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.

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.

Modelling the dynamics of an estuary and the evolution of its morphology requires a process-based description not only of the physical processes, but also of the influence of benthic fauna on sediment characteristics at ecosystem scale. A meta-analysis was tested as an approach for modelling the effect of bioturbation exerted by the cockle Cerastoderma edule on sediment erodibility. Six different erosion flume datasets were collected to ensure a broad range of experimental conditions including bed shear stress, population characteristics, and sediment composition. First, a model was built to describe the biogenic fluff layer created by C. edule activity in relation to (i) bioturbation activity using the population metabolic rate [mW·m-2] as a proxy for faunal metabolic energy, and (ii) the silt content [%] of the sediment. Second, different erosion models were compared by testing parameterization steps incorporating both erosion of the fluff layer and/or mass erosion of the sediment bed. Structural differences in the flumes and in the preparation of samples in the six different datasets makes it difficult to propose a single model that satisfactorily simulates all the data and encompasses both types of subsequent erosion, that of the fluff layer and that of the underlying consolidated bed. However, a generic model is proposed for the surficial fluff layer erosion covering a moderate range of bed shear stress (<1 Pa). This study shows that including several datasets covering a wide range of environmental conditions is a key to the robustness of this model, and that new insights can be gained by integrating the complexity of sediment features. We expect that this two-part model can be used in broad contexts in terms of cockle populations, estuarine habitats, and climatic conditions and can combined with various hydro-morpho-sedimentary models that include these biological effects.

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.

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.

As filter feeders, marine bivalves inhabiting estuarine and coastal areas are directly exposed to microplastics (MPs) in water. To assess whether MPs number, and their shape, size, colour, and polymer type present in mussels (Mytilus galloprovincialis) and cockles (Cerastoderma edule) varied over one year, bivalves were collected over the year of 2019 in the lower part of the coastal Aveiro lagoon, Portugal. After extraction from the bivalve\'s whole-body soft tissues, a subset of the visually inspected particles was randomly separated for identification using the Fourier-transform mid-infrared (FT-MIR) spectroscopy. A fraction of the inspected particles, 26-32% of particles >100 μm, and 59-100% of smaller ones were confirmed as MPs. Concentrations varied within the intervals of 0.77-4.3 items g-1 in mussels and 0.83-5.1 items g-1 in cockles, with the lowest values observed in January. In winter, the accumulation of large-sized fibers was composed of a mixture of plastic types, which contrasted against the most abundant MPs in summer consisting mainly of polyethylene of diverse size classes and shapes. Temperature decrease registered in winter might have triggered a lower filtration rate, resulting in lower MPs concentrations in the whole-soft body tissues of organisms. Different properties of MPs found in bivalves between January-February and August-September appear to reflect changes in the characteristics of MPs available in the Aveiro lagoon.

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.

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 recurrent emergence of infection outbreaks associated with shellfish consumption is of extreme importance for public health. The present study investigated the potential application of phages AH-1, AH-4, and AH-5 to inactivate Aeromonas hydrophila, a causative agent of infections in humans associated with bivalve shellfish consumption. The inactivation of A. hydrophila was assessed in vitro, using a liquid culture medium, and in vivo, using artificially contaminated cockles with A. hydrophila ATCC 7966. In the in vitro experiments, all phages were effective against A. hydrophila, but phage AH-1 (with a maximum reduction of 7.7 log colonies forming units CFU/mL) was more effective than phages AH-4 and AH-5 (with reductions of 4.9 and 4.5 log CFU/mL, respectively). The cocktails AH-1/AH-4, AH-1/AH-5, AH-4/AH-5, and AH-1/AH-4/AH-5 were slightly more effective than the single phage suspensions. The phages presented a low emergence rate of phage-resistant mutants. When artificially contaminated cockles were treated in static seawater with phage AH-1, around 44% of the added A. hydrophila (1.0 log CFU/g) was inactivated. The results of this study suggest that phage therapy can be an effective alternative to control human pathogenic bacteria during depuration.

Benthic organisms, in particular bioturbators, can influence erosion processes either by affecting sediment roughness through their mere presence and/or activities, or by modulating sediment characteristics (e.g., silt content, granulometry), thus altering its erodibility. To date, it was not possible to distinguish the influence of bioturbating species on sediment roughness from their impact on sediment erodibility. Consequently, uncertainties remain regarding the role played by benthic species on sediment dynamics. In this study, we used a canal flume which allows to record the bed shear stress at the surface of a non-cohesive sediment (4% of mud) during erosion experiments, thus allowing to disentangle the influence of bioturbators, here the common cockle Cerastoderma edule, on the two erosion mechanisms. In order to assess the influence of bioturbators on sediment stability in different environmental situations, we additionally tested for the effects of three factors, i.e. bivalve density, availability of suspended food (i.e. phytoplankton presence) and microphytobenthos (MPB) occurrence, which may modulate the behavior of cockles. We observed that cockles promote the erosion of the sediment surficial layer by increasing its roughness as a consequence of their sediment reworking activity and/or presence at the sediment surface (emerging shell). In contrast, we calculated similar critical bed shear stress for erosion with and without bivalves suggesting that cockles have a minor influence on the erodibility of non-cohesive substrates with a low silt content. The destabilizing effect of cockles increased with the bivalve density whereas it was attenuated by the presence of phytoplankton. We hypothesize that the magnitude of cockles\' bioturbation activity was lower when a high proportion of suspended food is available. High concentrations of suspended food may also have enhanced the filtration and biodeposition rates of cockles, thus rapidly leading to the \'muddification\' of the sediment bed and consequently counteracting with the own destabilizing effect of the bivalves. Finally, the sole presence of MPB did not significantly affect the resuspension dynamics of non-cohesive sediments with a low proportion of mud.