Research on interactions between grazers and toxigenic algae is fundamental for understanding toxin dynamics within aquatic ecosystems and developing biotic approaches to mitigate harmful algal blooms. The dinoflagellate Alexandrium minutum is a well-known microalga responsible for paralytic shellfish toxins (PSTs) contamination in many coastal regions worldwide. This study investigated the impact of the ciliate Euplotes balteatus on cell density and PSTs transfer in simulated A. minutum blooms under controlled conditions. E. balteatus exhibited resistance to the PSTs produced by A. minutum with a density of up to 10,000 cells/mL, sustaining growth and reproduction while eliminating algal cells within a few days. The cellular PSTs content of A. minutum increased in response to the grazing pressure from E. balteatus. However, due to the substantial reduction in density, the overall toxicity of the algal population decreased to a negligible level. Most PSTs contained within algal cells were temporarily accumulated in E. balteatus before being released into the water column, suggesting unclear mechanisms for PSTs excretion in unicellular grazers. In principle, the grazing of E. balteatus on A. minutum promotes the transfer of the majority of intracellular PSTs into extracellular portions, thereby mitigating the risk of their accumulation and contamination through marine trophic pathways. However, this process also introduces an increase in the potential environmental hazards posed by extracellular PSTs to some extent.

Dinoflagellates are responsible for most marine harmful algal blooms (HABs) and play vital roles in many ocean processes. More than 90% of dinoflagellates are vitamin B12 auxotrophs and that B12 availability can control dinoflagellate HABs, yet the genetic basis of B12 auxotrophy in dinoflagellates in the framework of the ecology of dinoflagellates and particularly HABs, which was the objective of this work. Here, we investigated the presence, phylogeny, and transcription of two methionine synthase genes (B12-dependent metH and B12-independent metE) via searching and assembling transcripts and genes from transcriptomic and genomic databases, cloning 38 cDNA isoforms of the two genes from 14 strains of dinoflagellates, measuring the expression at different scenarios of B12, and comprehensive phylogenetic analyses of more than 100 organisms. We found that 1) metH was present in all 58 dinoflagellates accessible and metE was present in 40 of 58 species, 2) all metE genes lacked N-terminal domains, 3) metE of dinoflagellates were phylogenetically distinct from other known metE genes, and 4) expression of metH in dinoflagellates was responsive to exogenous B12 levels while expression of metE was not responding as that of genuine metE genes. We conclude that most, hypothetically all, dinoflagellates have either non-functional metE genes lacking N-terminal domain for most species, or do not possess metE for other species, which provides the genetic basis for the widespread nature of B12 auxotrophy in dinoflagellates. The work elucidated a fundamental aspect of the nutritional ecology of dinoflagellates.

The escalating proliferation of cyanobacteria poses significant taste and odor (T/O) challenges, impacting freshwater ecosystems, public health, and water treatment costs. We examined monthly variations in four T/O compounds from September 2011 to August 2012 in Chaohu Lake\'s eastern drinking water source (DECL). More importantly, we compared the reported T/O occurrence and the related factors in freshwater bodies worldwide. The assessment of T/O issues indicated a severe and widespread problem, with many cases surpassing odor threshold values. Remarkably, China reported the highest frequency and severity of odor-related problems. A temporal analysis revealed variations in odor occurrences within the same water body across different years, emphasizing the need to consider high values in all seasons for water safety. Globally, T/O issues were widespread, demanding attention to variations within the same water body and across different layers. Algae were crucial contributors to odor compounds, necessitating targeted interventions due to diverse odorant sources and properties. A correlation analysis alone lacked definitive answers, emphasizing the essential role of further validation, such as algae isolation. Nutrients are likely to have influenced the T/O, as GSM and MIB correlated positively with nitrate and ammonia nitrogen in DECL, resulting in proposed control recommendations. This study offers recommendations for freshwater ecosystem management and serves as a foundation for future research and management strategies to address T/O challenges.

Beibu Gulf is an important semi-enclosed bay located in the northwestern South China Sea, and is famous for its high bio-productivity and rich bio-diversity. The fast development along the Beibu Gulf Economical Rim has brought pressure to the environment, and algal blooms occurred frequently in the gulf. In this study, surface water samples and micro-plankton samples (20-200 μm) were collected in the northern Beibu Gulf coast. Diversity and distribution of eukaryotic planktonic microalgae were analyzed by both metabarcoding and microscopic analyses. Metabarcoding revealed much higher diversity and species richness of microalgae than morphological observation, especially for dinoflagellates. Metabarcoding detected 144 microalgal genera in 8 phyla, while microscopy only detected 40 genera in 2 phyla. The two methods revealed different microalgal community structures. Dinoflagellates dominated in microalgal community based on metabarcoding due to their high copies of 18 s rRNA gene, and diatoms dominated under microscopy. Altogether 48 algal bloom and/or toxic species were detected in this study, 34 species by metabarcoding and 19 species by microscopy. Our result suggested a high potential risk of HABs in the Beibu Gulf. Microalgal community in the surface water samples demonstrated significantly higher OTU/species richness, alpha diversity, and abundance than those in the micro-plankton samples, although more HAB taxa were detected by microscopic observations in the micro-plankton samples. Furthermore, nano-sized taxa, such as those in chlorophytes, haptophytes, and chrysophyceans, occurred more abundantly in the surface water samples. This study provided a comprehensive morphological and molecular description of microalgal community in the northern Beibu Gulf.

Paralytic shellfish toxins (PSTs) produced by marine dinoflagellates significantly impact shellfish industries worldwide. Early detection on-farm and with minimal training would allow additional time for management decisions to minimize economic losses. Here, we describe and test a standardized workflow based on the detection of sxtA4, an initial gene in the biosynthesis of PSTs. The workflow is simple and inexpensive and does not require a specialized laboratory. It consists of (1) water collection and filtration using a custom gravity sampler, (2) buffer selection for sample preservation and cell lysis for DNA, and (3) an assay based on a region of sxtA, DinoDtec lyophilized quantitative polymerase chain reaction (qPCR) assay. Water samples spiked with Alexandrium catenella showed a cell recovery of >90% when compared to light microscopy counts. The performance of the lysis method (90.3% efficient), Longmire\'s buffer, and the DinoDtec qPCR assay (tested across a range of Alexandrium species (90.7-106.9% efficiency; r2 > 0.99)) was found to be specific, sensitive, and efficient. We tested the application of this workflow weekly from May 2016 to 30th October 2017 to compare the relationship between sxtA4 copies L-1 in seawater and PSTs in mussel tissue (Mytilus galloprovincialis) on-farm and spatially (across multiple sites), effectively demonstrating an ∼2 week early warning of two A. catenella HABs (r = 0.95). Our tool provides an early, accurate, and efficient method for the identification of PST risk in shellfish aquaculture.

The role of microorganisms in effectively terminating harmful algal blooms (HABs) is crucial for maintaining environmental stability. Recent studies have placed increased emphasis on bio-agents capable of inhibiting HABs. The bacterium Pseudoalteromonas sp. strain FDHY-MZ2 has exhibited impressive algicidal abilities against Karenia mikimotoi, a notorious global HAB-forming species. To augment this capability, cultures were progressively scaled from shake flask conditions to small-scale (5 L) and pilot-scale (50 L) fermentation. By employing a specifically tailored culture medium (2216E basal medium with 1.5% soluble starch and 0.5% peptone), under precise conditions (66 h, 20 °C, 450 rpm, 30 L/min ventilation, 3% seeding, and constant starch flow), a notable increase in algicidal bacterial biomass was observed; the bacterial dosage required to entirely wipe out K. mikimotoi within a day decreased from 1% to 0.025%. Compared to an unoptimized shake flask group, the optimized fermentation culture caused significant reductions in algal chlorophyll and protein levels (21.85% and 78.3%, respectively). Co-culturing induced increases in algal malondialdehyde and H2O2 by 5.98 and 5.38 times, respectively, leading to further disruption of algal photosynthesis. This study underscores the unexplored potential of systematically utilized microbial agents in mitigating HABs, providing a pathway for their wider application.

Microcystins (MCs) are toxins produced by cyanobacteria commonly found in harmful algal blooms (HABs). Due to their toxicity to humans and other organisms, the World Health Organization (WHO) sets a guideline of 1 μg L-1 for microcystin-leucine-arginine (MC-LR) in drinking water. However, current analytical techniques for the detection of MC-LR such as liquid chromatography-mass spectrometry (LC-MS) and ELISA are costly, bulky, time-consuming, and mostly conducted in a laboratory, requiring highly trained personnel. An analytical method that can be used in the field for rapid determination is essential. In this study, an anti-MC-LR/MC-LR/cysteamine-coated screen-printed carbon electrode (SPCE) biosensor was newly developed to detect MC-LR, bioelectrochemically, in water. The functionalization of the electrode surface was confirmed with surface characterization methods. The sensor performance was evaluated by electrochemical impedance spectroscopy (EIS), obtaining a linear working range of MC-LR concentrations between 0.1 and 100 μg L-1 with a limit of detection (LOD) of 0.69 ng L-1. Natural water samples experiencing HABs were collected and analyzed using the developed biosensor, demonstrating the excellent performance of the biosensor with a relative standard deviation (RSD) of 0.65%. The interference tests showed minimal error and RSD values against other common MCs and possible coexisting ions found in water. The biosensor showed acceptable functionality with a shelf life of up to 12 weeks. Overall, the anti-MC-LR/MC-LR/cysteamine/SPCE biosensors can be an innovative solution with characteristics that allow for in situ, low-cost, and easy-to-use capabilities which are essential for developing an overarching and integrated \"smart\" environmental management system.

Anthropogenic impacts are expected to increase the co-occurrence of stressors that can fundamentally alter ecosystem structure and function. To cope with stress, many organisms locally adapt, but how such adaptations affect the ability of an organism to manage co-occurring stressors is not well understood. In aquatic ecosystems, elevated temperatures and harmful algal blooms are common co-stressors. To better understand the role and potential trade-offs of local adaptations for mitigating the effects of stressors, Daphnia pulicaria genotypes that varied in their ability to consume toxic cyanobacteria prey (i.e., three tolerant and three sensitive) were exposed to five diets that included combinations of toxic cyanobacteria, Microcystis aeruginosa, and a green alga, Ankistrodesmus falcatus, under two temperatures (20 °C vs. 28 °C). A path analysis was conducted to understand how local adaptations affect energy allocation to intermediate life history traits (i.e., somatic growth, fecundity, survival) that maximize Daphnia fitness (i.e., population growth rate). Results from the 10-day study show that tolerant Daphnia genotypes had higher fitness than sensitive genotypes regardless of diet or temperature treatment, suggesting toxic cyanobacteria tolerance did not cause a decrease in fitness in the absence of cyanobacteria or under elevated temperatures. Results from the path analysis demonstrated that toxic cyanobacteria had a stronger effect on life history traits than temperature and that population growth rate was mainly constrained by reduced fecundity. These findings suggest that local adaptations to toxic cyanobacteria and elevated temperatures are synergistic, leading to higher survivorship of cyanobacteria-tolerant genotypes during summer cyanobacterial bloom events.

Alexandrium catenella is a harmful algal bloom (HAB)-forming dinoflagellate that causes significant damage to the cultivation and harvest of shellfish due to its synthesis of paralytic shellfish toxins.  To evaluate the potential for macroalgae aquaculture to mitigate A. catenella blooms, we determined the effects of three cultivable macroalgae - Saccharina latissima (sugar kelp), Chondrus crispus (Irish moss), and Ulva spp. - on A. catenella in culture- and field-based experiments.  Co-culture growth assays of A. catenella exposed to environmentally realistic concentrations of each macroalgae showed that all species except low levels of C. crispus caused cell lysis and significant reductions in A. catenella densities relative to control treatments of 17-74% in 2-3 days and 42-96% in ~one week (p<0.05 for all assays). In a toxin accumulation experiment, S. latissima significantly lessened (p<0.05) saxitoxin (STX) accumulation in blue mussels (Mytilus edulis), keeping levels (71.80±1.98 µg STX 100 g-1) below US closure limits (80 µg STX 100 g-1) compared to the untreated control (93.47±8.11 µg STX 100 g-1). Bottle incubations of field-collected, bloom populations of A. catenella experienced significant reductions in cell densities of up to 95% when exposed to aquaculture concentrations of all three macroalgae (p<0.005 for all). The stocking of aquacultured S. latissima within mesocosms containing a bloom population of A. catenella (initial density: 3.2 × 104 cells L-1) reduced the population of A. catenella by 73% over 48 h (p<0.005) while Ulva addition caused a 54% reduction in A. catenella over 96 h (p<0.01).  Among the three seaweeds, their ordered ability to inhibit A. catenella was S. latissima > Ulva spp. > C. crispus. Seaweeds\' primary anti-A. catenella activity were allelopathic, while nutrient competition, pH elevation, and macroalgae-attached bacteria may have played a contributory role in some experiments. Collectively, these results suggest that the integration of macroalgae with shellfish-centric aquaculture establishments should be considered as a non-invasive, environmentally friendly, and potentially profit-generating measure to mitigate A. catenella-caused damage to the shellfish aquaculture industry.

The Mechanisms driving phosphorus (P) release in sediment of shallow lakes is essential for managing harmful algal blooms (HABs). Accordingly, this study conducted field monitoring of labile P, iron (Fe), sulfur (S), and dissolved manganese (Mn) in different biomass of algae in Lake Taihu. The in-situ technique of ZrO-Chelex-AgI (ZrO-CA) diffusive gradients in thin-films (DGT) and high-resolution dialysis sampler (high resolution-Peeper (HR-Peeper)) were used to measure labile P, Fe, S, and dissolved Mn, as well as their apparent diffusion fluxes at the sediment-water interface (SWI). In addition, the distribution of iron-reducing bacteria (IRB) and sulfate-reducing bacteria (SRB) in sediments was also detected. Results showed that high HABs biomass promoted the reduction of sulfate into labile S, however, IRB is the dominant species. Thus, labile Fe concentrations greatly exceeded labile S concentrations across all sites, indicating that microbial iron reduction (MIR) is the principal pathway for ferric iron reduction. Furthermore, the simple relationship analysis revealed the principal influence P migration and transformation is the Fe-P in high algal biomass sites, while Fe and Mn redox reactions did not significantly influence labile P mobilization in low algal areas.