Synthetic phenolic antioxidants (SPAs), as representative emerging contaminants, have been widely detected across various environmental media and organisms, raising significant ecological and health concerns. However, the toxicological effects and underlying molecular mechanisms of SPAs in marine organisms remain poorly understood. By integrating transcriptomics, in silico simulations, and experimental validation, this study elucidates the "oxidative stress-macromolecular damage-cell death" mechanism in the Ruditapes philippinarum under BHT stress. Our results demonstrate that BHT exposure triggers significant ROS accumulation in the digestive glands. A time-dependent response was observed: an initial induction of antioxidant defenses (0-6 d) was followed by a late-stage (12-21 d) suppression of the NRF2-mediated pathway, culminating in compromised antioxidant capacity and severe macromolecular damage. Mechanistically, BHT-induced ROS disrupts the calcium pump, causing Ca2 + homeostasis imbalance and triggering the ER-mitochondria stress axis, which ultimately initiates apoptosis. Furthermore, BHT impairs iron homeostasis, causing Fe²⁺ overload, which drives ALOX5-mediated lipid peroxidation and may ultimately lead to ferroptosis. Simultaneously, activation of the NLRP3-CASPASE1 signaling cascade may also trigger pyroptosis. Collectively, this study provides comprehensive evidence of how BHT orchestrates oxidative stress-mediated multi-pathway programmed cell death (PCD) in aquatic invertebrates, offering crucial scientific insights for the ecological risk assessment of SPA pollution in marine ecosystems.

Polycyclic aromatic hydrocarbons (PAHs), as a typical type of important persistent organic pollutants (POPs) are hard to control since they are mainly derived from incomplete combustion of organic matters. This study investigates the occurrence, ecological and health risks, and tissue-specific biological effects of 16 priority PAHs in sediments and the clam Mactra veneriformis from Laizhou Bay and Haizhou Bay, China. Sediment PAH concentrations were moderate compared to other Chinese bays but lower than many international systems, with mean effect range-median quotient (M-ERM-Q) values indicating low ecological risk. Tissue-specific analysis revealed the digestive gland accumulated significantly higher levels of PAHs, resulting in elevated health risks (incremental lifetime cancer risk and margin of exposure) compared to gill, mantle, and adductor muscle. A novel tissue-specific integrated biomarker response (IBR) approach was developed, incorporating biomarker screening via Spearman’s correlation with PAHs levels. The digestive gland IBR demonstrated the strongest correlation with pollution and risk indices, supporting its utility as a sensitive monitoring tool for its significance as early warning indicators. This research provides a holistic framework for assessing PAHs contamination using tissue-specific bioaccumulation, biomarker screening, and risk evaluation, offering valuable insights for monitoring POPs in coastal ecosystems.

Dietary plant-derived bioactive compounds for enhancing physiological health are becoming a prevalent strategy for antibiotic alternatives. Our study revealed the effects and underlying mechanism of osthole (OST) and OST-tetramethylpyrazine (TMP) compound in Litopenaeus vannamei based on network pharmacology, molecular docking and a 42-d feeding trial verification. The results illustrated that OST and OST-TMP compound significantly improved the survival rate, weight gain rate, specific growth rate and feed conversion ratio, strengthening the growth performance of L. vannamei. Meanwhile, combining the predictive results from network pharmacology and molecular docking, we propose that OST and TMP synergistically enhance the antioxidant defence capacity of shrimp through the synergistic Nrf2 signalling pathway, thereby enhancing the expression of total antioxidant capacity, superoxide dismutase, catalase and glutathione peroxidase. Furthermore, OST-TMP exhibited a significant increase of the immune response in haemocyte and intestine of shrimp, increasing the expression of antimicrobial peptide and lysozyme and suppressing the inflammatory factors, via the synergistic (NF-κB) and complementary targets predicted by network pharmacology. Additionally, gut microbiota composition of L. vannamei was improved, and the dominant genera were correlated with intestinal immune in the compound groups. For the first time, we elucidated the mechanism of plant-derived bioactive compounds mediating physiological health in aquatic animals via a new strategy of network pharmacology-molecular docking-experimental verification and identified the optimal addition amount of OST-TMP in shrimp (150 mg/kg TMP + 20 mg/kg OST), providing a technical safeguard for the animal health and the safety of aquatic products.

Chlorothalonil is a widely used organochlorine fungicide frequently detected in aquatic ecosystems, yet its mechanisms of developmental toxicity in fish remain incompletely understood. This study employed an integrated phenotype-to-mechanism approach in zebrafish embryos to assess the acute and developmental toxicity of chlorothalonil and elucidate potential molecular pathways. The 96-hour median lethal concentration (LC₅₀) was determined to be 89.5 μg/L. Exposure to concentrations below the acute lethal threshold (20-80 μg/L) resulted in significant developmental impairments, including reduced heart rate and body length, along with increased incidences of pericardial edema, tail malformation, and scoliosis. Transcriptomic analysis of embryos exposed to 50 μg/L chlorothalonil at 24 h post-fertilization revealed coordinated upregulation of xenobiotic metabolism genes and dysregulation of genes essential for cardiac and skeletal development, notably the thyroid hormone receptor gene thrab. Subsequent functional analysis using a zebrafish-specific thyroid hormone receptor (TR) yeast two-hybrid assay demonstrated that chlorothalonil acts as a direct antagonist of both TRα and TRβ, dose-dependently inhibiting triiodothyronine (T3)-induced receptor activation. These findings establish disruption of thyroid hormone signaling as a key mechanistic pathway for chlorothalonil-induced developmental malformations, enhancing the mechanistic understanding of its risk to early-stage development of teleosts.

The farming yield of Penaeus vannamei has been steadily increasing in recent years,positioning it as the highest-producing economic crustacean in global aquaculture.However,with the ongoing depletion of fishmeal resources and the fluctuation of its prices,the need for alternative protein sources has become more urgent.Consequently,plant protein is a promising and sustainable replacement for fishmeal,with the potential to be the primary protein source in aquaculture feeds.Despite its potential,the absorption and utilization of plant protein by aquatic animals,including shrimp,has been challenging because they are not naturally adapted to efficiently digest plant-based proteins.To overcome this,probiotic fermentation of plant protein has become the primary method for enhancing the digestibility and bioavailability of plant-based protein for aquatic species,ultimately improving the overall feed efficiency and growth performance of aquaculture animals.In this study,a Bacillus subtilis strain that is Gram-positive bacterium,non-hemolytic,and sensitive to most antibiotics,named SQVGB8,was isolated and identified from the intestine of P.vannamei,with a 99.20％similarity to Bacillus subtilis subsp.subtilis strain 168.The characteristic analysis results showed that the strain can efficiently produce protease,lipase,amylase,cellulase,and phytase;inhibit Vibrio harveyi,V alginolyticus,and V.parahaemolyticus;secrete active substances such as short-chain fatty acids(SCFAs)and extracellular polymeric substances(EPS);and exhibit high intestinal adhesion properties. The whole genome analysis results showed that SQVGB8 can produce digestive enzymes,phytase,SCFAs,and EPS.Additionally,SQVGB8 can secrete various substances with antibacterial and antimicrobial potential,including surfactin,fengycin,and bacillibactin.These active substances have significant application prospects in antibacterial and pathogen control.The suitable fermentation conditions for SQVGB8 fermented plant meals(soybean meal∶cottonseed meal=3∶1)were determined using a single-factor experiment.The optimal conditions are as follows:inoculum amount 3％-12％,fermentation temperature 35-45 ℃,fermentation time 36-72 h,and solid-to-liquid ratio 1∶1.2-1∶0.6 g/mL.Therefore,a response surface methodology with a four-factor,three-level analysis was used,and the optimal fermentation conditions for SQVGB8 fermented plant meals were an inoculum amount of 7.70％,a fermentation temperature of 35.70 ℃,a fermentation time of 55.00 h,and a solid-to-liquid ratio of 1∶0.78 g/mL.The acid soluble protein and acidic polysaccharide contents were 10.30％and 12 mg/mL,respectively.Compared to that of the unfermented plant meals,the antinutritional factor content was significantly reduced.The trypsin inhibitor,soybean globulin,β-conglycinin,gossypol,and phytic acid contents were reduced by 91.67％,61.57％,76.75％,59.00％,and 48.72％,respectively.In conclusion,SQVGB8 is an efficient enzyme-producing,active substance-secreting,and gut-colonizing probiotic,with high efficacy in fermented plant meals.It has great potential for development and application in aquaculture feed.

Synthetic phenolic antioxidants (SPAs) are emerging contaminants (ECs) of growing concern in marine ecosystems, yet their detoxification and hepatotoxicity in marine invertebrates remain largely unknown. Here, we integrated transcriptomic profiling, in silico prediction, and experimental validation to characterize the detoxification of butylated hydroxytoluene (BHT) in Ruditapes philippinarum and to elucidate its hepatotoxicity. BHT metabolism was transcriptionally regulated by the aryl hydrocarbon receptor (AHR) and hormone receptor 96 (HR96) pathways. Key enzymes included Phase I cytochrome P450 isoforms (CYP1A1, CYP1A2, CYP3A4) and the Phase II enzyme UDP-glucuronosyltransferase (UGT). Metabolite profiling revealed the transformation products in the order: BHT-CHO > BHT-Q > BHT-OH > BHT-COOH, and a cascade detoxification pathway was subsequently proposed. BHT exposure impaired antioxidant defenses, induced oxidative stress, and caused macromolecular damage. Molecular dynamics simulations revealed a compromised DPPC bilayer integrity, suggesting structural membrane damage. In parallel, histopathological analysis exhibited characteristic hepatic lesions and cytoplasmic vacuolization, accompanied by significant increases in inflammatory cytokines (TNF, IL16) and transaminase levels (AST, ALT). Together, these results delineate a metabolism-oxidative stress-hepatotoxicity cascade for BHT in bivalves, providing novel mechanistic insights into SPA-induced toxicity. Our findings highlight the ecological risks posed by persistent SPAs in marine environments and underscore their potential to compromise the health of sentinel bivalves, supporting their inclusion in environmental monitoring and regulatory frameworks.

The growing global emphasis on food safety, coupled with projected increases in seafood consumption, underscores the need for rigorous health risk assessments associated with dietary intake of seafood products. This study investigates the distribution and ecological risk of PAHs in seawater and sediment, and further examines tissue-specific patterns of PAHs bioaccumulation in the clam Ruditapes philippinarum from typical bays of the Bohai Sea and the Yellow Sea. PAHs concentrations in environmental media were within globally reported ranges, with elevated ecological risks observed in a semi-enclosed, urban-adjacent bay (Jiaozhou Bay). Among tissues, the digestive gland consistently showed higher PAH accumulation and bioaccumulation factors than other organs. Health risk assessment approaches (ILCR and MOE) indicated generally acceptable risks from whole soft tissue consumption, while tissue-level scenarios identified the digestive gland as a dominant contributor to potential exposure. These findings provide mechanistic insight into tissue-specific PAHs accumulation and support targeted risk-reduction strategies in seafood consumption.

The ubiquitous presence of synthetic phenolic antioxidants (SPAs) has raised growing concern over their potential ecological and health risks. However, their bioaccumulative toxicity and underlying mechanisms remain poorly understood. Here, this study elucidated the bioaccumulative toxicity of butylated hydroxytoluene (BHT) in the marine invertebrate Ruditapes philippinarum. BHT exhibited pronounced tissue-specific accumulation, accompanied by elevated TG and PL levels. PP-LFER modeling, Spearman's correlation, and LMM analyses, revealed a significant positive association between BHT accumulation and TG/PL contents, suggesting preferential accumulation of BHT in lipid droplets enriched in TG and enclosed by PL. MD simulations demonstrated that BHT interacts with DPPC membranes and its tert-butyl substituents impede transmembrane diffusion. Moreover, BHT may induce ER stress and modulate mTOR-SREBP1 and PPARα/β signaling, together with altered expression of lipid metabolism-related genes, implying disruption of lipid metabolic homeostasis and enhanced lipid deposition. Target prediction and molecular docking further indicated stable binding of BHT to key lipid metabolic regulators. Meanwhile, BHT may activate NF-κB-related signaling, potentially mediated by lipotoxic, neurotransmitter-immune, and oxidative stress pathways, contributing to low-grade inflammation. Collectively, this study provides new insights into SPA-associated lipid metabolic disruption and inflammatory risk in aquatic invertebrates, supporting their health risk assessment.

The mechanisms by which Polycyclic Aromatic Hydrocarbons (PAHs) induce lipid metabolic disorder and inflammation in marine invertebrates remain poorly understood. This study utilized the clam Ruditapes philippinarum during its reproductive stage as a model organism, integrating high-throughput omics, computational simulation, and confocal microscopy to elucidate the accumulation characteristics and toxicological pathways of PAHs. The results demonstrated that PAHs significantly accumulated in the digestive gland and gonads, primarily sequestered within lipid droplets. This tissue distribution was found to be dependent on a lipid-dependent transport mechanism mediated by ApoB, FATP, and FABP4. Mechanistically, PAHs activated SREBP1 and PPARα, β nuclear receptors by interfering with the neuroendocrine system and endoplasmic reticulum stress pathways. This activation resulted in dysregulated lipid metabolism (favoring synthesis over degradation) and subsequent abnormal lipid (TG, PL) deposition. Furthermore, PAHs induced low-grade inflammation by synergistically activating the NF-κB and AP-1 pathways, a response driven by both lipotoxicity and cellular organelle stress. This finding provides important scientific evidence for contaminant risk assessment in aquatic organisms.

Synthetic phenolic antioxidants (SPAs) are ubiquitous marine contaminants with potential toxicity, yet their interference mechanisms on taste-active compound metabolism in aquatic organisms remain uncharacterized. This study employed Ruditapes philippinarum as a model to investigate the effects of butylated hydroxytoluene (BHT, a typical SPA) on taste compounds and their metabolic mechanisms. We quantified taste compound contents, evaluated taste indicators, and analyzed molecular pathways via transcriptomics and qRT-PCR. Results showed that BHT significantly altered the contents of umami (glutamate, GMP, succinic acid), sweet (glycine, betaine, glucose), and bitter (arginine) compounds, with taste activity values (TAV) and equivalent umami concentration (EUC) exhibiting corresponding changes. Mechanistically, BHT disrupted taste compound metabolism by regulating the mTOR and AMPK signaling pathways, as well as key metabolic cascades of specific taste compounds. This study clarifies the interference effects and mechanisms of SPAs on aquatic product taste quality, providing a scientific basis for ecological risk assessment of marine pollutants and quality control of aquatic products.

Synthetic phenolic antioxidants (SPAs) are emerging additive-derived contaminants widely detected in marine ecosystems, yet their source-specific health risks and potential source-oriented biological effects remain unclear. This study quantified 10 SPAs in Mactra veneriformis collected from the Bohai Sea and the Yellow Sea in May and August 2025 and integrated bioaccumulation, multi-pathway biomarker responses, and receptor modeling to establish a source-specific risk assessment framework. Total SPA concentrations ranged from 5802.21 to 14396.50 ng/g d.w. in May and increased to 13129.13 to 23676.85 ng/g d.w. in August. Biomarker analyses partly reflected coordinated activation of detoxification and antioxidant systems, accompanied by oxidative damage, neurotoxicity, and immunomodulation. Although most individual hazard quotients were below 1, assessment results support prioritizing AO1010 as a potential risk compound. Positive matrix factorization tentatively interpreted four sources: plastics (F1), rubber (F2), personal care products (F3), and petroleum/lubricants (F4). By coupling partial least squares regression with PMF, source-specific health risks (PMF-HQ) and potential source-oriented biological effects (PMF-IBR) were quantified alongside. By integrating the results of PMF-IBR and PMF-HQ, we ranked risk contributions as F2 > F1 ≈ F4 > F3, indicating that rubber-related inputs exert disproportionately high toxicological pressure. This study establishes a source-specific risk control priority framework for emerging contaminants in coastal ecosystems.

To enhance the feed utilization efficiency of Penaeus vannamei, six host-derived probiotic strains were selected to develop functional fermented feeds. Based on key bioactive compounds (acid-soluble protein, acidic polysaccharides, pH, SCFAs, and phytase), the optimal strain combination and ratio were determined (SQVGB8: SQVGM26: SQVGY6 = 3:2:1). Subsequently, three response surface methodology experiments (RSM-BBD) were conducted to optimize fermentation parameters using specific biochemical indicators as response values. Under optimized conditions, three fermented feeds with relative functional tendencies were obtained (F1, F2, and F3), which showed biased enrichment toward growth-related, bacteriostatic, and immune-associated properties, respectively. In a 56-day feeding trial, all functional feeds significantly improved shrimp growth performance, digestive enzyme activity, antioxidant capacity, and immune function. Specifically, F1 induced the highest protease activity in the hepatopancreas and intestine, while F3 markedly upregulated antioxidant levels. At the transcriptional level, functional fermented feeds upregulated the expression of key genes involved in the TLR4 and IMD immune defense pathways, while downregulating inflammation-related genes in the MAPK signaling pathway. Additionally, the fermented feeds reshaped the gut microbiota, increasing probiotic abundance and community stability. In conclusion, this work successfully established a multi-index RSM strategy to develop functional fermented feeds. While all three products shared a foundational efficacy in enhancing digestion and absorption to support growth and maintaining intestinal health, they exhibited relative functional tendencies in regulating growth performance, immune responses, and gut homeostasis, thereby providing a flexible framework for fermentation-based feed development in aquaculture.

With the advancement of global industrialization and urbanization, agricultural non-point source pollution, along with discharges from industrial and domestic sewage, has led to the proliferation of pathogenic bacteria in aquatic environments and a dramatic increase in nitrogen/sulfur pollutant loads, severely threatening aquatic ecological security and human health. In this study, a multifunctional safe probiotic strain, Lactobacillus paracasei SDVWA5, was screened from shrimp aquaculture environments. It exhibited inhibition rates exceeding 97.78% against five common aquatic pathogenic bacteria, and removal rates of more than 94.80% and 89.03% for nitrogen compounds (NH4+-N, NO2--N, and NO3--N) and H2S, respectively. Moreover, it demonstrated robust environmental tolerance (temperature 22-40 degrees C, pH 4.5-9.5, salinity 0-40, and C/N = 5-20). Genome sequencing and experimental validation revealed that SDVWA5 exerts antibacterial effects through the synergistic action of organic acids, class II bacteriocins, and exopolysaccharides. It achieves efficient nitrogen removal via ammonium assimilation, assimilatory nitrate reduction to ammonium, and denitrification pathways, and exhibits sulfide-removal potential potentially associated with rDsr-related genes. This strain maintained highefficiency multifunctional purification in aquaculture wastewater and domestic sewage. Thus, this study provides high-quality microbial resources and technological support for ecological disease control and composite nitrogen-sulfur pollution remediation in aquatic environments.

This study investigated the effects of glycerol monolaurate (GML) on the growth performance, physiological efficacy, intestinal structure, and microbiota of hybrid juvenile ♀ Epinephelus fuscoguttatus × ♂ E. lanceolatus. A total of 594 groupers were administered diets supplemented with 0, 2, 4, 6, 8, or 10 g/kg of GML over an 80-day period. The findings indicated that the group receiving 4 g/kg GML supplementation demonstrated significantly enhanced growth performance, as evidenced by increased weight gain, specific growth rate, and a reduced feed conversion ratio compared with the control and other experimental groups. This growth enhancement was associated with elevated catalase and glutathione levels, suggesting improved antioxidant capacity that may contribute to enhanced immune function. Furthermore, fish in the 4 g/kg group exhibited increased digestive enzyme activity and significantly improved intestinal morphology, characterized by thicker muscular layers and a greater fold width and height. Furthermore, compared with the control group, GML supplementation elevated the relative abundance of Proteobacteria while reducing that of Firmicutes at the phylum level. At the genus level, GML markedly suppressed the relative abundance of Mycoplasma and Vibrio. These compositional alterations are potentially linked to improved energy acquisition and a decreased risk of pathogenic infections. Collectively, these results reveal that GML acts as a promising functional feed additive to boost growth performance, optimize digestive capacity, and sustain intestinal structural integrity in hybrid grouper. Based on these results, the optimal GML supplementation dose was determined to be 4 g/kg, providing valuable insights into the role of GML in aquaculture practices for hybrid grouper.

This study investigated the distinct regulatory effects of a novel complex feed additive (quercetin: cinnamaldehyde: glycerol monolaurate = 1:1:2) on lipid metabolism, immune response, and antiviral defense in Penaeus vannamei (initial weight of 0.40 ± 0.13 g). During a 42-day feeding trial, both 0.1 % and 0.2 % complex feed additive (CFA)-supplemented groups modulated hepatopancreatic lipid metabolism by co-regulating the constitutive co-activator of PPARγ (CCPG) and the PI3K-AKT and PTGS2 pathways, promoting lipolysis, suppressing lipid synthesis, and significantly reducing lipid accumulation in the body and plasma (P < 0.05). CFA also activated the JAK-STAT pathway and specifically modulated the NF-κB and p38 MAPK-AP-1 pathways, significantly improving the hemocyte count (THC) and phagocytic activity (PA), and upregulating the expression of antimicrobial peptides and lysozyme, while suppressing inflammatory factors (TNFα, IL16) in both hemocytes and the intestine (P < 0.05). Moreover, CFA improved gut microbiota composition, with key differential genera positively correlating with host immune indices. Under white spot syndrome virus (WSSV) challenge, CFA significantly inhibited viral replication and infection (P < 0.05), thereby enhancing antiviral efficacy and improving shrimp survival, with the best results observed at the 0.2 % concentration. Collectively, this study demonstrates that dietary supplementation with 0.2 % CFA effectively regulates lipid metabolism and strengthens both innate and antiviral immunity in P. vannamei, providing mechanistic insights into host-pathogen interactions and the development of antibiotic alternatives for sustainable aquaculture.

Nitrogen (N) pollution has emerged as a critical environmental issue hindering the sustainable development of global aquatic ecosystems in the 21st century. Particularly, N accumulation in aquaculture water has become a major pollutant, severely restricting the green and healthy development of aquaculture. Here, two efficient and safe heterotrophic nitrification-aerobic denitrification (HN-AD) strains were isolated from shrimp aquaculture environments and identified as Yanghufangia pacifica HHVEN1 and Glutamicibacter nicotianae SDVEA2. Under single or mixed inorganic-N conditions, both strains achieved >82.29% removal of ammonia-N (NH4+-N) and nitrite-N (NO2--N), and maintained high performance across a wide range of temperature, pH, salinity, and C/N ratio. Notably, HHVEN1 completely removed NH4+-N within 18 h, whereas SDVEA2 showed particularly strong nitrate removal (99.43% within 36 h). Based on time-course profiles of N species, N-removal functional genes, enzyme activities, and inhibitor assays, HHVEN1 predominantly followed a canonical HN-AD route and showed evidence for an ammonia-to-nitrate shortcut and a potential direct ammonium-to-gaseous-N conversion. In contrast, SDVEA2 mainly converted ammonium via nitrate and nitrite to gaseous N and reduced nitrate to gaseous N, despite lacking detectable canonical amoA, hao, and nxr genes. Haldane kinetic models were fitted to describe the relationships between substrate concentration and growth/N removal, indicating adaptation to medium-low N levels. Using response surface methodology (RSM), a multi-factor coupling model linking salinity, N sources and inoculum size was established, and a calculation method for optimal inoculum in dynamic aquatic scenarios was proposed. This study provides scientific support for N pollution control in aquaculture and related aquatic environments.

Synthetic phenolic antioxidants (SPAs) are widely utilized in industrial production and consumer goods owing to their excellent antioxidant properties. However, their potential for multiple toxic effects, coupled with the increasing levels of environmental pollution, has heightened public concern. Currently, there is a relative scarcity of study on the reproductive toxicology of SPAs in hydrobios, with a particular dearth of studies in invertebrates. This study investigates the reproductive toxicity of the typical SPAs butylhydroxytoluene (BHT) in male clams Ruditapes philippinarum across distinct breeding period. The finding show that,the processes of meiosis and spermatogenesis were inhibited, leading to a decreased number of mature sperm and a reduced gonad index. Furthermore, an increase in reactive oxygen species (ROS) content was observed in the testes under BHT stress, causing biomacromolecular damage. BHT induces different types of cell death processes in testis cells, thereby damaging sperm development.Ultimately, this investigation clarifies the molecular basis for reproductive toxicity caused by BHT in invertebrates living in water. The research specifically analyzed contributing factors including endocrine disruption, epigenetic effects, and oxidative stress. The experimental results provide valuable data support for marine shellfish germplasm conservation and marine SPAs pollution detection.

The liver is a major organ of digestion and detoxification metabolism in animals, and the occurrence of most liver diseases is closely associated with environmental pollution. Besides, liver is a non-reproductive primary target organ regulated by sex steroid hormone signaling. In this study, we elucidated the detoxification metabolism pathways of B[a]P in the clam Ruditapes philippinarum and identified the hepatotoxicity mechanism of B[a]P using technical approaches such as transcriptomics, computer simulation and experimental validation. We found that the AhR signaling pathway and CYP450 family (CYP1A1, CYP2B1, CYP3A4) and FMO in the digestive gland of clam play important roles in the detoxification metabolism of B[a]P, but the performance varied between male and female clams. SOD1 and CAT, GPX, PRDX play antioxidant function but PRDX pathway did not function in females. The level of detoxification metabolism in reproductive clams under B[a]P stress was female < male, oxidative stress was female > male, and oxidative damage was female < male. Cell death (apoptosis, pyroptosis and ferroptosis) was aggravated in the digestive gland of both males and females, with a reduced level of hepatic function health and an increase in the level of inflammatory factors, but males presented a more pronounced tendency toward hepatic fibrosis. In summary, the results of this study enrich the research perspectives on the metabolic pathways of POPs in aquatic invertebrates and lay the foundation for the study of POPs-induced hepatotoxicity, which is of great significance for the conservation of marine biological resources and the monitoring of POPs pollution.

To address the limited environmental adaptability and instability of single heterotrophic nitrification-aerobic denitrification (HN-AD) bacteria in treating aquatic nitrogen (N) pollution, this study isolated five efficient probiotics from shrimp farming environments. These include two HN-AD strains, Pseudomonas stutzeri GMSN161 and Bacillus subtilis HHEB2, which effectively removed over 91.15 % of ammonia and nitrite from various N sources, demonstrating good environmental adaptability. Two lactic acid bacteria (LAB), Lactiplantibacillus plantarum SDEX1 and SDWX2, produced lactic and acetic acids, significantly inhibiting five aquatic pathogens. Pichia kudriavzevii HHWY1, a yeast, secreted its extracellular polysaccharides and digestive enzymes to provide nutritional supplementation for the synthetic microbial communities (SMCs). To overcome the limitations of single strains, three SMCs (NMY-1, NMY-2, and NMY-3) with N removal and antibacterial functions were constructed. The SMCs exhibited superior N removal rate (>97.51 %) across diverse N sources and broader environmental adaptability compared to single strain systems. Notably, the optimal SMC (NMY-3) maintained over 90 % N removal across a wide range of conditions: 10-40 degrees C, 0-50 ppt salinity, pH 4.5-9.5, and C/N ratios of 0-20. SMCs also effectively inhibited pathogens, with NMY-3 showing the strongest activity, followed by NMY-1 and NMY-2. When applied to real aquaculture wastewater, all three SMCs removed over 90 % of total nitrogen (TN). This innovative strain combination strategy provides a new paradigm for applying synthetic microbial communities in environmental remediation.

This study integrates network pharmacology, molecular docking, and a 56-day feeding trial to evaluate the effects of dietary pterostilbene (PTE) on growth, physiological status, and underlying regulatory pathways in Litopenaeus vannamei. The results indicate that dietary supplementation with a specific dose of PTE enhances shrimp growth, improves metabolic capacity, boosts antioxidant defenses, strengthens immune responses, and reduces inflammation. The regulatory mechanism of PTE involves targeting and binding to key proteins in lipid metabolism-related signaling pathways, including AKT, INSR, and SIRT1 within the insulin and SIRT1-AMPK pathways. This binding significantly increases the gene expression levels of these targets (P < 0.05), thereby activating the respective signaling pathways and enhancing glycolipid metabolism capacity. In terms of immune function, PTE targets and binds to key antioxidants and immune proteins, including Nrf2, Relish, and Dorsal, resulting in a significant increase in the expression levels of these genes (P < 0.05). Consequently, the expression of downstream antioxidant enzymes associated with Nrf2 (SOD, CAT, GPX) was significantly elevated (P < 0.05), enhancing antioxidant capacity. Additionally, the expression of antibacterial lysosomal factors (PEN3, CRU, ALF, and LYC) also increased (P < 0.05), thereby strengthening immune function. Moreover, dietary PTE reshapes the structure of gut microbiota and increases the abundance of beneficial bacteria. This study elucidates the molecular mechanisms and optimal dosage of dietary PTE in improving the physiological function of shrimp by combining molecular docking with in vivo findings, providing a theoretical basis for the development of efficient and safe aquaculture feed additives.