Combined effects of toxic Microcystis aeruginosa and high pH on antioxidant responses, immune responses, and apoptosis of the edible freshwater bivalve Corbicula fluminea.

Due to increasing anthropogenic perturbation and water eutrophication, cyanobacterial blooms (CYBs) have become a global ecological and environmental problem. Toxic CYBs and elevated pH are considered to be the two key stressors associated with eutrophication in natural waters, particularly in the event of CO2 depletion induced by dense blooms. However, previous research has been focused on investigating the impacts of toxic CYBs or pH changes in isolation, whereas the interactive effects of such stressors on edible bivalves that inhabit CYB waters still lack information. In this study, the combined effects of toxic Microcystis aeruginosa and pH shifts on the antioxidant responses, immune responses, and apoptosis of the edible freshwater bivalve Corbicula fluminea were explored. The results showed that the activity of antioxidant enzymes was significantly impacted by the interactive effects between toxic M. aeruginosa exposure and time course, yet pH shifts showed no significant effects on the activities of these antioxidant enzymes, implying that the antioxidant response in C. fluminea was mainly triggered by toxic M. aeruginosa exposure. Toxic M. aeruginosa also induced an increased production of reactive oxygen species and malondialdehyde in treated clams, particularly under high pH settings. The elevated lysosomal enzyme activity helped C. fluminea defend against toxic M. aeruginosa exposure under high pH conditions. The principal component analysis (PCA) and the integrated biomarker response (IBR) results suggested that the treated clams were subjected to the elevated toxicity of toxic M. aeruginosa in conditions of high pH. The heat shock proteins-related genes might be triggered to resist the oxidative damage in treated clams. Moreover, the upregulation of TNF and casp8 genes indicated the potential activation of the caspase8-mediated apoptotic pathway through TNF receptor interaction, potentially resulting in apoptosis. The TUNEL assay results further confirmed that apoptosis appeared in treated clams. These findings improve our understanding of the combined toxicological effects of harmful algae and pH shifts on bivalves, which will provide insights into a comprehensive ecological risk assessment of toxic CYBs to edible bivalve species.

Integrating regular and transcriptomic analyses reveal resistance mechanisms in Corbicula fluminea (Müller, 1774) in response to toxic Microcystis aeruginosa exposure.

The frequent occurrence of cyanobacterial blooms (CYBs) caused by toxic Microcystis aeruginosa poses a great threat to aquatic organisms. Although freshwater benthic bivalves have proven to be capable of uptake high levels of microcystins (MCs) due to their filter-feeding habits, there is a paucity of information concerning their systemic resistance mechanisms to MCs. In this study, the resistance mechanisms in Corbicula fluminea (O. F. Müller, 1774) in response to the exposure of toxic M. aeruginosa were explored through transcriptional analysis combined with histopathological and biochemical phenotypic analysis. Toxic M. aeruginosa exposure caused dose-dependent histological damage in the hepatopancreas. The conjugation reaction catalyzed by glutathione S-transferases was vulnerable to being activated by high concentrations of M. aeruginosa (10 ×105 cells mL-1). Additionally, reactive oxygen species scavenging processes mediated by superoxide dismutase and catalase were active in the initial stage of toxic M. aeruginosa exposure. The results of the integrated biomarker response index suggested that the biotransformation and antioxidant defense system in C. fluminea could be continuously activated after acute exposure to the high concentration of toxic M. aeruginosa. The eggNOG and GO analysis of the differentially expressed genes (DEGs) indicated that DEGs were significantly enriched in transporter activity, oxidant detoxification and response to oxidative stress categories, which were consistent with the alterations of biochemical indices. Besides, DEGs were significantly annotated in a few KEGG pathways involved in biotransformation (oxidation, cooxidation and conjugation) and immunoreaction (lysosome and phagosome responses), which could be responsible for the tolerance of C. fluminea to toxic M. aeruginosa. These findings improve our understanding of potential resistance mechanisms of freshwater bivalves to MCs.

Two machine learning approaches for predicting cyanobacteria abundance in aquaculture ponds.

Cyanobacteria blooms in aquaculture ponds harm the harvesting of aquatic animals and threaten human health. Therefore, it is crucial to identify key drivers and develop methods to predict cyanobacteria blooms in aquaculture water management. In this study, we analyzed monitoring data from 331 aquaculture ponds in central China and developed two machine learning models - the least absolute shrinkage and selection operator (LASSO) regression model and the random forest (RF) model - to predict cyanobacterial abundance by identifying the key drivers. Simulation results demonstrated that both machine learning models are feasible for predicting cyanobacterial abundance in aquaculture ponds. The LASSO model (R2 = 0.918, MSE = 0.354) outperformed the RF model (R2 = 0.798, MSE = 0.875) in predicting cyanobacteria abundance. Farmers with well-equipped aquaculture ponds that have abundant water monitoring data can use the nine environmental variables identified by the LASSO model as an operational solution to accurately predict cyanobacteria abundance. For crude ponds with limited monitoring data, the three environmental variables identified by the RF model provide a convenient solution for useful cyanobacteria prediction. Our findings revealed that chemical oxygen demand (COD) and total organic carbon (TOC) were the two most important predictors in both models, indicating that organic carbon concentration had a close relationship with cyanobacteria growth and should be considered a key metric in water monitoring and pond management of these aquaculture ponds. We suggest that monitoring of organic carbon coupled with phosphorus reduction in feed usage can be an effective management approach for cyanobacteria prevention and to maintain a healthy ecological state in aquaculture ponds.

Growth and photosynthesis responses of microcystin (MC)- and non-MC-producing Microcystis strains during co-culture with the submerged macrophyte Myriophyllum spicatum.

The growth and photosynthetic responses of microcystin (MC)- and non-MC-producing Microcystis to the submerged macrophyte Myriophyllum spicatum were investigated under plant-Microcystis co-culture conditions (PMC + , PMC-), in comparison with their corresponding mono-culture controls (SMC + , SMC-). The OJIP chlorophyll a fluorescence transient was recorded and analyzed using JIP-test parameters. In comparison with the corresponding mono-culture controls, the quantum efficiencies of the electron transport chain expressed as parameters Ψo and φEo decreased by more than 25% in the PMC- group, much higher than those in the PMC+ group (p < 0.05). The decreasing extent of the electron transport flux ratio ETo/RC was significantly larger in the PMC- group than in the PMC+ group (p < 0.05). The performance index (PIABS) decreased by 26.98% in the PMC- group and by 16.25% in the PMC+ group. These results indicated that the non-MC-producing Microcystis was more sensitive than the MC-producing strain when co-cultured with M. spicatum, and the efficiency and energy flux of electron transport might be the main targets of Photosystem II. Future research should focus on the intraspecific composition and toxigenic levels of Microcystis populations interacting with submerged macrophytes to guide the use of submerged vegetation restoration.

Effects of pyrogallic acid on Microcystis aeruginosa: oxidative stress related toxicity.

Pyrogallic acid (PA) is used in various industrial and consumer products. The molecular mechanisms underlying PA's toxicity was not fully understood. In this study, toxicity of PA on Microcystis aeruginosa with reactive oxygen species (ROS) generation as an end point was investigated. The results showed an increase in the percentage of cells with loss of membrane integrity and enhanced intracellular ROS production. Exposure to 50mgL(-1) PA for 48h caused the highest percentage of loss of membrane integrity (56.7%), and a 2.54-fold higher intracellular ROS level compared to control. Further investigation revealed that PA caused a dose-dependent increase in DNA strand breaks (DSB) of M. aeruginosa at exposure concentration from 2 to 50mgL(-1). The incubation of cells with ROS scavengers ascorbic acid, N-acetyl-l-cysteine (NAC) and tocopherol markedly alleviated the level of PA-induced DSB. Analysis of PA autoxidized products in culture solution showed that PA was quickly converted to purpurogallin (PG), and PG was further autoxidized to other polyphenolic compounds. PA and PG might participate a futile redox cycle, which mediated ROS production in M. aeruginosa. These results suggested DNA strands and cell membrane were two targets of ROS induced by PA, and oxidative damage was an important mechanism for the toxicity of PA against M. aeruginosa.

Analysis of genetic diversity and population structure of Bellamya quadrata from lakes of middle and lower Yangtze River.

As an endemic species of freshwater gastropods in China, Bellamya quadrata plays an important role in ecosystem service provision and commercial importance. However, the species is overharvested and its natural habitats are under severe threat due to fragmentation and loss. To estimate the genetic diversity and population structure of B. quadrata, 285 individuals from eight lake populations across middle and lower Yangtze River were sampled. Seven microsatellite loci were genotyped. Our results showed that (i) the genetic diversity of B. quadrata was high in most of the studied populations, yet effective population sizes appear to be rather small in some populations; (ii) low levels of genetic differentiation exists among populations but gene flow was generally high; (iii) no clear geographic or genetic structure was observed in the studied region, implying mechanisms (zoochoric dispersal and anthropogenic translocations) that enhance dispersal and gene flow have promoted population connectivity. However, the comparatively high genetic diversity of B. quadrata could be attributed to a lag phase, suggesting that the genetic diversity of this species may be lost in the future and the priorities for conservation of B. quadrata are necessary.

Laboratory-Simulated Inhibitory Effects of the Floating-Bed Plants on Microcystis aeruginosa and Their Microbial Communities' Responses to Microcystins.

Three common floating bed plants, Eichhornia crassipes, Pistia stratiotes, and Ipomoea aquatica, were selected in the present study to investigate their inhibitory effects on toxic Microcystis aeruginosa. The results showed that all three types of floating-bed plants could considerably inhibit the growth of M. aeruginosa and effectively remove the microcystins (MCs) from water systems, among which, E. crassipes and P. stratiotes were more effective in resisting M. aeruginosa, and the removal rate of the intracellular MCs could be up to 100%. In addition, the roots and leaves of the three plants were enriched with a large number of MCs and demonstrated significant antioxidant responses, as evidenced by the increase in the content of catalase (CAT), glutathione peroxidase (GSH-PX), superoxide dismutase (SOD), and malondialdehyde (MDA) in the roots, stems, and leaves of the plants. Furthermore, this study also showed that Proteobacteria, Bacteroidota, Myxococcota, Verrucomicrobiota, and Actinobacteriota dominated the root microorganisms of the three plants. Moreover, a variety of MC-degrading bacteria, including Sphingomonas, Acinetobacter, Novosphingobium, and Pseudomonas, were found at the genus level, which further provides important basic data for the regulation of eutrophic water bodies and the removal of MCs.

The Combined Effects of Toxic Microcystis aeruginosa and Thermal Stress on the Edible Clam (Corbicula fluminea): Insights into Oxidative Stress Responses and Molecular Networks.

Cyanobacterial blooms (CYBs) have become a global environmental issue, posing risks to edible bivalves. Toxic cyanobacteria and thermal stress represent the two key co-occurring stressors to bivalves experiencing CYBs. To investigate the combined effects of these stressors on the edible bivalve Corbicula fluminea, the responses to oxidative stress and the molecular mechanisms of physiological adaptations in C. fluminea were examined under co-exposure to toxic Microcystis aeruginosa and thermal stress. The activity of antioxidant enzymes, including GST, SOD, CAT, GPx and GR, was significantly influenced by the interaction between temperature and M. aeruginosa (p < 0.05). A positive correlation was observed between toxic M. aeruginosa exposure and elevated SOD and GPx activities at 30 °C, demonstrating that SOD and GPx may help C. fluminea defend effectively against MCs under thermal stress. Furthermore, significant interactive effects between toxic M. aeruginosa and temperature were also observed in ROS and MDA (p < 0.05). The results of the PCA and IBR index also evidenced the apparent influence of toxic M. aeruginosa and thermal stress on oxidative stress responses of C. fluminea. The eggNOG and GO annotations confirmed that a substantial portion of differentially expressed genes (DEGs) exhibited associations with responses to oxidative stress and transporter activity. Additionally, KEGG analysis revealed that abundant DEGs were involved in pathways related to inflammatory responses, immune functions and metabolic functions. These findings improve our understanding of the mechanism of the physiological adaptation in bivalves in response to cyanotoxins under thermal conditions, potentially enabling the evaluation of the viability of using bivalves as a bioremediation tool to manage CYBs in eutrophic waters.

Feeding behavior, microcystin accumulation, biochemical response, and ultramicrostructure changes in edible freshwater bivalve Corbicula fluminea exposed to Microcystis aeruginosa.

As filter-feeders, bivalves naturally come into direct contact with microcystins (MCs) in eutrophic water bodies suffering from cyanobacteria blooms. To date, however, no studies have quantified the dynamics of microcystin accumulation and depuration in the edible freshwater bivalve Corbicula fluminea when exposed to dense bloom concentrations of Microcystis aeruginosa, while considering dynamic changes of biochemical indexes and feeding structure. In the present study, the bioaccumulation and detoxification of microcystin-LR (MC-LR) in C. fluminea were investigated. Our results showed that C. fluminea would graze equally efficiently on green algae and M. aeruginosa, irrespective of whether the M. aeruginosa strains were toxic or non-toxic. MCs could be accumulated and depurated by C. fluminea efficiently. In addition, linear and exposure time-dependent MC-LR accumulation patterns were observed in C. fluminea. Activities of biotransformation (glutathione S-transferase, GST) and antioxidant enzymes (superoxide dismutase, SOD, and catalase, CAT) and malondialdehyde (MDA) contents in various tissues of treated clams were stimulated by MCs in a tissue-specific manner. Our findings indicated that C. fluminea hepatopancreas was the primary target organ for MC-LR detoxification processes, as evidenced by a significant increase in GST activity. Besides, gills and mantle were more sensitive than the other tissues to oxidative stress in the initial microcystin exposure period with a significant increase in SOD activity. The scanning electron microscopy (SEM) observations revealed that the lateral cilia in the gill aperture were well developed during the MCs exposure period, which could perform the filter-feeding function instead of the damaged frontal cilium. This study provides insight into the possible tolerance of C. fluminea exposed to dense bloom concentrations of M. aeruginosa.

Higher resistance of a microcystin (MC)-producing cyanobacterium, Microcystis, to the submerged macrophyte Myriophyllum spicatum.

Outbreaks of Microcystis blooms can affect growth of submerged plants, which in turn can inhibit cyanobacterial growth. Microcystin (MC)-producing and non-MC-producing Microcystis strains typically coexist in Microcystis-dominated blooms. However, the interaction between submerged plants and Microcystis at strain level is not clear. This study was aimed at assessing the effects of a submerged macrophyte Myriophyllum spicatum on one MC-producing versus one non-MC-producing strains of the cyanobacterium Microcystis using plant-Microcystis co-culture experiments. The impacts of Microcystis on M. spicatum were also examined. It showed that the MC-producing Microcystis strain had a higher resistance to negative impacts by the cocultured submerged plant M. spicatum than the non-MC-producing strain. By contrast, the plant M. spicatum was impacted more by the MC-producing Microcystis than the non-MC-producer. The associated bacterioplankton community was affected more by the MC-producing Microcystis than the cocultured M. spicatum. The MC cell quotas were significantly higher in the coculture treatment (the PM + treatment, p < 0.05), indicating that the production and release of MCs might be a key factor responsible for the reduced impact of M. spicatum. The higher concentrations of dissolved organic and reducing inorganic compounds might eventually exacerbate the recovering capacity of coexisting submerged plants. Overall, this study indicated that the capacity to produce MCs, as well as the density of Microcystis, should be taken into account when attempting to reestablish submerged vegetation to undertake remediation works.

[Temporal-spatial distribution of functional feeding groups of macroinvertebrate and biological evaluation of water quality in Xinyang section of the Huaihe River main stream]. / æ·®æ²³å¹²æµä¿¡é˜³æ®µå¤§åž‹åº•æ –åŠ¨ç‰©åŠŸèƒ½æ‘„é£Ÿç±»ç¾¤æ—¶ç©ºåˆ†å¸ƒåŠæ°´è´¨è¯„ä»·.

From November 2021 to September 2022, we conducted four field surveys on macroinvertebrates and water environmental factors in Xinyang section of the Huaihe River main stream. We analyzed the structure and spatiotemporal distribution characteristics of the functional feeding groups of macroinvertebrates, and evaluated river water quality. A total of 73 macroinvertebrate species were collected in the basin, belonging to 42 families, 7 classes, and 3 phyla. The dominant species of macroinvertebrates changed significantly in different months, with Exopalaemon modestus being the absolute advantage species in the basin in July and September 2022. In different sampling months, the functional feeding group of macroinvertebrates was mainly dominated by shredders, accounting for 35.9%. The results of redundancy analysis showed that the main environmental factors affecting the distribution of functional feeding groups of macroinvertebrates varied across different months, with conductivity in February, temperature in July, and oxidation-reduction potential in September and November. The evaluation based on the biolo-gical index and Shannon index of macroinvertebrates indicated that water quality in the investigated section was at a light pollution level.

Responses of submerged macrophyte Ceratophyllum demersum to the gradient concentrations of microcystin-LR (MC-LR).

The responses of Ceratophyllum demersum to gradient concentrations (0, 0.8, 3.2, and 10 µg/L) of microcystin-LR (MC-LR) were comprehensively investigated by laboratory simulation experiments. The high reduction and accumulation efficiency of MC-LR by C. demersum were verified in this study. Results showed that the reduction ratio of MC-LR in the cultivation medium was up to 99% after 14 days of exposure, and the accumulation of MC-LR in C. demersum was highest at an exposure concentration of 10 µg/L, the value of which was 0.9 ng/g fresh weight (FW). Meanwhile, a series of negative effects on C. demersum was detectable, accompanied by a significant biomass reduction of the plant and changes in microbial community composition. In particular, this study indicated that the amount of Flavobacteria was elevated under the stress of MC-LR, provoking great threats to aquatic ecosystems. Moreover, oxidative damage was evidenced by the changes in total antioxidant capacity, superoxide dismutase, and glutathione. The results also demonstrated significant increases in sugar (0.025 mg/g FW), protein (0.3 mg/g FW), and carotenoids (0.6 mg/g FW) in C. demersum stressed by 10 µg/L of MC-LR, compared with the control without microcystins, which were among the defense strategies for dealing with adverse conditions. These results verified the good potential of submerged macrophytes as an eco-friendly strategy for controlling cyanobacterial blooms. However, the negative effects of MC-LR on the macrophytes themselves were also demonstrated, which would be considered in future practice and management.

Enhanced resistance of co-existing toxigenic and non-toxigenic Microcystis aeruginosa to pyrogallol compared with monostrains.

Cyanobacteria species are sensitive to many plant allelochemicals, such as pyrogallol. However, little attention has been paid to the relative effects of these xenobiotics on co-occurring toxigenic and non-toxigenic cyanobacterial strains, despite their co-existence in blooms. Hence, the responses of one toxigenic (TS2) and two non-toxigenic (NS1, NS2) Microcystis aeruginosa strains to pyrogallol were tested under three conditions: mono-culture and co-cultured either directly or separately by dialysis membrane. The study showed that the inhibitory effects of pyrogallol on the growth and photosynthetic yield (Fv/Fm) of either toxigenic or non-toxigenic M. aeruginosa strains were lower in direct and dialysis co-culture conditions than those in mono-culture conditions. This result indicated that chemical-mediated reciprocal effects occur between the co-existing toxigenic and non-toxigenic strains. The toxigenic M. aeruginosa strain was more sensitive to pyrogallol than the non-toxigenic strains in both mono- and co-culture systems, though whether this outcome is due to the former's toxigenic status is unclear. Intracellular microcystin-LR (MC-LR) concentrations of the toxigenic strain decreased after pyrogallol addition in both mono- and co-culture systems, whereas extracellular MC-LR concentrations increased. This finding may reflect the cell damage of M. aeruginosa because of the pyrogallol. At the same initial number of cells, the extracellular MC-LR concentration released from the same amount of TS2 cells in mono-culture was slightly higher than that in dialysis co-culture conditions. Overall, this study shows that plant allelochemicals may have the potential to reduce bloom toxicity by reducing the proportion of toxigenic cyanobacterial strains, and the effects of co-existing strains must be considered when assessing the effects of plant allelochemicals on target strains.

Generation of reactive oxygen species in cyanobacteria and green algae induced by allelochemicals of submerged macrophytes.

Inhibition of phytoplankton by allelochemicals released by submerged macrophytes is reported to be one of the mechanisms that maintain a clear-water state in shallow lakes. In order to elucidate this mechanism, the ability of six polyphenols and two long-chain fatty acids to induce the generation of reactive oxygen species (ROS) in phytoplankton was studied using the ROS sensitive probe 2',7'- dichlorodihydrofluorescein diacetate (DCFH-DA). The results showed that only (+)-catechin (CA) and pyrogallic acid (PA) could induce ROS formation in Microcystis aeruginosa and Pseudokirchneriella subcapitata. 25 mg L⁻¹ CA caused 1.2, 1.4 and 1.8 times increase of ROS levels in M. aeruginosa at 1, 2 and 4h exposure, respectively, and, correspondingly in P. subcapitata cells, these values were 3.7, 6.2 and 7.7, respectively. PA also significantly increased the levels of intracellular ROS in P. subcapitata (P < 0.01); however, significant ROS generation in M. aeruginosa was observed at only 4h exposure (P < 0.01). Light enhanced ROS generation in CA treated cells, but not in the cells treated with PA. CA and PA may act as redox cyclers after uptake by test organisms and produce ROS successively. These results suggest that the oxidative stress induced by the redox cycling property of allelochemicals may be one of the important causes for the inhibitory effect of some submerged macrophytes towards undesired phytoplankton in natural aquatic ecosystems.

The management of undesirable cyanobacteria blooms in channel catfish ponds using a constructed wetland: Contribution to the control of off-flavor occurrences.

An exploratory study on the management of undesirable cyanobacteria blooms with respect to off-flavor problems using an integrated vertical-flow constructed wetland (CW) was performed at a small commercial-scale channel catfish farm from 2004 to 2007. The results of the three-year experiment indicated that water treatment by the CW could reduce the possibility of dominance by undesirable cyanobacteria species that often cause off-flavor problems. A detailed investigation in 2007, showed that the concentrations of geosmin, MIB (2-methylisoborneol), and ß-cyclocitral in the water of the recirculating pond (4.3ngL(-1), U.D. (undetected) and 0.2ngL(-1), respectively) treated by the CW were significantly lower than those in the control pond (152.6ngL(-1), 63.3ngL(-1) and 254.8ngL(-1), respectively). In addition, the relationships among the cyanobacteria species, the off-flavor compounds and ten environmental variables were explored by canonical correspondence analysis (CCA). The results showed that Oscillatoria sp., Oscillatoria kawamurae and Microcystis aeruginosa were the main sources of off-flavor compounds in the catfish ponds. The successful manipulation of undesirable cyanobacteria species potentially resulted in lower concentrations of odorous compounds in the water of the recirculating pond. An investigation of the concentrations of geosmin and MIB in catfish fillets showed that the levels of odorous compounds were below the OTC (odor threshold concentration) values in the recirculating pond but were above the OTC values from July to October in the control pond. Water recycling by the CW could potentially be one of the best management practices to control off-flavor occurrences in aquaculture.

[Allelopathic influence of Myriophyllum spicatum on the photosynthetic efficiency of Microcystis aeruginosa].

The allelopathic influence of Myriophyllum spicatum on chlorophyll content and chlorophyll fluorescence parameters of Microcystis aeruginosa was studied in coexistence condition. Chlorophyll fluorescence parameters included q(N) (non-photochemical quenching), Y II (effective quantum yield), F(v)/F(m) (maximum quantum yield), F'(v)/F'(m) (effective quantum yield of photosystem II photochemistry) and ETR (electron transport rate). During the three days under coexistence condition, chlorophyll content and chlorophyll fluorescence parameters of M. aeruginosa were affected by M. spicatum and presented different sensitivities. Chlorophyll content of M. aeruginosa was significantly inhibited by 20.80% on the second day at 10.0 g/L of M. spicatum (P < 0.05). However, chlorophyll fluorescence parameters of M. aeruginosa decreased earlier and rapider than chlorophyll content. On the first day, q(N) and Y II of M. aeruginosa were significantly inhibited by 15.59% and 13.00% at 5.0 g/L of M. spicatum (P < 0.05), and F(v)/F(m) and F'(v) /F'(m) were declined by 15.87% and 12.07% at 10.0 g/L (P < 0.05), respectively. On the third day, ETR and three parameters based on ETR were affected at all levels of M. spicatum (P < 0.05). The inhibition effects on the photosynthetic activity of M. aeruginosa might be considered as one of the target sites of M. spicatum and chlorophyll fluorescence parameters were more sensitive parameters than chlorophyll content, especially q(n).

Study on the mechanism of allelopathic influence on cyanobacteria and chlorophytes by submerged macrophyte (Myriophyllum spicatum) and its secretion.

For revealing the mechanism of allelopathic influence on phytoplankton by aquatic macrophytes, the growth and photosynthetic activities of cyanobacteria Microcystis aeruginosa and the chlorophyte Selenastrum capricornutum were investigated when they coexisted with submerged macrophyte Myriophyllum spicatum and were exposed to allelopathic polyphenols: pyrogallic acid (PA), gallic acid (GA), ellagic acid (EA) and (+)-catechin (CA). According to the results of coexistence assays, the non-photochemical quenching (NPQ) and effective quantum efficiency (YII) of M. aeruginosa were affected earlier and more rapidly than the cell density. However, the influence of M. spicatum on S. capricornutum was not found. When the Toxicity Index (TI) was applied to evaluate the combined effects of binary and multiple mixtures of polyphenols, it was found that the four tested polyphenols with the proportion identified in the M. spicatum-cultured solution were observed to present synergistic effect (0.36-0.49) according to the cell density, NPQ and YII of M. aeruginosa. With the combined effects of polyphenols on S. capricornutum, only additive action (0.52-1.62) was found. On the other hand, PA (2.97mgL(-1)), GA (2.65mgL(-1)) caused significant reductions of photosystem II (PSII) and whole electron transport chain activities of M. aeruginosa by 71.43 and 18.37%, 70.95 and 40.77% (P<0.05), respectively, after 24-h exposure, but no inhibition effect was found in S. capricornutum. The dark respiration and photosystem I (PSI) activities of M. aeruginosa were significantly increased by exposure to PA and GA (P<0.05). Nevertheless, EA and CA had no influence on the electron transport activities of the tested organisms. These results indicate that the reduction in photosynthetic activity of M. aeruginosa and the synergistic effect of allelochemicals may be two important causes for the inhibition of undesired phytoplankton by submersed macrophytes in natural aquatic ecosystems, and PSII in cyanobacteria is considered to be one of the target sites attacked by allelopathic polyphenols.