sediment reworking activity and also deeply influence geochemical properties of sediments via frequent bioirrigation events. The influence that mud shrimp have on the environment is related to the magnitude of bioturbation processes and subsequently depends on their physiological condition. In natural environments, several factors act together and influence the well-being of organisms. Among them, the deleterious role of parasites on the physiology and the behavior of their host is well established. Aquatic organisms are also subject to pollutants released by anthropogenic activities. However, the effect of both stressors on the fitness and bioturbation activity of mud shrimp has never been investigated yet. We conducted a 14-day ex-situ experiment to evaluate the influence of trace metal contamination (cadmium Cd) and parasitism infestation on the gene expression (molecular endpoint) and sediment reworking activity (behavioral endpoint) of the mud shrimp Upogebia cf. pusilla. At completion, mud shrimp exhibited substantial Cd bioaccumulation, with parasitized organisms showing a significantly lower contaminant burden than unparasitized specimens. Cadmium contamination induces modifications of gene expression in both unparasitized and parasitized organisms. We report an antagonistic effect of both stressors on gene expression, which cannot be fully explained by a lower Cd bioaccumulation. At the behaviour level, parasitism seems to reduce the sediment reworking activity of mud shrimp, while Cd contamination appears to stimulate this activity. This study highlights that the effects of multiple stressors may be quite different from the effects of each stressor considered individually. It should also motivate for more studies evaluating the influence of multiple stressors on different endpoints encompassing various levels of organization., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Resulting mainly from atmospheric carbon dioxide (CO 2 ) build-up, seawater temperature rise is among the most important climate change related factors affecting costal marine ecosystems. Global warming will have implications on the water cycle, increasing the risk of heavy rainfalls and consequent freshwater input into the oceans but also increasing the frequency of extreme drought periods with consequent salinity increase. For Europe, by the end of the century, projections describe an increase of CO 2 concentration up to 1120 ppm (corresponding to 0.5 pH unit decrease), an increase in the water temperature up to 4 °C and a higher frequency of heavy precipitation. These changes are likely to impact many biotic interactions, including host-parasite relationships which are particularly dependent on abiotic conditions. In the present study, we tested the hypothesis that the edible cockle, Cerastoderma edule, exposed to different salinity, temperature and pH levels as proxy for climate change, modify the infection success of the trematode parasite Himasthla elongata, with consequences to cockles biochemical performance. The results showed that the cercariae infection success increased with acidification but higher biochemical alterations were observed in infected cockles exposed to all abiotic experimental stressful conditions tested. The present study suggested that changes forecasted by many models may promote the proliferation of the parasites infective stages in many ecosystems leading to enhanced transmission, especially on temperate regions, that will influence the geographical distribution of some diseases and, probably, the survival capacity of infected bivalves., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Marine sediments are an important source of contaminants since they are susceptible to be remobilized to the water column. By modifying the physical and biogeochemical characteristics of sediments, bioturbation can influence contaminants remobilization. Within bioturbators, mud shrimp are considered as among the most influential organisms in marine soft-bottom environments. The physiological state of mud shrimp can be impaired by bopyrid parasites. The present study aims to evaluate the influence of bopyrid-uninfested and bopyrid-infested mud shrimp on sediment contaminants resuspension. Through a 14-days ex-situ experiment we showed a moderate effect of mud shrimp on contaminant remobilization compared to molecular diffusion without the bioturbator. Conversely, parasite presence significantly impaired contaminant bioaccumulation in mud shrimp and interfered with genetic expression. The weak effect of mud shrimp on contaminant remobilization may be due to trace metal accumulation and thus we suggested to evaluate the influence of contaminants on activities of bioturbating species., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Cerastoderma edule (edible cockle) is among the most exploited bivalves in Europe playing an important socio-economic role. Cockles live in estuaries and lagoons where their population is controlled by several environmental factors including parasitism. Parasites represent an important part of the world known biodiversity but are often neglected. Trematodes are the most prevalent macroparasites of cockles being able to exert an impact both at the individual and population levels. Therefore, it is of prime relevance to recognize and understand the parasite-host system dynamics in order to better predict potential conservation threats to bivalve populations and to maximize the success of stock and disease episodes management. Cockle monitoring was conducted in 2012 and 2016, in six and eight stations, respectively, at the Ria de Aveiro coastal lagoon, Portugal. Cockles were sampled in one single occasion in 2012 and seasonally in 2016. The tested hypothesis is that the trematode community in cockles was spatially and seasonally heterogeneous but stable over time. The main result showed that despite a relative homogeneity of the parasite community structure in cockles, the among-years heterogeneity of trematode communities was higher than among-stations and among-seasons heterogeneity rejecting the postulated hypothesis. Results demonstrated that trematode communities from the Ria de Aveiro are characterized by low abundance, which resulted in a spatial and seasonal trematode homogeneity (despite an overall channel difference and a slight downstream-upstream gradient). The interannual analysis showed a worrisome loss of trematode diversity and prevalence which consequently indicates an important loss of overall diversity and/or environmental conditions reflecting the negative effects of global change (mean temperature rise and overharvesting, among others). The present study highlighted the importance of trematodes in characterising their associated environment and respective biodiversity which might be helpful to assess ecosystem ecological status and to identify threatened areas., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Bivalve populations are controlled by several biotic and abiotic factors. Parasitism is among the biotic factors but is often neglected. In the present study, we focused on the transcriptomic and biochemical responses of Cerastoderma edule when parasitized as first intermediate host by the trematode Bucephalus minimus (sporocyst, the most damaging stage), and taking into account seasonal patterns. In order to test the hypothesis that the presence of B. minimus compromises cockle regular gene expression and biochemical performance and increases their vulnerability to other parasite species infection, cockles were sampled every other month during one year in Arcachon Bay (French Atlantic coast). Overall, results showed that B. minimus induced its first intermediate host defence mechanism against oxidative stress (mainly at gene level), increased host metabolism and energy demand especially in summer (revealed at both gene and biochemical level, although without significant differences) and was accompanied by a higher metacercariae abundance. Results allowed to accept the posted hypothesis and to conclude that transcriptomic and biochemical markers can provide additional and ecologically relevant information about parasite effects on their hosts, reflecting the invasion effects of pathogens but also the environmental conditions that animals experience., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Marine invertebrate microbiota has a key function in host physiology and health. To date, knowledge about bivalve microbiota is poorly documented except public health concerns. This study used a molecular approach to characterize the microbiota associated with the bivalve Manila clam (Ruditapes philippinarum) by determining (1) the difference among organs either or not under the influence of host habitat, (2) small-scale variability of microbiota, and (3) the experimental response of the Manila clam microbiota submitted to different lateral transmissions. These questions were investigated by sampling two groups of individuals living in contrasting habitats and carrying out a transplant experiment. Manila clam microbiota (i.e., bacterial community structure) was determined at organ-scale (gills, gut, and a pool of remaining tissues) by capillary electrophoresis DNA fingerprinting (CE fingerprinting). The Manila clam microbiota structure differed among organs indicating a selection of Manila clam microbiota at organ scale. Habitat strongly influenced gill and gut microbiota. In contrast, microbiota associated with remaining tissues was similar between group individuals suggesting that these communities are mostly autochthonous, i.e., Manila clam specific. Transplant experiment showed that improving living condition did not induce any change in microbiota associated with remaining tissues. In contrast, the reduction in individual habitat quality led to individuals in declining health as strongly suggested by the increase in phagocytosis activity and decrease in condition index together with the change in internal organ microbiota. This study provides a first description of the Manila clam holobiont which can withstand disturbance and respond opportunistically to improved environmental conditions.

Trematodes are the most prevalent and abundant macroparasites in coastal waters. They display a complex life cycle with alternation of free-living and parasitic stages generally involving three host species. The most deleterious stage is in the first intermediate host (a mollusc) where the parasite penetrates as miracidium larvae and asexually multiplicates in sporocysts/rediae to provide cercariae larvae. However, due to basic low prevalence in ecosystems, this system remains difficult to study. Taking the example of the cockle (Cerastoderma edule), an exploited bivalve along North-Eastern Atlantic coasts, and Bucephalus minimus, its most prevalent parasite as first intermediate host, we summarised the 51 most relevant papers (1887-2015). Besides, a 16-year monthly monitoring was performed at Banc d'Arguin (Atlantic coast of France), and allowed to obtain a sufficient number of infected cockles (276 out of 5,420 individuals) in order to provide new information concerning this parasite/host system. Sporocysts (diameter 80-500 μm) and developing cercariae (length 300-500 μm) are not visible before cockle reaches 16-mm shell length and then prevalence increases with host size. Seasonality of infection was not observed but variation of prevalence was significant among years and negatively correlated to the temperature of the former year, which could correspond to the period of infection by miracidium. Seven other species of trematode were identified in cockles as second intermediate host. For six of them, metacercariae abundance per individual was 2 to 12 folds higher in B. minimus-infected cockles, exacerbating the potential negative impact on host. From the parasite point of view, metacercariae can be considered as hitchhikers, taking advantage of the abnormal migration of B. minimus-infected cockles to the sediment surface where they become more vulnerable to predators that are also the final hosts of many of these parasites.

Wedge clams Donax trunculus inhabit high-energy environments along sandy coasts of the northeastern Atlantic Ocean and the Mediterranean Sea. Two sites were sampled monthly, one in Morocco (Mehdia), where the density was normal, and one in France (Biscarosse), where the density was very low. We tested the hypothesis that the difference in density between the sites was related to infection by the trematode parasite Bacciger bacciger. Identity of both the parasite and the host were verified using anatomical and molecular criteria. Parasite prevalence (i.e. the percentage of parasitized clams) was almost 3 times higher at Biscarosse. At this site, overall prevalence reached 32% in July and was correlated with the migration of several individuals (with a prevalence of 88%) to the sediment surface. After this peak, prevalence decreased rapidly, suggesting death of parasitized clams. The deleterious effect of B. bacciger on wedge clams was also supported by our calculations indicating that the weight of the parasite made up to 56% of the total weight of the parasitized clams. However, condition indices of trematode-free clams were also lower in Biscarosse than in Mehdia or other sites, suggesting that other factors such as pollutants or microparasites (Microcytos sp.) may alter wedge clam population fitness in Biscarosse.

Manila clam stock from Arcachon Bay, France, is declining, as is commercial harvest. To understand the role of environmental biotic interactions in this decrease, effects of a toxic dinoflagellate, Alexandrium ostenfeldii, which blooms regularly in Arcachon bay, and the interaction with perkinsosis on clam physiology were investigated. Manila clams from Arcachon Bay, with variable natural levels of perkinsosis, were exposed for seven days to a mix of the nutritious microalga T-Iso and the toxic dinoflagellate A. ostenfeldii, a producer of spirolides, followed by seven days of depuration fed only T-Iso. Following sacrifice and quantification of protozoan parasite Perkinsus olseni burden, clams were divided into two groups according to intensity of the infection ("Light-Moderate" and "Moderate-Heavy"). Hemocyte and plasma responses, digestive enzyme activities, antioxidant enzyme activities in gills, and histopathological responses were analyzed. Reactive oxygen species (ROS) production in hemocytes and catalase (CAT) activity in gills increased with P. olseni intensity of infection in control clams fed T-Iso, but did not vary among A. ostenfeldii-exposed clams. Exposure to A. ostenfeldii caused tissue alterations associated with an inflammatory response and modifications in hemocyte morphology. In the gills, superoxide dismutase (SOD) activity decreased, and an increase in brown cell occurrence was seen, suggesting oxidative stress. Observations of hemocytes and brown cells in tissues during exposure and depuration suggest involvement of both cell types in detoxication processes. Results suggest that exposure to A. ostenfeldii disrupted the pro-/anti-oxidant response of clams to heavy P. olseni intensity. In addition, depressed mitochondrial membrane potential (MMP) in hemocytes of clams exposed to A. ostenfeldii suggests that mitochondrial functions are regulated to maintain homeostasis of digestive enzyme activity and condition index., (Copyright © 2014 Elsevier B.V. All rights reserved.)

This study assessed the effect of Brown Muscle Disease (BMD) on Manila clam Venerupis philippinarum fitness. BMD was discovered in 2005. It affects the posterior adductor muscle and leads to clam gaping and eventually death. Three statuses of clams were compared: buried individuals with no signs of BMD (BUR); clams at the surface of the sediment with no signs of BMD (SURF) and clams at the surface of the sediment exhibiting signs of brown muscle disease (BMD). Physiological (condition index), immune (hemocyte parameters) and molecular (gene expressions) parameters collected seasonally were analyzed and compared. Results demonstrated a seasonal pattern in condition index (CI) with peaks in spring/summer and decreases in autumn/winter. At each season, the highest CI was observed in BUR and the lowest CI was observed in BMD. In terms of immune response, phagocytosis rate and capacity were higher in clams with BMD whereas the health status of the clams did not influence the total hemocyte count. Genes involved in the immune system (comp, tnf, inter) were upregulated in clams with BMD. The molecular analysis of gill and posterior muscle showed higher mitochondrial metabolism (cox-1, 16S) in cells of infected clams, suggesting a stronger energetic demand by these cells. Finally, genes involved in oxidative stress response (cat, sod), detoxification (mt) and DNA repair (gadd45) were also overexpressed due to reactive oxygen species production. Most of the studied parameters underlined a cause-effect correlation between Manila clam health status (BUR, SUR, BMD) and physiological parameters. An important stress response was observed in BMD-infected clams at different scales, i.e. condition index, immune parameters and stress-related gene expression., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Perkinsosis is one of the most widespread diseases affecting commercially important species of molluscs globally. We examined the impact of Perkinsus spp. on shell growth at the individual scale in 2 clam species: Ruditapes decussatus from Mundaka Estuary (Spain) and R. philippinarum from Arcachon Bay (France). At Arcachon, 2 contrasting sites in terms of environment and Perkinsus olseni presence were chosen: Arguin (disease-free) and Ile aux Oiseaux (infected site). We monitored the dynamics of perkinsosis over the course of the experiment at Mundaka and Ile aux Oiseaux. Prevalences were high (>70%), and intensities were around 105 cells g-1 wet gills at Ile aux Oiseaux, and 106 cells g-1 at Mundaka. No significant differences in prevalence or intensity were observed over time. A 2 yr field growth experiment of tagged-recaptured clams was performed to determine individual clam growth rate, condition index (CI), and Perkinsus spp. infection intensity. Clams were collected at Ile aux Oiseaux and transplanted to Arguin. The growth rate was always significantly and negatively correlated with Perkinsus spp. infection, and positively correlated with CI. CI and Perkinsus spp. infection explained 19% and 7% of the variability of the growth rate at Mundaka and Ile aux Oiseaux, respectively. In experimental clams at Arguin, P. olseni infection explained 26% of the variability of the growth rate at the lower tidal level. Our results suggest that at a concentration of between 105 and 106 cells g-1, perkinsosis affects the physiological functions of the clams, highlighted by its impact on the growth rate.

In 2005, dredging activities in Arcachon Bay (France) led in burying 320,000 m(2) of Zostera noltii intertidal seagrass. Recovery by macrobenthos and seagrass was monitored. Six months after works, seagrass was absent and macrobenthos drastically different from surrounding vegetated stations. Rapidly and due to sediment dispersal, disposal area was divided into a sandflat with a specific benthic community which maintained its difference until the end of the survey (2010), and a mudflat where associated fauna became similar to those in adjacent seagrass. Macrobenthic community needs 3 years to recover while seagrass needs 5 years to recover in the station impacted by mud. The secondary production loss due to works was low. In this naturally carbon enriched system, univariate biotic indices did not perform well to detect seagrass destruction and recovery. Multivariate index MISS gave more relevant conclusions and a simplified version was tested with success, at this local scale., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Metallothionein (MT) genes encode crucial metal-binding proteins ubiquitously expressed in living organisms and which play important roles in homeostasis of essential metals and detoxification processes. Here, the molecular organization of the first metallothionein gene of the edible cockle Cerastoderma edule and its expression after cadmium (Cd) or mercury (Hg) exposures were determined. The resulting sequence (Cemt1) exhibits unusual features. The full length cDNA encodes a protein of 73 amino acids with nine classical Cys-X((1-3))-Cys motifs, but also one Cys-Cys not generally found in molluscan MT. Moreover, characterization of the molecular organization of the Cemt1 gene revealed two different alleles (A1 and A2) with length differences due to large deletion events in their intronic sequences involving direct Short Interspersed repeated Elements (SINE), while their exonic sequences were identical. To our knowledge, such large excision mechanisms have never before been reported in a bivalve gene sequence. After 10 days of Cd exposure at environmentally relevant doses, quantitative real-time PCR revealed a strong induction of Cemt1 in gills of C. edule. Surprisingly, neither induction of the Cemt1 gene nor of MT protein was shown after Hg exposure, despite the fact that this organism is able to bioaccumulate a high amount of this trace metal which is theoretically one of the most powerful inducers of MT biosynthesis., (Copyright © 2011. Published by Elsevier Inc.)

The bacterial communities associated with the cockle (Cerastoderma edule) were investigated at the individual level through a 10-month monitoring programme. Temporal changes and those changes associated with a common parasite of the cockle, Bucephalus minimus, were investigated by monthly sampling of individuals, selected based on their shell length (cohort monitoring). Cockle bacterial community abundance (CBCA) and diversity (CBCD) were estimated by epifluorescence microscopy counts and automated ribosomal intergenic spacer analysis, respectively. CBCA showed a temporal pattern peaking at 30 × 10(6) cells per gram of cockle flesh and intervalval liquid in October and a significant 1.8-fold increase linked with B. minimus occurrence. CBCD was characterized by 112 ± 26 intergenic transcribed spacer (ITS) per individual and showed a relative homology between individuals (52 ± 6%, Jaccard similarity) in spite of more than 30% of rare ITS. Consistent with an undisturbed evolution of the condition index of the studied cohort individuals as an estimate of their physiological state, neither temporal nor parasite-induced change in CBCA has been related to marked changes in CBCD.

In natural environment, marine organisms are concomitantly exposed to pollutants and multiple disease agents resulting in detrimental interactions. The present study evaluated interactive effects of metal contamination (cadmium) and pathogenic organisms (trematode parasites Himasthla elongata and pathogenic bacteria Vibrio tapetis) singularly and in combination on the bivalve Ruditapes philippinarum, an introduced species to Europe, under laboratory controlled conditions. After 7 days, metal bioaccumulation and pathogen load were analyzed as well as metallothionein (MT) response and hemocyte concentrations and activities. Results showed that infection by opportunistic pathogens affects metal accumulation, leading to maximal Cd accumulation in co-infected clams. Among stressors only V. tapetis induced significant effects on immune parameters whereas a particular interaction "trematode-bacteria" was shown on MT responses. Despite low trematode infection in agreement with the resistant status of R. philippinarum to these macroparasites, significant interaction with bacteria and metal occurred. Such results highlight the necessity of taking pathogens into account in ecotoxicological studies., (Copyright © 2010. Published by Elsevier Ltd.)

Pathogens belonging to the genus Perkinsus infect many bivalve molluscan species around the world, including the Manila clam Ruditapes philippinarum. We investigated the spatial distribution of this parasite at 34 stations throughout Arcachon Bay (SW France). Prevalence of perkinsosis was 93% and mean infection abundance was 96 x 10(3) cells g(-1) wet gill. Lowest mean abundances were found close to the Leyre River mouth and a significant negative correlation was observed between mean abundance and salinity. Perkinsosis was rare at the oceanic site where salinities and other environmental parameters were stable. A second aim of this study was to survey perkinsosis during annual cycles at 4 sites within Arcachon Bay. Prevalence and intensities (+/- SE) of the disease were high, on average between 70 and 100%, and 130 x 10(3) +/- 6.7 x 10(3) cells g(-1) wet gill. No seasonal cycle was evident. Clams were infected at 9 mm shell length and infection increased with clam size. The third objective was to determine the disinfection and infection kinetics through a 21 mo reciprocal transplantation between a nearly Perkinsus sp.-free area and a highly affected site. Disinfection appeared to be a very slow process and was similar at the site with favorable conditions for Perkinsus sp. as at the site with unfavorable conditions. Conversely, infection acquisition appeared to be episodic with spatially defined areas. Consequently, the overall lack of a clear seasonal infection pattern is interpreted as the combination of episodic infection events and slow disinfection kinetics.

This study investigated some stress (metals, parasites) and response (immunity, metallothionein) factors in two cockle and two Manila clam populations. Data from eight seasons were averaged to obtain global baseline values. Stress/response characteristics of each population were compared to population health status that was determined through population dynamics parameters. Four different scenarios were discussed: (1) a lightly stressed cockle population with correct population health but with a risk of deterioration (hot spot); (2) a lightly stressed introduced cockle population threatened of extinction. In this case ecological factors were suspected; (3) a moderately stressed clam population with moderate adaptative response. The population was sustainable but the level of stress should not increase (hotspot); and (4) a stressed clam population and unfavourable ecological conditions preventing clam settlement. This monitoring highlighted that the discrepancy between population health and stress levels could be due to insufficient response by bivalves and/or by unfavourable ecological factors., (Copyright 2010 Elsevier Ltd. All rights reserved.)

The present report presents the first evidence of disseminated neoplasia (DN) in cockles Cerastoderma edule from Arcachon Bay (France). Aneuploidy of neoplastic cells allowed the use of flow cytometry (FCM) to diagnose and stage DN. A 1year survey (2007) of the prevalence and intensity (% of aneuploid circulating cells in neoplastic cockles) was conducted. Prevalences ranged from 2.2% (June) to 13.6% (May), and disease intensity ranged from 18.7% (June) to 95.5% (September). These percentages were not correlated with seawater temperature, but rather showed unexplained oscillations over the year. Prevalence and intensity of DN were higher in cockles found at the surface of sediment compared to those buried normally (11.8% vs. 6.7% and 53.0% vs. 40.6%, respectively, p<0.05). DN could thus be one mechanism leading to unexplained presence of cockles at the surface of the sediment in Arcachon Bay. Ploidy characteristics of neoplastic cells were also investigated using FCM, revealing an unusual, broad continuum of ploidy distribution from 1.6 to 9.6n. Ploidy values were not in whole numbers in contrast to the rounded values reported in other studies. Ploidy varied according to DN intensity, with the ploidy distribution of neoplastic cells from lightly-diseased cockles being unimodal (3.7n median). In contrast, highly-diseased cockles showed a bimodal ploidy distribution (3.0n and 4.7n medians). This suggests that, in cockles from Arcachon Bay, mechanisms leading to aneuploidy are complex, developing during disease progression.

Metallothioneins (MT) are essential metal binding proteins involved in metal homeostasis and detoxification in living organisms. Numerous studies have focused on MT response to metal exposure and showed an important variability according to species, metal, concentration and time of exposure. In this study, the expression of one isoform of MT gene (Cemt1) and associated MT protein synthesis were determined after 1, 3, 9, 24, 72 and 168h of cadmium (Cd) or mercury (Hg) exposures in gills of the cockle Cerastoderma edule. This experiment, carried out in laboratory conditions, revealed that in Cd-exposed cockles, induction of Cemt1 is time-dependent following a "pulse-scheme" with significant upregulation at 24h and 168h intersected by time point (72h) with significant downregulation. MT protein concentration increases with time in gills of exposed cockles in relation with the progressive accumulation of Cd in soluble fraction. On contrary, Hg exposure does not lead to any induction of Cemt1 mRNA expression or MT protein synthesis compared to control, despite a higher accumulation of this metal in gills of cockles compared to Cd. The localization of Hg (85-90%) is in insoluble fraction, whereas MT was located in the cytoplasm of cells. This gives us a first clue to understand the inability of Hg to activate MT synthesis. However, other biochemical processes probably occur in gills of C. edule since the remaining soluble fraction of Hg exceeds MT sequestration ability. Finally, since one of the first main targets of metal toxicity in cells was the mitochondria, some genes involved in mitochondria metabolism were also analyzed in order to assess potential differences in cellular damages between two metal exposures. Indeed, until T(168), no impact on mitochondrial genes was shown following Hg exposure, despite the complete lack of MT response. This result indicated the presence of other effective cellular ligands which sequester the cytosolic fraction of this metal and consequently inhibit metal reactivity. Such competition mechanisms with other cytosolic ligands more sensitive to Hg were particularly argued in the discussion.

Introduction: Within the last decade, numerous studies have investigated the role of environmental history on tolerance to stress of many organisms. This study aims to assess if Manila clams Ruditapes philippinarum may react differently to cadmium exposure and trematode parasite infection (Himasthla elongata) depending on their origin and environmental history in Arcachon Bay (France)., Materials and Methods: Clams were exposed to Cd (15 microg L(-1)) and parasites (25 cercariae per clam), alone or in combination, at 15 degrees C under controlled laboratory conditions for 7 days. Metal accumulation and success of parasite infestation were examined, also physiological parameters such as metallothionein response and hemocyte counts and activities (phagocytosis, oxidative burst, viability, and adhesion)., Results and Discussion: Sensitivity of Manila clams to both stressors differed from one site to another, suggesting local adaptation of populations. Clams from the more parasitized site presented better resistance to trematodes than the others in terms of first line defense, i.e., avoidance of infection. On the other hand, clams that adapted to chronic Cd contamination showed better detoxification mechanisms, both in a faster transfer of metal from gills to visceral mass and in a higher metallothionein baseline, than clams which had never experienced Cd contamination. Finally, hemocyte concentration and viability differed between clam origin site, highlighting the fact that populations living in different environments may adapt their physiological and biochemical responses to environmental stressors., Conclusion: It is therefore important to be cautious when extrapolating results from field studies of one species and one site, if the life history of the organisms is not taken into account.

The present study evaluated the interactive effects of cadmium contamination and pathogenic organisms (trematodes Himasthla elongata and bacteria Vibrio tapetis) singularly and in combination during 7 days on the bivalve Cerastoderma edule. Some defense-related activities were analyzed such as genetic expression, metallothionein and immune responses. Trematode metacercarial infection, similar whatever the treatment, induced the strongest responses of immune parameters. Particularly, the interaction between cadmium and parasite exposures induced unusual responses on gene expression and immune responses. No effect of bacterial challenge appeared on bivalve responses, nevertheless a strong mortality of V. tapetis infected cockles occurred between 7 and 14 days. Cadmium bioaccumulation was significantly modulated by both pathogenic organisms. Furthermore, an antagonistic effect of trematodes and bacteria was shown on metal bioaccumulation of co-infected cockles. These results highlighted the importance of considering the multiplicity of perturbation sources in coastal ecosystems to assess the health status of organisms., ((c) 2009 Elsevier Ltd. All rights reserved.)

Recently, Manila clam, Ruditapes philippinarum, populations have suffered mortalities in Arcachon Bay (SW France). Mortality was associated with extensive lesions of the posterior adductor muscle, which become progressively brown and calcified. Ultrastructural observations by transmission electron microscopy revealed tissue degradation with necrotized muscle fibres and granulocytomas. Unenveloped virus-like particles (VLPs) were detected in muscle, granulocytic, epithelial and rectal cells. VLPs were abundant in the extracellular space, in the cytoplasm (free or enclosed in vesicles) and in the nucleoplasm of granulocytes. Nuclei and mitochondria of granulocytes displayed changes which suggested reactive oxygen species production and apoptosis induction. VLPs exhibited an icosahedral structure with a diameter of 25 to 35 nm. These observations suggest that the VLPs could belong to the family Picornaviridae or the Parvoviridae.

The similarity in species composition between two communities generally decays as a function of increasing distance between them. Parasite communities in vertebrate definitive hosts follow this pattern but the respective relationship in intermediate invertebrate hosts of parasites with complex life cycles is unknown. In intermediate hosts, parasite communities are affected not only by the varying vagility of their definitive hosts (dispersing infective propagules) but also by the necessary coincidence of all their hosts in environmentally suitable localities. As intermediate hosts often hardly move they do not contribute to parasite dispersal. Hence, their parasite assemblages may decrease faster in similarity with increasing distance than those in highly mobile vertebrate definitive hosts. We use published field survey data to investigate distance decay of similarity in trematode communities from three prominent coastal molluscs of the Eastern North-Atlantic: the gastropods Littorina littorea and Hydrobia ulvae, and the bivalve Cerastoderma edule. We found that the similarity of trematode communities in all three hosts decayed with distance, independently of local sampling effort, and whether or not the parasites used the mollusc as first or second intermediate host in their life cycle. In H. ulvae, the halving distance (i.e. the distance that halves the similarity from its initial similarity at 1 km distance) for the trematode species using birds as definitive hosts was approximately two to three times larger than for species using fish. The initial similarities (estimated at 1 km distance) among trematode communities were relatively higher, whereas mean halving distances were lower, compared to published values for parasite communities in vertebrate hosts. We conclude that the vagility of definitive hosts accounts for a high similarity at the local scale, while the strong decay of similarity across regions is a consequence of the low probability that all necessary hosts and suitable environmental conditions coincide on a large scale.

The stable carbon and nitrogen isotopic composition of digenean trematode parasites and their marine mollusc hosts was investigated to describe the potential influence of parasites on their host and its different tissues, and to obtain further insight into their trophic relationships. Four parasite-host systems were studied: Labratrema minimus-Cerastoderma edule, Monorchis parvus-C. edule, Lepocreadiidae parasites-Nassarius reticulatus and Zoogonidae parasites-N. reticulatus. Among the 4 sampling occasions reported here and corresponding to the 4 parasite-host systems, isotopic shifts from pathologic (i.e. linked to disturbances in host metabolism) and mass-balance (i.e. linked to significant differences between host and parasite isotopic signatures) origins were observed only once. Both corresponded to delta 13C measurements of the L. minimus-C. edule system when the infestation load (percentage parasite dry weight compared to total flesh dry weight) was highest (9 to 25%, mean = 16%) over the sampling period. Overall, measurements indicate that digenean trematode parasitism induced low or no shifts in isotopic signatures of C. edule and N. reticulatus tissues. The 2 endoparasites L. minimus and M. parvus appeared to be slightly depleted in 13C compared to C. edule digestive gland and gonads, which were the most parasitized tissues. In contrast, no fractionation or low 15N trophic enrichments occurred in the parasites. These results highly contrast with the classical trophic enrichment reported in prey-predator systems but are in agreement with the scarce literature regarding other parasite-host systems. Our results indicate that (1) digenean trematodes mainly feed on digestive glands (the cockle tissue with which they are mainly associated) with a possible slight preference for lipids, and (2) fractionation due to parasite metabolism should be low due to abbreviated metabolic pathways and/or slight loss of materials through excretion, tegument diffusion and respiration.

We describe an emerging pathology, brown muscle disease (BMD), which specifically affects the Manila clam Ruditapes philippinarum in Arcachon Bay (France). BMD induces a transformation of the posterior adductor muscle, which becomes infused by conchiolin and calcified, reducing the ability of clams to bury. The disease affects both types of muscular tissue, with striated muscle becoming affected to a higher degree than smooth muscle. Two indices were created to quantify the symptoms: the Muscle Print Index, used for empty and live shells, and the Final Disease Index, utilized for live clams only. Histological sections were made and observed under light microscopy to examine the muscular damage and to investigate a causal agent. Sections revealed an important inflammatory response with a large invasion of hemocytes into tissues and a heavy necrosis of muscular fibers. Additionally, molecular biology analyses were carried out to search for bacteria and protozoan agents using generic primers. In both histological and molecular assays, bacteria and protozoans were discounted. We monitored 4 sites scattered around the bay over 2 yr. The mean prevalence was <12% without seasonal variation in 3 sites against 30% and a winter peak in 1 site. The latter site was accurately surveyed and revealed that clams at the sediment surface (abnormal position) were affected 3 times more frequently than buried clams (normal position).

The aim of the present study was to analyse the relation between parasitism and subsequent metallothioneins (MT) in the case of metal contamination. Experimental exposure of parasitized and unparasitized cockles (Cerastoderma edule) to cadmium (Cd) was performed, with the cockle as first or second intermediate host of 2 digenean species. After 7 days of Cd exposure in microcosms, cockles infected as first intermediate host by Labratrema minimus exhibited metal concentrations in tissues double that in uninfected cockles. Jointly, MT concentrations of parasitized cockles were not modified in comparison with uninfected individuals in which concentrations were increased 4.3-fold compared with controls. In cockles experimentally infected as the second intermediate host by Himasthla elongata, cadmium concentrations significantly increased again in parasitized cockles compared with uninfected individuals in contaminated conditions. Simultaneously, MT concentrations in healthy cockles increased, whereas they significantly decreased in parasitized individuals. Therefore, the presence of digenean parasites in Cd-exposed cockles leads to a maintenance or a decrease in MT concentrations compared with healthy individuals, whereas Cd accumulation in tissues is significantly increased. These experiments indicate a significant alteration of the protective effect of metallothioneins towards metals which could consequently enhance cockle vulnerability. Moreover, these results highlight the limit of the use of MT as a biomarker of metal pollution in field monitoring if parasitism is not taken into account.

A new Himasthlinae species, Curtuteria arguinae, is described as metacercariae from the cockle Cerastoderma edule (L.), collected at Banc d'Arguin (southwestern France). These metacercariae encysted preferentially in the mantle and also in the foot of cockles. Encysted and chemically excysted metacercariae were studied by light microscopy and scanning electron microscopy, respectively. Excysted metacercariae were elongated and curved ventrally. They bore a 33-spine circumoral collar. Sensory papillae were arranged around the oral sucker and also symmetrically along the ventral surface body, from the collar to the acetabulum. The dorsal and ventral tegument surfaces were densely packed with similar pointed spines. The posterior end of the body was without any spines. Among the Curtuteria species described previously, only Curtuteria haematopodis Smogorjewskaja and Iskova, 1966 had the same number of circumoral collar spines. A 6-yr field survey showed that the cockle population at Banc d'Arguin was subjected to a summer infection of C. arguinae. Curtuteria arguinae phenology of infection is characterized by interannual variability and seasonality (beginning in July-August and maximum in autumn). The first intermediate and final hosts remain unknown.

Metallothioneins (MT) are proteins that play an important role in metabolism of essential metals and detoxification of trace metals from living organisms. Their synthesis is induced by metal pollution but can also be exacerbated by other factors such as reproduction processes. In this context, we monitored MT concentrations in a cockle Cerastoderma edule (marine bivalve) population and highlighted the effect of a castrating digenean parasite, Labratrema minimus. In spent cockles, MT levels were low (ca. 5 nmol sites g(-1), fresh weight) but slightly higher in parasitized individuals. During gametogenesis, MT synthesis increased in all cockles, but concentrations were lower in parasitized individuals (18 against 27 nmol sites g(-1), fw in unparasitized cockles) in relation with gonad damage by parasites. Therefore, it is suggested that parasite infection in cockles can modulate MT synthesis that could consequently interfere with the response of these protective proteins in case of metal contamination.