Interactions between bacteria and cyanobacteria influence the occurrence and development of harmful cyanobacterial blooms (HCBs). Bloom-forming cyanobacteria and cyanotoxin-degrading bacteria are essential in HCBs, nonetheless, their interactions and the underlying mechanisms remain unclear. To address this gap, a typical microcystin-LR (MC-LR)-degrading bacterium and a toxic Microcystis aeruginosa strain were co-cultivated to investigate their interactions. The cyanobacterial growth was enhanced by 24.8 %-44.3 % in the presence of the bacterium in the first 7 days, and the cyanobacterium enhanced the bacterial growth by 59.2 %-117.5 % throughout the growth phases, suggesting a mutualistic relationship between them. The presence of the bacterium increased cyanobacterial intracellular MC-LR content on days 4, 8, and 10 while reducing the extracellular MC-LR concentration, revealing the dual roles of the bacterium in enhancing cyanotoxin production and degrading cyanotoxins. The bacterium alleviated the oxidative stress, which may be crucial in promoting cyanobacterial growth. Critical functional genes related to cyanobacterial photosynthesis and MC-LR synthesis, and bacterial MC-LR degradation were up-regulated in the presence of the bacterium and cyanobacterium, respectively. Moreover, extracellular polymeric substances (EPS) were produced at the cell interface, implying EPS play a role in cyanobacterial-bacterial interactions. This study is the first to unveil the interaction mechanisms between cyanotoxin-degrading bacteria and bloom-forming cyanobacteria, shedding light on the dynamics of HCBs.

Cyanobacteria proliferate in warm, nutrient-rich environments and release toxic secondary metabolites into natural waters. Using cyanotoxin-contaminated water to irrigate crops could expose humans and biota, but the risk may be low if agricultural soils can sorb and retain cyanotoxins. In this report, we compared the sorption and desorption capacities of multi-class cyanotoxins/anabaenopeptins in soils of variable properties with a batch sorption procedure. The target compounds were anabaenopeptin-A, anabaenopeptin-B, anatoxin-a, cylindrospermopsin, and microcystins -LR, -RR, -LA, -LY, -LW, and -LF. Based on solid-liquid distribution coefficients (Kd), we classified cylindrospermopsin and microcystin-LA/-LY as \"very low sorptivity\", anabaenopeptin-A, -B and microcystin-LR, -LF, and -LW as \"low sorptivity\", and anatoxin-a and microcystin-RR as \"medium sorptivity\". We remain concerned about irrigating agricultural land with water contaminated with high levels of CYN and MC-LA/-LY because of their relatively low affinity and high desorption proportion in soils. The results also suggest that soil sorption can be an effective immobilization pathway for anatoxin-a and microcystin-RR. The generated data will be useful for prioritizing research on the most bioavailable cyanotoxins/anabaenopeptins that are likely to be released by the soil matrix, for environmental risk assessment.

With global warming and increasing eutrophication of water bodies, a variety of algal toxins, including microcystin (MC), released into water by cyanobacterial blooms pose a serious threat to the survival of aquatic organisms. To investigate the mechanism of the Nrf2/Keap1a pathway on resisting MC-induced oxidative stress and apoptosis in Cristata plicata, we cloned the full-length cDNA of CpBcl-2. The cDNA full-length of CpBcl-2 was 760 bp, encoded a 177 amino acid peptide, and contained a highly conserved Bcl-2-like superfamily domain. MC stimulation increased the expression and activity levels of related antioxidant enzymes. After CpNrf2 knockdown, the transcription levels of NAD(P)H quinone redox Enzyme-1 (NQO1) and related antioxidant enzymes activity in the gills and kidney of C. plicata were significantly down-regulated upon MC stress, but that was significantly upregulated after knockdown of CpKeap1a. Additionally, Upon MC stress, the mRNA levels of CpBcl-2 were increased in the gills and kidney after knockdown of CpNrf2 at 24 h, and that of CpBcl-2 were decreased at 72 and 96 h in the CpKeap1a-siRNA+MC group. Moreover, MC stimulation significantly inhibited CpJNK expression in the gills and kidney, but which regulated the Nrf2/Keap1a pathway in C. plicata. However, the JNK inhibitor SP600125 promoted the expression of CpNrf2 and related enzymes with antioxidant response element (ARE-driven enzyme) in the gills and kidney. Then, we speculated that CpKeap1a was a negative regulator of CpNrf2, and C. plicata resisted MC-induced oxidative damage and apoptosis by inhibiting JNK transcription via the Nrf2/Keap1a pathway.

Algal blooms can threaten human health if cyanotoxins such as microcystin are produced by cyanobacteria. Regularly monitoring microcystin concentrations in recreational waters to inform management action is a tool for protecting public health; however, monitoring cyanotoxins is resource- and time-intensive. Statistical models that identify waterbodies likely to produce microcystin can help guide monitoring efforts, but variability in bloom severity and cyanotoxin production among lakes and years makes prediction challenging. We evaluated the skill of a statistical classification model developed from water quality surveys in one season with low temporal replication but broad spatial coverage to predict if microcystin is likely to be detected in a lake in subsequent years. We used summertime monitoring data from 128 lakes in Iowa (USA) sampled between 2017 and 2021 to build and evaluate a predictive model of microcystin detection as a function of lake physical and chemical attributes, watershed characteristics, zooplankton abundance, and weather. The model built from 2017 data identified pH, total nutrient concentrations, and ecogeographic variables as the best predictors of microcystin detection in this population of lakes. We then applied the 2017 classification model to data collected in subsequent years and found that model skill declined but remained effective at predicting microcystin detection (area under the curve, AUC ≥ 0.7). We assessed if classification skill could be improved by assimilating the previous years\' monitoring data into the model, but model skill was only minimally enhanced. Overall, the classification model remained reliable under varying climatic conditions. Finally, we tested if early season observations could be combined with a trained model to provide early warning for late summer microcystin detection, but model skill was low in all years and below the AUC threshold for two years. The results of these modeling exercises support the application of correlative analyses built on single-season sampling data to monitoring decision-making, but similar investigations are needed in other regions to build further evidence for this approach in management application.

The toxicity of triclosan (TCS) to various aquatic organisms has been demonstrated at environmental concentrations. However, the effects and mechanisms of TCS on toxic cyanobacteria remains largely unexplored. This study investigated the physiological and molecular variations in two representative toxic Microcystis species (M. aeruginosa and M. viridis) under exposure to TCS for 12 d. Our findings demonstrated that the median effective concentration (EC50) of TCS for both Microcystis species were close to the levels detected in the environment (M. aeruginosa: 9.62 μg L-1; M. viridis: 27.56 μg L-1). An increased level of reactive oxygen species (ROS) was observed in Microcystis, resulting in oxidative damage when exposed to TCS at concentrations ranging from 10 μg L-1 to 50 μg L-1. The photosynthetic activity of Microcystis had a certain degree of recovery capability at low concentrations of TCS. Compared to M. aeruginosa, the higher recovery capability of the photosynthetic system in M. viridis would be mainly attributed to the increased ability for PSII repair and phycobilisome synthesis. Additionally, the synthesis of microcystins in the two species and the release rate in M. viridis significantly increased under 10-50 μg L-1 TCS. At the molecular level, exposure to TCS at EC50 for 12 d induced the dysregulation of genes associated with photosynthesis and antioxidant system. The upregulation of genes associated with microcystin synthesis and nitrogen metabolism further increased the potential risk of microcystin release. Our results revealed the aquatic toxicity and secondary ecological risks of TCS at environmental concentrations, and provided theoretical data with practical reference value for TCS monitoring.

Harmful algal bloom (HAB) formation leads to the eutrophication of water ecosystems and may render recreational lakes unsuitable for human use. We evaluated the applicability and comparison of metabarcoding, metagenomics, qPCR, and ELISA-based methods for cyanobacteria/cyanotoxin detection in bloom and non-bloom sites for the Great Lakes region. DNA sequencing-based methods robustly identified differences between bloom and non-bloom samples (e.g., the relative prominence of Anabaena and Planktothrix). Shotgun sequencing strategies also identified the enrichment of metabolic genes typical of cyanobacteria in bloom samples, though toxin genes were not detected, suggesting deeper sequencing or PCR methods may be needed to detect low-abundance toxin genes. PCR and ELISA indicated microcystin levels and microcystin gene copies were significantly more abundant in bloom sites. However, not all bloom samples were positive for microcystin, possibly due to bloom development by non-toxin-producing species. Additionally, microcystin levels were significantly correlated (positively) with microcystin gene copy number but not with total cyanobacterial 16S gene copies. In summary, next-generation sequencing-based methods can identify specific taxonomic and functional targets, which can be used for absolute quantification methods (qPCR and ELISA) to augment conventional water monitoring strategies.

This study explored the regulatory role of bta-miR-149-3p in the inflammatory response induced by microcystin-leucine arginine (MC-LR) exposure in bovine Sertoli cells. The research endeavored to enhance the comprehension of the epigenetic mechanisms underlying MC-LR-induced cytotoxicity in Sertoli cells and establish a foundation for mitigating these effects in vitro. In this study, we elucidated the regulatory mechanism of bta-miR-149-3p in the MC-LR-induced inflammatory response by verifying the target gene of bta-miR-149-3p through luciferase assays and treating the cells with a bta-miR-149-3p inhibitor for 24 h. The results demonstrate that nuclear factor κB (NF-κB) acts as a downstream target gene of bta-miR-149-3p, which inhibits the MC-LR-induced inflammatory response in bovine Sertoli cells. This inhibition occurs by regulating the downregulation of tight junction constitutive proteins of the blood-testis barrier (BTB) through the suppression of the TLR-4/NF-κB signaling pathway (p < 0.05) and the up-regulation of the adhesion junction protein β-catenin (p < 0.05). Notably, MC-LR exposure resulted in the up-regulation (p < 0.05) of inflammatory cytokines (IL-6, IL-1β, and NLRP3) and the down-regulation (p < 0.05) of BTB tight junction constitutive proteins (ZO-1, Occludin) in Sertoli cells. Furthermore, the BTB constitutive protein ZO-1 exhibited significant down-regulation in Sertoli cells pretreated with the bta-miR-149-3p inhibitor compared to controls (p < 0.05), while Occludin showed no significant difference from CTNNB1 (p > 0.05). In summary, our findings suggest that bta-miR-149-3p suppresses the MC-LR-induced inflammatory response and alterations in the expression of BTB proteins in bovine Sertoli cells by inhibiting the TLR-4/NF-κB signaling pathway.

Cyanobacterial blooms occur at increasing frequency and intensity, notably in freshwater. This leads to the introduction of complex mixtures of their products, i.e., cyano-metabolites, to drinking water treatment plants. To assess the fate of cyano-metabolite mixtures during ozonation, a novel multicompound ozone (O3) competition kinetics method was developed. Sixteen competitors with known second-order rate constants for their reaction with O3 ranging between 1 and 108 M-1 s-1 were applied to cover a wide range of the O3 reactivity. The apparent second-order rate constants (kapp,O3) at pH 7 were simultaneously determined for 31 cyano-metabolites. kapp,O3 for olefin- and phenol-containing cyano-metabolites were consistent with their expected reactivity (0.4-1.7 × 106 M-1 s-1) while kapp,O3 for tryptophan- and thioether-containing cyano-metabolites were significantly higher than expected (3.4-7.3 × 107 M-1 s-1). Cyano-metabolites containing these moieties are predicted to be well abated during ozonation. For cyano-metabolites containing heterocycles, kapp,O3 varied from <102 to 5.0 × 103 M-1 s-1, giving first insights into the O3 reactivity of this class of compounds. Due to lower O3 reactivities, heterocycle- and aliphatic amine-containing cyano-metabolites may be only partially degraded by a direct O3 reaction near circumneutral pH. Hydroxyl radicals, which are formed during ozonation, may be more important for their abatement. This novel multicompound kinetic method allows a high-throughput screening of ozonation kinetics.

Cyanobacteria harmful algal blooms (CHABs) are a growing water quality problem in the upper San Francisco Estuary (California), also known as the Sacramento-San Joaquin Delta. We conducted cyanobacteria and cyanotoxin monitoring from 2020 to 2023, which spanned California\'s driest consecutive 3-year period and one of the wettest years on record (2023). To assess the impact of CHABs over this range of hydrologic conditions, we monitored invasive Asian Clams (Corbicula fluminea) for microcystin contamination and used molecular tools (qPCR and sequencing) to characterize cyanobacteria in the water column. We also used solid phase adsorption toxin tracking (SPATT) samplers to track microcystins (MCs) and other cyanotoxins in 2023. During the drought years, record breaking MCs, in excess of 1000 μg/L, were documented in water grab samples and Asian clams also accumulated higher MCs relative to the wet year. However, MCs were present in Asian clams during the entire study period. SPATT\'s confirmed MC presence during wet 2023 and sequencing results corroborated the integrative sampler findings. Yet, no MC was detected in water grab samples at our primary sampling sites during the drought year of 2022 or the wet year of 2023. This highlights the importance of using multiple sampling modalities to provide a more accurate assessment of MC contamination, especially in large estuaries where traditional discrete monitoring can easily miss episodic and transient CHAB events.

Luteolin has shown great potential in inhibiting Microcystis-dominated cyanobacterial blooms. However, widespread microplastics (MPs) in natural aquatic systems often serve as substrates for cyanobacterial growth, which could impact cyanobacterial resistance to external stresses and interfere with luteolin\'s algicidal effect. This study explored the influence of virgin and diversely-aged polystyrene microplastics (PS-MPs) on inhibitory effect of luteolin on Microcystis growth and its microcystins (MCs) production/release. Moreover, the underlying mechanisms were also revealed by jointly analyzing SEM image, antioxidant response, exopolymeric substances (EPSs) production, and functional gene expression. Results suggested that 0.5, 5, and 50 mg/L virgin and diversely-aged PS-MPs almost weakened growth inhibition and oxidative damage of two doses of luteolin against Microcystisby stimulating its EPSs production and inducing self-aggregation of Microcystis cells and/or hetero-aggregation between Microcystis cells and PS-MPs. Compared to virgin PS-MPs, photo-aged PS-MPs possessed rougher flaky surfaces, and hydrothermal-aged PS-MPs showed internal cracking. These characteristics led to greater stimulation of EPS production and exhibited more significant protective effects on Microcystis. Notably, PS-MPs also decreased MCs content in aqueous phase, likely because they adsorbed some MCs. Such toxigenic hetero-aggregates formed by MCs, MPs, and Microcystis cells would directly poison grazing organisms that consume them and create more pathways for MCs into food web, posing greater eco-risks. This is the first study to clarify the influence and mechanisms of virgin and diversely-aged MPs on allelopathic algicidal effects from the perspective of microalgal inherent adaptive strategies.