Search

Objectives: The two oyster species studied hold considerable economic importance for artisanal harvest (Crassostrea rhizophorae) and aquaculture (Crassostrea gasar). Their draft genomes will play an important role in the application of genomic methods such as RNAseq, population-based genomic scans aiming at addressing expression responses to pollution stress, adaptation to salinity and temperature variation, and will also permit investigating the genetic bases and enable marker-assisted selection of economically important traits like shell and mantle coloration and resistance to temperature and disease., Data Description: The draft assembly size of Crassostrea gasar is 506 Mbp, and of Crassostrea rhizophorae is 584 Mbp with scaffolds N50 of 11,3 Mbp and 4,9 Mbp, respectively. The general masked bases by RepeatMasker in both genomes were highly similar using different datasets. The masked bases varied from 9.41% in C. gasar to 10.05% in C. rhizophorae and 42.85% in C. gasar to 44.44% in C. rhizophorae using Dfam and RepeatModeler datasets, respectively. Functional annotation with eggNog resulted in 34,693 annotated proteins in C. rhizophorae and 26,328 in C. gasar. BUSCO analysis shows that almost 99% of genes (5,295) are complete in relation to the mollusk orthologous genes dataset (mollusca_odb10)., (© 2024. The Author(s).)

Microplastic ingestion by marine organisms presents a challenge to both ecosystem functioning and human health. We characterized microplastic abundance, shape, size, and polymer types ingested by the West African mangrove oyster, Crassostrea tulipa (Lamarck, 1819) sampled from estuaries and lagoons from the Gambia, Sierra Leone, Ghana, Benin, and Nigeria using optical microscopy and Fourier transform infrared (FTIR) techniques. A total of 780 microplastics were isolated in the whole tissues of the 250 oysters (n = 50 oysters per country). The abundance and distribution of microplastics in the oysters followed the pattern: the Gambia > Ghana > Sierra Leone > Nigeria > Benin. The Tanbi wetlands in the Gambia recorded the highest average of 10.50 ± 6.69 per oyster while the Ouidah lagoon in Benin recorded the lowest average of 1.80 ± 1.90 per oyster. Overall, microplastic numbers varied significantly (p < 0.05) among the five countries. Microfibers, particularly those within 1001-5000 μm size, dominated the total microplastic count with a few fragments and films. No spherical microplastics were isolated in the oysters. In the Sierra Leone and Benin oysters, fragments and films were absent in the samples. Microplastic between the 1001 and 5000 μm size class dominated the counts, followed by 501-1000 μm, 101-500 μm, and 51-100 μm. Five polymer groups namely polyethylene, polyester, nylon, polypropylene, and polyamide were identified across the five countries, with polyethylene occurring in oysters from all five countries and polyester occurring in all but the oysters from Nigeria. This diversity of polymers suggests varied sources of microplastics ingested by the studied oysters. The absence of microspheres across the five supports findings from other studies that they are the least ingested and highly egested by the oysters., (© 2024. The Author(s).)

Microplastics (MP) have repeatedly been found in commercially cultured species of bivalves. There are concerns regarding the amount of MP released into the environment by aquaculture activities, and questions regarding possible higher MP loads in farm-grown shellfish compared to levels in shellfish collected from recreational beds. To explore this concept, seawater, aquaculture gear, and eastern oysters (Crassostrea virginica) were sampled from an aquaculture site in Niantic Bay, CT, USA, and a 2-week transplantation experiment was performed in which oysters were transplanted between the aquaculture site and a plastic-free cage off the dock at the University of Connecticut-Avery Point campus. The digestive gland-stomach complex (gut) was dissected from the oysters and MP were extracted from the adjacent seawater and oyster gut samples using previously validated extraction methods. Extensive quality assurance and control measures were taken to reduce MP contamination. Particles in all samples were isolated, imaged under a stereomicroscope, and characterized (size, shape, polymer) using ImageJ software and micro-Fourier transform infrared spectroscopy. Water samples contained 0-0.3 MP/L and oyster gut samples contained 0-1.3 MP/g wet weight indicating very low concentrations of MP at the farm (0-2 MP/individual) or away from the farm (0-3 MP/individual). Aquaculture gear in this area is not contributing to MP ingestion in farmed oysters or elevated MP levels in the surrounding water., 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 © 2024 Elsevier Ltd. All rights reserved.)

Tetrodotoxin (TTX) is a potent neurotoxin causing human intoxications from contaminated seafood worldwide and is of emerging concern in Europe. Shellfish have been shown to contain varying TTX concentrations globally, with concentrations typically higher in Pacific oysters Crassostrea gigas in Europe. Despite many decades of research, the source of TTX remains unknown, with bacterial or algal origins having been suggested. The aim of this study was to identify potential source organisms causing TTX contamination in Pacific oysters in French coastal waters, using three different techniques. Oysters were deployed in cages from April to September 2021 in an estuary where TTX was previously detected. Microscopic analyses of water samples were used to investigate potential microalgal blooms present prior or during the peak in TTX. Differences in the bacterial communities from oyster digestive glands (DG) and remaining flesh were explored using metabarcoding, and lastly, droplet digital PCR assays were developed to investigate the presence of Cephalothrix sp., one European TTX-bearing species in the DG of toxic C. gigas. Oysters analysed by liquid chromatography-tandem mass spectrometry contained quantifiable levels of TTX over a three-week period (24 June-15 July 2021), with concentrations decreasing in the DG from 424 μg/kg for the first detection to 101 μg/kg (equivalent to 74 to 17 μg/kg of total flesh), and trace levels being detected until August 13, 2021. These concentrations are the first report of the European TTX guidance levels being exceeded in French shellfish. Microscopy revealed that some microalgae bloomed during the TTX peak, (e.g., Chaetoceros spp., reaching 40,000 cells/L). Prokaryotic metabarcoding showed increases in abundance of Rubritaleaceae (genus Persicirhabdus) and Neolyngbya, before and during the TTX peak. Both phyla have previously been described as possible TTX-producers and should be investigated further. Droplet digital PCR analyses were negative for the targeted TTX-bearing genus Cephalothrix., 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 © 2024 Elsevier Ltd. All rights reserved.)

Introduction and establishment of non-indigenous species (NIS) has been accelerated on a global scale by climate change. NIS Magallana gigas' (formerly Crassostrea gigas') global spread over the past several decades has been linked to warming waters, specifically during summer months, raising the specter of more spread due to predicted warming. We tracked changes in density and size distribution of M. gigas in two southern California, USA bays over the decade spanning 2010-2020 using randomly placed quadrats across multiple intertidal habitats (e.g., cobble, seawalls, riprap) and documented density increases by 2.2 to 32.8 times at 7 of the 8 sites surveyed across the two bays. These increases in density were coincident with 2-4° C increases in median monthly seawater temperature during summer months, consistent with global spread of M. gigas elsewhere. Size frequency distribution data, with all size classes represented across sites, suggest now-regular recruitment of M. gigas. Our data provide a baseline against which to compare future changes in density and abundance of a globally-spread NIS of significant concern., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Wolfe et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

LPS induced TNF-α Factor (LITAF) is a transcription factor widely involving in activation of Tumor Necrosis Factor (TNF) and other cytokines in the inflammatory response. In the present study, a homologue of LITAF with a conserved LITAF domain was identified from the Pacific oyster Crassostrea gigas. The transcripts of CgLITAF were detected in all examined tissues with highest expression in hepatopancrease. The immunofluorescence assay and Western blot showed that LPS stimulation induced an obvious nucleus translocation of CgLITAF protein in haemocytes. While the mRNA level of CgLITAF changed slightly after LPS stimulation. When the siRNA of CgLITAF was injected to inhibit its expression, the apoptotic level of haemocytes decreased observably after LPS stimulation. Consistently, the transcripts of CgTNF3 and CgTNF4 (LOC105343080, LOC105341146), the apoptotic-related molecules including CgBax, CgCytochrome c, CgCaspase9 and CgCaspase3, were significantly suppressed in the CgLITAF-RNAi oysters. While the mRNA expression level of CgBcl was enhanced significantly in the CgLITAF-RNAi oysters. These results indicated that CgLITAF promoted haemocyte apoptosis by regulating the expression of apoptotic-related factors, suggesting its important role in the immune response of oysters., Competing Interests: Declaration of competing interest The authors declare that they have no conflict of interests., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Pacific oysters ( Magallana gigas, a.k.a. Crassostrea gigas ), the most widely farmed oysters, are under threat from climate change and emerging pathogens. In part, their resilience may be affected by their microbiome, which, in turn, may be influenced by ocean warming and acidification. To understand these impacts, we exposed early-development Pacific oyster spat to different temperatures (18°C and 24°C) and p CO 2 levels (800, 1,600, and 2,800 µatm) in a fully crossed design for 3 weeks. Under all conditions, the microbiome changed over time, with a large decrease in the relative abundance of potentially pathogenic ciliates ( Uronema marinum ) in all treatments with time. The microbiome composition differed significantly with temperature, but not acidification, indicating that Pacific oyster spat microbiomes can be altered by ocean warming but is resilient to ocean acidification in our experiments. Microbial taxa differed in relative abundance with temperature, implying different adaptive strategies and ecological specializations among microorganisms. Additionally, a small proportion (~0.2% of the total taxa) of the relatively abundant microbial taxa were core constituents (>50% occurrence among samples) across different temperatures, p CO 2 levels, or time. Some taxa, including A4b bacteria and members of the family Saprospiraceae in the phyla Chloroflexi (syn. Chloroflexota ) and Bacteroidetes (syn. Bacteroidota ), respectively, as well as protists in the genera Labyrinthula and Aplanochytrium in the class Labyrinthulomycetes , and Pseudoperkinsus tapetis in the class Ichthyosporea were core constituents across temperatures, p CO 2 levels, and time, suggesting that they play an important, albeit unknown, role in maintaining the structural and functional stability of the Pacific oyster spat microbiome in response to ocean warming and acidification. These findings highlight the flexibility of the spat microbiome to environmental changes.IMPORTANCEPacific oysters are the most economically important and widely farmed species of oyster, and their production depends on healthy oyster spat. In turn, spat health and productivity are affected by the associated microbiota; yet, studies have not scrutinized the effects of temperature and p CO 2 on the prokaryotic and eukaryotic microbiomes of spat. Here, we show that both the prokaryotic and, for the first time, eukaryotic microbiome of Pacific oyster spat are surprisingly resilient to changes in acidification, but sensitive to ocean warming. The findings have potential implications for oyster survival amid climate change and underscore the need to understand temperature and p CO 2 effects on the microbiome and the cascading effects on oyster health and productivity., Competing Interests: The authors declare no conflict of interest.

Global climate change drives species redistribution, threatening biodiversity and ecosystem heterogeneity. The Kumamoto oyster, Crassostrea sikamea (Amemiya, 1928), one of the most promising aquaculture species because of its delayed reproductive timing, was once prevalent in southern China. In this study, an ensemble species distribution model was employed to analyze the distribution range shift and ecological niche dynamics of C. sikamea along China's coastline under the current and future climate scenarios (RCP 2.6-8.5 covering 2050 s and 2100 s). The model results indicated that the current habitat distribution for C. sikamea consists of a continuous stretch extending from the coastlines of Hainan Province to the northern shores of Jiangsu Province. By the 2050 s, the distribution range will stabilize at its southern end along the coast of Hainan Province, while expanding northward to cover the coastal areas of Shandong Province, showing a more dramatic trend of contraction in the south and invasion in the north by the 2100 s. In RCP8.5, the southern end retracts to the coasts of Guangdong, whereas the northern end covers all of China's coastal areas north of 34°N. C. sikamea can maintain relatively stable ecological niche characteristics, while it may occupy different ecological niche spaces under future climate conditions. Significant niche expansion will occur in lower temperature. We concluded C. sikamea habitats are susceptible to climate change. The rapid northward expansion of C. sikamea may open new possibilities for oyster farming in China, but it will also have important consequences for the ecological balance and biodiversity of receiving areas. It's imperative that we closely examine and strategize to address these repercussions for a win-win situation., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Haiyan Wang reports financial support was provided by National Natural Science Foundation of China (42076092, 41776179). Haiyan Wang reports financial support was provided by National Key Research and Development Program of China (2022YFD2401301, 2022FY100304). Haiyan Wang reports financial support was provided by Strategic Priority Research Program of the Chinese Academy of Sciences (XDB42000000). Haiyan Wang reports financial support was provided by Earmarked Fund for Modern Agro-industry Technology Research System (CARS-47). If there are other authors, they 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 © 2024. Published by Elsevier B.V.)

The kleptoparasitic pea crab Zaops ostreum lives within the gills of bivalves, including the economically important eastern oyster Crassostrea virginica. Previous research along the east coast of central Florida has found an average of 2.3 pieces of plastic per oyster. The goals of our research were to determine if filter-feeding oysters transfer microfibers to Z. ostreum via the crab: 1) actively consuming plastic particles, or 2) passively becoming entangled in microfibers. Our results show that both occur. While only 11.6 % of Z. ostreum (total n = 122) consumed microfibers, those that did had up to 14 pieces in their soft tissues. Similarly, only 7.4 % of Z. ostreum had microfibers entangled around their appendages. Mean lengths of consumed and entangled fibers were similar, 1.9 and 2.7 mm, respectively. Additional research is needed to understand the positive and negative impacts of microfibers associated with pea crabs on both species., 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 © 2024 Elsevier Ltd. All rights reserved.)

The present study investigated the contamination of oysters farmed in Amazonian estuaries by microplastics (MPs). A total of 120 adult oysters (Crassostrea gasar) were collected from four sites along the Mangrove Coast of Pará/Brazil: S1, S2, S3 and S4, with 30 oyster for each. Overall, 58.33 % of the oyster samples contained microplastics, with mean concentrations of 0.23 MPs/g and 1.9 MPs/ind. The concentration of microplastics varied among the four sites, where S1 and S3 had the highest values while S4 had the lowest. PA fibers were the majority of particles (91 %), followed by PS fragments (9 %). The hepatopancreas and the gonad concentrated more microplastics than the rest of the body. As an important species for aquaculture in Amazon, we recommend additional regulation to reduce human exposure to microplastics, such as the installation of depuration facilities and constant monitoring of the contamination of oysters from farms in the region., 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 © 2024 Elsevier Ltd. All rights reserved.)

Understanding the metal concentrations in oysters is important because of its relevance to human health and biomonitoring. However, metal concentrations in oysters are highly variable in nature and not well explained by metal exposure. This study examined the metal contamination in farm oysters Crassostrea hongkongensis grown in Qinzhou Bay, south China. Cadmium (Cd), zinc (Zn), nickel (Ni), and copper (Cu) concentrations in the oysters varied between 7.9 and 72.2, 282-17003, 0.37-47.7 and 37-4012 μg g -1 , respectively, showing large metal variability among different individuals. Oyster metal concentrations decreased with increasing body size and significantly higher levels were observed in wet season. Low salinity and slower oyster growth due to inferior growth conditions could be responsible for the elevated metal concentrations in the wet season. Biokinetic modeling showed that the coupling of ingestion rate and growth can cause 2.8-4.2 folds differences in the oyster Cd and Zn concentrations, respectively, suggesting the significant role of oyster bioenergetics in contributing to the metal variability. Modeling data revealed that Cd and Zn concentrations in oyster tissues reach maximum levels when oysters have their lowest growth efficiency. This suggests that any factors influencing the energy budget in oysters could simultaneously alter their metal concentrations, which might be the reason why oyster metal concentrations are so variable in the natural environment., 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 © 2024 Elsevier Ltd. All rights reserved.)

In estuarine food webs, bivalve molluscs transfer nutrients and pollutants to higher trophic levels. Mercury (Hg) pollution is ubiquitous, but it is especially elevated in estuaries historically impacted by industrial activities, such as those in the U.S. Northeast. Monomethylmercury (MeHg), the organic form of Hg, is highly bioaccumulative and transferable in the food web resulting in the highest concentrations in the largest and oldest marine predators. Patterns of Hg concentrations in marine bivalve molluscs, however, are poorly understood. In this study, inorganic Hg (iHg), MeHg, and the total Hg (THg) in soft tissues of the northern quahogs (Mercenaria mercenaria), eastern oysters (Crassostrea virginica), and ribbed mussels (Geukensia demissa) from eastern Long Island sound, a temperate estuary of the western North Atlantic Ocean was investigated. In all three species, concentrations of THg remained similar between the four sampling months (May, June, July, and September), and were mostly independent of animal size. In quahogs, MeHg and iHg displayed significant (p < 0.05) positive (iHg in May and June) and negative (MeHg in July and September) changes with shell height. Variability in concentrations of THg, MeHg, and iHg, both inter- and intra-specifically was high and greater in quahogs and oysters (THg: 37, 39%, MeHg: 28, 39%, respectively) than in mussels (THg: 13%, MeHg: 20%). The percentage of THg that was MeHg (%MeHg) was also highly variable in the three species (range: 10-80%), highlighting the importance of measuring MeHg and not only THg in molluscs., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

The Negombo Lagoon is a coastal lagoon influenced by local communities that introduce waste into its ecosystem. This study examined seven sewage entry points, out of which five sites were chosen for oyster sampling based on availability. Physicochemical and microbiological parameters of water (measured in triplicate at each site, n = 84) and oyster samples (total length, TL > 6 cm, n = 30) were assessed. Variation in regional coliform contamination was analyzed employing a one-way analysis of variance (ANOVA). Results indicated that the northern part of the lagoon exceeded recommended coliform thresholds for swimming (total coliform concentration (TCC) < 126 most probable number (MPN)) and seafood consumption (TCC < 100 MPN/g), indicating the presence of Escherichia coli. Water quality indices affirmed fecal pollution, except in the southern part of the lagoon. Furthermore, the study found high oyster consumption (76.7 %), elucidating that oysters from the northern part of Negombo Lagoon pose health risks., 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 © 2024 Elsevier Ltd. All rights reserved.)

The discharge of sanitary sewage into the bays of the Florianópolis Metropolitan Area (Southern Brazil), has led to the contamination of oyster farms. Consequently, linear alkylbenzenes (LABs) were quantified in the sediment, and the biochemical responses in gills and digestive gland of oysters from six farms were assessed. Our findings revealed elevated levels of LABs in the sediment of the Imaruim and Serraria farms. Additionally, alterations were observed in the antioxidant enzymes: catalase, glutathione peroxidase and superoxide dismutase in both oyster tissue from the Serraria, Santo Antonio de Lisboa and Sambaqui farms. Furthermore, correlation analyses indicated strong and moderate associations between biochemical responses, organic contaminants, and certain physicochemical parameters. Consequently, our results demonstrated the activation of the antioxidant system in oysters, representing a protective response to the presence of sanitary sewage and other contaminants. Therefore, we propose the utilization of biochemical biomarkers for monitoring the environmental quality of farms., 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 © 2024 Elsevier Ltd. All rights reserved.)

Background: Lycopene (LYC), a carotenoid found in abundance in ripe red fruits, exhibits higher singlet oxygen quenching activity than other carotenoids. However, the stability of LYC is extremely poor due to its high double-bond content. In this paper, a nano-encapsulation strategy based on highly stable marine-derived ferritin GF1 nanocages was used to improve the thermal stability and oxidation resistance of LYC, thereby boosting its functional effectiveness and industrial applicability., Results: The preparation of GF1-LYC nanoparticles benefited from the pH-responsive reversible self-assembly of GF1 to capture LYC molecules into GF1 cavities with a LYC-to-protein ratio of 51 to 1. After the encapsulation of the LYC, the reassembled GF1 nanocages maintained intact morphology and good monodispersity. The GF1-LYC nanoparticles incorporated the characteristic LYC peaks in spectrograms, and their powder form contained the crystalline form of LYC. Molecular docking revealed that LYC bound with the inner triple-axis channel areas of GF1, interacting with VAL139, LYS72, LYS65, TYR69, PHE129, HIS133, HIS62, and TYR134 amino acids through hydrophobic bonds. Fourier transform infrared spectroscopy also demonstrated the bonding of GF1 and LYC. In comparison with free LYC, GF1 reduced the thermal degradation of encapsulated LYC at 37 °C significantly and maintained the 2,2-Diphenyl-1-picrylhydrazyl (DPPH)-scavenging ability of LYC., Conclusion: As expected, the water solubility, thermal stability, and antioxidant capacity of encapsulated LYC from GF1-LYC nanoparticles was notably improved in comparison with free LYC, indicating that the shell-like marine ferritin nanoplatform might enhance the stable delivery of LYC and promote its utilization in the field of food nutrition and in other industries. © 2023 Society of Chemical Industry., (© 2023 Society of Chemical Industry.)

Ostreid herpesvirus 1 (OsHV-1) and its microvariants (µVars) cause economically devastating mass mortalities of oysters and pose a threat to the shellfish aquaculture industry globally. OsHV-1 outbreaks can cause up to 100% mortality in the Pacific oyster Crassostrea gigas. However, OsHV-1 and its variants have a broad host range and can infect at least 7 bivalve species, including bay scallops Argopecten irradians and eastern oysters C. virginica. Determining the susceptibility of economically and ecologically important bivalve species to OsHV-1 is critical for improving biosecurity and disease management to protect the aquaculture industry. Surveys of eastern oysters were conducted in June to August 2021 in the Maryland portion of the Chesapeake Bay to determine the prevalence and viral load of OsHV-1 at 5 aquaculture farms. Using quantitative PCR, OsHV-1 was not detected at any sites. Experiments examined the susceptibility of single stocks of eastern oysters and hard clams Mercenaria mercenaria to the virus and their ability to horizontally transmit it using OsHV-1 µVar SD (San Diego, California) and OsHV-1 µVar FRA (Marennes-Olreon, France). Results showed that OsHV-1 µVars did not cause mortality or symptomatic infection in the single stocks of eastern oysters and hard clams used in these experiments using natural infection pathways. However, the eastern oyster stock, when injected with OsHV-1, did transmit the virus to naïve Pacific oysters. Further experimentation using additional stocks and lines and establishment of surveillance programs along the east and Gulf coasts of the USA are necessary to prepare for the potential spread and impact of OsHV-1 related disease.

Microplastics (MPs) and polychlorinated biphenyls (PCBs) are pervasive pollutants in the marine environment, exerting adverse effects on marine organisms. While it is suggested that their exposure may compromise the immune responses of marine organisms, the cumulative immunotoxic effects remain uncertain. Additionally, the intricate mechanisms underlying the immunotoxicity of PCBs and MPs in marine organisms are not yet fully comprehended. To illuminate their combined biological impacts, Crassostrea gigas were exposed to 50 μg/L MPs (30-μm porous) alone, as well as 10 or 100 ng/L PCBs individually or in combination with 50 μg/L of MPs for 28 days. Our data demonstrated that oysters treated with the pollutants examined led to decreased total haemocyte count, inhibited phagocytosis of haemocytes, enhanced the intracellular contents of reactive oxygen species, lipid peroxidation and DNA damage, reduced lysozyme concentration and activity, gave rise to superoxide dismutase. Catalaseand glutathione S-transferaseactivity. The expression of three immune-related genes (NF-κB, TNF-α, TLR-6) was drastically suppressed by the PCBs and MPs treatment, while the apoptosis pathway-related genes (BAX and Caspase-3) showed a significant increase. In addition, compared to oysters treated with a single type of pollutant, coexposure to MPs and PCBs exerted more severe adverse impacts on all the parameters investigated, indicating a significant synergistic effect. Therefore, the risk of MPs and PCBs chemicals on marine organisms should be paid more attention., 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 © 2024. Published by Elsevier Ltd.)

This study focused on spatial-temporal variations in the composition and structure of polychaete communities associated with two coexisting oyster species in a southern Gulf of Mexico coastal lagoon, showed that Crassostrea rhizophorae hosted 23 polychaete species and C. virginica only four. The observed variability was mainly explained by differences in salinity and sedimentation rate, two factors that also influenced the host species distribution. The highest values of richness and diversity occurred at C. rhizophorae sites during the rainy season, coinciding with increased salinity and a decrease in sedimentation rate. The shell-borer species Nereis garwoodi and Polydora websteri occurred in both oysters, but showing higher frequencies in C. virginica during the Nortes season (i.e., the cold fronts season). Our results underscore the contrasting diversity of associated polychaetes with two coexisting oyster species and emphasize the need for considering additional environmental parameters to allow better understanding of the factors that are driving the structure of these communities., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Araujo-Leyva Osmar Roberto reports a relationship with Consejo Nacional de Humanidades, Ciencias y Tecnologías that includes: funding grants., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

An increasing number of evidences have shown that invertebrate taxa can be primed to produce immune memory to resist the secondary infection of pathogens, which was considered as a viable option to protect invertebrates from pathogens. In this work, we compared the protective effect of several different immune priming methods on the Vibrio alginolyticus secondary infection of the Crassostrea gigas. The results showed that C. gigas primed with live V. alginolyticus had higher ROS level, which led to hemocytes necrosis and higher mortality rate in the later stage. Low-dose of formalin-inactivated V. alginolyticus (including 5 × 10 4  CFU/mL and 5 × 10 5  CFU/mL) elicited appropriate immune response in C. gigas, protecting C. gigas from V. alginolyticus infection. Immersion with 5 × 10 4  CFU/mL formalin-inactivated V. alginolyticus was performed to prime C. gigas immunity in the trans-generational immune priming. Trans-generational immune priming significantly increased the resistance of larvae to various Vibrio species. Overall, these results suggested that low-dose of formalin-inactivated V. alginolyticus can protect C. gigas from secondary infection and confer broad-spectrum Vibrio resistance on offspring. This work provided valuable information toward a new direction for the protection of C. gigas from Vibrio infection., Competing Interests: Declaration of competing interest The authors declare that there are no conflicts of interest., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Muscle atrophy is a detrimental and injurious condition that leads to reduced skeletal muscle mass and disruption of protein metabolism. Oyster ( Crassostrea nippona ) is a famous and commonly consumed shellfish in East Asia and has become a popular dietary choice worldwide. The current investigation evaluated the efficacy of C. nippona against muscle atrophy, which has become a severe health issue. Mammalian skeletal muscles are primarily responsible for efficient metabolism, energy consumption, and body movements. The proteins that regulate muscle hypertrophy and atrophy are involved in muscle growth. C. nippona extracts were enzymatically hydrolyzed using alcalase (AOH), flavourzyme (FOH), and protamex (POH) to evaluate their efficacy in mitigating dexamethasone-induced muscle damage in C2C12 cells in vitro. AOH exhibited notable cell proliferative abilities, promoting dose-dependent myotube formation. These results were further solidified by protein expression analysis. Western blot and gene expression analysis via RT-qPCR demonstrated that AOH downregulated MuRF-1, Atrogin, Smad 2/3, and Foxo-3a, while upregulating myogenin, MyoD, myosin heavy chain expression, and mTOR, key components of the ubiquitin-proteasome and mTOR signaling pathways. Finally, this study suggests that AOH holds promise for alleviating dexamethasone-induced muscle atrophy in C2C12 cells in vitro, offering insights for developing functional foods targeting conditions akin to sarcopenia.

The Olympia oyster, Ostrea lurida, is the target of many restoration projects along estuaries on the North American Pacific coast, while the non-native Pacific oyster, Magallana gigas, dominates oyster aquaculture globally. Both species provide filtration functions that were investigated in three California bays using a whole-habitat, in situ approach, a laboratory particle selection experiment, and a regional physiological comparison. Measurements of chlorophyll α, temperature, salinity, and turbidity upstream and downstream, as well as point samples of seston total particulate matter and organic content to estimate habitat clearance rates (HCR, L hr -1  m -2 ) were collected. From February 2018 to June 2019, twenty-two trials were conducted across four sites. HCRs were highly variable within and among sites, ranging from site averages of -464 to 166 L hr -1  m -2 , and not significantly different among sites, indicating field filtration performance of O. lurida habitat and M. gigas aquaculture is similar. Using a random forest regression, site was the most important predictor of HCR, with a variable importance score of 25.7 % (SD = 4.6 %). O. lurida and M. gigas had significantly different particle size selection preferences, likely affecting the quality of their filtration. This study's findings suggest that restoring O. lurida habitat may provide similar filtration benefits as M. gigas aquaculture, but the unique hydrodynamics and food quality of individual bays, as well as regional differences in filter feeder communities, must be considered in managing oyster habitat for filtration functions., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Althea Marks reports financial support was provided by the California Sea Grant. Althea Marks reports financial support was provided by the National Geographic Society. Althea Marks reports travel was provided by the National Shellfisheries Association. Danielle Zacherl reports financial support was provided by the California State Coastal Conservancy. Danielle Zacherl reports financial support was provided by the Honda Marine Science Foundation. Althea Marks reports travel was provided by the Pacific Shellfish Grower Association., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

The present study aimed to evaluate concentrations of 16 priority PAHs in the mussel Perna perna and oyster Crassostrea rhizophorae of the Santos Estuary (Brazil) and the potential risk to human health associated with the consumption of these species. The levels of ΣPAHs in the tissues of bivalves ranged from 96.94 to 988.76 μg/kg for mussels and from 88.38 to 138.62 μg/kg for oysters. A general trend of higher concentrations of PAHs was observed according to the increase in log K ow values in both species. The estimated amount of ΣPAHs via mussel and oyster intake ranged from 1.0 to 3.2 ng/kg and from 0.4 to 1.2 ng/kg of body weight per day, respectively. The calculated THQs, HI and CRs for the regular consumption of raw bivalves followed the internationally acceptable limits and represent a low risk to human health., 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.)

Akanda National Park (ANP) is composed of mangrove ecosystems bordering Libreville, Gabon's capital. The contamination of aquatic resources from the ANP by persistent organic pollutants (POPs) and trace metals (TMs) was never evaluated. To provide a basis for their monitoring in the ANP, five species (two fish, two mollusks, and one crustacean) were analyzed from three sampling sites in 2017. Contamination levels for POPs and TMs were below maximum acceptable limits for seafood, including Cd and Pb. No DDT was found in any sample. Inter-specific differences were more obvious than the differences among sites, although the results may be biased by an unbalanced sampling design. The oyster Crassostrea gasar was the most contaminated species, making this species a good candidate to assess environmental contamination in the area. The studied species also contained essential elements, such as Fe, Zn and Mn at interesting levels in a nutritional point of view., 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 © 2024 Elsevier Ltd. All rights reserved.)

Thioester-containing proteins (TEPs) play a vital role in the innate immune response to biotic and abiotic stresses. In this study, the TEPs in C. gigas were identified, and their gene structure, phylogenetic relationships, collinearity relationships, expression profiles, sequence diversity, and alternative splicing were analyzed. Eight Tep genes were identified in C. gigas genome. Functional analysis and evolutionary relationships indicated a high level of homology to other mollusks TEPs. The transcriptome quantitative analysis results showed that the Tep genes in C. gigas respond to heat stress and Vibrio stress. Alternative splicing analysis revealed four Tep genes (designated A2M&#95;1, CD109&#95;3, CD109&#95;5, complement C3) encode multiple alternative splice variants. Analysis of gene structure and multiple alignments revealed that seven CD109&#95;5 variants are produced through the alternative splicing of the 19th exon, which encodes the highly variable central region. Sequence diversity analysis revealed thirteen missense variants within the 19th exon region of these seven CD109&#95;5 alternative splice variants. Furthermore, the differential alternative splicing analysis showed significant induction of CD109&#95;5, A2M&#95;1 and A2M&#95;2 variants after infection with V. parahaemolyticus. This study explores the Tep genes of C. gigas, providing insights into the molecular mechanisms underlying the involvement of C. gigas TEPs in innate immunity., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

This study aimed to carry out a general diagnosis of the contamination of the coastal marine environment of the Santa Catarina state (SC, Brazil) by different classes of environmental pollutants, as well as to evaluate possible adverse effects of the contaminants on biochemical biomarkers of oysters, Crassostrea gasar and Crassostrea rhizophorae. 107 chemicals were evaluated in water, sediment and oyster samples from nine sites along the coastline of SC. We also examined various biochemical biomarkers in the oysters' gills and digestive glands to assess potential effects of contaminants. In general, the northern and central regions of the littoral of SC presented higher occurrences and magnitudes of contaminants than the southern region, which is probably related to higher urbanization of center and northern areas of the littoral. The biomarker analysis in the oysters reflected these contamination patterns, with more significant alterations observed in regions with higher levels of pollutants. Our results may serve as a first baseline for future and more extensive monitoring actions and follow-up of the degree of contamination in the state, allowing for inspection actions and management of areas most affected by marine pollutants., 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.)

The tremendous loss of microplastics from the sea surface and the low density of microplastics found in the water column and sediments indicate that the oceans have mechanisms capable of transporting microplastics from the surface to the seafloor. These include physicochemical processes and biological influences from marine organisms that drive the vertical migration of microplastics. Little is known, however, about the biological processes involved in the deposition of plastics in the marine environment. A considerable number of mariculture filter-feeding organisms can consume substantial amounts of suspended substances in the water column, and these organisms are ideal candidates for depositing microplastics. In this study, we analyzed microplastic abundance in typical mariculture filter feeders, i.e., ascidians (Halocynthia roretzi), oysters (Crassostrea gigas), scallops (Chlamys farreri) and clams (Ruditapes philippinarum), quantified the number and characteristics of the microplastics they deposited in situ, and further compared microplastic biodeposition rates. Microplastics were present in feces and pseudofeces and sank to form biodeposits rather than accumulating to significant levels in organisms. Microplastics were found in significantly higher numbers in the biodeposits of mariculture organisms than in the control deposits (p < 0.01). The shape and color of the microplastics in the sediments were not impacted by the presence of organisms (p > 0.05), but the deposition of <1000 μm and positive-buoyancy (less dense than seawater) microplastics was significantly increased in the biodeposits (p < 0.05). The highest microplastic biodeposition rate was found in scallops (1.14 ± 0.07 items·ind -1 ·d -1 or 0.5 ± 0.03 items·g -1 ·d -1 ). These results suggest that mariculture filter-feeding organisms have important biodepositional functions that influence the fate of microplastics through the transfer of microplastics from the surface to the seafloor. This study could contribute to a better understanding of the biological plastic pump mechanisms in oceans., 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 B.V. All rights reserved.)

The current study investigated the effects of heat treatment (85 °C or 100 °C for 5-20 min) on human norovirus (HuNoV) GII.4's capsid stability in fresh oysters. In addition, propidium monoazide (PMA) was used in viral samples to distinguish infectious viruses and evaluated using real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR). Further, we explored the effect of the heat treatment on oyster quality (Hunter color and hardness). The titer of HuNoV for oysters significantly ( p < 0.05) decreased to 0.39-1.32 and 0.93-2.27 log 10 copy number/μL in the non-PMA and PMA-treated groups, respectively, after heat treatment. HuNoV in oysters not treated with PMA showed a decrease of <1.5 - log 10 , whereas in PMA-treated oysters, a decrease of >1 - log 10 was observed after treatment at 85 °C for 10 min. Treatments for both 15 min and 20 min at 100 °C showed a >99% log 10 reduction using PMA/RT-qPCR. In the Hunter color, an increase in heat temperature and duration was associated with a significant decrease in 'L' (brightness+, darkness-) and an increase in 'a' (redness+, greenness-) and 'b' (yellowness+, blueness-) ( p < 0.05). Our findings confirmed that the hardness of oyster meat significantly increased with increasing temperature and time ( p < 0.05). This study demonstrated that PMA/RT-qPCR was effective in distinguishing HuNoV viability in heat-treated oysters. The optimal heat treatment for oysters was 10 min at 85 °C and 5 min at 100 °C.

MnO 2 is a nanozyme that inhibits the decomposition of hydrogen peroxide (H 2 O 2 ) into a hydroxyl radical (OH • ), thus preventing its conversion into reactive oxygen species (ROS). Oyster ferritin (GF1) is a macromolecular protein that provides uniform size and high stability and serves as an excellent template for the biomineralization of nanozyme. This study presents a unique method in which MnO 2 is grown in situ in the GF1 cavity, yielding a structurally stable ferritin-based nanozyme (GF1@Mn). GF1@Mn is demonstrated to be stable at 80 °C and pH 4-8, exhibiting a higher affinity with H 2 O 2 than many other catalases (CAT) with a Michaelis constant (Km) of 25.45 mmol/L. In vitro experiments have demonstrated the potential of GF1@Mn to enhance cell survival by reducing nitric oxide (NO) production while mitigating macrophage damage from ROS. The findings are essential to developing ferritin-based nanozymes and hold great potential for applications in functional food development.

In this study, novel umami peptides were prepared from oyster ( Crassostrea gigas ) hydrolysates, and their umami mechanisms were investigated. Umami fractions G2 and G3 were isolated by gel filtration chromatography (GFC) and sensory evaluation. The umami scores of the G2 and G3 fractions were 7.8 ± 0.12 and 7.5 ± 0.18, respectively. 36 potential umami peptides with molecular weights below 1500 Da, E and D accounting for >30% of the peptides and iUmami-SCM > 588 were screened by peptidomics. Peptide source analysis revealed that myosin, paramyosin, and sarcoplasmic were the major precursor proteins for these peptides. The electronic tongue results demonstrated that the synthetic peptides DPNDPDMKY and NARIEELEEE possessed an umami characteristic, whereas SIEDVEESRNK and ISIEDVEESRNK possessed a saltiness characteristic. Additionally, molecular docking results indicated that the umami peptide (DPNDPDMKY, NARIEELEEE, SIEDVEESRNK, and ISIEDVEESRNK) binds to H145, S276, H388, T305, Y218, D216, and Q389 residues in the T1R3 taste receptor via a conventional hydrogen bond and a carbon-hydrogen bond. This research provides a new strategy for the screening of umami peptides.

Doublesex and Mab-3-related transcription factor (Dmrt) is a type of transcription factor with a zinc-finger DM structural domain, which plays a significant role in sex determination and differentiation in animals. Although Dmrt has been reported in many vertebrates and invertebrates, it has rarely been studied in bivalves. In this study, a total of three members of the Dmrt gene family were identified and characterized in Crassostrea gigas, and all these CgDmrt genes contained a conserved DM domain. Analysis of the phylogenetic tree and gene structure revealed that Dmrt genes clustered on one branch may have similar functions in bivalves. Expression profiling of CgDmrt mRNA in different tissues and stages of gonad development indicated that CgDmrt3 exhibited sexually dimorphic expression and played an important role in the development of the male gonad in C. gigas. Furthermore, analysis of CgDmrt mRNA expression between fertile triploids and sterile triploids showed that CgDmrt3 may be involved in sperm production. Collectively, the systematic analysis of the CgDmrt genes will provide potential insights into the function of these genes in gonadal development., Competing Interests: Declaration of Competing Interest The authors declare that they have no conflicts of interest with the contents of this article., (Copyright © 2023 Elsevier Inc. All rights reserved.)

The pollution of seawater by both biotic (bacteria, viruses) and abiotic contaminants (biocides, pharmaceutical residues) frequently leads to economic losses in aquaculture activities mostly mortality events caused by microbial infection. Advanced Oxidation Processes (AOPs) such as heterogeneous photocatalysis allow the removal of all organic contaminants present in water and therefore could reduce production losses in land-based farms. Oysters in land-based farms such as hatcheries and nurseries suffer from a large number of mortality events, resulting in significant losses. If photocatalysis has been widely studied for the decontamination, its application for disinfection is still overlooked, especially on seawater for viruses. We therefore studied seawater disinfection using the photocatalysis (UV 365 /TiO 2 ) method in the context of Pacific oyster mortality syndrome (POMS). POMS has been defined as a polymicrobial disease involving an initial viral infection with Ostreid Herpes Virus 1, accompanied by multiple bacterial infections. We investigated the impact of treatment on Vibrio harveyi, a unique opportunistic pathogenic bacterium, and on a complex microbial community reflecting a natural POMS event. Viral inactivation was monitored using experimental infections to determine whether viral particles were still infectious after. Changes in the total bacterial community in seawater were studied by comparing UV 365 /TiO 2 treatment with UV 365 -irradiated seawater and untreated seawater. In the case of OsHV-1, a 2-h photocatalytic treatment prevents POMS disease and oyster mortality. The same treatment also inactivates 80% of viable Vibrio harveyi culture (c.a. 1.5 log). Since OsHV-1 and Vibrio harveyi are effectively inactivated without long-term destabilization of the total bacterial microbiota in the seawater, photocatalysis appears to be a relevant alternative for disinfecting seawater in land-based oyster beds., 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. Published by Elsevier Ltd.)

Vibrio is an important group of aquatic animal pathogens, which has been identified as the main pathogenic factor causing mass summer mortality of Crassostrea gigas in northern China. This study aims to investigate the potential pathogenic mechanisms of Vibrio Cg5 isolate in C. gigas. We sequenced and annotated the genome of Vibrio Cg5 to analyze potential virulence factors. The gentamicin protection assays were performed with C. gigas primary cells to reveal the cell-invasive behavior of Cg5. The genome analysis showed that Cg5 was a strain of human disease-associated pathogen with multiple antibiotic resistance, and four virulence factors associated with intracellular survival were present in the genome. The gentamicin protection assays showed that Cg5 could potentially invade the cells of C. gigas, indicating that Cg5 could be a facultative intracellular pathogen of C. gigas. These results provide insights into the pathogenic mechanism of V. diabolicus, an emerging pathogenic Vibrio on aquatic animals, which would be valuable in preventing and controlling diseases in oysters., Competing Interests: Declaration of competing interest No potential conflict of interest was reported by the authors., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

The DM9-containing proteins have been identified as pattern recognition receptors (PRRs) to recognize invading pathogens and subsequently mediate downstream signal pathways, playing essential roles in innate immune responses of molluscs. In the present study, a novel DM9-containing protein (named as CgDM9CP-7) was identified from Pacific oyster Crassostrea gigas, which contained two tandem DM9 repeats similar to the previously identified CgDM9CPs. The mRNA transcripts of CgDM9CP-7 were found to be constitutively expressed in all the tested tissues including haemolymph, gill, hepatopancreas, mantle, adductor muscle and labial palp. The expression level of CgDM9CP-7 mRNA in haemocytes significantly up-regulated at 3 and 6 h after Vibrio splendidus stimulation, which was 5.67-fold (p < 0.01) and 4.71-fold (p < 0.05) of that in the control group, respectively, and it also increased significantly at 6 h (3.08-fold, p < 0.01) post lipopolysaccharide (LPS) stimulation. The protein of CgDM9CP-7 was mainly detected in membrane and cytoplasm of oyster haemocytes after V. splendidus stimulation. The recombinant CgDM9CP-7 protein (rCgDM9CP-7) displayed binding activities to MAN, LPS, PGN, Poly (I:C) as well as gram-negative bacteria (V. splendidus and Escherichia coli), gram-positive bacteria (Staphylococcus aureus and Micrococcus luteus) and fungi (Pichia pastoris and Yarrowia lipolytica). rCgDM9CP-7 was able to agglutinate Bacillus subtilis, V. splendidus, E. coli and S. aureus, inhibit their growth, and bind the recombinant protein CgMyd88-2 (K D  = 5.98 × 10 -6  M) and CgMyd88s (K D  = 8.5 × 10 -7  M) in vitro as well. The transcripts of CgIL17-1 (0.45-fold of the control group, p < 0.01), CgIL17-2 (0.19-fold, p < 0.05), CgIL17-3 (0.54-fold, p < 0.05), CgIL17-5 (0.36-fold, p < 0.05) and CgIL17-6 (0.24-fold, p < 0.01) in CgDM9CP-7-siRNA oysters decreased significantly at 6 h after V. splendidus stimulation. These results collectively indicated that CgDM9CP-7 was involved in the regulation of CgMyD88 and CgIL-17 expression in the immune response of oyster., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Lead can induce oxidative stress and increase lipid peroxidation in biofilms, leading to liver damage and physiological dysfunction. This study aimed to investigate how oyster ferritin (GF1) attenuates lead-induced oxidative damage to the liver in vitro and in vivo. Animal experiments have confirmed that lead exposure can lead to oxidative damage and lipid peroxidation of the liver, and ferritin can regulate the activity of antioxidant enzymes and alleviate pathological changes in the liver. At the same time, oyster ferritin can regulate the expression of oxidative stress-related genes and reduce the expression of inflammasome-related genes. In addition, lead can induce apoptosis and mitophagy, leading to overproduction of reactive oxygen species and cell death, which can be effectively alleviated by oyster ferritin. Overall, this study provides a theoretical foundation for the use of oyster ferritin as a means of mitigating and preventing lead-induced damage., 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. Published by Elsevier B.V.)

Artificial Light at Night (ALAN) is a fast-spreading threat to organisms, especially in coastal environments, where night lighting is increasing due to constant anthropization. Considering that ALAN affects a large diversity of coastal organisms, finding efficient solutions to limit these effects is of great importance but poorly investigated. The potential benefit of one strategy, in particular, should be studied since its use is growing: part-night lighting (PNL), which consists in switching off the lights for a few hours during nighttime. The aim of this study is to investigate the positive potential of the PNL strategy on the daily rhythm of the oyster Crassostrea gigas, a key species of coastal areas of ecological and commercial interest. Oysters were exposed to a control condition and three different ALAN modalities. A realistic PNL condition is applied, recreating a strategy of city policy in a coastal city boarding an urbanized bay (Lanton, Arcachon Bay, France). The PNL modality consists in switching off ALAN direct sources (5 lx) for 4 h (23-3 h) during which oysters are in darkness. Then, a PNL + skyglow (PNL + S) modality reproduces the previous one mimicking a skyglow (0.1 lx), an indirect ALAN source, during the direct lighting switch off, to get as close as possible to realistic conditions. Finally, the third ALAN condition mimics full-night direct lighting (FNL). Results revealed that PNL reduces some adverse effects of FNL on the behavioral daily rhythm. But, counterintuitively, PNL + S appears more harmful than FNL for some parameters of the behavioral daily rhythm. PNL + S modality is also the only one that affect oysters' clock and melatonin synthesis gene expression, suggesting physiological consequences. Thus, in realistic conditions, the PNL mitigation strategy might not be beneficial in the presence of skyglow, seeing worse for a coastal organism such as the oysters., 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 B.V. All rights reserved.)

Copper (Cu) is hyperaccumulated in oyster hemocytes and is an essential trace metal indispensable for diverse innate immune functions. However, the roles of Cu in oyster immune defense are still unclear. In this study, Cu exposure enhanced the phagocytosis of zymosan by increasing the number and length of filopodia, as well as mitochondrial ROS (mitoROS) production mainly in granulocytes, followed by semigranulocytes and agranulocytes. The intracellular calcium level increased to promote the phagosome-lysosome fusion after Cu exposure. The enhancement of phagosomal acidification and mitochondrion-phagosome juxtaposition were also found in granulocytes after Cu exposure. These results indicated that Cu could regulate the phagolysosomal system to enhance the antimicrobial ability of oyster hemocytes with the assistance of mitoROS. Furthermore, Cu(I) and Cu(II) were predominately located in lysosomes, and degranulation may provide a mechanism for exposing Cu to bacteria to prevent their survival and proliferation. Specifically, we showed that the newly formed Cu(I) arising from lysosomal Cu(II) moved to lysosomes and mitochondria in activated hemocytes to induce strong immune responses. The ability of the transformation of Cu(I) from Cu(II) followed granulocytes > semigranlocytes > agranulocytes, indicating that granulocytes played important roles in immune functions of oysters. Our results provided new insights into the understanding of antimicrobial effects of Cu in oyster hemocytes.

Glycation offers a promising potential to improve protein gelling properties in food industries. Therefore, the study was aimed to illustrate the effect of five monosaccharides (erythrose-aldotetrose, xylose-aldopentose, glucose-aldohexose, galactose-aldohexose, and fructose-ketohexose) with different carbon numbers and structure on the structure-gelling relationship of myofibrillar protein (MP) from oyster (Crassostrea gigas). Results showed that monosaccharides significantly increased the glycation degree of MP by increasing sulfhydryl content, forming stable tertiary conformation and decreasing surface hydrophobicity. Moreover, the gel properties of MP like gel strength, water holding capacity, water mobility were improved by alleviating aggregation including the increase of solubility and the decrease of particle sizes. Oyster MP glycated by glucose (aldohexose) possessed the optimal gel properties. Molecular docking simulation showed that hydrogen bonds and hydrocarbon bonds were the mainly non-covalent binding modes. The study will provide a theoretical basis for oyster protein glycation and expand its application on food gel., 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.)

Members of the nuclear factor-kappa B (NF-κB) family are crucial regulators of physiological processes such as apoptosis, inflammation, and the immune response, acting as vital transcription factors to perform their function. In this study, we identified a NF-κB homologous gene (CfRel1) in Zhikong scallops. The 3006-bp-long open reading frame encodes 1001 amino acids. The N-terminus of the CfRel1 protein harbors a conserved Rel homology domain (RHD) that contains a DNA-binding domain and a dimerization domain. According to the multiple sequence alignment results, both the DNA-binding and dimerization domains are highly conserved. Phylogenetic analysis indicated that CfRel1 is closely related to both the Dorsal protein of Pinctada fucata and the Rel2 protein of Crassostrea gigas. CfRel1 mRNA was expressed in all tissues tested in the quantitative reverse transcription PCR experiments, with hepatopancreatic tissue expressing the highest levels. Furthermore, after stimulation with lipopolysaccharide, peptidoglycan, or polyinosinic:polycytidylic acid, the mRNA expression level of CfRel1 was markedly increased. The co-immunoprecipitation test results showed that CfRel1 interacted with scallop IκB protein through its RHD DNA-binding domain, suggesting that IκB may regulate the activity of Rel1 by binding to this domain. Dual-luciferase reporter gene assays revealed that CfRel1 overexpression in HEK293T cells activated the activator protein 1 (AP-1), NF-κB, interferon (IFN)α, IFNβ, and IFNγ reporter genes, indicating the diverse functions of the protein. In summary, CfRel1 is capable of responding to attacks from pathogen-associated molecular patterns, participating in immune signaling, and activating NF-κB and IFN reporter genes. Our findings contribute to the advancement of invertebrate innate immunity theory, enrich the theory of comparative immunology, and serve as a reference for the future screening of disease-resistant strains in scallops., (Copyright © 2023. Published by Elsevier Ltd.)

Municipal wastewater treatment plant (WWTP) effluent is a primary source of pharmaceuticals and personal care products (PPCPs) to the marine environment, as most of these compounds are not fully removed during the treatment process. Continual discharge from WWTPs into coastal areas may act as a stressor by continually exposing organisms to a suite of PPCPs. To quantify organismal exposure to PPCP mixtures, we conducted a 12-week lab experiment that exposed Pacific oysters to effluent from two Oregon coastal WWTPs of different discharge capacities (permitted as <1 million gallons/day and >1 million gallons/day; or < or >3.785 million liters/day) at a dilution of 25 %. Composite samples of weekly collected effluent and a subset of freeze-dried oysters from experiment week 12 were analyzed for PPCPs. Though challenges with food availability inhibited our ability to confidently identify effects of the contaminants on growth and fitness, the experiment allowed us to examine uptake of contaminants from effluent into an estuarine bivalve of commercial importance. We detected 30 PPCPs and three alkylphenols in effluent and 13 PPCPs and four alkylphenols in oyster tissue, indicating high rates of release from secondary treatment and significant potential for marine organism exposure to and uptake of PPCPs in rural coastal areas., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Elise Granek reports financial support was provided by USGS-PSU Partnership. Amy Ehrhart reports financial support was provided by Sigma Xi Grants in Aid of Research. Amy Ehrhart reports financial support was provided by Edward D. and Olive C. Bushby Scholarship from Portland State University., (Copyright © 2023 Elsevier B.V. All rights reserved.)

This study investigates the impact of various zinc supplementation methods on anemia in rats induced by phenylhydrazine (PHZ) and in 5/6-nephrectomized anemic rats. We compare oral zinc sulfate (ZnSO 4 ) supplementation, oyster Crassostrea gigas supplementation, and hard clam Meretrix lusoria supplementation on red blood cell (RBC) levels. Oral zinc-rich oyster supplementation (2.70 mg Zn (30 g oyster)/day/rat) effectively corrects anemia in both experimental groups. Rats orally fed oysters for four days exhibit similar effectiveness as those receiving a single ZnSO 4 injection (0.95 mg Zn (4.18 mg ZnSO 4 ⋅7H 2 O)/rat). In contrast, oral ZnSO 4 supplementation (2.70 mg Zn (11.88 mg ZnSO 4 ⋅7H 2 O)/day/rat) does not significantly increase RBC levels, suggesting better zinc absorption from oysters. A placebo group of anemic rats supplemented with hard clams, similar in composition to oysters but much lower in zinc, did not change RBC counts. This supports oysters' high zinc content as the key to correcting anemia. Oysters also contain high iron levels, offering a potential solution for iron-deficiency anemia while supporting bone marrow erythropoiesis. In summary, oral oyster supplementation emerges as an effective strategy to correct anemia in rats with added zinc and iron support for erythropoiesis.

Dynamic extraction of edible tissues of Jinjiang oyster (Crassostrea rivularis) and Flower clam (Paphia undulata) was conducted using a fully biomimetic digestion (in vitro) method. The impact of different cooking methods on the bioavailability of Zn and Cd in the edible shellfish tissues was analyzed, and the human health risk of Zn and Cd was evaluated. The results show that the gastric biomimetic extractions of Zn and Cd in unheated samples of C. rivularis and P. undulata were higher than those in the intestinal biomimetic extraction. The extraction patterns of cooked samples were consistent with those of raw samples. The bioavailability of Zn and Cd in cooked samples of C. rivularis was 94.9% and 82.5%, respectively, indicating increased Zn bioavailability but decreased Cd bioavailability compared to the raw samples. The bioavailability of Zn and Cd in cooked samples of P. undulata was 85.1% and 83.0%, respectively, both of which decreased compared to the raw samples. Consumption of C. rivularis can provide 21.0% to 34.2% of the daily required Zn intake, while consumption of P. undulata can provide 3.8% to 6.4%. The intake of Cd from both shellfish species is below the monthly tolerable intake recommended by FAO/WHO. Consuming cooked C. rivularis can increase the intake of Zn and decrease the intake of Cd., 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.)

The study was conducted to assess the health risks of selected metals in cultured and wild Crassostrea madrasensis in Sri Lanka over 13 months. Metal concentrations varied monthly, and Zn and Cu concentrations significantly varied between cultured and wild oysters. The mean metal concentrations (mg kg -1 ) were accumulated in order of Zn (1220.60) > Mg (496.97) > Fe (356.30) > Cu (114.95) > Pb (23.20) > Cd (6.15) > As (3.09) > Hg (0.12). The metal concentrations were well below the standard guidelines given by international standards except for Pb, Cd, As and Zn. The weekly intakes of Hg, Cu, Fe, and Zn from C.madrasensis were below the Provisional Tolerable Weekly Intake while Cd exceeded the established limit. The calculated non-carcinogenic and carcinogenic risk indices for Hg, As, Pb and Zn were within safe levels and the values for Cd exceeded the limit revealing carcinogenic results with long-term consumption. Hence, creating proper awareness and a suitable depuration system ensures human health and supports export-oriented markets., Competing Interests: Declaration of competing interest The authors declare that they have no competing interests in the content of this article., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Mangroves represent a challenge in monitoring studies due to their physical and chemical conditions under constant marine and anthropogenic influences. This study investigated metals/metalloids whole-body bioaccumulation (soft tissues) and the risk associated with their uptake, biochemical and morphological detoxification processes in gills and metals/metalloids immobilisation in shells of the neotropical sentinel oyster Crassostrea rhizophorae from two Brazilian estuarine sites. Biochemical and morphological responses indicated three main mechanisms: (1) catalase, superoxide dismutase and glutathione played important roles as the first defence against reactive oxygen species; (2) antioxidant capacity against peroxyl radicals, glutathione S-transferase, metallothionein prevent protein damage and (3) metals/metalloids sequestration into oyster shells as a mechanism of oyster detoxification. However, the estimated daily intake, target hazard quotient, and hazard index showed that the human consumption of oysters would not represent a human health risk. Among 14 analysed metals/metalloids, chemometrics indicate that Mn, As, Pb, Zn and Fe overload the antioxidant system leading to morphological alterations in gills. Overall, results indicated cellular vacuolization and increases in mucous cell density as defence mechanisms to prevent metals/metalloids accumulation and the reduction in gill cilia; these have long-term implications in respiration and feeding and, consequently, for growth and development. The integration of data from different sites and environmental conditions using chemometrics highlights the main biological patterns of detoxification from a neotropical estuarine bivalve, indicating the way in which species can cope with metals/metalloids contamination and its ecological consequences., 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. Published by Elsevier Ltd.)

Neuroendocrine-immune system is an evolution-conserved regulatory network in maintaining the homeostasis of animals. While knowledge on the roles of neuroendocrine-immune system in the disease and stress responses of organisms is growing, the ecological roles of neuroendocrine-immune system, especially how it shapes the unique lifestyle of organisms remain insufficiently investigated. As an endemic and dominant mollusc in intertidal region, oysters have evolved with a primitive neuroendocrine-immune system and with a sessile lifestyle. Recently, a novel neuroendocrine-immune pathway, Ca 2+ /calmodulin (CaM)-nitrite oxide synthase (NOS)/nitrite oxide (NO)-tumor necrosis factor (TNF) pathway, is identified in oysters and found altered dynamically during aerial exposure, one common but challenging stresses for intertidal organisms and a decisive factor shaping their habitat. Since the pathway proves fatal in prolonged aerial exposure, we hypothesized that the activation/deactivation of pathway could be strictly modulated in adaptation to the sessile lifestyle of oysters. Here, a synergistic modulation on the Ca 2+ /CaM-NOS/NO-TNF pathway by four members of miR-92 family and two oyster-specific miRNAs was identified, which further hallmarks the resilience and survival strategy of oysters to aerial exposure. Briefly, these six miRNAs were down-regulating CgCaM24243 post-transcriptionally and deactivating the pathway during the early-stage of stress. However, a robust recession of these miRNAs occurred at the late-stage of stress, resulting in the reactivation of pathway and overwhelming accumulation of cytokines. These results demonstrated a complicated interaction between miRNAs and ancient neuroendocrine-immune system, which facilitates the environmental adaptation of intertidal oysters and provides novel insight on the function and evolution of neuroendocrine-immune system in ecological context., Competing Interests: Declaration of competing interest The authors declare no competing interests., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Oysters, which are flavourful edible marine products, have been utilised to produce Maillard reaction products (MRPs), which contribute to saltiness enhancement. Here, the molecular weight distribution, free amino acids, and taste characteristics of MRPs were analysed, while ultraviolet light was used to observe the Maillard reaction. Both thermal degradation and cross-linking reactions occur during the Maillard reaction. When the Maillard reaction time was 90 min, the saltiness, umami, and richness of the MRPs peaked, however bitterness reached its lowest value. Moreover, at an MRP concentration of 1.5 mg/mL, salts were reduced by 35.71% in a 3 mg/mL sodium chloride solution without reducing saltiness, based on sensory evaluation. Glycation sites of the MRPs, which are crucial for saltiness enhancement and derived from a variety of protein sources, were determined using nano-HPLC-MS/MS analysis. Our study establishes the foundation for preparing salt-enhancing peptides, accelerating the popularisation of oyster-derived flavouring agents., 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.)

Background: Host genetics influences the development of infectious diseases in many agricultural animal species. Identifying genes associated with disease development has the potential to make selective breeding for disease tolerance more likely to succeed through the selection of different genes in diverse signaling pathways. In this study, four families of Pacific oysters (Crassostrea gigas) were identified to be segregating for a quantitative trait locus (QTL) on chromosome 8. This QTL was previously found to be associated with basal antiviral gene expression and survival to ostreid herpesvirus 1 (OsHV-1) mortality events in Tomales Bay, California. Individuals from these four families were phenotyped and genotyped in an attempt to find candidate genes associated with the QTL on chromosome 8., Results: Genome-wide allele frequencies of oysters from each family prior to being planting in Tomales Bay were compared with the allele frequencies of oysters from respective families that survived an OsHV-1 mortality event. Six significant unique QTL were identified in two families in these genome-wide allele frequency studies, all of which were located on chromosome 8. Three QTL were assigned to candidate genes (ABCA1, PIK3R1, and WBP2) that have been previously associated with antiviral innate immunity in vertebrates., Conclusion: The identification of vertebrate antiviral innate immunity genes as candidate genes involved in molluscan antiviral innate immunity reinforces the similarities between the innate immune systems of these two groups. Causal variant identification in these candidate genes will enable future functional studies of these genes in an effort to better understand their antiviral modes of action., (© 2023. BioMed Central Ltd., part of Springer Nature.)

Pollutant exposure is considered an important factor responsible for the decline in marine biodiversity of Latin American coastal ecosystems. This threat has been detected in an estuarine system in southern Brazil, which prompted an investigation into the long-term biological effects of a chronic metal contamination on resident oysters from the Laguna Estuarine System (LES). Here, we present the species- and size-specific variations of biomarker responses (catalase, glucose-6-phosphate dehydrogenase, glutathione S-transferase, and protein carbonylation) in the gills and digestive gland of Crassostrea gigas and Crassostrea gasar. In parallel, concentrations of eight metals (Al, Cd, Cr, Cu, Fe, Mn, Pb, Zn) in soft tissues were measured. Our analyses revealed that the metal levels exhibited decreasing order in both species: Zn > Fe > Al > Cu > Mn > Cd. Except for Cu and Al, metal concentrations did not differ between oyster species. Biomarker results highlighted that C. gasar presented higher antioxidant responses, whereas C. gigas showed increased biotransformation upon exposure to LES pollutants, which varied according to the tissue. However, C. gasar showed a significant higher content of protein carbonylation but was not related to metals. In our research approach, the observation of metals presence and biomarkers-related responses are considered biologically relevant from an ecotoxicological perspective and serve as a baseline for future pollution studies in estuaries of Latin America. Finally, we recommend adopting a suite of biomarkers in both C. gasar and C. gigas, regardless their size and weight, as sentinel organisms in future regional biomonitoring studies in southern Brazil., 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.)

Toll-like receptors (TLRs) are expressed on various immune cells as key elements of innate and adaptive immunity, and they also play significant roles in regulating cell proliferation and differentiation. In the present study, the binding activity of CgTLR3 to PAMPs and CgMyD88-2, and its role in mediating the proliferation of haemocytes was investigated. The recombinant proteins of the extracellular six LRR domains (rCgTLR3-LRR) and intracellular TIR domain (rCgTLR3-TIR) of CgTLR3 were obtained respectively. rCgTLR3-LRR exhibited binding activity to lipopolysaccharide (LPS), peptidoglycan (PGN), mannan (MAN) and Poly (I:C), with the highest affinity for LPS. While rCgTLR3-TIR displayed binding activity to the recombinant protein of rCgMyD88-2, with K D value of 7.22 × 10 -7  M. The CgTLR3 mRNA and protein were detected in three subpopulations of oyster haemocytes, and they were mainly concentrated in granulocytes, which was 7.27-fold (p < 0.05) of that in semi-granulocytes and 8.51-fold (p < 0.01) of that in agranulocytes. The percentage of CgTLR3 positive cells (FITC + haemocytes) in granulocytes was 4.45-fold (p < 0.01) and 2.57-fold (p < 0.05) of that in agranulocytes and semi-granulocytes, respectively. After Vibrio splendidus stimulation, the mRNA expression level of CgTLR3 in haemocytes significantly upregulated at 6 h and 12 h, which was 2.93-fold (p < 0.05) and 4.15-fold (p < 0.05) of that in the control group. After the expression of CgTLR3 was inhibited by the injection of si-CgTLR3, the expression levels of transcription factors associated with hematopoiesis (CgGATA, CgRunx), cell cycle-related genes (CgPCNA, CgCDC-45, CgCDK-2), the agranulocyte marker CgCD-9, the granulocyte marker CgAATase, and the inflammatory factor CgIL17-1 significantly decreased (p < 0.05) after the V. splendidus stimulation, which were 0.43-fold, 0.83-fold, 0.48-fold, 0.44-fold, 0.53-fold, 0.7-fold, 0.62-fold, and 0.47-fold of that in NC + V. s group in vivo, respectively. Meanwhile, the percentage of EdU + haemocytes in si-CgTLR3+V. s group was significantly reduced by 0.54-fold (p < 0.05) compared to the control group (2.7%). These results collectively indicated that CgTLR3 was involved in modulating the proliferation of haemocytes by regulating the expression of proliferation-related genes and inflammatory factor in oyster C. gigas., (Copyright © 2023. Published by Elsevier Ltd.)

Autophagy related 16-like (ATG16L) protein is a core autophagy protein, which promotes the extension of autophagosome membrane through microtubule-associated protein light chain 3 (LC3). In the present study, an ATG16L was identified from oyster Crassostrea gigas (defined as CgATG16L1). The full-length cDNA of CgATG16L1 was of 3184 bp with an open reading frame of 1650 bp that encoded a polypeptide of 549 amino acids. There was an ATG5-interacting motif (AFIM) domain, a coiled-coil (CC) domain and seven tryptophan-aspartic acid 40 (WD40) repeats in CgATG16L1. ATG16L1 mRNA was expressed in all the examined tissues with the highest expression in haemolymph (11.22-fold of that in hepatopancreas, p < 0.05). The mRNA expressions of CgATG16L1 in haemocytes increased significantly at 3, 6, 12, 24 and 72 h after lipopolysaccharide (LPS) stimulation, which were 81.15-fold, 24.95-fold, 6.02-fold, 3.90-fold and 5.97-fold (p < 0.05) of that in control group, respectively. The green positive signals of CgATG16L1 protein and the red positive signals of CgLC3 protein were dotted in the cytoplasm of agranulocytes, semi-granulocytes and granulocytes. The co-localization of CgATG16L1 and CgLC3 was observed in haemocytes after Vibrio splendidus stimulation. In CgATG16L1-RNAi oysters, the number of autophagosomes and autolysosomes in haemocytes was reduced. All these results suggested that CgATG16L1 participated in the bacteria-induced autophagy process in the haemocytes of oyster response to bacteria invasion., 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. Published by Elsevier Ltd.)