The molecular mechanisms and environmental effects of modified clay control algal blooms in aquacultural water.

Modified clay (MC) technology is an effective method for controlling harmful algal blooms (HABs). Based on field experience, a bloom does not continue after treatment with MC, even though the residual HAB biomass accounts for 20-30% of the initial biomass. Laboratory studies using unialgal cultures have found that MC could inhibit the growth of the residual algal cells to prevent HABs. Nevertheless, the phytoplankton in field waters is diverse. Therefore, unclassified complex mechanisms may exist. To illustrate the molecular mechanisms through which MC controls HABs in the field and verify the previous laboratory findings, a series of experiments and bioinformatics analyses were conducted using bloom waters from aquacultural ponds. The results showed that a 72.29% removal efficiency of algal biomass could effectively control blooms. The metatranscriptomic results revealed that the number of downregulated genes (131,546) was greater than that of upregulated genes (24,318) at 3 h after MC addition. Among these genes, several genes related to DNA replication were downregulated; however, genes involved in DNA repair were upregulated. Metabolism-related pathways were the most significantly upregulated (q < 0.05), including photosynthesis and oxidative phosphorylation. The results also showed that MC reduced most of the biomass of the dominant phytoplankton species, likely by removing apical dominance, which increased the diversity and stability of the phytoplankton community. In addition to reducing the pathogenic bacterial density, MC reduced the concentrations of PO43- (96.22%) and SiO32- (66.77%), thus improving the aquaculture water quality, altering the phytoplankton community structure (the proportion of Diatomea decreased, and that of Chlorophyta increased), and inhibiting phytoplankton growth. These effects hindered the rapid development of large phytoplankton biomasses and allowed the community structure to remain stable, reducing HAB threats. This study illustrates the molecular mechanisms through which MC controls HABs in the field and provides a scientific method for removing HABs in aquacultural waters.

The synthesis of an acrylamide copolymer and its synergistic effects on clay flocculation of red tide organisms.

The modified clay (MC) method is a common emergency treatment technology for red tides, and the selection of surface modifiers is the key to the MC technology. A cationic polymeric modifier, the copolymer of dimethyl diallyl ammonium chloride and acrylamide (P (DMDAAC-co-AM), PDA) was optimized via a visible-light-induced polymerization technique. The PDA-modified clay (PDAMC) was prepared with strong salt tolerance and achieved efficiencies of 86% at the concentration of 50 mg L-1, and the dose was 90% lower than that of aluminum polychloride-modified clay (PACMC). While polyacrylamide and commercial PDA can achieve efficiencies of only 25 and 67%, respectively, but high doses were required. This is because PDA changed the surface charges of clay particles from negative to positive, which promotes the formation of the polymer-chains bridging network to overcome the difficulties of curling in seawater. According to the analysis of flocculation parameters and spatial conformation of PDAMC, the high salinity tolerance of the PDAMC was attributed to the synergistic processes of charge neutralization and the three-dimensional network bridging. Therefore, this study has developed a highly effective flocculant material used in seawater and provided an important reference for the management of red tide organisms.

Long-term IgE immunological tolerance to peanut allergens: An alternative to Noon's daily desensitization paradigm.

Herein, we show that profound afferent long-term peanut-allergen-specific IgE immunological tolerance for 3 to 9 months induced sustained unresponsiveness (SU) in naïve or peanut-sensitized rodents after peanut allergen immunization. Rodents were vaccinated sublingually with a peanut allergen extract or recombinant peanut allergen in chenodeoxycholate (CDCA), a fanesoid X receptor (FXR, NR1H4) agonist that downregulates SREBP-1c (sterol regulatory element binding protein-1c) and upregulates SHP in bone marrow-derived tolerogenic dendritic cells (DCs). Approximately 90 âˆ¼ 95 % of the total circulating PE-potentiated IgE and Ara h1, Ara h 2, and Ara h 6 peanut allergen-specific IgE responses were suppressed by recombinant peanut allergen-conjugated solid magnetic beads (sensitivity of 0.2 IU/ml). In contrast, peanut allergen-specific IgG production was not affected. Similarly, oleoylethanolamine (OEA), a peroxisome proliferator-activator receptor alpha (PPARα) agonist, and GW9662, a PPARγ antagonist, induced long-term peanut-specific IgE tolerance when administered via the sublingual, oral or i.p. route. Prophylactic Ara h2 DNA immunization with caNRF2 and IL-35 coexpression induced Ara h2 IgE tolerance. In summary, peanut allergen vaccination with select natural molecular ligands of nuclear receptors induced long-term peanut allergen-specific IgE tolerance via the afferent limb, which indicates that vaccination is an immune tolerance-promoting strategy that is effective at the DC level and that differs from Noon's daily desensitization program, which is effective at the mast cell level.

Programmed cell death induced by modified clay in controlling Prorocentrum donghaiense bloom.

Modified clay (MC), an effective material used for the emergency elimination of algal blooms, can rapidly reduce the biomass of harmful algal blooms (HABs) via flocculation. After that, MC can still control bloom population through indirect effects such as oxidative stress, which was initially proposed to be related to programmed cell death (PCD) at molecular level. To further study the MC induced cell death in residual bloom organisms, especially identifying PCD process, we studied the physiological state of the residual Prorocentrum donghaiense. The experimental results showed that flocculation changed the physiological state of the residual cells, as evidenced by growth inhibition and increased reactive oxygen species production. Moreover, this research provides biochemical and ultrastructural evidence showing that MC induces PCD in P. donghaiense. Nuclear changes were observed, and increased caspase-like activity, externalization of phosphatidylserine and DNA fragmentation were detected in MC-treated groups and quantified. And the mitochondrial apoptosis pathway was activated in both MC-treated groups. Besides, the features of MC-induced PCD in a unicellular organism were summarized and its concentration dependent manner was proved. All our preliminary results elucidate the mechanism through which MC can further control HABs by inducing PCD and suggest a promising application of PCD in bloom control.

Profiles of and variations in aluminum species in PAC-MC used for the removal of blooming microalgae.

Polyaluminum chloride modified clay (PAC-MC) is a safe and efficient red tide control agent that has been studied and applied worldwide. Although it is well known that the distribution of hydrolytic aluminum species in PAC affects its flocculation, little is known about the influence of particulars aluminum species on the microalgae removal efficiency of PAC-MC; this lack of knowledge creates a bottleneck in the development of more efficient MCs based on aluminum salts. The ferron method was used in this study to quantitatively analyze the distributions of and variations in different hydrolytic aluminum species during the process of microalgae removal by PAC-MC. The results showed that Ala, which made up 5%-20% of the total aluminum, and Alp, which made up 15%-55% of the total aluminum, significantly affected microalgae removal, with Pearson's correlation coefficients of 0.83 and 0.89, respectively. Most of the aluminum in the PAC-MC sank rapidly into the sediments, but the rate and velocity of settlement were affected by the dose of modified clay. The optimal dose of PAC-MC for precipitating microalgae was determined based on its aluminum profile. These results provide guidance for the precise application of PAC-MC in the control of harmful algal blooms.

Molecular Mechanism of Modified Clay Controlling the Brown Tide Organism Aureococcus anophagefferens Revealed by Transcriptome Analysis.

The data and experiences in mitigating harmful algal blooms (HABs) by modified clay (MC) show that a bloom does not continue after the dispersal of the MC, even though the density of the residual cells in the water is still high, at 20-30% of the initial cell density. This interesting phenomenon indicates that in addition to flocculation, MC has an additional control mechanism. Here, transcriptome sequencing technology was used to study the molecular mechanism of MC in controlling HABs. In residual cells treated with MC, the photosynthetic light reaction was the most affected physiological process. Some genes related to the light harvesting complex, photosystem (PS) I and PS II, were significantly up-regulated ( p < 0.05), and several transcripts increased by as much as 6-fold. In contrast, genes associated with the dark reaction did not significantly change. In addition to genes associated with photosynthesis, numerous genes related to energy metabolism, stress adaptation, cytoskeletal functioning, and cell division also responded to MC treatment. These results indicated that following treatment with MC, the normal physiological processes of algal cells were disrupted, which inhibited cell proliferation and growth. Thus, these findings provide scientific proof that HABs are controlled by MC.

Phycosphere bacterial composition and function in colony and solitary Phaeocystis globosa strains providing novel insights into the algal blooms.

Phycosphere bacteria can regulate the dynamics of different algal blooms that impact marine ecosystems. Phaeocystis globosa can alternate between solitary free-living cells and colonies and the latter morphotype is dominate during blooms. The mechanisms underlying the formation of these blooms have received much attention. High throughput sequencing results showed that the bacterial community composition differed significantly between colony and solitary strains in bacterial composition and function. It was found that the genera SM1A02 and Haliea were detected only among the colony strains and contribute to ammonium accumulation in colonies, and the genus Sulfitobacter was abundant among the colony strains that were excellent at producing DMS. In addition, the bacterial communities of the two colony strains exhibited stronger abilities for carbon and sulfur metabolism, energy metabolism, vitamin B synthesis, and signal transduction, providing inorganic and organic nutrients and facilitating tight communication with the host algae, thereby promoting growth and bloom development.

BTXs removals by modified clay during mitigation of Karenia brevis bloom: Insights from adsorption and transformation.

Harmful algal blooms (HABs), especially those caused by toxic dinoflagellates, are spreading in marine ecosystems worldwide. Notably, the prevalence of Karenia brevis blooms and potent brevetoxins (BTXs) pose a serious risk to public health and marine ecosystems. Therefore, developing an environmentally friendly method to effectively control HABs and associated BTXs has been the focus of increasing attention. As a promising method, modified clay (MC) application could effectively control HABs. However, the environmental fate of BTXs during MC treatment has not been fully investigated. For the first time, this study revealed the effect and mechanism of BTX removal by MC from the perspective of adsorption and transformation. The results indicated that polyaluminium chloride-modified clay (PAC-MC, a typical kind of MC) performed well in the adsorption of BTX2 due to the elevated surface potential and more binding sites. The adsorption process was a spontaneous endothermic process that conformed to pseudo-second-order adsorption kinetics (k2 = 6.8 × 10-4, PAC-MC = 0.20 g L-1) and the Freundlich isotherm (Kf = 55.30, 20 °C). In addition, detailed product analysis using liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) indicated that PAC-MC treatment effectively removed the BTX2 and BTX3, especially those in the particulate forms. Surprisingly, PAC-MC could promote the transformation of BTX2 to derivatives, including OR-BTX2, OR-BTX3, and OR-BTX-B5, which were proven to have lower cytotoxicity.

Insights into how characteristics of dissolved algal organic matter affect the efficiency of modified clay in controlling harmful algal blooms.

Dissolved algal organic matter (dAOM) originating from harmful algal blooms (HABs) can deteriorate the quality of municipal water supplies, threaten the health of aquatic environments, and interfere with modified clay (MC)-based HABs control measures. In this study, we explored the composition of dAOM from Prorocentrum donghaiense, a typical HAB organism, and assessed the influence of dAOM on MC flocculation. Our results suggested that dAOM composition was complex and had a wide molecular weight (MW) distribution. MW and electrical properties were important dAOM characteristics affecting flocculation and algal removal efficiency of MC. Negatively charged high-MW components (>50 kDa) critically affected algal removal efficiency, reducing the zeta potential of MC particles and leading to small and weak flocs. However, the effect of dAOM depended on its concentration. When the cell density of P. donghaiense reached HAB levels, the high-MW dAOM strongly decreased the algal removal efficiency of MC.

Regulation of proteolytic cleavage of retinoid X receptor-α by GSK-3ß.

We recently reported that an N-terminally truncated retinoid X receptor-α (tRXRα) produced in cancer cells acts to promote cancer cell growth and survival through AKT activation. However, how RXRα is cleaved and how the cleavage is regulated in cancer cells remain undefined. In this study, we demonstrated that calpain II could cleave RXRα protein in vitro, generating two truncated RXRα products. The cleavage sites in RXRα were mapped by Edman N-terminal sequencing to Gly(90)↓Ser(91) and Lys(118)↓Val(119). Transfection of the resulting cleavage product RXRα/90, but not RXRα/118, resulted in activation of AKT in cancer cells, similar to the effect of tRXRα. In support of the role of calpain II in cancer cells, transfection of calpain II expression vector or its activation by ionomycin enhanced the production of tRXRα, whereas treatment of cells with calpain inhibitors reduced the levels of tRXRα. Co-immunoprecipitation assays also showed an interaction between calpain II and RXRα. In studying the regulation of tRXRα production, we observed that treatment of cells with lithium chloride or knockdown of glycogen synthase kinase-3ß (GSK-3ß) significantly increased the production of tRXRα. Conversely, overexpression of GSK-3ß reduced tRXRα expression. Furthermore, we found that the inhibitory effect of GSK-3ß on tRXRα production was due to its suppression of calpain II expression. Taken together, our data demonstrate that GSK-3ß plays an important role in regulating tRXRα production by calpain II in cancer cells, providing new insights into the development of new strategies and agents for the prevention and treatment of tRXRα-related cancers.

Novel insights into toxin changes associated with the growth of Alexandrium pacificum: Revealing active toxin-secretion ability and toxin cell quota variation.

Paralytic shellfish toxins (PSTs) are widely distributed globally and are produced by Alexandrium pacificum in marine system. However, the characteristics of toxins producing and secreting associated with growth phases are still unclear, especially whether A. pacificum has the ability to actively secrete PSTs is controversial. In this study, variation characteristics of intracellular and extracellular PSTs contents associated with A. pacificum growth phases were investigated thoroughly. The results showed that intracellular and extracellular PSTs contents increased sharply during the exponential phase. But during the stationary phase, the intracellular PSTs content increased by only 26 %, and the extracellular PSTs content did not increase significantly. Since the increase in extracellular PSTs content mainly occurred at the exponential phase, when most cells were living, we speculated that active PSTs secretion of living cells might be an important production pathway of extracellular toxins besides leakage from dead cells. Furthermore, toxin cell quota variation associated with the growth phase was analysed. In the exponential phase, the toxin cell quota first increased and then decreased, with a maximum of 19.02 ± 1.80 fmol/cell at 6 d. However, after entering the stationary phase, this value slowly increased again, suggesting that vigilance should be raised for the plateau of Alexandrium blooms. In addition, cells in the exponential phase mainly produced O-sulfated components such as GTX1&4, cells in the stationary phase mainly produced O-sulfate-free components such as GTX5. In this study, the toxigenic rules of A. pacificum were comprehensively uncovered, which provided theoretical guidance for the prevention and mitigation of A. pacificum blooms.

Metabarcoding analysis identifies high diversity of harmful algal bloom species in the coastal waters of the Beibu Gulf.

Harmful algal blooms (HABs) have occurred more frequently in recent years. In this study, to investigate their potential impact in the Beibu Gulf, short-read and long-read metabarcoding analyses were combined for annual marine phytoplankton community and HAB species identification. Short-read metabarcoding showed a high level of phytoplankton biodiversity in this area, with Dinophyceae dominating, especially Gymnodiniales. Multiple small phytoplankton, including Prymnesiophyceae and Prasinophyceae, were also identified, which complements the previous lack of identifying small phytoplankton and those unstable after fixation. Of the top 20 phytoplankton genera identified, 15 were HAB-forming genera, which accounted for 47.3%-71.5% of the relative abundance of phytoplankton. Based on long-read metabarcoding, a total of 147 OTUs (PID > 97%) belonging to phytoplankton were identified at the species level, including 118 species. Among them, 37 species belonged to HAB-forming species, and 98 species were reported for the first time in the Beibu Gulf. Contrasting the two metabarcoding approaches at the class level, they both showed a predominance of Dinophyceae, and both included high abundances of Bacillariophyceae, Prasinophyceae, and Prymnesiophyceae, but the relative contents of the classes varied. Notably, the results of the two metabarcoding approaches were quite different below the genus level. The high abundance and diversity of HAB species were probably due to their special life history and multiple nutritional modes. Annual HAB species variation revealed in this study provided a basis for evaluating their potential impact on aquaculture and even nuclear power plant safety in the Beibu Gulf.

Mechanisms of Phaeocystis globosa blooms in the Beibu Gulf revealed by metatranscriptome analysis.

The haptophyceae Phaeocystis globosa is a species responsible for harmful algal blooms in the global ocean, forming blooms in the Beibu Gulf annually since 2011. This species can alternate between solitary free-living cells and colonies. Colonies are the dominant morphotype during blooms. To date, the underlying mechanism of P. globosa blooms in the Beibu Gulf is poorly understood. After combining results of ecological surveys, laboratory studies, and metatranscriptome and bioinformatics analyses, it was found that low temperatures, high nitrate, and low organic phosphorus induced P. globosa blooms in the Beibu Gulf. Additionally, the unique genetic and physiological characteristics that allow P. globosa to stand out as a dominant species in such an environment include (1) several genes encoding high-affinity nitrate transport proteins that could be highly expressed under sufficient nitrate conditions; (2) energy metabolism genes involved in photosynthesis and oxidative phosphorylation that were actively expressed at low temperatures to carry out carbon and energy reversion and produce sufficient ATP for various life activities, individually; (3) abundant glycan synthesis genes that were highly expressed at low temperatures, thus synthesizing large quantities of proteoglycans to construct the mucilaginous envelope forming the colony; (4) cells in colonies exhibited active gene expression in DNA replication contributing to a faster growth rate, which could help P. globosa occupy niches quickly; and (5) the energy and material expenditure was redistributed in colonial cells accompanied with chitin filaments and flagella degraded, more expenditure was used for the synthesis of the mucilaginous envelope and the rapid proliferation.

Regulation of Nur77 expression by ß-catenin and its mitogenic effect in colon cancer cells.

The orphan nuclear receptor Nur77 is an immediate-early response gene whose expression is rapidly induced by various extracellular stimuli. The aims of this study were to study the role of Nur77 expression in the growth and survival of colon cancer cells and the mechanism by which Nur77 expression was regulated. We showed that levels of Nur77 were elevated in a majority of human colon tumors (9/12) compared to their nontumorous tissues and that Nur77 expression could be strongly induced by different colonic carcinogens including deoxycholic acid (DCA). DCA-induced Nur77 expression resulted in up-regulation of antiapoptotic BRE and angiogenic VEGF, and it enhanced the growth, colony formation, and migration of colon cancer cells. In studying the mechanism by which Nur77 was regulated in colon cancer cells, we found that ß-catenin was involved in induction of Nur77 expression through its activation of the transcriptional activity of AP-1 (c-Fos/c-Jun) that bound to and transactivated the Nur77 promoter. Together, our results demonstrate that Nur77 acts to promote the growth and survival of colon cancer cells and serves as an important mediator of the Wnt/ß-catenin and AP-1 signaling pathways.

Effects of modified clay on the formation of Phaeocystis globosa colony revealed by physiological and transcriptomic analyses.

The harmful algal bloom (HAB) species Phaeocystis globosa is commonly observed in global temperate and tropical oceans, and colonies of P. globosa exhibit a dominant morphotype during blooms. The use of polyaluminium chloride modified clay (PAC-MC) is an effective mitigation strategy for P. globosa blooms. Although previous studies have found that PAC-MC can stimulate P. globosa colony formation at low concentrations and inhibit it at higher concentrations, the underlying mechanisms of these effects are poorly understood. Here, we comprehensively compared the physiochemical indices and transcriptomic response of residual P. globosa cells after treatment with two concentrations of PAC-MC. The results showed that PAC-MC induced oxidative stress, photosynthetic inhibition, and DNA damage in residual cells. Moreover, it could activate antioxidant responses and enhance the repair of photosynthetic structure and DNA damage in cells. The biosynthesis of polysaccharides was enhanced and genes associated with cell motility were down-regulated after treatment with PAC-MC, resulting in the accumulation of colonial matrixes. After treatment with a low concentration of PAC-MC (0.1 g/L), the residual cells were slightly stressed, including physical damage, oxidative stress and other damage, and polysaccharide synthesis was enhanced to promote colony formation to alleviate environmental stress. Moreover, the damage to residual cells was slight; thus, normal cell function provided abundant energy and matter for colony formation. After treatment with a high concentration of PAC-MC (0.5 g/L), the residual cells suffered severe damage, which disrupted normal physiological processes and inhibited cell proliferation and colony formation. The present study elucidated the concentration-dependent mechanism of PAC-MC affecting the formation of P. globosa colonies and provided a reference for the application of PAC-MC to control P. globosa blooms.

Effects of Modified Clay on Phaeocystis globosa Growth and Colony Formation.

Phaeocystis globosa is a globally distributed harmful algal blooms (HABs) species dominated by the colonial morphotype, which presents dramatic environmental hazards and poses a threat to human health. Modified clay (MC) can effectively flocculate HAB organisms and prevent their subsequent growth, but the effects of MC on colony-dominated P. globosa blooms remain uncertain. In this paper, a series of removal and incubation experiments were conducted to investigate the growth, colony formation and colony development of P. globosa cells after treatment with MC. The results show that the density of colonies was higher at MC concentrations below 0.2 g/L compared to those in the control, indicating the role of P. globosa colonies in resistance to environmental stress. Concentrations of MC greater than 0.2 g/L could reduce the density of solitary cells and colonies, and the colony diameter and extracellular polysaccharide (EPS) content were also decreased. The adsorption of MC to dissolved inorganic phosphorus (DIP) and the cell damage caused by collision may be the main mechanisms underlying this phenomenon. These results elucidate that the treatment with an appropriate concentration of MC may provide an effective mitigation strategy for P. globosa blooms by preventing their growth and colony formation.

Mechanism by Which MC Controls Harmful Algal Blooms Revealed by Cell Morphology of Aureococcus anophagefferens.

On the basis of field experience, a bloom does not continue after treatment with modified clay (MC), even though the residual harmful algal bloom (HAB) biomass accounts for 20-30% of the initial cells. This interesting phenomenon indicates that, in addition to causing flocculation, MC can inhibit the growth of residual cells. Here, from a cell morphology perspective, Aureococcus anophagefferens was used as a model organism to explore this scientific issue and clarify the mechanism by which MC mitigates harmful algal blooms (HABs). The results showed that, at an ~70% removal efficiency, neutral clay (NC) could not effectively inhibit the growth of residual cells, although it caused various forms of damage to residual cells, such as cell deformation, cell breakage, decreased extracellular polysaccharides (EPS), increased cell membrane permeability, and increased cytoplasmic granularity, due to physical collisions. After modification, some physical and chemical properties of the clay particle surface were changed; for example, the surface electrical properties changed from negative to positive, lamellar spacing increased, hardness decreased, adhesion chains increased, adhesion improved, and the number of absorption sites increased, enhancing the occurrence of chemical and electrochemical effects and physical collisions with residual cells, leading to severe cell deformation and chemical cell breakage. Thus, MC effectively inhibited the growth of residual cells and controlled HABs.

Effects of soluble organics on the settling rate of modified clay and development of improved clay formulations for harmful algal bloom control.

For many years, the dispersal of modified clay (MC) has been used to control harmful algal blooms (HABs) in coastal waters of China. MC flocculation efficiency can be influenced by many factors in variable and complex natural environments, including high concentrations of dissolved organic matter (DOM) in the water to be treated. Since many HABs occur in nearshore waters where DOM concentrations are high, this is a significant problem that requires urgent attention. This study involved the use of humic acid as a representative form of DOM to study the influence of organic matter on the MC flocculation process. At high concentrations, humic acid was adsorbed onto MC particles, resulting in a decrease in surface potential and an increase in electrostatic repulsion between the clay particles; this decreased the MC settling rate and increased the water clarification time. Flocs were characterized by their relatively small particle size, high particle concentration, and low collision efficiency, which together resulted in slow clarification of the water after MC spraying. Based on the mechanism of the DOM-MC interaction and combined with the Derjaguin-Landau-Verwey-Overbeek theory and theoretical considerations of clay surface modification, the "ionic atmosphere compression" method was used to improve MC flocculation efficiency in high-organic water. This method increased the ionic strength of the clay stock solution by adding salt, thereby compressing the ionic atmosphere of MC particles and lowering the potential barrier, allowing the MC particles in the treated water to flocculate rapidly and form large flocs, followed by further floc growth and rapid settling via differential sedimentation. The settling rate of MCs improved by a factor of two and the removal efficiency of the HAB cells increased by 7-28%. This study provides important baseline information that will extend the application of MC to HAB control in water bodies with high organic loadings.

Healthier Communities of Phytoplankton and Bacteria Achieved via the Application of Modified Clay in Shrimp Aquaculture Ponds.

The composition and stability of microbial communities in aquaculture water are crucial for the healthy growth of shrimp and present considerable risk to aquatic ecosystems. The modified clay (MC) method has been proposed as an efficient and safe solution for the mitigation of harmful algal blooms (HABs). Currently, the effects of MC on microbial communities in aquaculture water remain unknown. Here, we adopted the MC method to regulate shrimp-culture water quality and evaluated the effects of MC on the composition and stability of phytoplankton together with bacteria communities through high-throughput sequencing. On the one hand, a prominent change in the composition of microbial community was observed, with green algae becoming the most abundant genera and pathogens being infrequent in the MC-treated pond, which was more conducive to the growth of shrimp than that in the control pond. Moreover, MC could increase the diversity and stability of the microbial community structure in the water column, which had a higher anti-interference ability, as demonstrated by the analysis of the diversity and molecular ecological network. Taken together, MC could reduce the possibility for the occurrence of HABs and maintain a stable microbial community, which is beneficial for the health and high yield of shrimp.

Heterogeneity of the sediment oxygen demand and its contribution to the hypoxia off the Changjiang estuary and its adjacent waters.

The severe hypoxia off the Changjiang estuary (CE) has a dual-core structure, and the two hypoxic zones exhibit behavioural, physical and biochemical differences. Currently, few studies have revealed straightforward differences regarding the key biochemical processes between these two hypoxic zones. In this study, the phytoplankton sinking rate (PSR) and sediment oxygen demand (SOD) were measured by field experiments and compared between the two hypoxic regions. PSR and SOD ranged from 0.75-3.34 m day-1 and 5.67-16.19 mmol m-2 day-1, respectively. Interestingly, PSR and SOD were higher in the southern region than in the northern region, implying stronger pelagic-benthic biogeochemical coupling in the southern region. SOD accounted for approximately 44% and 51% of DO net consumption in the northern and southern regions, respectively, from July to August. The southern hypoxic region appeared to exhibit intense DO consumption and fast DO supplementation, while the northern hypoxic region seemed to exhibit slow DO consumption and supplementation.