Algicidal activity of Salvia miltiorrhiza Bung on Microcystis aeruginosa—Towards identification of algicidal substance and determination of inhibition mechanism

The present study was to isolate and identify a potent algicidal compound from extract of Salvia miltiorrhiza and study the potential inhibition mechanism on Microcystis aeruginosa. Column chromatography and bioassay-guided fractionation methods were carried out to yield neo-przewaquinone A, which was identified by spectral analysis. The EC50 of neo-przewaquinone A on M. aeruginosa were 4.68 mg L⁻¹. In addition, neo-przewaquinone A showed relatively higher security on Chlorella pyrenoidosa and Scenedesmus obliquus, with the EC50 values of 14.78 and 10.37 mg L⁻¹, respectively. For the potential inhibition mechanisms, neo-przewaquinone A caused M. aeruginosa cells morphologic damage or lysis, increased malondialdehyde content and decreased the soluble protein content, total antioxidant and superoxide dismutase activity, and significantly inhibited three photosynthesis-related genes (psaB, psbD, and rbcL). The results demonstrated the algicidal effect of neo-przewaquinone A on M. aeruginosa and provided the possible inhibition mechanisms.     

Effects of allelochemical extracted from water lettuce (Pistia stratiotes Linn.) on the growth, microcystin production and release of Microcystis aeruginosa

This study explored the optimisation of a method of extracting allelochemicals from Pistia stratiotes Linn., identified the optimal dose range for the allelochemicals’ anti-algal effect and investigated their impact on the growth of Microcystis aeruginosa, as well as the production and release of microcystin-LR (MC-LR). Based on measured changes in algal cell density and chlorophyll a (Chl-a) content, the allelochemicals were confirmed to have the strongest anti-algal effect with the lowest half-effect concentration of 65 mg L^?1 when they were extracted using ethyl acetate as the extraction solvent, 1:20 g mL^?1 as the extraction ratio and 1 h as the extraction time. The allelochemicals extracted from P. stratiotes using this optimal method exhibited the strongest inhibitory effect on the growth of algae when used within a dose range of 60–100 mg L^?1; the relative inhibitory ratio reached 50–90 %, and Chl-a content reduced 50–75 % in algae cell cultures within 3–7 days. In addition, the extracted allelochemical compounds demonstrated no significant impact on the extracellular release of MC-LR during the culturing period. The amount of intracellular MC-LR per 10⁶ algal cells increased depending on the increasing dose of allelochemicals from P. stratiotes after 7 days of culturing and maintained stability after 16 days. There was no increase in the total amount of MC-LR in the algal cell culture medium. Therefore, the application of allelochemicals from P. stratiotes to inhibit M. aeruginosa has a high degree of ecological safety and can be adopted in practical applications for treating water subjected to algae blooms because the treatment can effectively inhibit the proliferation of algal cells without increasing the release of cyanotoxin.     

Mechanisms of photosynthetic inactivation on growth suppression of Microcystis aeruginosa under UV-C stress

This study aims to investigate the effects of UV-C irradiation on photosynthetic processes of Microcystis aeruginosa to unravel the mechanism(s) involved in how and in what ways UV-C mediates growth suppression and cellular recovery. Changes in the concentration of photosynthetic pigments, photochemical efficiency, PS II core protein (D1) content, and the coding genes expressions were measured. The results indicate that UV-C doses at 20–200 mJ cm⁻² lead to rapid reduction in gene expression of both psbA (for D1) and cpc (for phycocyanin), but the suppression was short term and recoverable within 3 d of post-UV incubation. Conversely, UV-C doses at ?50 mJ cm⁻² could induce marked decline in photochemical efficiency (represented by the optimal PS II quantum yield, F_V/F_M, and the effective PS II quantum yield, Y) as well as decreases in D1 content and water soluble pigments (phycoerythrins, phycocyanins, allophycocyanins) in M. aeruginosa during the post UV-C incubation period. The results suggest that interruption of both the light energy harvesting apparatus (especially the water soluble pigments) and the photochemical process mainly accounted for the growth suppression effect in UV-C irradiated M. aeruginosa.     

Antioxidant and metabolism responses to polyphenol stress in cyanobacterium Microcystis aeruginosa

Three common polyphenol compounds Gallic Acid (GA), Pyrogallic Acid (PA) and Catechol (CA) are known to have allelochemical-exhibiting inhibitory effects on the growth of the cyanobacterium Microcystis aeruginosa (M. aeruginosa). Metabolism and antioxidant responses in M. aeruginosa were investigated to elucidate the mechanism by which the three polyphenols inhibit algal growth. The inhibition effects of polyphenols were in the order of CA > PA > GA. The GA and CA exposures increased protein contents, superoxide dismutase (SOD) activity, catalase (CAT) activity and soluble sugar, especially for exposure to GA of 25 mg L^?1. Soluble sugar content increased significantly especially when exposed to CA for 72 h. When exposed to PA, protein content, and SOD and CAT activities initially increased but over longer treatment time the activities decreased, in contrast to sugar content. Our results suggest that PA exposure for longer periods of time may inhibit catabolism action, while CA exposure could induce more oxide stress than GA or PA. The overall study showed that polyphenol-induced oxidative damage might be responsible for polyphenol inhibition on M. aeruginosa growth. The increases in cellular antioxidant enzymes and soluble sugar may have been to counteract the oxidative stress.     

Polystyrene nanoplastics affect growth and microcystin production of Microcystis aeruginosa

Nanoplastics are widely distributed in freshwater environments, but few studies have addressed their effects on freshwater algae, especially on harmful algae. In this study, the effects of polystyrene (PS) nanoplastics on Microcystis aeruginosa (M. aeruginosa) growth, as well as microcystin (MC) production and release, were investigated over the whole growth period. The results show that PS nanoplastics caused a dose-dependent inhibitory effect on M. aeruginosa growth and a dose-dependent increase in the aggregation rate peaking at 60.16% and 46.34%, respectively, when the PS nanoplastic concentration was 100 mg/L. This caused significant growth of M. aeruginosa with a specific growth rate up to 0.41 d^?1 (50 mg/L PS nanoplastics). After a brief period of rapid growth, the tested algal cells steadily grew. In addition, the increase in PS nanoplastics concentration promoted the production and release of MC. When the PS nanoplastic concentration was 100 mg/L, the content of the intracellular (intra-) and extracellular (extra-) MC increased to 199.1 and 166.5 μg/L, respectively, on day 26, which was 31.4% and 31.1% higher, respectively, than the control. Our results provide insights into the action mechanism of nanoplastics on harmful algae and the potential risks to freshwater environments.

     

Salicylic acid involved in the regulation of nutrient elements uptake and oxidative stress in Vallisneria natans (Lour.) Hara under Pb stress

In this study, the alterations in nutrient elements content, reactive oxygen species level and antioxidant response were studied in leaves of Vallisneria natans (Lour.) Hara exposed to salicylic acid (SA, 10 or 100 μM), or Pb (50 μM) or their combinations for 4 d. No significant alterations in Mn and Ca content were observed but content of Cu, Zn, Fe and P decreased in plants exposed to SA alone. SA application inhibited the uptake of Pb and partially reversed Pb-induced the alterations in Mn, Ca and Fe content in leaves of V. natans exposed to 50 μM Pb. The decreased chlorophyll (a + b) and increased malondialdehyde and O²⁻ and H₂O₂ content were detected in plants exposed to 100 μM SA, 50 μM Pb, 10 μM SA + 50 μM Pb or 100 μM SA + 50 μM Pb. Application SA partially inhibited Pb-induced the increase of malondialdehyde, O²⁻ and H₂O₂ content. 100 μM SA decreased the activity of NADH oxidase and the content of non-protein thiols, carotenoids and ascorbic acid and increased the content of dehydroascorbate in plants treated with or without Pb. SA alone decreased the ascorbate peroxidase activity and increased the catalase and peroxidase activity, while SA application increased catalase activity but had no significant effect on ascorbate peroxidase and peroxidase activity in V. natans exposed to Pb. The results indicate that SA involves in the regulation of Pb uptake, nutrient balance and oxidative stress.     

Distribution of Se and its species in Myriophyllum spicatum and Ceratophyllum demersum growing in water containing Se (VI)

The uptake of Se (VI) by two aquatic plants, Myriophyllum spicatum L. and Ceratophyllum demersum L., and its effects on their physiological characteristics have been studied. Plants were cultivated outdoors under semi-controlled conditions and in two concentrations of Na selenate solution (20 μg Se L⁻¹ and 10 mg Se L⁻¹). The higher dose of Se reduced the photochemical efficiency of PSII in both species, while the lower dose had no effect on PSII. Addition of Se had no effect on the amounts of chlorophyll a and b. The concentration of Se in plants grown in 10 mg Se L⁻¹, averaged 212 ± 12 μg Se g⁻¹ DM in M. spicatum (grown from 8-13 d), and 492 ± 85 μg Se g⁻¹ DM in C. demersum (grown for 31 d). Both species could take up a large amount of Se. The amount of soluble Se compounds in enzyme extracts ranged from 16% to 26% in control, and in high Se solution from 48% to 36% in M. spicatum and C. demersum, respectively. Se-species were determined using HPLC-ICP-MS. The main soluble species in both plants was selenate (∼37%), while SeMet and SeMeSeCys were detected at trace levels.     

不同质量浓度黄菖蒲和狭叶香蒲对铜绿微囊藻的化感作用简

分别采用黄菖蒲（Iris pseudacorus L.）、狭叶香蒲（Typha angustifolia L.）和铜绿微囊藻（Microcystis aeruginosa）共生培养的实验方法研究了不同质量浓度黄菖蒲、狭叶香蒲对铜绿微囊藻的化感作用。结果表明，黄菖蒲在质量浓度大于10 g/L时对初始密度为1.0×10⁷ ind/mL的铜绿微囊藻具有较好的抑制作用，表现为黄菖蒲质量浓度为10、20和40 g/L时，第15天对铜绿微囊藻的抑制率分别为30.1%、51.8%和84.0%；狭叶香蒲在质量浓度大于20 g/L时对铜绿微囊藻有明显的抑制作用，表现为狭叶香蒲质量浓度为20 g/L和40 g/L时，第15天对铜绿微囊藻的抑制率分别为34.2%和77.7%，实验过程中，铜绿微囊藻叶绿素a含量逐渐减少，而藻密度、SOD活性及MDA含量先增加后逐渐降低，表明经过一段时间持续地化感胁迫，黄菖蒲和狭叶香蒲可以诱导铜绿微囊藻产生氧化胁迫，导致细胞结构严重损伤和叶绿素大量分解，从而强烈抑制铜绿微囊藻的生长。     

Converging hazard assessment of gold nanoparticles to aquatic organisms

The gold nanoparticles (Au-NPs) are being increasingly used because of their huge diversity of applications, and consequently, elevated levels in the environment are expected. However, due to their physico-chemical properties and functionalization a high variety of Au-NPs can be found, and complete toxicological information for each type of Au-NPs still lacks, and even, the toxicological information for the same species is sometimes contradictory. Therefore, hazard assessment should be done case by case. Hence, the objective of this study was to obtain ecotoxicological information of the same Au-NPs in aquatic organisms and to find a rationale for Au-NPs toxicity. For such a purpose, bare and hyaluronic acid capped Au-NPs (12.5 nm) along with Au-NPs bulk material were tested on freshwater algae, Daphnia and zebrafish. Results showed that while gold nanoparticles were found to be harmless to the tested organisms, the soluble gold showed to be toxic to algae and Daphnia, with an LC₅₀ between 1 and 2 mg L⁻¹. Comparing our results with those gathered in the literature, it appears that a common hazard assessment of Au-NPs on the studied organisms can be elucidated.     

Bioaccumulation and depuration of anthracene in Penaeus monodon (Fibricius) through food ingestion

Understanding on the bioaccumulation and depuration of PAHs (polycyclic aromatic hydrocarbons) in Penaeus monodon is important in seafood safety because it is one of the most popular seafood consumed worldwide. In this study, we used anthracene as the precursor compound for PAHs accumulation and depuration in the shrimp. Commercial feed pellets spiked with anthracene were fed to P. monodon. At 20 mg kg⁻¹ anthracene, P. monodon accumulated 0.1% of the anthracene from the feed. P. monodon deputed the PAH two times faster than its accumulation. The shrimp reduced its feed consumption when anthracene content in the feed exceeded 20 mg kg⁻¹. At 100 mg kg⁻¹ anthracene, P. monodon started to have necrosis tissues on the posterior end of their thorax. The bioaccumulation factor (BAF), uptake rate constant (k₁) and depuration rate constant (k₂) of anthracene in P. monodon were 1.15 × 10⁻³, 6.80 × 10⁻⁴ d⁻¹ and 6.28 × 10⁻¹ d⁻¹, respectively. The depuration rate constant is about thousand times higher than the uptake rate constant and this indicated that this crustacean is efficient in depurating hydrocarbons from their tissue.     

Bioadsorption and bioaccumulation of chromium trivalent in Cr(III)-tolerant microalgae: A mechanisms for chromium resistance

Anthropogenic activity constantly releases heavy metals into the environment. The heavy metal chromium has a wide industrial use and exists in two stable oxidation states: trivalent and hexavalent. While hexavalent chromium uptake in plant cells has been reported that an active process by carrying essential anions, the cation Cr(III) appears to be taken up inactively. Dictyosphaerium chlorelloides (Dc1M), an unicellular green alga is a well-studied cell biological model organism. The present study was carried out to investigate the toxic effect of chromium exposures on wild-type Cr(III)-sensitive (Dc1M^wt) and Cr(III)-tolerant (Dc1M^Cr(III)R30) strains of these green algae, and to determine the potential mechanism of chromium resistance. Using cell growth as endpoint to determine Cr(III)-sensitivity, the IC₅₀₍₇₂₎ values obtained show significant differences of sensitivity between wild type and Cr(III)-tolerant cells. Scanning electron microscopy (SEM) showed significant morphological differences between both strains, such as decrease in cell size or reducing the coefficient of form; and transmission electron microscopy (TEM) revealed ultrastructural changes such as increased vacuolization and cell wall thickening in the Cr(III)-tolerant strain with respect to the wild-type strain. Energy dispersive X-ray analysis (SEM/XEDS) revealed that Cr(III)-tolerant D. chlorelloides cells are able to accumulate considerable amounts of chromium distributed in cell wall (bioadsorption) as well as in cytoplasm, vacuoles, and chloroplast (bio-accumulation). Morphological changes of Cr(III)-tolerant D. chlorelloides cells and the presence of these electron-dense bodies in their cell structures can be understood as a Cr(III) detoxification mechanism.     

Kinetics and equilibrium properties of the biosorption of Cu2+ by algae

The purpose of this study was to examine the kinetics and equilibrium properties of freshwater algae with Cu²⁺. This was a model system to explore using algae as biosensors for water quality. Methods included making luminescence measurements (fluorescence) and copper ion-selective electrode (CuISE) measurements vs. time to obtain kinetic data. Results were analyzed using a pseudo-first-order model to calculate the rate constants of Cu²⁺ uptake by algae: k _p(Cu?Calgae)?=?0.0025?±?0.0006?s^?1 by CuISE and k _p(Cu?Calgae)?=?0.0034?±?0.0011?s^?1 by luminescence. The binding constant of Cu?Calgae, K _Cu?Calgae, was 1.62?±?0.07?×?10⁷?M^?1. Fluorescence results analyzed using the Stern?CVolmer relationship indicate that algae have two types of binding sites of which only one appears to affect quenching. The fluorescence-based method was found to be able to detect the reaction of algae with Cu²⁺ quickly and at a detection limit of 0.1?mg?L^?1.     

Toxic risk associated with sporadic occurrences of Microcystis aeruginosa blooms from tidal rivers in marine and estuarine ecosystems and its impact on Artemia franciscana nauplii populations

Microcystis aeruginosa is a species of freshwater cyanobacteria which can form harmful algal blooms in freshwater water bodies worldwide. However, in spite its sporadic occurrences for short periods of time in estuarine waters, their influence on zooplankton populations present in these ecosystems has not been extensively studied. In this work, Artemia franciscana was used as test organism model, studying mortality against several strains of M. aeruginosa with different degrees of toxigenicity, measuring whole-live cells and homogenate extracts. Results were compared with microcystin-LR equivalent content, measured by immunoassay. The results show that there were no significant differences between both exposure models (whole cells and extracts), and there are significant differences respect to the toxigenicity of cyanobacterial blooms depending of the M. aerugionosa strain involved in the process. Analysis of microcystin-LR equivalent concentration test immediately below the lowest significant concentration in all M. aerugionosa strains was used to determine the potential risk associated with the cell densities during a bloom. Comparison among the selected M. aerugionsa strains show that these factors have influence in the results obtained, and thus, several differences have been evidenced depending of the microcystin-LR equivalent production and the strain type involved.     

Identification of microcystins from three collection strains of Microcystis aeruginosa

Microcystins (MCs) are toxic cyclic heptapeptides produced by various cyanobacteria genera, especially Microcystis. We identified 10 out of 12 MCs produced by three Microcystis aeruginosa strains from cyanobacteria collections, UTEX 2666, UTEX 2670 and UAM 1303, by using two analytical methods: Matrix-assisted Laser Desorption Ionization Time of Flight Mass Spectrometry (MALDI-TOF/MS) and HPLC Photodiode Array Detector coupled to a hybrid Quadrupole Time of Flight Mass Spectrometry (HPLC-PDA-QTOF/MS). MALDI-TOF/MS failed to detect non-polar MCs, such as MC-LY and MC-LW. HPLC-QTOF/MS permitted the accurate identification of most MCs present in methanolic extracts. Besides, three new MCs, namely: [D-Glu(OCH₃)⁶, D-Asp³] MC-LAba, MC-YL and MC-YM were detected by HPLC-QTOF/MS.     

Physiological and biochemical responses of Microcystis aeruginosa to phosphite

Phosphorus (P) is a key biological element and limiting nutrient in aquatic environments. Phosphate (+5) is traditionally associated with the P nutrient supply. However, phosphite (+3) has recently generated a great deal of interest, because of the possibility that it is a P source based on recognition of its vital role in the original life of the early earth. This study investigated whether phosphite can be an alternative P source for Microcystis aeruginosa PCC 7806, one of the predominant bloom species in freshwater systems. The results indicated that M. aeruginosa could not utilize phosphite as a sole P-nutrient directly for cell growth at any concentration, but that phosphite could boost cell numbers and chlorophyll a (Chl-a) content as long as phosphate was provided simultaneously. Specifically, Chl-a production increased sharply when 5.44 mg P L⁻¹ phosphite was added to 0.54 mg P L⁻¹ phosphate medium. Analysis of the maximum yield of PSII indicated that phosphite may stimulate the photosynthesis process of cells in phosphate-phosphite medium. In addition, phosphite failed to support cell growth, even though it more readily permeated the cells in P-deficient medium than in P-sufficient medium. Alkaline phosphatase activity (APA) analysis indicated that, unlike organic P, phosphite inhibits the response of cells to deficient P status, especially under P-deprived conditions.     

Accumulation, transformation, and release of inorganic arsenic by the freshwater cyanobacterium Microcystis aeruginosa

Arsenic (As) as a major hazardous metalloid was affected by phytoplankton in many aquatic environments. The toxic dominant algae Microcystis aeruginosa was exposed to different concentrations of inorganic arsenic (arsenate or arsenite) for 15 days in BG11 culture media. Arsenic accumulation, toxicity, and speciation in M. aeruginos as well as the changes of As species in media were examined. M. aeruginosa has a general well tolerance to arsenate and a definite sensitivity to arsenite. Additionally, arsenate actively elevated As methylation by the algae but arsenite definitely inhibited it. Interestingly, the uptake of arsenite was more pronounced than that of arsenate, and it was correlated to the toxicity. Arsenate was the predominant species in both cells and their growth media after 15 days of exposure to arsenate or arsenite. However, the amount of the methylated As species in cells was limited and insignificantly affected by the external As concentrations. Upon uptake of the inorganic arsenic, significant quantities of arsenate as well as small amounts of arsenite, DMA, and MMA were produced by the algae and, in turn, released back into the growth media. Bio-oxidation was the first and primary process and methylation was the minor process for arsenite exposures, while bioreduction and the subsequent methylation were the primary metabolisms for arsenate exposures. Arsenic bioaccumulation and transformation by M. aeruginosa in aquatic environment should be paid more attention during a period of eutrophication.     

Physiological effects of tetracycline antibiotic pollutants on non-target aquatic Microcystis aeruginosa

This study aimed to evaluate the aquatic toxicity of three typical tetracycline antibiotics, including tetracycline, oxytetracycline, and chlortetracycline, on the cyanobacterium Microcystis aeruginosa. The cell density, chlorophyll a content, protein content, and enzymatic antioxidant activities were determined. The results showed that the cell growth was significantly inhibited by the three compounds at a low concentration. The chlorophyll a and protein content decreased significantly after exposure to 0.05 mg L^?1 of each compound for 9 d. When exposed to 0.2–1 mg L^?1 of tetracycline, the superoxide dismutase (SOD) activity increased, but peroxidase (POD) and catalase (CAT) activities decreased. In contrast, when exposed to oxytetracycline and chlortetracycline at different concentrations ranging from 0.2 to 1 mg L^?1 and from 0.01 to 0.05 mg L^?1, the SOD activity decreased, but the POD and CAT activities increased. These findings indicate that tetracycline antibiotics influence cell growth and protein synthesis, and they also induce oxidative stress in M. aeruginosa at environmentally similar concentrations. Thus, this study may provide further insights into the toxic effects of tetracycline antibiotics and the controlled use of antibiotics.     

Response of two terrestrial green microalgae (Chlorophyta, Trebouxiophyceae) isolated from Cu-rich and unpolluted soils to copper stress

Some algae inhabit Cu-polluted soils. Intracellular Cu-accumulation and production of non-protein thiols in response to copper stress were compared in Stichococcus minor and Geminella terricola isolated from Cu-polluted and unpolluted soils, respectively. Cu-exposed (0.5 μM) S. minor accumulated lower amounts of copper (0.38 mM) than G. terricola (4.20 mM) and maintained 8.5-fold higher level of glutathione (GSH) than G. terricola. The ratio GSH/0.5 GSSG in the Cu-treated S. minor (7.21) was 7-times higher than in G. terricola. Reduced and oxidized forms of phytochelatins were found in both algae. Under copper stress (5 μM) the ratio -SH_total/Cu_{intracellular} in S. minor ranged from 2.3 to 6.2, while it was lower than 1.0 in G. terricola. Low intracellular Cu-accumulation and maintenance of high GSH level concomitant with PCs production seem to be responsible for a higher Cu-resistance of S. minor than G. terricola.     

An evaluation of the toxicity and bioaccumulation of cisplatin in the marine environment using the macroalga, Ulva lactuca

The cytotoxic drug, cisplatin (cis-PtCl₂(NH₃)₂), has been added to cultures of the marine macroalga, Ulva lactuca, under various experimental conditions. Both accumulation and internalisation over a 48 h period was greater when cisplatin was added to coastal sea water (salinity = 33) from a distilled water solution than when added to either sea water or estuarine water (salinity = 16.5) from a saline solution. This effect is attributed to the greater abundance of the more reactive monoaqua complex (cis-PtCl(OH₂)(NH₃)₂⁺) in the distilled water solution and kinetic constraints on its conversion back to cis-PtCl₂(NH₃)₂ in sea water. Despite its mode of action at the cellular level, cisplatin added up to concentrations of 150 nM did not incur a measurable reduction in the efficiency of photochemical energy conversion under any of experimental conditions tested.