Physiological and molecular mechanisms of the inhibitory effects of artemisinin on Microcystis aeruginosa and Chlorella pyrenoidosa.

Artemisinin, a novel plant allelochemical, has attracted attention for its potential selective inhibitory effects on algae, yet to be fully explored. This study compares the sensitivity and action targets of Microcystis aeruginosa (M. aeruginosa) and Chlorella pyrenoidosa (C. pyrenoidosa) to artemisinin algaecide (AMA), highlighting their differences. Results indicate that at high concentrations, AMA displaces the natural PQ at the QB binding site within M. aeruginosa photosynthetic system, impairing the D1 protein repair function. Furthermore, AMA disrupts electron transfer from reduced ferredoxin (Fd) to NADP+ by interfering with the iron-sulfur clusters in the ferredoxin-NADP+ reductases (FNR) domain of Fd. Moreover, significant reactive oxygen species (ROS) accumulation triggers oxidative stress and interrupts the tricarboxylic acid cycle, hindering energy acquisition. Notably, AMA suppresses arginine synthesis in M. aeruginosa, leading to reduced microcystins (MCs) release. Conversely, C. pyrenoidosa counters ROS accumulation via photosynthesis protection, antioxidant defenses, and by regulating intracellular osmotic pressure, accelerating damaged protein degradation, and effectively repairing DNA for cellular detoxification. Additionally, AMA stimulates the expression of DNA replication-related genes, facilitating cell proliferation. Our finding offer a unique approach for selectively eradicating cyanobacteria while preserving beneficial algae, and shed new light on employing eco-friendly algicides with high specificity.

Community Structure and Water Quality Assessment of Benthic Macroinvertebrates in Hongze Lake.

This study investigated the species, density, biomass and physicochemical factors of benthic macroinvertebrates in Hongze Lake from 2016 to 2020. Redundancy analysis (RDA) was used to analyze the relationship between physicochemical parameters and the community structure of macroinvertebrates. Macroinvertebrate-based indices were used to evaluate the water quality conditions in Hongze Lake. The results showed that a total of 50 benthic species (10 annelids, 21 arthropods and 19 mollusks) were collected. The community structure of benthic macroinvertebrates varied in time and space. The dominant species were Limnodrilus hoffmeisteri (L.hoffmeisteri), Corbicula fluminea (C.fluminea), Nephtys oligobranchia (N.oligobranchia). In 2016, arthropods such as Grandidierella sp. were the dominant species of benthos in Hongze Lake while annelids and mollusks dominated from 2017 to 2020, such as L.hoffmeisteri, N.oligobranchia, C.fluminea. The benthic fauna of Chengzi Lake and Lihewa District were relatively abundant and showed slight variation, while the benthic macroinvertebrates of the Crossing the water area were few and varied greatly. RDA showed that changes in benthic macroinvertebrate structure were significantly correlated with dissolved oxygen (DO), Pondus Hydrogenii (pH) and transparency (SD). The Shannon Wiener, Pielou, and Margalef indices indicate that Hongze Lake is currently in a moderately polluted state. Future studies should focus on the combined effects of various physicochemical indicators and other environmental factors on benthic communities.

Decoding Microcystis aeruginosa quorum sensing through AHL-mediated transcriptomic molecular regulation mechanisms.

Acyl-homoserine lactone (AHL) serves as a key signaling molecule for quorum sensing (QS) in bacteria. QS-related genes and physiological processes in Microcystis aeruginosa remain elusive. In this study, we elucidated the regulatory role of AHL-mediated QS in M. aeruginosa. Using AHL activity extract and transcriptomic analysis, we revealed significant effects of the AHL on growth and photosynthesis. AHL significantly increased chlorophyll a (Chl-a) content and accelerated photosynthetic rate thereby promoting growth. Transcriptome analysis revealed that AHL stimulated the up-regulation of photosynthesis-related genes (apcABF, petE, psaBFK, psbUV, etc.) as well as nitrogen metabolism and ribosomal metabolism. In addition, AHL-regulated pathways are associated with lipopolysaccharide and phenazine synthesis. Our findings deepen the understanding of the QS system in M. aeruginosa and are important for gaining insights into the role of QS in Microcystis bloom formation. It also provides new insights into the prevalence of M. aeruginosa in water blooms.

Quorum sensing regulation in Microcystis aeruginosa: Insights into AHL-mediated physiological processes and MC-LR production.

Quorum sensing (QS) is a widespread regulatory mechanism in Gram-negative bacteria, primarily involving the secretion of N-acyl homoserine lactone (AHL) to facilitate population density sensing. However, the existence of QS in blue-green algae, a subset of photoautotrophic Gram-negative bacteria forming high-density communities in water blooms, remains elusive. This study delves into the unexplored realm of QS in Microcystis aeruginosa (M. aeruginosa) by investigating AHL-related regulatory mechanisms and their impact on various physiological processes. Utilizing high-performance liquid chromatography coupled with mass spectrometry (HPLC-MS) and biosensors, a hitherto unknown long-chain AHL exhibiting a mass-to-charge ratio of 318 was identified in sterile M. aeruginosa cultures. Our investigation focused on discerning correlations between AHL activity fluctuations and key parameters such as microcystin (MC-LR) production, algal density, photosynthesis, buoyancy, and aggregation. Furthermore, the AHL extract was introduced during the logarithmic stage of M. aeruginosa cultures to observe the response in physiological processes. The results revealed that AHL, functioning as an autoinducer (AI), positively influenced algal growth and photosynthesis, as evidenced by the upregulated photosynthetic conversion efficiency of PSI and chlorophyll synthesis gene (psbA). AI also played a crucial role in altering surface characteristics through the synthesis of polysaccharides and proteins in EPS, subsequently promoting cell aggregation. Concomitantly, AI upregulated mcyD, enhancing the synthesis of MC-LR. Notably, our investigation pinpointed the initiation of QS in Microcystis at a density of approximately 1.22 × 10^7 cells/mL. This groundbreaking evidence underscores the regulatory role of AI in governing the physiological processes of growth, aggregation, buoyancy, and MC-LR production by activating pertinent gene expressions. This study significantly expands the understanding of QS in AHL, providing crucial insights into the regulatory networks operating in blue-green algae.

Characterization of a Novel Artemisinin Algicidal Particle and Its Inhibitory Effect on Microcystis aeruginosa.

A new artemisinin sustained-release particle (ASP) was developed that significantly inhibits Microcystis aeruginosa (M. aeruginosa) growth. The physical and chemical properties of ASPs were characterized using scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD), differential scanning calorimetry, and thermogravimetry (DSC-TG). The results demonstrated that ASPs are thermally stable and have sustained-release properties. On the sixth day, the ASPs (0.2 g L-1) inhibited M. aeruginosa with an inhibition rate (IR) greater than 70%. Additionally, ASPs inhibited M. aeruginosa without increasing microcystin-LR release (MC-LR). This research offers a novel approach to the management of cyanobacterial blooms.

Response of cytotoxin production ability to gene expression and cell molecular structure of Microcystis aeruginosa FACHB-905.

To investigate the inhibitory mechanism of artemisinin sustained-release microspheres (ASMs) on Microcystis aeruginosa (M. aeruginosa) from the molecular level, prx, psbA, fabZ, and mcyD were studied, and the cell death mode were also explored. The results showed that expression of prx was slightly up-regulated, while the expression of psbA, fabZ, and mcyD was significantly reduced. It can infer that oxidant damage and photic damage are the main mechanisms for the algicidal effect of ASMs on M. aeruginosa. It can be seen from the changes in cell morphology and structure that microspheres stress triggers apoptosis-like cell death, and the cell membrane is intact effectively preventing the leakage of microcystin-LR (MC-LR). Moreover, the down-regulation of mcyD gene also played major role in less extracellular MC-LR than intracellular MC-LR. It was concluded that the ASMs will not cause secondary ecological hazards while killing algae cells and have good application prospects.

An innovative strategy to control Microcystis growth using tea polyphenols sustained-release particles: preparation, characterization, and inhibition mechanism.

Allelochemicals have been shown to inhibit cyanobacterial blooms for several years. In view of the disadvantages of "direct-added" mode, natural and pollution-free tea polyphenolic allelochemicals with good inhibitory effect on cyanobacteria were selected to prepare sustained-release particles by microcapsule technology. Results showed that the encapsulation efficiency of tea polyphenols sustained-release particles (TPSPs) was 50.6% and the particle size ranged from 700 to 970 nm, which reached the nanoscale under optimum preparation condition. Physical and chemical properties of TPSPs were characterized to prove that tea polyphenols were well encapsulated and the particles had good thermal stability. The optimal dosage of TPSPs was determined to be 0.3 g/L, at which the inhibition rate on Microcystis aeruginosa in logarithmic growth period could be maintained above 95%. Simultaneous decrease in algal density and chlorophyll-a content indicated that the photosynthesis of algal cells was affected leading to cell death. Significant changes of antioxidant enzyme activities suggested that Microcystis aeruginosa's antioxidant systems had been disrupted. Furthermore, TPSPs increased the concentration of O2- which led to lipid peroxidation of cell membrane and a subsequent increase in malondialdehyde (MDA) content. Meanwhile, the protein content, nucleic acid content, and electrical conductivity in culture medium rose significantly indicating the cell membrane was irreversibly damaged. This work can provide a basis for the utilization of environmentally friendly algal suppressants.

Integrated transcriptomic and metabolomic analysis of Microcystis aeruginosa exposed to artemisinin sustained-release microspheres.

Artemisinin sustained-release microspheres (ASMs) have been shown to inhibit Microcystis aeruginosa (M. aeruginosa) blooms. Previous studies have focused on inhibitory mechanism of ASMs on the physiological level of M. aeruginosa, but the algal inhibitory mechanism of ASMs has not been comprehensively and profoundly revealed. The study proposed to reveal the toxicity mechanism of ASMs on M. aeruginosa based on transcriptomics and metabolomics. After exposure to 0.2 g·L-1 ASMs for 7 days, M. aeruginosa biomass was significantly inhibited, with an inhibition rate (IR) of 47 % on day 7. Transcriptomic and metabolomic results showed that: (1) 478 differentially expressed genes (DEGs) and 251 differential metabolites (DMs) were obtained; (2) ASMs inhibited photosynthesis by blocking photosynthetic pigment synthesis, destroying photoreaction centers and photosynthetic carbon reactions; (3) ASMs reduced L-glutamic acid content and blocked glutathione (GSH) synthesis, leading to an imbalance in the antioxidant system; (4) ASM disrupted nitrogen metabolism and the hindered synthesis of various amino acids; (5) ASMs inhibited glyoxylate cycle and TCA cycle. This study provides an important prerequisite for the practical application of ASMs and a new perspective for the management of harmful algal blooms (HABs).

Effect of artemisinin sustained-release algaecide on the growth of Microcystis aeruginosa and the underlying physiological mechanisms.

The aim of the study was to determine the effect of phycobiliprotein and esterase activity of Microcystis aeruginosa cells on the effect of artemisinin slow-release algaecide. We analyzed the sustained stress of artemisinin slow-release algaecide and the associated changes in density, phycobiliprotein, and esterase activity in Microcystis aeruginosa (M. aeruginosa) and monitored changes in the physical and chemical properties of the algae during the process. The results showed that the cumulative release concentration of artemisinin sustained-release algaecide in different media was similar. When the total amount of artemisinin was kept at 5.00-5.30 mg L-1, the effect of artemisinin on algal cells and the release amount of slow-release algicides reached a dynamic balance, and the equilibrium concentration could inhibit the growth of M. aeruginosa. Artemisinin slow-release algaecide slowly released artemisinin and inhibited the content of phycobiliprotein in M. aeruginosa. The esterase activity recovered after 15 days and continued to increase. Artemisinin showed no harmful effect on M. aeruginosa and increased the metabolic activity of algal cells. M. aeruginosa may undergo programmed cell death, keeping the cell membrane structure intact. The use of micro-nano materials can increase the effect of allelochemicals on Microcystis aeruginosa. The slow release of allelopathic active substances from the algae inhibitor reduces the algal density of Microcystis aeruginosa cells. However, the enhanced metabolic activity of algal cells may be due to artemisinin causing PCD in Microcystis cells, keeping the cell membrane structure intact, thereby preventing algal cell rupture and release of a large amount of algal toxins.

Effects of Cinnamomum camphora Leaves Extracts-Flocculants Composite Algaecide on Microcystis aeruginosa Growth and Microcystins Release.

In this study, a composite algaecide containing flocculants and Cinnamomum. camphora leaves extracts (CCCLE) were synthesized. The inhibition and flocculation effects on Microcystis aeruginosa (M. aeruginosa) were investigated, and the release of microcystin-LR (MC-LR) was determined. Results showed that the CCLEC composite algaecide was effective for the inhibition and flocculation of M. aeruginosa, and the optimal dose of CCLEC composite algaecide was 1.8%, which resulted in an algae inhibition ratio of 98.00% and a flocculation efficiency of 99.44% within 5 days of M. aeruginosa culturing. Besides, the total amount of MC-LR decreased by 80.04% on day 20 compared with the control group, while the concentration of intracellular MC-LR on day 5 was 36.69 µg L-1, which was related to a portion of cells underwent apoptosis-like cell death under CCLEC composite algaecide stress. The results of this study may improve our understanding of the M. aeruginosa control by CCCLE composite algaecide.

Stress of Artemisinin Sustained-Release Granules on Photosystem II, Reactive Oxygen Species and Metabolic Activity of Microcystis aeruginosa Cells.

The inhibitory mechanisms of artemisinin anti-algae sustained-release granules (AASG) on algal cells at cytoplasmic level were investigated. The results showed that 0.2 g L-1 AASG could effectively inhibit the growth of Microcystis aeruginosa (M.aeruginosa). The stress of 0.2 g L-1 AASG changed the excitation energy distribution pattern of Photosystem II (PSII) of algal cells, which showed the increase of heat dissipation share and the inhibition of physiological activities related to PSII. At the same time, AASG induced a large amount of reactive oxygen species (ROS), which aggravated the membrane lipid peroxidation and caused serious damage to algae cell membrane. AASG also resulted in the decrease of esterase activity and alkaline phosphatase activity (APA) in algal cells. Results showed that AASG inhibited algal growth by exerting adverse effects on PSII, ROS and metabolic activity of M.aeruginosa.

Effects of allelochemical artemisinin in Artemisia annua on Microcystis aeruginosa: growth, death mode, and microcystin-LR changes.

To investigate the effects of an allelochemical artemisinin extracted from Artemisia annua (A. annua) on cell growth, death mode, and microcystin-LR (MC-LR) changes of Microcystis aeruginosa (M. aeruginosa), a series of morphological and biochemical characteristics were studied. The results showed that artemisinin could inhibit the growth of M. aeruginosa and reduce the content of phycobiliprotein. Under the allelopathy of artemisinin, algae cells deformed due to swelling, which caused cell membranes to rupture and cell contents to leak. FDA/PI double-staining results showed that 15.10-94.90% of algae cells experienced the death mode of necrosis-like. Moreover, there were 8.35-14.50% of algae cells undergoing programmed cell death, but their caspase-3-like protease activity remained unchanged, which may mean that algae cells were not experiencing caspase-dependent apoptosis under artemisinin stress. Attacked by artemisinin directly, both intracellular and extracellular MC-LR increased sharply with the upregulation of mcyB, mcyD, and mcyH. The upregulation multiple of mcyH suggested that M. aeruginosa could accelerate transportation of algal toxin under adverse conditions of artemisinin. Artemisinin not only can inhibit the growth of M. aeruginosa but it also causes the accelerated release and increase of microcystin-LR. These imply that the application of artemisinin should be reconsidered in practical water bodies.