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The present study was conducted to investigate the distribution of pit organ in the critically endangered Chinese sturgeon (Acipenser sinensis), the similarities and differences in the microstructure of pit organ between Hybrid sturgeon and Chinese sturgeon, and to provide a histological basis for studying the pit organ in sturgeon species. We used hematoxylin-eosin (HE) staining, nissl staining, multi-layer fine scanning, and scanning electron microscopy to compare and analyse the microstructure of pit organ between Hybrid sturgeon and Chinese sturgeon. The results showed that the structure of pit organ of Hybrid sturgeon and Chinese sturgeon are similar. The distribution density of pit organ in different parts of Chinese sturgeon was different, and the density of pit organ on the ventral surface of the snout was the highest. Scanning electron microscopy revealed that the pit organ of Hybrid sturgeon and Chinese sturgeon were densely packed and about 500–1000 μm wide. Nissl staining revealed that more peripheral nerves are distributed under the sensory and support cells of the pit organ. Based on previous research and our results, the increase in electrical receptivity of sturgeons with individual development is not only due to the increase in the number of inductive receptors, but may also be related to the increase in the width of the pit organ. [ABSTRACT FROM AUTHOR]

Microplastics (MPs) and organophosphate flame retardants (OPFRs) have recently become ubiquitous and cumulative pollutants in the oceans. Since OPFRs are added to or adsorbed onto MPs as additives, it is necessary to study the composite contamination of OPFRs and MPs, with less focus on bio-based PLA. Therefore, this study focused on the ecotoxicity of the biodegradable MP polylactic acid (PLA) (5 μm, irregular fragments, 102 and 106 particles/L), and a representative OPFRs tris(1-chloro-2-propyl) phosphate (TCPP, 0.5 and 50 μg/L) at environmental and high concentrations. The mussel Mytilus coruscus was used as a standardised bioindicator for exposure experiments. The focus was on examining oxidative stress (catalase, CAT, superoxide dismutase, SOD, malondialdehyde, MDA), immune responses acid (phosphatase, ACP, alkaline phosphatase, AKP, lysozyme, LZM), neurotoxicity (acetylcholinesterase, AChE), energy metabolism (lactate dehydrogenase, LDH, succinate dehydrogenase, SDH, hexokinase, HK), and physiological indices (absorption efficiency, AE, excretion rate, ER, respiration rate, RR, condition index, CI) after 14 days exposure. The results of significantly increased oxidative stress and immune responses, and significantly disturbed energy metabolism and physiological activities, together with an integrated biomarker response (IBR) analysis, indicate that bio-based PLA MPs and TCPP could cause adverse effects on mussels. Meanwhile, TCPP interacted significantly with PLA, especially at environmental concentrations, resulting in more severe negative impacts on oxidative and immune stress, and neurotoxicity. The more severe adverse effects at environmental concentrations indicate higher ecological risks of PLA, TCPP and their combination in the real marine environment. Our study presents reliable data on the complex effects of bio-based MP PLA, TCPP and their combination on marine organisms and the environment. [Display omitted] • The bio-based MP PLA can cause adverse effects similar to traditional MPs. • PLA and TCPP could cause oxidative stress and immune responses. • Disturbances in energy metabolism activities after exposure to PLA and TCPP. • Adverse effects on marine organisms were more severe at environmental concentrations. • The combination of TCPP and PLA can exacerbate the adverse impacts. [ABSTRACT FROM AUTHOR]

Pollutants often exist as mixtures in environmental settings, creating a challenge in selecting the most effective combination of biomarkers for routine monitoring. This study was conducted seasonally in Victoria Harbour, Hong Kong, to compare the responses of nine biomarkers in the green-lipped mussel Perna viridis with respect to its tissue levels of persistent organic pollutants and heavy metals. Multivariate statistical techniques were utilised to determine the single best predictor and optimal subset of biomarkers in P. viridis for each of the four scenarios: representing overall biomarker responses in the dry season, and wet season, as well as correlating tissue levels of mixed pollutants in the dry season, and wet season. Our findings recommend lysosomal destabilisation, and the nucleic acid ratio of RNA to DNA, as the core biomarkers in P. viridis for marine pollution monitoring. The non-specificity of these biomarkers allows effective identification of pollution hotspots and guides further detailed assessment. • Mussels were used to evaluate nine biomarkers for pollution monitoring • Lysosomal destabilisation and RNA:DNA ratio are recommended predictors • Effective screening tools for pollution's biological impacts are presented [ABSTRACT FROM AUTHOR]

In the marine environment, nanoparticles play a role in adsorbing and catalytically degrading organic pollutants, thereby mitigating their toxic effects on aquatic organisms. This study aimed to investigate the impact of nano titanium dioxide (nTiO 2) and tris (2-chloropropyl) phosphate (TCPP) on the hemolymph and digestive function of the thick-shell mussel Mytilus coruscus. Mussels were divided into a control group, a group exposed to TCPP alone, a group exposed to a combination of TCPP and 0.5 mg/L nTiO 2 , and a group exposed to a combination of TCPP and 1 mg/L nTiO 2. After 14 days of exposure, oxidative stress responses, including superoxide dismutase (SOD) activity and malondialdehyde (MDA) content, immune defense responses, including acid phosphatase (ACP) and alkaline phosphatase (AKP) activities, and gene expression, including HSP70 expression, were measured in the hemolymph and digestive glands of the mussels. Compared to the control group, mussels solely exposed to 100 μg/L TCPP exhibited a significant reduction in SOD activity in the hemolymph. When TCPP was co-exposed with 0.5 mg/L nTiO 2 , there were significant increases in MDA content and AKP activity in both the digestive gland and hemolymph compared to the control group. Upon co-exposure of TCPP with 1 mg/L nTiO 2 , MDA content and AKP activity in the digestive gland significantly decreased, while SOD, ACP, and AKP activity in the hemolymph significantly increased and MDA content significantly decreased, returning to the control group levels. Furthermore, in the combined exposure, HSP70 gene expression significantly decreased as the nTiO 2 concentration increased from 0.5 mg/L to 1 mg/L. In summary, TCPP impacted the hemolymph and digestive function of mussels, whereas a concentration of 1 mg/L nTiO 2 effectively alleviated the toxic effects of TCPP. This study is crucial for assessing the ecological risks of nanoparticles and emerging organic pollutants in marine environments, and provides new insights into the interaction between nTiO 2 and TCPP, as well as the influence of nTiO 2 concentration on mitigating TCPP toxicity. [Display omitted] • Low concentration of nTiO 2 enhances the toxic effects of TCPP on mussels. • High concentration of nTiO 2 reduces the toxic effects of TCPP on mussels. • Mussels exhibit tissue-specific responses under combined low-concentration nTiO 2 and TCPP stress. [ABSTRACT FROM AUTHOR]

The green-lipped mussel Perna viridis was utilised for pollution biomonitoring in Victoria Harbour and its adjacent aquaculture area in Hong Kong. P. viridis was collected from a reference site and redeployed at five study sites for five weeks during the dry and wet seasons of 2019. Our study found various polycyclic aromatic hydrocarbons (PAHs) and heavy metals in the mussel tissue, while polychlorinated biphenyls (PCBs) and organochlorine pesticides (OCPs) were not detected. P. viridis at the reference site generally displayed lower levels of pollutants. Comparing with previous research in the 1980s and 2000s, we observed substantial reduction in the tissue levels of PAHs, PCBs, OCPs and heavy metals in P. viridis. The human health risks associated with consuming these mussels were determined to be insignificant. Our findings imply that the Harbour Area Treatment Scheme has been effective in improving the water quality in Victoria Harbour and its adjacent aquaculture area. • Mussel biomonitoring to indicate bioavailability of pollutants • Water quality improvement in Victoria Harbour and its eastern aquaculture area • Minimal human health risks of consuming mussels from the study sites • Effectiveness of the chemically enhanced primary treatment [ABSTRACT FROM AUTHOR]

Coastal habitats are exposed to increasing pressure of nanopollutants commonly combined with warming due to the seasonal temperature cycles and global climate change. To investigate the toxicological effects of TiO 2 nanoparticles (TiO 2 NPs) and elevated temperature on the intestinal health of the mussels (Mytilus coruscus), the mussels were exposed to 0.1 mg/L TiO 2 NPs with different crystal structures for 14 days at 20 °C and 28 °C, respectively. Compared to 20 °C, the agglomeration of TiO 2 NPs was more serious at 28 °C. Exposure to TiO 2 NPs led to elevated mortality of M. coruscus and modified the intestinal microbial community as shown by 16S rRNA sequence analysis. Exposure to TiO 2 NPs changed the relative abundance of Bacteroidetes, Proteobacteria and Firmicutes. The relative abundances of putative mutualistic symbionts Tenericutes and Fusobacteria increased in the gut of M. coruscus exposed to anatase, which have contributed to the lower mortality in this group. LEfSe showed the combined stress of warming and TiO 2 NPs increased the risk of M. coruscus being infected with potential pathogenic bacteria. This study emphasizes the toxicity differences between crystal structures of TiO 2 NPs, and will provides an important reference for analyzing the physiological and ecological effects of nanomaterial pollution on bivalves under the background of global climate change. [Display omitted] • Exposure to TiO 2 NPs led to elevated mortality of M. coruscus and modified the gut microbial community. • The response mechanisms of gut microbiota to TiO 2 NPs with different crystal structure were different. • TiO 2 -induced changes of gut microbiota were correlated with community function and host metabolism alterations. • The agglomeration behaviour of TiO 2 NPs and gut microbiota was modified by temperature. • An interaction between TiO 2 NPs and temperature on the gut microbiota of M. coruscus was found. [ABSTRACT FROM AUTHOR]

Pollution with complex mixtures of contaminants including micro- and nano-plastics (MNPs) and organic pollutants like polycyclic aromatic hydrocarbons (PAH) poses a major threat to coastal marine ecosystems. Toxic mechanisms of contaminant mixtures are not well understood in marine organisms. We studied the effects of single and combined exposures to polycyclic aromatic hydrocarbon phenanthrene (Phe) and MNPs mixture with sizes of 70 nm, 5 μm and 100 μm on the immune health and oxidative stress parameters in the thick-shell mussel Mytilus coruscus. Immune cells (hemocytes) were more sensitive to the pollutant-induced oxidative stress than the gills. In hemocytes of co-exposed mussels, elevated mortality, lower lysosomal content, high production of reactive oxygen species (ROS) and decrease mitochondrial were found. Disparate responses of antioxidant enzymes in the hemolymph (e.g. increased superoxide dismutase (SOD) activity without a corresponding increase in catalase (CAT) in Phe exposures and an increase in CAT without a change in SOD in MNPs exposures) suggests misbalance of the antioxidant defense in the pollutant-exposed mussels. Gill lacked pronounced oxidative stress response showing a decline in ROS and antioxidant levels. Tissue-specific single and combined effects of Phe and MNPs suggest variation in bioavailability and/or different sensitivity to these pollutants in the studied tissues. Notably, the combined effects of MNPs and Phe were additive or antagonistic, showing that MNPs do not enhance and occasionally mitigate the toxic effects of Phe on the hemocytes and the gills of the mussels. Overall, our study sheds light on the impact of long-term exposure to MNPs and Phe mixtures on mussels, showing high sensitivity of the immune system and modulation of the Phe toxicity by MNPs co-exposure. These findings that may have implications for understanding the impacts of combined PAH and MNPs pollution on the health of mussel populations from polluted coastal habitats. [Display omitted] • Single pollutant exposures to phenanthrene (Phe) and MNPs disrupted metabolic and redox status of M. coruscus hemocytes. • Antioxidant defense was stimulated by pollutants in the hemocytes and suppressed in the gills. • Integrated biomarker response showed concentration-dependent cellular stress in Phe-exposed mussels. • Combined effects and interaction reflected the mitigating effects of co-exposures. [ABSTRACT FROM AUTHOR]

Microplastics have been found to adhere to the surface of specific tissues or organs other than being ingested by the organisms. To further test the hypothesis that microplastics might get into specific body parts of organisms, mussel byssus was chosen as a target subject in the present study. In the field investigation, microplastics were found in mussel byssus, and the abundance of microplastics was 0.85–1.02 items/individual mussel and 3.69–9.16 items/g byssus, but the location of microplastics in byssus was not easily determined. Therefore, we simulated environmental conditions in the laboratory for mussels to form fresh byssus in the presence of microplastics. Three types of man-made microplastics (Polystyrene beads, Polyamide fragments, and Polyester fibers) were found in newly formed byssus of mussels after exposure to these test materials. We observed that microplastics not only adhered to the surface but also fused into the byssus of mussels. Since byssus is important for the well-being of mussels, the incorporation of microplastics into the byssus might impair the function of byssus. To the authors' best knowledge, this is the first study to show that microplastics can contact and fuse with the byssus of mussels during their formation, suggesting possible alternations for mussels to grip and interact with microplastics in the aquatic environments. Image 1 • Microplastic pollution was ubiquitous in mussel byssus in the field. • Microplastics were found to accumulate in newly formed mussel byssus. • Three types of microplastics were found to attach to the byssus in laboratory observations. • Polystyrene microbeads were observed to be incorporated into the structure of byssal threads. Microplastics in mussel byssus. [ABSTRACT FROM AUTHOR]

The effects of short-term (7 days) experimental ocean acidification (−0.4 pH units) and warming (+5 °C) on anti-predator defenses of two sympatric Mytilus species from China, M. coruscus and M. edulis , in the presence and absence of predator cues were investigated. Results suggested species-specific independent negative effects of acidification and warming on the number and weight of byssal threads, the force of thread attachment, and total thread plaque area. Similar negative effects were observed for clustering behaviour, with acidification and warming independently increasing the number of solitary individuals and decreasing the percentage of mussels in clusters. Acidification effects on byssus were strongly exacerbated when predators were present. Ultimately, this study suggests that short-term exposure to experimental warming and acidification can negatively impact anti-predator defense strategies in mussels with potential ramifications for predator-prey interactions and ecological functioning in systems where mussel beds play a key ecological role. Unlabelled Image • Predator, pH and temperature affected the mussels' anti-predator behaviour. • Species specific effects were observed between Mytilus coruscus and Mytilus edulis. • Low pH and high temperature reduced the anti-predation performance of mussels. • Predator induced mussel clustering while low pH and warming had negative effects. [ABSTRACT FROM AUTHOR]

Anthropologic activities caused frequent eutrophication in coastal and estuarine waters, resulting in diel-cycling hypoxia. Given global climate change, extreme weather events often occur, thus salinity fluctuation frequently breaks out in these waters. This study aimed to evaluate the combined effects of salinity and hypoxia on intestinal microbiota and digestive enzymes of Crassostrea hongkongensis. Specifically, we sequenced 16 S rRNA of intestinal microbiota and measured the digestive enzymes trypsin (TRS), lipase (LPS) and amylase (AMY) in oysters exposed for 28 days to three salinities (10, 25 and 35) and two dissolved oxygen conditions, normoxia (6 mg/L) and hypoxia (6 mg/L for 12 h, 2 mg/L for 12 h). Oysters in normoxia and salinity of 25 were treated as control. After 28-day exposure, for microbial components, Fusobacteriota, Firmicutes, Bacteroidota, Proteobacteria and Actinobacteriota comprised the majority for all experimental groups. Compared with the control group, the diversity and structure of intestinal microbiota tended to change in all treated groups. The species richness in C. hongkongensis intestine also changed. It was the most significant that high salinity increased Proteobacteria proportion while low salinity and hypoxia increased Fusobacteriota but decreased Proteobacteria, respectively. Additionally, Actinobacteriota was sensitive and changed under environmental stressor (P < 0.01). The prediction results on intestinal microbiota showed that, all functions of oysters were up-regulated to distinct degrees under low/high salinity with hypoxia. According to the KEGG prediction, cellular processes were more active and energy metabolism upregulated, indicating the adaptation of C. hongkongensis to environmental change. Periodical hypoxia and low/high salinity had complex effect on the digestive enzymes, in which the activity of TRS and LPS decreased while AMY increased. High/low salinity and periodical hypoxia can change the secretion of digestive enzymes and influence intestinal microbial diversity and species richness of C. hongkongensis , deducing the chronic adverse effects on the digestive physiology in long-term exposure. [Display omitted] • The combined effects of salinity and hypoxia on oyster were studied. • The intestinal health of oysters was affected by the combined stress. • Hypoxia and low/high salinity decreased diversity and richness of intestinal flora. • Both stressors increased amylase but decreased trypsin and lipase. • Oyster adjusted digestive activity and enhanced energy metabolism to resist stress. [ABSTRACT FROM AUTHOR]

Anthropologic activities caused frequent eutrophication in coastal and estuarine waters, resulting in diel-cycling hypoxia. Given global climate change, extreme weather events often occur, thus salinity fluctuation frequently breaks out in these waters. This study aimed to evaluate the combined effects of salinity and hypoxia on intestinal microbiota and digestive enzymes of Crassostrea hongkongensis. Specifically, we sequenced 16 S rRNA of intestinal microbiota and measured the digestive enzymes trypsin (TRS), lipase (LPS) and amylase (AMY) in oysters exposed for 28 days to three salinities (10, 25 and 35) and two dissolved oxygen conditions, normoxia (6 mg/L) and hypoxia (6 mg/L for 12 h, 2 mg/L for 12 h). Oysters in normoxia and salinity of 25 were treated as control. After 28-day exposure, for microbial components, Fusobacteriota, Firmicutes, Bacteroidota, Proteobacteria and Actinobacteriota comprised the majority for all experimental groups. Compared with the control group, the diversity and structure of intestinal microbiota tended to change in all treated groups. The species richness in C. hongkongensis intestine also changed. It was the most significant that high salinity increased Proteobacteria proportion while low salinity and hypoxia increased Fusobacteriota but decreased Proteobacteria, respectively. Additionally, Actinobacteriota was sensitive and changed under environmental stressor (P < 0.01). The prediction results on intestinal microbiota showed that, all functions of oysters were up-regulated to distinct degrees under low/high salinity with hypoxia. According to the KEGG prediction, cellular processes were more active and energy metabolism upregulated, indicating the adaptation of C. hongkongensis to environmental change. Periodical hypoxia and low/high salinity had complex effect on the digestive enzymes, in which the activity of TRS and LPS decreased while AMY increased. High/low salinity and periodical hypoxia can change the secretion of digestive enzymes and influence intestinal microbial diversity and species richness of C. hongkongensis , deducing the chronic adverse effects on the digestive physiology in long-term exposure. [Display omitted] • The combined effects of salinity and hypoxia on oyster were studied. • The intestinal health of oysters was affected by the combined stress. • Hypoxia and low/high salinity decreased diversity and richness of intestinal flora. • Both stressors increased amylase but decreased trypsin and lipase. • Oyster adjusted digestive activity and enhanced energy metabolism to resist stress. [ABSTRACT FROM AUTHOR]

We examined prey selection and foraging behaviors of the crab Charybdis japonica exposed to four combinations of pH (7.3 and 8.1) and temperature (18 °C and 25 °C). The order of prey selection by C. japonica was Potamocorbula laevis , Ruditapes philippinarum , Tegillarca granosa and Mactra veneriformis . Under high pCO 2 , times for searching, breaking, eating and handling were all significantly longer than those at the normal pCO 2 , and the prey profitability and predation rate under high pCO 2 were significantly lower than normal pCO 2 . Moreover, temperature significantly influenced the foraging behaviors, but its effects were not as strong as those of pH; times for searching, eating and handling under high temperature were significantly lower than the low temperature, and the prey predation rates under high temperature was significantly higher than low temperature. In conclusion, high pCO 2 negatively affected the foraging behavior, but high temperature actively stimulated the foraging behaviors of crabs. [ABSTRACT FROM AUTHOR]

Ocean acidification (OA) and hypoxic events are increasing worldwide problems, their interactive effects have not been well clarified, although their co-occurrence is prevalent. The East China Sea (the Yangtze River estuary area) suffers from not only coastal hypoxia but also pH fluctuation, representing an ideal study site to explore the combined effect of OA and hypoxia on marine bivalves. We experimentally evaluated the antioxidant response of the mussel Mytilus coruscus exposed to three pH levels (8.1, 7.7 and 7.3) at two dissolved oxygen (DO) levels (2.0 mg L −1 and 6.0 mg L −1 ) for 72 h. Activities of superoxide dismutase, catalase, glutathione peroxidase, acid phosphatase, and alkaline phosphatase and levels of malondialdehyde were measured in gills and hemolymph. All enzymatic activities in hemolymph and gills followed a similar pattern throughout the experiment duration. Generally, low DO showed greater effects on enzyme activities than elevated CO 2 . Significant interactions between DO, pH and time were only observed at superoxide dismutase and catalase in both tissues. PCA revealed positive relationships between most enzyme activities in both gills and hemolymph with the exception of alkaline phosphatase activity and the level of malondialdehyde in the hemolymph. Overall, our results suggested that decreased pH and low DO induced similar antioxidant responses in the hard shelled mussel, and showed an additive effect on most enzyme activities. The evaluation of multiple environmental stressors, a more realistic scenario than single ones, is crucial to predict the effect of future global changes on coastal species and our results supply some insights on the potential combined effects of reduced pH and DO on marine bivalves. [ABSTRACT FROM AUTHOR]

The impacts of high-density microplastics, namely polyamine 6,6 (nylon), polymethyl methacrylate (PMMA) and polyethylene terephthalate (PET), on growth and behaviour of the endangered tri-spine horseshoe crab Tachypleus tridentatus were investigated for 100 days. Negative changes in wet weight and prosomal width of the juveniles were observed in all treatments of microplastics, but significant difference was only detected in prosomal width between control and PMMA. T. tridentatus became significantly less active upon exposure to nylon and PET. The extent of burrowing by T. tridentatus did not significantly differ among the treatments but was overall significantly reduced towards day 100. T. tridentatus exposed to PET significantly showed the lowest survival probability (30 %), compared to the other treatments (70–90 %). In conclusion, high-density microplastics compromised growth and behaviour of juvenile horseshoe crabs. Among the polymers that were tested, PET was considered more harmful and associated with higher mortality. [Display omitted] • Exposure to microplastics of nylon, PMMA and PET for 100 days • Growth, locomotion and burrowing behaviour compromised • PET more harmful to horseshoe crabs leading to higher mortality [ABSTRACT FROM AUTHOR]

The prevalence of microplastics in the marine environment poses potential health risks to humans through seafood consumption. Relevant data are available but the diverse analytical approaches adopted to characterise microplastics have hampered data comparison among studies. Here, the techniques for extraction and identification of microplastics are summarised among studies of marine mussels and fish, two major groups of seafood. Among the reviewed papers published in 2018–2021, the most common practice to extract microplastics was through tissue digestion in alkaline chemicals (46 % for mussels, 56 % for fish) and oxidative chemicals (28 % for mussels, 12 % for fish). High-density solutions such as sodium chloride could be used to isolate microplastics from other undigested residues by flotation. Polymer analysis of microplastics was mainly carried out with Fourier-transform infrared (FTIR) spectroscopy (58 % for both mussels and fish) and Raman spectroscopy (14 % for mussels, 8 % for fish). Among these methods, we recommend alkaline digestion for microplastic extraction, and the automated mapping approach of FTIR or Raman spectroscopy for microplastic identification. Overall, this study highlights the need for a standard protocol for characterising microplastics in seafood samples. [Display omitted] • Common methods for extraction and identification of microplastics in seafood reviewed • Alkaline chemicals, followed by oxidative chemicals, usually used in biomass digestion to extract microplastics • FTIR, followed by Raman spectroscopy, mainly used to identify the extracted microplastics • The need for a standard protocol for the assessment of microplastics in seafood and other biological samples [ABSTRACT FROM AUTHOR]

Hypoxia often intensifies with rising dissolved CO 2 , but the concurrent effects of hypoxia and acidification on bivalves are largely unknown. In this study, immune responses of hemocytes in the mussel Mytilus coruscus were examined under six combinations of pH (7.3, 7.7 and 8.1) and dissolved oxygen (DO) concentrations (2 mg L − 1 , 6 mg L − 1 ) for 72 h. Generally, total hemocyte account, phagocytosis, esterase and lysosomal content were reduced under low DO and pH conditions, whereas hemocyte mortality and reactive oxygen species production increased under low DO and pH. Both hypoxia and low pH have negative effects on mussels, but the effects of pH are not as strong as DO. Moreover, significant interactions between DO and pH occurred. However, acidification generally doesn't aggravate the effects induced by hypoxia. Acidification and hypoxia may increase disease risk and impact the aquaculture of this species. [ABSTRACT FROM AUTHOR]

Ocean acidification may increase the risk of disease outbreaks that would challenge the future persistence of marine organisms if their immune system and capacity to produce vital structures for survival (e.g., byssus threads produced by bivalves) are compromised by acidified seawater. These potential adverse effects may be exacerbated by microplastic pollution, which is forecast to co-occur with ocean acidification in the future. Thus, we evaluated the impact of ocean acidification and microplastics on the health of a mussel species (Mytilus coruscus) by assessing its physiological performance, immunity and byssus properties. We found that ocean acidification and microplastics not only reduced hemocyte concentration and viability due to elevated oxidative stress, but also undermined phagocytic activity of hemocytes due to lowered energy budget of mussels, which was in turn caused by the reduced feeding performance and energy assimilation. Byssus quality (strength and extensibility) and production were also reduced by ocean acidification and microplastics. To increase the chance of survival with these stressors, the mussels prioritized the synthesis of some byssus proteins (Mfp-4 and Mfp-5) to help maintain adhesion to substrata. Nevertheless, our findings suggest that co-occurrence of ocean acidification and microplastic pollution would increase the susceptibility of bivalves to infectious diseases and dislodgement risk, thereby threatening their survival and undermining their ecological contributions to the community. [Display omitted] • Ocean acidification reduced phagocytic activity and hence immunity of mussels. • The reduced phagocytic activity was associated with lowered energy budget. • Ocean acidification also reduced byssus strength, extensibility and production. • Microplastics can aggravate these negative effects of ocean acidification. • Mussels would be more prone to diseases and dislodgement in future oceans. [ABSTRACT FROM AUTHOR]

Abstract: Flow cytometry was used to examine immune responses in haemocytes of the green-lipped mussel Perna viridis under six combinations of oxygen level (1.5mg O2 l−1, 6.0mg O2 l−1) and temperature (20°C, 25°C and 30°C) at 24h, 48h, 96h and 168h. The mussels were then transferred to normoxic condition (6.0mg O2 l−1) at 20°C for further 24h to study their recovery from the combined hypoxic and temperature stress. Esterase (Est), reactive oxygen species (ROS), lysosome content (Lyso) and phagocytosis (Pha) were reduced at high temperatures, whereas hypoxia resulted in higher haemocyte mortality (HM) and reduced phagocytosis. For HM and Pha, changes were observed after being exposed to the stresses for 96h, whereas only a 24h period was required for ROS and Lyso, and a 48h one for Est. Recovery from the stresses was observed for HM and Pha but not other immune responses. [Copyright &y& Elsevier]

Abstract: Mussels were maintained for 4weeks under different combinations of dissolved oxygen concentration (1.5, 3.0 and 6.0mgO2 l−1) and salinity (15, 20, 25 and 30) in a 3×4 factorial design experiment. Clearance rate (CR), absorption efficiency (AE), respiration rate (RR) and scope for growth (SFG) decreased with decreasing salinity and dissolved oxygen concentration (DO), while excretion rate (ER) increased with decreasing salinity and increasing DO. The O:N ratio was <10 at salinities of 15 and 20, irrespective of DO levels. SFG was negative in most of the treatments, except for those under 6.0mgO2 l−1 or at a salinity of 30 when DO was lower. The results may help explain the distribution pattern of Perna viridis in Hong Kong waters and provide guidelines for mussel culture site selection. [Copyright &y& Elsevier]

Microplastics represent an emerging environmental issue and have been found almost everywhere including seafood, raising a great concern about the ecological and human health risks they pose. This study addressed the common technical challenges in the assessment of microplastics in seafood by developing an improved protocol based on Raman spectroscopy and using the green-lipped mussel Perna viridis and the Japanese jack mackerel Trachurus japonicus as the test models. Our findings identified a type of stainless-steel filter membranes with minimal Raman interference, and a combination of chemicals that achieved 99–100% digestion efficiency for both organic and inorganic biomass. This combined chemical treatment reached 90–100% recovery rates for seven types of microplastics, on which the surface modification was considered negligible and did not affect the accuracy of polymer identification based on Raman spectra, which showed 94–99% similarity to corresponding untreated microplastics. The developed extraction method for microplastics was further combined with an automated Raman mapping approach, from which our results confirmed the presence of microplastics in P. viridis and T. japonicus collected from Hong Kong waters. Identified microplastics included polypropylene, polyethylene, polystyrene and poly(ethylene terephthalate), mainly in the form of fragments and fibres. Our protocol is applicable to other biological samples, and provides an improved alternative to streamline the workflow of microplastic analysis for routine monitoring purposes. [Display omitted] • An improved method to extract microplastics from organic and inorganic biomass. • An automated mapping approach to characterise the extracted microplastics. • The protocol for seafood also applicable to other biological samples. [ABSTRACT FROM AUTHOR]

There are many toxicological studies on microplastics, but little is known about the effect of nanoplastics (NPs). Here, we evaluated the oxidative stress responses induced by NPs (10, 104 and 106 particles/l) in juvenile Larimichthys crocea during 14-d NPs exposure followed by a 7-d recovery. After exposure, the activities of antioxidant enzymes (SOD, CAT, GPx) and MDA levels increased in the liver of fish at the highest NPs concentration. SOD and CAT activities remained elevated above the baseline after recovery under high-concentration NPs but returned to the baseline in two other NP treatments. Although lipid peroxidation in liver was reversible, juvenile fish in NPs treatments exhibited a lower survival rate than the control during both exposure and recovery. Furthermore, IBR value and PCA analysis showed the potential adverse effects of NPs. Considering that NPs can reduce the survival of fish juveniles, impacts of NPs on fishery productivity should be considered. [Display omitted] • Nanoplastics induced oxidative stress in the liver of juvenile fish. • Nanoplastics showed latent effects on antioxidant responses in fish liver. • Nanoplastics resulted in low survival rate of juvenile fish. • After depuration antioxidant performance showed a recovery trend but not survival. [ABSTRACT FROM AUTHOR]

Ocean acidification and hypoxia have become increasingly severe in coastal areas, and their co-occurrence poses emerging threats to coastal ecosystems. Here, we investigated the combined effects of ocean acidification and hypoxia on the reproductive capacity of the thick-shelled mussel Mytilus coruscus. Our results demonstrated low pH but not low oxygen induced decreased gonadosomatic index (GSI) in mussels. Male mussels had a lower level of sex steroids (estradiol, testosterone, and progesterone) when kept at low pH. Expression of genes related to reproduction were also impacted by low pH with a downregulation of genes involved in gonad development in males (β-catenin and Wnt-7b involved in males) and an upregulation of testosterone synthesis inhibition-related gene (Wnt-4) in females. Overall, our results suggest that ocean acidification has an impact on the gonadal development through an alternation of gene expression and level of steroids while hypoxia had no significant effect. [Display omitted] • Ocean acidification adversely affects the gonadal performance of mussels. • Hypoxia does not have effects on fecundity of mussels. • Steroids and Wnt/β-catenin gene expressions have weak correlations with fecundity in mussels. [ABSTRACT FROM AUTHOR]

In the present study, the combined effects of pH, dissolved oxygen (DO) and temperature levels on the antioxidant responses of the mussel Mytilus coruscus were evaluated. Mussels were exposed to two pH (8.1, 7.7-acidification), two DO (6 mg L−1, 2 mg L−1-hypoxia) and two temperature levels (20 °C, 30 °C-warming) for 30 days. SOD, CAT, MDA, GPx, GSH, GST, TAOC, AKP, ACP, GPT, AST levels were measured in the gills of mussels. All tested biochemical parameters were altered by these three environmental stressors. Values for all the test parameters except GSH first increased and then decreased at various experimental treatments during days 15 and 30 as a result of acidification, hypoxia and warming. GSH content always increased with decreased pH, decreased DO and increased temperature. PCA showed a positive correlation among all the measured biochemical indexes. IBR results showed that M. coruscus were adversely affected by reduced pH, low DO and elevated temperature. Unlabelled Image • Low pH, low DO and high temperature have adverse effects on the antioxidant parameters. • Exposure time of stressors has significant effects on the antioxidant parameters. • Combined stressors exert more severe effects on the antioxidant indicators than single ones. [ABSTRACT FROM AUTHOR]

In recent years, antibiotics have been widely detected in coastal waters of China, which raising concerns for coastal biodiversity and aquaculture. This study evaluated the effects of short-term exposure of norfloxacin (NOR) on oxidative stress and intestinal health of the large yellow croaker Pseudosciaena crocea. Juvenile fish were exposed to four concentrations of NOR (0.1, 10, 100 and 1000 μg/L) for 14 days. The results showed that NOR inhibited growth and threatened the survival of juveniles. According to the changes of intestinal microbiota, we found that NOR led to a significant decrease in intestinal microbiota diversity, with the decreased relative abundance of Proteobacteria , but the increased Tenericutes. From the perspective of microbial function, NOR inhibited metabolism, cellular defence mechanism and information transduction process. In terms of biochemical indicators, NOR caused an increase in malondialdehyde (MDA) level and inhibited superoxide dismutase (SOD) and acetyl cholinesterase (AChE) activities. Catalase (CAT) activity was activated at low concentration but significantly inhibited at high concentration of NOR. Moreover, there was a high correlation between change in biochemical indicators and change in the microbial community. Overall, environmentally relevant concentrations (0.1 μg/L) and high concentrations (10, 100 and 1000 μg/L) of NOR have negative effects on the defence function and intestinal health of large yellow croaker juveniles. Image 1 • Norfloxacin harms intestinal health of fish. • Norfloxacin inhibits the antioxidant enzyme activity of fish. • Norfloxacin is neurotoxic to large yellow croaker juveniles. • Environmentally relevant norfloxacin concentrations have toxic effects on fish. Environmentally relevant concentrations of norfloxacin disrupted the homeostasis of the intestine microflora and caused oxidative stress and neurotoxicity in fish. [ABSTRACT FROM AUTHOR]

Microplastic pollution in estuarine and coastal environments has recently been characterised in several countries but few researchers have addressed the influence of different forms of coastal zone use on the distribution of microplastic. Here, microplastic particles were sampled in Hangzhou Bay, which is heavily influenced by a range of human activities, and their abundance, size, and polymer type characterised. The abundance of microplastics was 0.14 ± 0.12 items/m3 in water, 84.3 ± 56.6 items/kg dry weight of sediment, and between 0.25 ± 0.14 and 1.4 ± 0.37 items/individual in biota. These results show that Hangzhou Bay has a low level of microplastic contamination compared to other coastal systems in China, although abundance was spatially variable within the bay; relatively higher microplastic abundances were found in the southern area of the bay, which has adjacent industrial and urban land-use zones, while lower abundances were observed in the central and northern bay areas where mariculture, fisheries, and mineral and energy industries are most common. The relatively low microplastic abundance observed in the biota samples is consistent with the generally low values for the seawater and sediment samples. Pellets were the most common of four particle-shape classes (fibres, fragments, films, and pellets) in surface seawater, while fibres were most abundant in sediment and biota. Smaller-sized microplastics (<1.0 mm) were dominant in all samples. Microplastics in the surface seawater were dominated by low-density polypropylene and polyethylene particles, while rayon was dominant in the sediment and biota samples. Our results demonstrate that regional variability in anthropogenic activity and land-use are important controls on the spatial pattern of microplastic pollution in Hangzhou Bay. Image 1 • Microplastics (MPs) pollution in Hangzhou bay is lower than in adjacent coastal systems overall. • MP pollution in the bay is dominated by small (<1 mm) MP. • The distribution, size, and shape of MPs are closely linked to regional land-use. Human activities associated with different land-use types influence the spatial distribution of microplastics in Hangzhou Bay. [ABSTRACT FROM AUTHOR]

In the marine environment, microplastic contamination and acidification may occur simultaneously, this study evaluated the effects of ocean acidification and microplastics on oxidative stress responses and digestive enzymes in mussels. The thick shell mussels Mytilus coruscus were exposed to four concentrations of polystyrene microspheres (diameter 2 μm, 0, 10, 104 and 106 particles/L) under two pH levels (7.7 and 8.1) for 14 days followed by a 7-day recovery acclimation. Throughout the experiment, we found that microplastics and ocean acidification exerted little oxidative stress to the digestive gland. Only catalase (CAT) and glutathione (GSH) showed a significant increase along with increased microplastics during the experiment, but recovered to the control levels once these stressors were removed. No significant effects of pH and microplastics on glutathione peroxidase (GPx) and superoxide dismutase (SOD) were observed. The responses of digestive enzymes to both stressors were more pronounced than antioxidant enzymes. During the experiment, pepsin (PES), trypsin (TRS), alpha-amylase (AMS) and lipase (LPS) were significantly inhibited under microplastics exposure and this inhibition was aggravated by acidification conditions. Only PES and AMS tended to recover during the recovery period. Lysozyme (LZM) increased significantly under microplastic exposure conditions, but acidification did not exacerbate this effect. Therefore, combined stress of microplastics and ocean acidification slightly impacts oxidative responses but significantly inhibits digestive enzymes in mussels. Image 1 • The immunity and digestion of mussels were impaired by MPs and OA. • OA can enhance the toxicity of MPs to mussels. • Digestion was more severely affected by OA and MPs than antioxidant system. • The negative effects on PES and AMS showed a reversible trend. [ABSTRACT FROM AUTHOR]

Apart from ocean acidification, hypoxia is another stressor to marine organisms, especially those in coastal waters. Their interactive effects of elevated CO 2 and hypoxia on the physiological energetics in mussel Mytilus edulis were evaluated. Mussels were exposed to three pH levels (8.1, 7.7, 7.3) at two dissolved oxygen levels (6 and 2 mg L−1) and clearance rate, absorption efficiency, respiration rate, excretion rate, scope for growth and O: N ratio were measured during a14-day exposure. After exposure, all parameters (except excretion rate) were significantly reduced under low pH and hypoxic conditions, whereas excretion rate was significantly increased. Additive effects of low pH and hypoxia were evident for all parameters and low pH appeared to elicit a stronger effect than hypoxia (2.0 mg L−1). Overall, hypoxia can aggravate the effects of acidification on the physiological energetics of mussels, and their populations may be diminished by these stressors. Unlabelled Image • Combined effects of ocean acidification and hypoxia are investigated in mussels. • Physiological activities of mussels are inhibited by low pH and hypoxia. • OA and hypoxia exert additive effects on the physiological metabolism of mussels. [ABSTRACT FROM AUTHOR]