Atrazine (ATZ), a widely used triazine herbicide frequently detected in aquatic environments, poses ecological risks, yet its toxicity mechanisms in estuarine/marine organisms remain unclear. The present study employed juvenile Oryzias melastigma (marine medaka) as the model organism, and the 96-h median lethal concentration (LC50) of ATZ was determined to be 31.65 mg/L. Juvenile marine medaka were exposed to 0, 0.09, 0.36, 1.07, and 3.28 mg/L ATZ for 30 days. Histopathology revealed severe gill alterations (hyperplasia, apical enlargement, disintegration of the lamellae and filament congestion) and hepatic injuries (sinusoidal dilation, cytoplasmic vacuolization, acidophilic body formation and nuclear pyknosis). Biochemically, ATZ altered Na+-K+-ATPase and Ca2+-ATPase activities in the gills, reduced the activity of catalase (CAT) in the liver. ATZ at concentrations of 0.09 and 0.36 mg/L could markedly decrease the activity of total superoxide dismutase (T-SOD), and increase the malondialdehyde (MDA) content in medaka liver. Additionally, ATZ exposure upregulated the expression levels of ucp-2 and decreased the expression levels of cox-2, ccl20, il-1β, cat and sod. Transcriptome results showed 1,078 differentially expressed genes (DEGs) in the liver, with 436 upregulated and 642 downregulated DEGs. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis revealed that ATZ exposure disturbed the expression of genes associated with the HIF-1 and PI3K-Akt signaling pathways, suggesting that ATZ may induce hypoxia, oxidative damage and apoptosis. These results corroborate the histological and enzymatic evidence, providing insights into the toxicological impacts and mechanisms of chronic ATZ exposure in estuarine and marine fish.

Arsenic, a toxic metalloid widely distributed in nature, stands as one of the most hazardous environmental pollutants in estuarine and coastal ecosystems. The biotoxicity of arsenic is highly dependent on its chemical speciation and environmental conditions, where salinity acts as a critical factor that markedly influences its toxicological effects on aquatic organisms. To date, few studies have reported the combined effects of arsenic and salinity on fish, and the mechanisms of arsenic-induced toxicity on early fish development under varying salinity conditions remain poorly understood. In our study, the toxic effects of arsenic at different concentrations (0, 0.1, 1, and 10 mg/L) on the embryonic development of marine medaka (Oryzias melastigma) under three salinities (10, 20, and 30 ppt) were investigated. Our research results showed that arsenic exposure reduced the hatching rate of medaka embryos. At salinities of 10 and 20 ppt, the exposure to arsenic induced additional developmental toxicities, including decreased heart rate, shortened hatching time, increased malformation rate, and reduced body length of newly hatched larvae. Additionally, arsenic exposure altered the expression levels of cardiac development-related genes and immune-related genes, with varying expression changes observed across different salinity levels. These findings, taken together, demonstrate that salinity influences the arsenic-induced toxicity (cardiotoxicity, oxidative damage) on the embryonic development of medaka. Our study provides additional data support for the toxicity research and ecological risk assessment of arsenic in estuarine and coastal ecosystems.

The aquaculture industry's expansion necessitates sustainable strategies to enhance aquatic animal health and production. This study evaluates Mychonastes afer-a microalga rich in nervonic acid-as a novel aquafeed supplement for marine medaka (Oryzias melastigma), focusing on growth, intestinal health, and reproduction. Fish fed high-concentration M. afer exhibited 10.04 % polyunsaturated fatty acid content, surpassing the 8.80 % in controls, alongside improved growth metrics. While M. afer enhanced gut microbial diversity, its primary reproductive benefits included an 8.86-9.33 % higher embryo fertilization rate compared to C. sorokiniana and blank groups. Under nickel exposure (5-20 mg/L), larvae from M. afer-supplemented environments demonstrated superior resilience: at 10 mg/L Ni, hatching rates in M. afer and C. sorokiniana groups remained stable, while controls dropped from 73 % to 52.84 %. Abnormalities were lowest in M. afer larvae (26 % vs. 30-37 % in other groups), with a 1-day delay in irreversible developmental damage. These findings highlight M. afer's ability to address nutritional gaps in conventional feeds, promoting growth, stress tolerance, and reproductive success in medaka. Nervonic acid in M. afer may further mitigate neurotoxicity and support neural regeneration, underscoring its potential as an eco-friendly aquafeed alternative to enhance aquaculture productivity under heavy metal stress.

Artificial reefs with sufficient vertical relief can support secondary fish production by attracting pelagic fishes and/or providing both shelter and food resources for them. Although vertical relief is a widely investigated factor in reef design, little is known about how the shelter-seeking and foraging processes of benthic fishes respond to relief changes. With a laboratory mesocosm experiment using hollow blocks to construct different combinations of reef volume and relief, we tested the effect of vertical relief of artificial reefs on a benthic juvenile fish (fat greenling, Hexagrammos otakii). The volume of the experimental reef depended on the number of blocks, and the reef relief was changed by the stacking pattern of the blocks at each volume. We measured the aggregation, sheltering, and foraging effects of each reef structure, respectively, based on fish distribution around the reef, their shelter-seeking behavior under disturbance, and their foraging rate. Our results demonstrate that increasing the vertical relief while maintaining the reef volume had a positive effect on the aggregating juvenile fat greenling but decreased the sheltering effect of the reef. The relationship between the foraging effect and relief was nonlinear, and high relief would also limit the foraging rate of juvenile fat greenling. We suggest that a reef needs to have sufficient relief to provide habitat for juvenile fat greenling, as well as sufficient horizontal cover to shelter the fish during a disturbance. Not only limited resources (e.g., food and shelter) but also the shelter-seeking and foraging processes of the benthic fish could have a potential impact on secondary production. Our findings highlight the importance of considering the aggregation, sheltering, and foraging effects of relief on benthic fishes in the field observation and the design of artificial reefs.

Litopenaeus vannamei is one of the important crustaceans in aquaculture in China. In recent years, the shrimp culture industry has gradually developed into an intensive and high-density model, which can lead to environmental deterioration and large-scale outbreak of diseases. The widespread use of antibiotics has led to the increase of pathogen drug resistance, environmental pollution, and ecological imbalance, resulting in secondary pollution in the water. Probiotics are live microbial additives that promote good health and are environmentally sustainable and safe for use. They are often used as important substitutes for antibiotics. Brevibacillus laterosporu, as a biocontrol probiotic, is commonly used for disease control of crops, animals, and poultry, but its use is rarely reported in aquaculture. A 28-day breeding experiment was carried out with L. vannamei weighing (1.00±0.08) g, to study the effects of adding B. laterosporu FAS05 in the feed on the growth, disease resistance, and immunity of L. vannamei. The experiment was divided into four groups with three replicates in each group, and 50 shrimps in each replicate were fed with the experimental feed supplemented with 0 CFU/g (group C, as the control group), 105 CFU/g (group BL1), 107 CFU/g (group BL2), and 109 CFU/g (group BL3) of B. laterosporu FAS05. B. laterosporu FAS05 was isolated from the aquaculture pond with a low number of Vibrio in summer. The bacterium was added into the basic feed with fish meal, soybean meal, and corn meal as the main protein source, fish oil and phospholipid oil were added as the fat source, wheat flour as the main sugar source, and inorganic salts and vitamins as supplements. L. vannamei was purchased from a prawn farm in Weihai City, Shandong Province. The temperature was 25–28 ℃ and the salinity was 27–30. During the breeding experiment, eight shrimps were randomly selected in each tank every two weeks and their body length, weight, and plumpness were measured. The phagocytic activity assay was modified on the basis of the method reported by Delaporte et al (2003). Fl-1 channel flow cytometry was used to detect and analyze the offset of respiratory burst peak. The blood lymphocytes, serum, and hepatopancreas of shrimp were collected 24 hours after the experiment. The activities of immune related enzymes in hepatopancreas, such as superoxide dismutase (SOD), catalase (CAT), acid phosphatase (ACP), and alkaline phosphatase (ALP) levels, and the activities of immune related enzymes in serum, phenol oxidase (PO), and lysozyme (LZM), were determined by kits. The phagocytic activity was measured, and the assay was modified on the basis of Delaporte et al (2003). Fl-1 channel flow cytometry was used to detect and analyze the offset of respiratory burst peak. After the feeding test, a one-week infection test of Vibrio parahaemolyticus was carried out to determine disease resistance. The experimental results were expressed as Mean±SD. One-way ANOVA analysis was conducted for all data using statistical software SPSS 16.0, and significance level was defined as P < 0.05. LSD homogeneity of variance test was used to compare the differences between experimental treatment groups and control groups. The results showed that there was no significant difference in survival rate among all groups (P > 0.05), and the body length, body weight, and specific growth rate of shrimps in the BL1 and BL2 groups were significantly higher than those in group C (P < 0.05). Compared to that in group C, the abundance of Vibrio in the aquaculture water in the BL1 to BL3 groups was significantly decreased (P < 0.05). After infection with V. parahemolyticus, the survival rate of group C was about 45%, while the survival rate of the BL1 to BL3 groups was more than 80%; therefore, the survival rate of the BL1 to BL3 groups was significantly higher than that in group C (P < 0.05), and no significant difference was found among the BL1 to BL3 groups (P > 0.05). Compared to that of group C, the phagocytizing rate of shrimp blood cells in the BL1 to BL3 groups increased significantly (P < 0.05), while the production of ROS in blood cells decreased significantly (P < 0.05). The LZM, CAT and ACP activities of shrimps in the BL1 to BL3 groups were significantly higher than those in group C (P < 0.05). There was no significant difference in LZM and ACP activities among the BL1 to BL3 groups (P > 0.05). The PO of shrimps in the BL2 group was significantly higher than those in other groups (P < 0.05). The ALP and SOD in the BL1 and BL2 groups were significantly higher than those in group C and BL3 (P < 0.05). The above results showed that B. laterosporu FAS05 as a feed additive could promote the growth of shrimp, activate the immune system, improve disease resistance, and inhibit the growth of Vibrio in the surrounding environment. The reference dosage was 105 CFU/g. When the infection is serious, the dosage can be increased to 107 CFU/g to further improve the non-specific immunity of shrimp. The results of this study can provide basic data for the application of B. laterosporu FAS05 in shrimp culture and production.

Manganese (Mn) is an essential metal for organisms, but high levels can induce serious toxicity. To date, the toxic mechanism of Mn to marine fish is still poorly understood. In the present study, Oryzias melastigma embryos were exposed to different concentrations of MnCl2 (0-152.00 mg/L) to investigate its effect on early development. The results showed that exposure to MnCl2 caused developmental toxicity to embryos, including increased heart rate, delayed hatching time, decreased hatching rate and increased malformation rate. MnCl2 exposure could induce oxidative stress in O. melastigma embryos, as indicated by increased the contents of malondialdehyde (MDA) and the activities of the antioxidant enzymes (superoxide dismutase (SOD), glutathione peroxidase (GPx) and catalase (CAT)). The heart might be an important target organ for MnCl2 because of cardiac malformations and disruption in the expression of cardiac development-related genes (ATPase, epo, fg8g, cox1, cox2, bmp4 and gata4). In addition, the expression levels of stress- (omTERT and p53) and inflammation-related genes (TNFα and il1β) were significantly up-regulated, suggesting that MnCl2 can trigger stress and inflammatory response in O. melastigma embryos. In conclusion, this study demonstrated that MnCl2 exposure can induce developmental toxicity, oxidative stress and inflammatory response in O. melastigma embryos, providing insights into the toxic mechanism of Mn to the early development of marine fish.

海水青鳉是研究微塑料对海洋生物毒理作用的理想模式生物.文章围绕微塑料在海水青鳉中毒性研究,通过查阅相关文献,对已发表文献年度变化和地区分布进行了统计分析;通过分析实验所用微塑料种类、颗粒大小和实验浓度、暴露时间和方式以及不同生长阶段海水青鳉实验对象选择,总结归纳微塑料在海水青鳉体内代谢、肠道损伤、生长和发育毒性、生殖毒性、遗传毒性以及对其他海洋污染物的放大或缓解效应 6 个方面的研究结果,并以分析结果为基础,建议在后期研究海洋微塑料对海水青鳉毒性效应,应重点考虑实验时间长短、实验材料选择和实验设计、毒理作用机制分析等因素.

Identifying the relationship between fish aggregations and artificial reefs (ARs) is important for optimizing reef structures and protecting marine resources subjected to external disturbance. Yet, knowledge remains limited of how the distribution of fish is affected by shelter availability provided by different AR structures. Here, we tested the effects of two structural attributes on the distribution of a benthic juvenile reef fish (fat greenling, Hexagrammos otakii). We used a laboratory mesocosm experiment with a simplified reef unit that was made of covered structure and non-covered structure. The covered structure was defined as the area inside ARs that provided effective shelter. The non-covered structure was defined as the area along the edge of ARs, which attracts fish but has lower sheltering effects. Four scenarios of two orthogonal structural attributes contained in a reef unit were implemented: size of covered structure (small shelter versus large shelter) and size of non-covered structure (small edge versus large edge), forming three size ratios of shelters to edges (low, medium, and high). The sheltering effects of the four scenarios were evaluated based on changes to the distribution patterns of fish under disturbance. We found that the reef with a large shelter had a better sheltering effect than the reef with a small shelter, but was limited by its small edge, especially when fish density was high. In contrast, the sheltering effect of the reef with a small shelter was limited by its large edge compared to the small edge. Thus, a moderate shelter-edge ratio enhanced the ability of juvenile fat greenling to elude external disturbance. Our findings highlight the importance of quantifying how the structural composition of reefs affects fish distributions, providing guidance to optimize AR structures.

As the only semi-enclosed sea area in China, the Bohai Sea is the strategic support and key support for the vigorous development of Beijing-Tianjin-Hebei Urban Agglomeration. The stability of its ecological environment is closely related to the well-being of coastal human beings, and its ecological environment restoration has become a research hotspot in recent years. This study focused on the analysis of the impact of global climate change and human activities on the ecological environment of the Bohai Sea, and comprehensively compared and analyzed the successful cases of ecological restoration in similar waters at home and abroad. Based on the characteristics and present situation of the Bohai Sea, some ecological restoration principles such as dynamic thinking and holistic preservation, habitat connectivity restoration principles, and different spatio-temporal scale restoration principle were proposed. It also pointed out that the ecological restoration of the Bohai Sea required rational planning of fishery economic activities, and overall consideration of the relationship between the Bohai Sea ecology and flushing water. It was recommended to use a forward-looking thinking model and concept, reasonably predict, plan, locate, configure, implement and maintain the stability of the Bohai Sea ecosystem, and focus on exploring the best adaptive management strategies to deal with global climate change.

Artificial reefs play a key functional role in marine ecosystem recovery and particularly in the rebuilding of fish populations. However, improving the reef design for viscous-egg-spawning fish is very challenging. Viscous eggs must adhere to a substrate to continue the hatching process, i.e., increasing the egg adhesion rate of the reef makes it a better fish spawning habitat. Therefore, the movement characteristics of viscous fish eggs in an artificial reef are a vital issue in marine ecology. In this study, a vertical two-dimensional lattice Boltzmann method was developed to simulate the complex hydraulic field in a reef zone and to predict the movement of fish eggs as affected by the hydrographic environment. A particle image velocimetry experiment on fish egg transportation was carried out to validate the model. These results revealed the effects of different hydrodynamic conditions on the migration of sticky eggs in the reef zone, to promote the sustainable development of fisheries and restore the marine ecology.

To investigate the effect of reduced salinity on diatoms’ capacity to cope with changing ultraviolet radiation (UVR) and photosynthetically active radiation (PAR), Skeletonema costatum was grown in a range of salinity (15, 25, and 35). The photosystem II (PSII) function was analyzed by increasing PAR and UVR to mimic a mixing event in turbulent waters. The results show that high UVR exposure significantly reduced PSII activity, especially in cells grown at low salinity. UVR, but not salinity, stimulated the ‘removal’ rate of PSII protein PsbA. Salinity alone, in the range of 15 to 35, did not regulate PSII acceptor region; however, the low salinity+UVR treatment decreased the energy flux for electron transport per PSII reaction center in S. costatum. It showed that low salinity exacerbated the damaging effect of UVR on PSII function in S. costatum by suppressing PsbA protein synthesis and modifying the photochemistry of PSII. Although higher catalase (CAT) activity and NPQs were induced, they were unable to prevent the combined damage effect of low salinity+UVR. Our findings indicate that reduced salinity and increased UVR potentially affect the abundance and distribution of S. costatum with the escalation of climate disturbances.

随着人类对生态环境和生物多样性保护意识的增强,基于食物网结构和物种间相互作用的关键种(Key-stone species)这一概念一经提出就引起了广泛关注和讨论,相关研究亦成为国际生态学研究的热点.本文从关键种概念起源出发,探讨了关键种定义中丰度与作用的不成比例性、在实际应用中存在的模糊性和无法量化等问题,综述了不同类型海洋生态系统中关键种的特点和共性问题,探讨人类作为超级关键种(Hyperkeystone species)对海洋的影响,并尝试提出人类应当如何理性看待关键种及其在海洋生态系统中的功能和作用.

通过对绿鳍马面鲀早期发育的连续观察,研究绿鳍马面鲀体色的发育过程.结果表明,绿鳍马面鲀仔鱼黑色素和黄色素在胚胎时期发生;虹彩色素在仔鱼3日龄出现,仅限于眼球,预示视觉功能化的完善;处于躯干腹部消化腔上侧浅褐色的色素团在4日龄后分化为黄色素;腹部虹彩色素集中发生在8～10日龄;稚鱼发育至40日龄,躯干部体色主要表现为横向黑色暗纹,属于过度体色模式;200日龄幼鱼,除了偶鳍绿色偏淡外,体色模式与3龄成鱼一致.绿鳍马面鲀体色的形成与形态分布,是判断其早期发育时序的主要特征之一.

本试验旨在研究饲料中添加枯草芽孢杆菌(Bacillus subtilis)HSP05对凡纳滨对虾(Litopenaeus vannamei)生长性能、抗病力和非特异性免疫力的影响.挑选大小一致的1200尾凡纳滨对虾虾苗,随机分为4组,每组3个重复,每个重复100尾.各组对虾分别投喂枯草芽孢杆菌HSP05添加量为0(对照组)、105(BS1组)、107(BS2组)、109 CFU/g(BS3组)的试验饲料.试验期4周.结果表明:1)BS1、BS2和BS3组的终末体重、终末体长、增重率和特定生长率显著高于对照组(P<0.05).2)BS1、BS2和BS3组的肝胰腺蛋白酶、淀粉酶活性显著高于对照组(P<0.05),BS1、BS2和BS3组的肝胰腺谷丙转氨酶和谷草转氨酶活性均显著低于对照组(P<0.05).3)BS1、BS2和BS3组的血清酚氧化酶和溶菌酶活性显著高于对照组(P<0.05).BS1、BS2和BS3组的肝胰腺过氧化氢酶活性显著高于对照组(P<0.05),BS2和BS3组的肝胰腺超氧化物歧化酶活性显著高于对照组(P<0.05).4)BS1、BS2和BS3组的血细胞吞噬率显著高于对照组(P<0.05),BS1、BS2和BS3组的呼吸爆发活性显著低于对照组(P<0.05).5)副溶血弧菌侵染持续7 d时,对照组的存活率为46.67％,而BS1、BS2和BS3组的存活率均达到82.67％以上.由此可见,饲料中添加枯草芽孢杆菌HSP05可促进凡纳滨对虾生长,增强免疫力,提高抗病力.枯草芽孢杆菌HSP05的添加量为107 CFU/g时,凡纳滨对虾的生长表现最优,非特异性免疫得到有效提高.

Triphenyltin (TPT) is an endocrine contaminant that is often detected in the environment. However, the mechanism of the effects of TPT on biological systems is not fully understood. Here we exposed marine medaka (Oryzias melastigma) to TPT for 21 days. Brain transcriptome, intestinal content metabolism group, and behavior analysis were carried out. Through the comprehensive analysis of multiomics for the in-depth understanding of the ways related to health improvement, we determined that the glycine-serine-threonine metabolic axis was most perturbed by TPT. Through behavioral analysis, it was found that there was behavioral hyperactivity in the exposed group; behavioral hyperactivity may be caused by the interference of TPT with the neuroendocrine system. In order to gain a full understanding of the impacts of TPT on human health, transcriptomic screening of differential genes and an impartial attitude based on bioinformatics were used. Gene-disease interaction analysis using the Comparative Toxicogenomics Database (CTD) revealed the possible effects of TPT on human health. Finally, based on these findings, the relevant adverse outcome pathway (AOP), which is the "epigenetic modi-fication of PPARG leading to adipogenesis," was identified from AOP Wiki. Further research is required to validate the potential AOP of TPT.

针对对虾养殖过程水处理的研究现状和瓶颈问题,文章综述了国际和国内重要的水处理管理理念与策略,主要包括:养殖用水"减法"管理理念、投入品"减法"管理理念、基于微生物的水处理策略和基于不同营养级的生物水处理策略等.较全面地探讨了养殖节水、生态调水、精准投喂、补偿生长、生物饵料、生物过滤器、水动力等对于水处理的重要性.并建议建立基于对虾福利生物学的水处理评价体系,联合生物行为变化、生态生理指标、水体理化指标进行综合评价.该综述期望为对虾产业和水产养殖的可持续发展提供新思路和可行路径.

Triphenyltin (TPT), an organic compound with a wide range of applications, is often detected in water bodies and aquatic animals. However, the mechanism underlying the biological metabolic health problems caused by longterm exposure to environment concentrations of TPT remains unclear. The morphology and gene expression in the gut and liver were investigated; and 16SrRNA gene amplification sequencing and non-targeted LC-MS/MS metabonomics were investigated after marine medaka (Oryzias melastigma) was treated with 1, 10, and 100 ng/L TPT for 21 days. During prolonged exposure to TPT, the adaptation mechanism maximized the energy of absorption, increased the length of intestinal microvilli, reduced the number of rough endoplasmic reticulum in the liver, and caused loss of weight. TPT exposure significantly changed the intestinal microbiome of marine medaka, thereby resulting in a significant decrease in microbial diversity. Following exposure to 100 ng/L TPT, the metabolic profiles were significantly changed and the altered metabolites were mainly concentrated in the lipid metabolic pathway. Finally, based on comprehensive network analysis, the association between the significantly changed bacteria and metabolites contributed further to the prediction of the impact of TPT on the host. This study provides a novel insight into the underlying mechanisms of host metabolic diseases caused by TPT and emphasizes the importance of monitoring pollutants in the environment.

随着对虾产业的不断壮大,对虾类水产养殖过程中疾病频发、水环境污染等问题日益突出,其品种和养殖模式的多样化使得这一矛盾更加难以解决.本文回顾了中国对虾产业50年发展过程中的疾病伴生史,并从生态学视角重新审视了对虾病原体的发生、进化以及与对虾免疫系统的关系,进而提出避免药物滥用、防重于治、注重对虾选育工作、基于生态系统方法开展对虾养殖工作等建议,以期将来人们能够更加重视生态系统健康,把One Health理念应用到水产养殖的生产实践中,真正实现对虾养殖的健康可持续发展.

Oceans provide unique ecosystem services due to their unique ecological, environmental, and biological resources. In recent years, the dramatic impact of humans on the oceans, especially in coastal waters, has led to changes in ecosystem services that are vital to human well-being. In this study, we focused on China's Yellow Sea ecosystem services and compared the composition and value of ecosystem services in the South and North Yellow Sea between 2000 and 2010. Our results showed that the total service values of the South and North Yellow Sea in 2000 were 199–322 billion Chinese Yuan (CNY) and 111–177 billion CNY, respectively. In 2010, the respective total service values were 386–555 billion CNY and 218–292 billion CNY. The ecosystems of the South and North Yellow Sea are mainly supported by food production. However, compared to 2000, percentage of food production contributing to total service value in 2010 was reduced by 21–23% and 22–27% in the South and North Yellow Sea, respectively. Meanwhile, the coastal tourism service values increased significantly by 15–20% and 25–30%. Changes in the structural composition of ecosystem services in the Yellow Sea are possibly related to the reduction of biodiversity and the decrease of wetland area. The rapid urbanization has created the necessary conditions for tourism cultural services and is a reasonable explanation for the rapid rise of cultural service values in the South and North Yellow Sea. The significance of ecosystem service evaluation to marine fisheries management in China was also discussed, pointing that long-term evaluation of marine ecosystem service is needed for coastal management decisions.

Extracellular traps (ETs) have been found to be an important strategy of mammals to immobilize and kill invading microorganisms. In the present study, we observed the formation of ETs in the hemocytes of marine mollusks Ruditapes philippinarum in response to challenge from bacteria Vibrio anguillarum, and examined the potential factors and signaling pathways underling this process. We detected an increase of reactive oxygen species (ROS) and myeloperoxidase (MPO) production during ETosis, accompanied by significantly up-regulated expression of ROS-related and MPO genes. The suppression of ETs structures by nicotinamide adenine dinucleotide phosphate (NADPH) oxidase inhibitor (diphenyleneiodonium chloride, DPI) and MPO inhibitor (aminobenzoic acid hydrazide, ABAH) further confirmed the essential roles ROS and MPO played in ETosis. Furthermore, ET production could be inhibited by phosphotidylinsitol-3-kinase (PI3K) inhibitor (LY294002) and extracellular regulated protein kinase (ERK) inhibitor (U0126), suggesting the idea that both the PI3K and ERK pathways were suggested to function during ETosis. In addition, the ETosis process was accompanied by enhancement of glycolysis-related enzymatic activities, e.g., pyruvate kinase (PK) and hexokinase (HK), and over-expression of the glycolysis-related genes, e.g., PK, HK and glucose transport protein (GLUT), indicating high involvement of glycolysis in the ETosis process. Furthermore, our scanning electron microscopy (SEM) observation and antibacterial activities test successfully showed the patterns how clam ETs entrapped and killed the invading V. anguillarum. Taken together, our results revealed that ETosis with bactericidal effect increased ROS, MPO and glycolysis level and carried out in a ROS-, MPO-, PI3K-ERK-dependent manner.