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Toxic algae in eutrophic lakes produce cyanotoxic microcystins. Prior research on the effect of microcystin-LR in the kidney utilized intraperitoneal injections, which did not reflect natural exposure. Oral microcystin-LR research has focused on renal function and histopathology without examining the molecular mechanisms. The present study aimed to evaluate the mechanism of microcystin-LR in the kidneys via oral administration in WKAH/HkmSlc rats over 7 weeks, alongside stimulation of the proximal tubular cells. Although there were no differences in the concentrations of plasma albumin, blood urea nitrogen, and creatinine, which are parameters of renal function, between the control and microcystin-LR-administrated rats, prorenin expression was significantly increased in the renal cortex of the rats administered microcystin-LR and the microcystin-LR-treated proximal tubular cells. The expression levels of (pro)renin receptor (PRR), transforming growth factor-β1 (TGFβ1), and α-smooth muscle actin (α-SMA) in the renal cortex did not differ significantly between the control and microcystin-LR-administered rats. However, the expression levels of prorenin were significantly positively correlated with those of PRR, TGFβ1, and α-SMA in the renal cortex of rats administered microcystin-LR. Additionally, a significant positive correlation was observed between the expression levels of TGFβ1 and α-SMA. Collectively, increased prorenin expression caused by the long-term consumption of microcystin-LR may initiate a process that influences renal fibrosis and abnormal renal function by regulating the expression levels of PRR, TGFβ1, and α-SMA. [ABSTRACT FROM AUTHOR]

Key Takeaways: Flow imaging microscopy (FIM) combines a flow cytometer's fluidic architecture with a microscope's spatial resolution to allow morphometric analysis of objects in suspensions such as water. FIM's real‐time analysis and identification of potential cyanotoxin producers help utilities understand population dynamics to maintain healthier source waters and reduce response times when a bloom is indicated. Case studies show FIM's advantages and limitations in harmful algal bloom monitoring programs for reservoir management and water treatment. [ABSTRACT FROM AUTHOR]

Abstract Spirulina (Arthrospira platensis) production dominates the global microalgae market due to its high nutritional value. Spirulina can be cultivated in ponds with quality control due to the possible growth of cyanotoxins like Microcystis sp. that produce microcystins. The Enzyme-Linked Immunosorbent Assay (ELISA) method was used and validated with linearity, precision, reproducibility, and accuracy parameters that met the criteria for microcystin analysis. The regression curve gave the linear regression, the value of the coefficient of correlation (r) 0.99 showed a very strong correlation, and coefficient determination (R2) was 0.988 (R2 > 0.90). The precision analysis in repeatability (RSDanalysis < 2/3RSD Horwitz) and reproducibility (RSDanalysis < RSD Horwitz) showed an acceptable value with a significance value of 0.191 (p-value

Harmful Algal Blooms (HABs) are outbreaks of aquatic toxic microalgae emerging as a global problem driven by nutrient enrichment, global climate change and invasive species. We uniquely describe a HAB of unprecedented duration, extent and magnitude during 2023 in Lough Neagh; the UK and Ireland’s largest freshwater lake, using an unparalleled combination of satellite imagery, nutrient analysis, 16S rRNA gene sequencing and cyanotoxin profiling. The causative agent Microcystis aeruginosa accounted for over a third of DNA in water samples though common bacterioplankton species also bloomed. Water phosphate levels were hypertrophic and drove local algal biomass. The HAB pervaded the entire ecosystem with algal mats accumulating around jetties, marinas and lock gates. Over 80 % of bacterial DNA isolated from algal mat samples consisted of species associated with wildfowl or livestock faeces and human-effluent wastewater including 13 potential pathogens that can cause serious human illness including: E. coli, Salmonella, Enterobacter and Clostridium among others. Ten microcystins, nodularin and two anabaenopeptin toxins were confirmed as present (with a further microcystin and four anabaenopeptins suspected), with MC-RR and -LR in high concentrations at some locations (1,137–18,493 μg/L) with MC-LR exceeding World Health Organisation (WHO) recreational exposure guidelines in all algal mats sampled. This is the first detection of anabaenopeptins in any waterbody on the island of Ireland. Notwithstanding the ecological impacts, this HAB represented an environmental and public health risk, curtailing recreational activities in-and-around the lake and damaging local businesses. Reducing agricultural runoff and discharge from human-effluent wastewater treatment to manage nutrient loading, and the public health risk, should be the top priority of stakeholders, especially government. Key recommendations include Nature-based Solutions that avoid conflict with the productivity and profitability of the farming sector enhancing sustainability. We hope this stimulates real-world action to resolve the problems besetting this internationally important ecosystem.

There is a growing concern regarding the adverse risks exposure to cylindrospermopsin (CYN) might exert on animals and humans. However, data regarding the toxicity of this cyanotoxin to neotropical fish species are scarce. Using the fish species Poecilia reticulata, the influence of CYN concentrations equal to and above the tolerable for drinking water may produce on liver was determined by assessing biomarkers of antioxidant defense mechanisms and correlated to qualitative and semiquantitative histopathological observations. Adult females were exposed to 0.0 (Control); 0.5, 1 and 1.5 µg/L pure CYN for 24 or 96 hr, in triplicate. Subsequently the livers were extracted for biochemical assays and histopathological evaluation. Catalase (CAT) activity was significantly increased only by 1.5 µg/L CYN-treatment, at both exposure times. Glutathione -S-transferase (GST) activity presented a biphasic response for both exposure times. It was markedly decreased after exposure by 0.5 µg/L CYN treatment but significantly elevated by 1.5 µg/L CYN treatment. All CYN treatments produced histopathological alterations, as evidenced by hepatocyte cords degeneration, steatosis, inflammatory infiltration, melanomacrophage centers, vessel congestion, and areas with necrosis. Further, an IORG >35 was achieved for all treatments, indicative of the presence of severe histological alterations in P. reticulata hepatic parenchyma and stroma. Taken together, data demonstrated evidence that CYN-induced hepatotoxicity in P. reticulata appears to be associated with an imbalance of antioxidant defense mechanisms accompanied by histopathological liver alterations. It is worthy to note that exposure to low environmentally-relevant CYN concentrations might constitute a significant risk to health of aquatic organisms. [ABSTRACT FROM AUTHOR]

• The non-toxigenic M. wesenbergii FACHB-908 was more sensitive to C 14 -benzalkonium chloride (BAC-14) than the two toxigenic M. aeruginosa. • Photosynthetic and metabolic responses showed higher sensitivity to BAC-14 than growth inhibition. • BAC-14 specific responses in glycerophospholipid metabolism, synchronized by dysregulation of energy metabolism and amino acid biosynthesis. • BAC-14 induced elevation of extracellular microcystin contents in the absence of growth inhibition of the two toxigenic Microcystis. Quaternary ammonium compounds (QACs) are commonly used in a variety of consumer and commercial products, typically as a component of disinfectants. During the COVID-19 pandemic, QACs became one of the primary agents utilized to inactivate the SARS-CoV-2 virus on surfaces. However, the ecotoxicological effects of QACs upon aquatic organisms have not been fully assessed. In this study, we examined the effects of a widely used QAC (benzalkonium chloride-C 14 , BAC-14) on two toxigenic Microcystis strains and one non-toxigenic freshwater Microcystis strain and carried out an analysis focused on primary, adaptive and compensatory stress responses at apical (growth and photosynthesis) and metabolic levels. This analysis revealed that the two toxic Microcystis strains were more tolerant than the non-toxic strain, with 96 hr-EC 50 values of 0.70, 0.76, and 0.38 mg/L BAC-14 for toxigenic M. aeruginosa FACHB-905, toxigenic M. aeruginosa FACHB-469, and non-toxigenic M. wesenbergii FACHB-908, respectively. The photosynthetic activities of the Microcystis , assessed via F v /F m values, were significantly suppressed under 0.4 mg/L BAC-14. Furthermore, this analysis revealed that BAC-14 altered 14, 12, and 8 metabolic pathways in M. aeruginosa FACHB-905, M. aeruginosa FACHB-469, and M. wesenbergii FACHB-908, respectively. It is noteworthy that BAC-14 enhanced the level of extracellular microcystin production in the toxigenic Microcystis strains , although cell growth was not significantly affected. Collectively, these data show that BAC-14 disrupted the physiological and metabolic status of Microcystis cells and stimulated the production and release of microcystin, which could result in damage to aquatic systems. [Display omitted] [ABSTRACT FROM AUTHOR]

With the rapid advancement of nanotechnology and its widespread applications, increasing amounts of manufactured and natural nanoparticles (NPs) have been tested for their potential utilization in treating harmful cyanobacterial blooms (HCBs). NPs can be used as a photocatalyst, algaecide, adsorbent, flocculant, or coagulant. The primary mechanisms explored for NPs to mitigate HCBs include photocatalysis, metal ion-induced cytotoxicity, physical disruption of the cell membrane, light-shielding, flocculation/coagulation/sedimentation of cyanobacterial cells, and the removal of phosphorus (P) and cyanotoxins from bloom water by adsorption. As an emerging and promising chemical/physical approach for HCB mitigation, versatile NP-based technologies offer great advantages, such as being environmentally benign, cost-effective, highly efficient, recyclable, and adaptable. The challenges we face include cost reduction, scalability, and impacts on non-target species co-inhabiting in the same environment. Further efforts are required to scale up to real-world operations through developing more efficient, recoverable, reusable, and deployable NP-based lattices or materials that are adaptable to bloom events in different water bodies of different sizes, such as reservoirs, lakes, rivers, and marine environments. [ABSTRACT FROM AUTHOR]

Globally, estuaries are undergoing profound hydrological alterations due to changes in climate and flood management coincident with the increasing frequency of harmful algal blooms (HABs). In coastal Louisiana, the introduction of nutrient-rich Mississippi River water into Lake Pontchartrain Estuary supports the proliferation of toxic cyanobacterial blooms (CyanoHABs). In 2019, over 28 km3 of Mississippi River water was diverted into the estuary (volume of 6 km3) to prevent flooding of downstream communities. This study characterized the impact of this river input containing 38,000 + metric tons of bioavailable N and 2,300 + metric tons of bioavailable P on monthly biological measures of the water column including biomass of pelagic cyanobacteria and associated toxin production. The estuary was freshened for months due to the river pulses, which initially limited phytoplankton response due to high total suspended solids. There was a cyanobacterial bloom once the river pulse stopped, related to increased water residence time, clarity, and nutrient availability. Particulate toxin concentration correlated to cyanobacterial biomass but was non-linear. Once bioavailable N decreased, nitrogen-fixing Dolichospermum spp. became dominant. This research on CyanoHABs in Lake Pontchartrain Estuary can help provide a basis to predict the response of estuaries to large nutrient-loading events. [ABSTRACT FROM AUTHOR]

A new microcystin-producing mangrove cyanobacterium, Aliinostoc bakau sp. nov., was isolated from a tropical mangrove in Penang, Malaysia, and characterized using combined morphological and phylogenetic approaches. Cultures were established in liquid media of different salinities (0, 7, 14, 21, 28, and 35 ppt). Optimal growth observed at both 7 and 14 ppt was consistent with the origin of the strain from an estuarine mangrove environment. Phylogenetic analysis based on the 16S rRNA gene strongly indicated that the strain is a member of the genus Aliinostoc and is distinct from other currently sequenced species in the genus. The sequences and secondary structure of the 16S–23S ITS region D1–D1' and Box–B helices provided further confirmation that the new species is clearly distinct from previously described Aliinostoc species. Amplification of the mcyE gene fragment associated with the production of microcystin in A. bakau revealed that it is identical to that in other known microcystin-producing cyanobacteria. Analysis of the extracts obtained from this strain by HPLC-MS/MS confirmed the presence of microcystin variants (MC-LR and -YR) at concentrations of 0.60 μg/L and MC-RR at a concentration of 0.30 μg/L. This is the first record of microcystin production from Aliinostoc species in tropical mangrove habitats. [ABSTRACT FROM AUTHOR]

Cyanobacterial blooms are increasing in frequency and intensity globally, and impacting recreational waters as well as waters used for drinking water provisioning. They are sources of bioactive metabolites including retinoids and the neurotoxin anatoxin-a. Here, we investigated the effects of anatoxin-a on a differentiating in vitro human neural stem cell model previously characterised with retinoic acids. Effects on protein and gene expression upon exposure for 9 or 18 days to anatoxin-a alone or in co-exposure with all- trans retinoic acid were evaluated using a panel of neural and glial differentiation biomarkers. Anatoxin-a did not cause distinct developmental neurotoxicity alone, or in co-exposure with retinoic acid. However, in line with its excitotoxicity, in co-exposure with 200 nM all- trans retinoic acid it reduced the differentiation of acetylcholinergic neuron subtypes in the culture at 1000 nM (highest tested concentration). While this could have substantial functional implications for the developing nervous system, there is no indication for developmental neurotoxicity beyond its (excito-)toxicity to acetylcholinergic neurons, which only occurred in co-exposure to all- trans retinoic acid. • ATX does not alter in vitro neurodifferentiation of pluripotent stem cells-derived Neural Stem Cells. • No developmental neurotoxicity was evident. • 1000 nM ATX suppressed differentiation of acetylcholinergic neurons in co-exposure with atRA. [ABSTRACT FROM AUTHOR]

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Background: Microcystin-leucine arginine (MC-LR) is a cyanobacterial hepatotoxic toxin found in water sources worldwide, including in northeastern Thailand, where opisthorchiasis-associated cholangiocarcinoma (CCA) is most prevalent. MC-LR is a potential carcinogen; however, its involvement in liver fluke-associated CCA remains ambiguous. Here, we aimed to evaluate the effect of MC-LR on the progression of CCA via the Wnt/β-catenin pathway in vitro. Methods: Cell division, migration, cell cycle transition, and MC-LR transporter expression were evaluated in vitro through MTT assay, wound healing assay, flow cytometry, and immunofluorescence staining, respectively. Following a 24-h treatment of cultured cells with 1, 10, 100, and 1,000 nM of MC-LR, the proliferative effect of MC-LR on the Wnt/β-catenin signaling pathway was investigated using immunoblotting and qRT-PCR analysis. Immunohistochemistry was used to determine β-catenin expression in CCA tissue compared to adjacent tissue. Results: Human immortalized cholangiocyte cells (MMNK-1) and a human cell line established from opisthorchiasis-associated CCA (KKU-213B) expressed the MC-LR transporter and internalized MC-LR. Exposure to 10 nM and 100 nM of MC-LR notably enhanced cells division and migration in both cell lines (P

Toxic algae in eutrophic lakes produce cyanotoxic microcystins. Prior research on the effect of microcystin-LR in the kidney utilized intraperitoneal injections, which did not reflect natural exposure. Oral microcystin-LR research has focused on renal function and histopathology without examining the molecular mechanisms. The present study aimed to evaluate the mechanism of microcystin-LR in the kidneys via oral administration in WKAH/HkmSlc rats over 7 weeks, alongside stimulation of the proximal tubular cells. Although there were no differences in the concentrations of plasma albumin, blood urea nitrogen, and creatinine, which are parameters of renal function, between the control and microcystin-LR-administrated rats, prorenin expression was significantly increased in the renal cortex of the rats administered microcystin-LR and the microcystin-LR-treated proximal tubular cells. The expression levels of (pro)renin receptor (PRR), transforming growth factor-β1 (TGFβ1), and α-smooth muscle actin (α-SMA) in the renal cortex did not differ significantly between the control and microcystin-LR-administered rats. However, the expression levels of prorenin were significantly positively correlated with those of PRR, TGFβ1, and α-SMA in the renal cortex of rats administered microcystin-LR. Additionally, a significant positive correlation was observed between the expression levels of TGFβ1 and α-SMA. Collectively, increased prorenin expression caused by the long-term consumption of microcystin-LR may initiate a process that influences renal fibrosis and abnormal renal function by regulating the expression levels of PRR, TGFβ1, and α-SMA.

The CYN degradation kinetics of each bacterial species were studied using High Performance Liquid Chromatography. The greatest CYN degradation (28.22 ± 0.24%) was shown by the bacterium B. cereus-S in 5.0 mg/L CYN within 14 days. The CYN degradation by the other strains was lower than 10% under the same conditions. Further studies employing different initial concentrations of CYN revealed that B. cereus-S could degrade lower CYN concentrations at a higher percentage (1.0 mg/L, 2.5 mg/L, and 5.0 mg/L of CYN removal percentages were 36.83 ± 2.43%, 32.25 ± 1.25%, and 24.72 ± 0.40%, respectively, after 14 days of incubation at 28℃ and pH 7). The maximum average degradation rates were recorded for 1.0 mg/L, 2.5 mg/L, and 5.0 mg/L CYN on the 6th (0.05 ± 0.00 mg/L/day), 8th (0.04 ± 0.01 mg/L/day), and 12th (0.02 ± 0.01 mg/L/day) days of incubation, respectively. The study showed the potentiality of the bacterium B. cereus-S on the application for degrading CYN among the tested bacteria species. [ABSTRACT FROM AUTHOR]

As source waters for many drinking water treatment plants are increasingly subject to intermittent blooms of cyanobacteria, utilities must adapt treatment methods to remove associated cyanotoxins. There is concern that treatment methods applied to remove dissolved extracellular toxins may damage whole cells, leading to the release of intracellular toxins and other secondary metabolites. The objective of this study was to determine whether sodium hypochlorite and potassium permanganate preoxidation treatments cause increased susceptibility of cyanobacterial cells to mechanical shear using a method that can be easily adapted by utilities. Bench-scale simulations examined Microcystis sp., Planktothrix sp., and Dolichospermum sp., which are three types of microcystin-producing cyanobacteria that are prominent in many bloom-impacted waters. We found that chlorine led to increased susceptibility to shear in Dolichospermum and permanganate led to increased susceptibility to shear in Planktothrix, each for shear rates of 750 s−1 (a typical rate for in-line mixers used in drinking water treatment). We found that shear does not cause immediate critical damage to cells, regardless of preoxidation conditions. However, shear appears to cause a stress response in cyanobacteria, indicating the possibility of delayed toxin release. The results of this study will help water treatment utilities to manage stressors within the treatment process that could lead to release of cellular compounds during cyanobacterial blooms. Additionally, utilities can adapt the applied method of intracellular phycocyanin analysis to independently evaluate real-time cell response to their treatment processes. [ABSTRACT FROM AUTHOR]

Cyanobacterial harmful algal blooms (cyanoHABs) are assumed to be increasing in abundance and toxicity, but comprehensive analysis of change through time is limited, in part, because some key taxa (e.g., Microcystis) leave ambiguous evidence of historical abundance and toxicity. Sedimentary DNA (sedDNA) can allow the reconstruction of the cyanobacteria community as well as the frequency of genes specific to cyanotoxin production, enabling us to determine which taxa are present and their potential for toxin‐production.Using a combination of droplet digital polymerase chain reaction (ddPCR) and high‐throughput sequencing (HTS), we quantified the abundance of cyanobacterial genes of known function and changes in cyanobacteria taxa from sedDNA over the last century in nine lakes along a gradient of lake size, depth and trophic state in Minnesota, U.S.A. Using ddPCR, we quantified genes associated with microcystin toxin‐producing potential (mcyE), total cyanobacteria (CYA, 16S rRNA) and the genus Microcystis (MICR, 16S rRNA). Using HTS on a subset of lakes, we investigated how the abundance of this toxin‐producing gene covaried with the cyanobacteria community composition. We also compared ddPCR and HTS data to fossil pigments, a well‐established palaeolimnological method used to track changes in primary producers over time.Our results showed a significant correlation between MICR and the quantity of mcyE gene and cyanobacterial taxa with known toxin‐production potential. The abundance of both genes likewise increased concomitantly through time.Community analyses of HTS data showed significant change in cyanobacterial communities commencing c. 1950 when major land‐use change in this region led to increased lake productivity, and c. 1990 when Dolichospermum and Microcystis genera increased in abundance, and the subtropical exotic cyanobacteria Raphidiopsis raciborskii and Sphaerospermopsis aphanizomenoides became abundant. Cyanobacteria pigment data reflected these changes only in deeper lakes, suggesting issues related to benthic production or biomarker preservation in shallower systems.This study provides evidence for historical development of increasingly toxic cyanoHABs across a diverse set of lakes and illustrates how sedDNA may help link changes in the cyanobacteria community to the expression of potentially toxic genes. [ABSTRACT FROM AUTHOR]

Cyanobacteria are infamous producers of toxins. While the toxic potential of planktonic cyanobacterial blooms is well documented, the ecosystem level effects of toxigenic benthic and epiphytic cyanobacteria are an understudied threat. The freshwater epiphytic cyanobacterium Aetokthonos hydrillicola has recently been shown to produce the "eagle killer" neurotoxin aetokthonotoxin (AETX) causing the fatal neurological disease vacuolar myelinopathy. The disease affects a wide array of wildlife in the southeastern United States, most notably waterfowl and birds of prey, including the bald eagle. In an assay for cytotoxicity, we found the crude extract of the cyanobacterium to be much more potent than pure AETX, prompting further investigation. Here, we describe the isolation and structure elucidation of the aetokthonostatins (AESTs), linear peptides belonging to the dolastatin compound family, featuring a unique modification of the C-terminal phenylalanine-derived moiety. Using immunofluorescence microscopy and molecular modeling, we confirmed that AEST potently impacts microtubule dynamics and can bind to tubulin in a similar matter as dolastatin 10. We also show that AEST inhibits reproduction of the nematode Caenorhabditis elegans. Bioinformatic analysis revealed the AEST biosynthetic gene cluster encoding a nonribosomal peptide synthetase/polyketide synthase accompanied by a unique tailoring machinery. The biosynthetic activity of a specific N-terminal methyltransferase was confirmed by in vitro biochemical studies, establishing a mechanistic link between the gene cluster and its product. [ABSTRACT FROM AUTHOR]

Background: Amyotrophic lateral sclerosis (ALS) is a progressive and often fatal neurodegenerative disease characterized by the loss of Motor Neurons (MNs) in spinal cord, motor cortex and brainstem. Despite significant efforts in the field, the exact pathogenetic mechanisms underlying both familial and sporadic forms of ALS have not been fully elucidated, and the therapeutic possibilities are still very limited. Here we investigate the molecular mechanisms of neurodegeneration induced by chronic exposure to the environmental cyanotoxin L-BMAA, which causes a form of ALS/Parkinson’s disease (PD) in several populations consuming food and/or water containing high amounts of this compound. Methods: In this effort, mice were chronically exposed to L-BMAA and analyzed at different time points to evaluate cellular and molecular alterations and behavioral deficits, performing MTT assay, immunoblot, immunofluorescence and immunohistochemistry analysis, and behavioral tests. Results: We found that cyanotoxin L-BMAA determines apoptotic cell death and a marked astrogliosis in spinal cord and motor cortex, and induces neurotoxicity by favoring TDP-43 cytoplasmic accumulation. Conclusions: Overall, our results characterize a new versatile neurotoxic animal model of ALS that may be useful for the identification of new druggable targets to develop innovative therapeutic strategies for this disease.

Anatoxins (ATXs) are a potent class of cyanobacterial neurotoxins for which only a handful of structural analogues have been well characterized. Here, we report the development of an LC–HRMS/MS method for the comprehensive detection of ATXs. Application of this method to samples of benthic cyanobacterial mats and laboratory cultures showed detection of several new ATXs. Many of these result from nucleophilic addition to the olefinic bond of the α,β-unsaturated ketone functional group of anatoxin-a (ATX) and homoanatoxin-a (hATX), analogous to the conjugation chemistry of microcystins, which contain similar α,β-unsaturated amide functionality. Conjugates with glutathione, γ-glutamylcysteine, methanethiol, ammonia, methanol and water were detected, as well as putative C-10 alcohol derivatives. Structural confirmation was obtained by simple and selective analytical-scale semisynthetic reactions starting from available ATX standards. Methanol, water and ammonia conjugates were found to result primarily from sample preparation. Reduction products were found to result from enzymatic reactions occurring primarily after cell lysis in laboratory cultures of Kamptonema formosum and Cuspidothrix issatschenkoi. The relative contributions of the identified analogues to the anatoxin profiles in a set of 22 benthic-cyanobacterial-mat field samples were estimated, showing conjugates to account for up to 15% of total ATX peak area and 10-hydroxyanatoxins up to 38%. The developed methodology, new analogues and insight into the chemical and enzymatic reactivity of ATXs will enable a more comprehensive study of the class than possible previously. [ABSTRACT FROM AUTHOR]

Nutrient-driven cyanobacteria blooms are an increasingly common issue in freshwater environments, particularly in anthropogenically altered landscapes. As stormwater runoff is one of the largest sources of nutrients for freshwater bodies, stormwater retention ponds in urban and suburban areas are likely environments for harmful cyanobacteria blooms and were thus targeted for an in-depth investigation assessing taxonomic composition, bloom morphological composition, toxicity, and impact of nutrients and other environmental drivers. Eighty-seven algal blooms were sampled from 2019 to 2022 in the greater Wilmington, North Carolina, area. Physicochemical parameters were recorded, and blooms were classified by type (defined as surface mat, surface scum, water column distribution, or benthic mat) and dominant taxa. Blooms of potentially toxic cyanobacteria genera in the water column of stormwater retention ponds were most prevalent. Dissolved inorganic phosphorus was significantly related to chlorophyll-α, Microcystis bloom formation, and the production of microcystin. Seventeen potentially toxic cyanobacteria genera were identified in retention ponds, some of whose blooms demonstrated detectable microcystin. Monoclonal cultures isolated from some blooms were found to produce anabaenopeptin and saxitoxin. The results demonstrate a higher incidence of potentially toxic cyanobacteria over other bloom-forming taxa (chlorophytes, euglenoids, chrysophytes, dinoflagellates, and diatoms) in the 39 water bodies sampled. The frequency of blooms occurring in stormwater ponds and the diversity of potentially toxic cyanobacteria identified suggest such harmful blooms are likely widespread in similar freshwater environments across multiple urbanizing areas. The blooms sampled in this study were all within residential, commercial, or recreational areas easily accessible to people, presenting serious hazards to both environmental and public health. [ABSTRACT FROM AUTHOR]

Cylindrospermopsin (CYN) is a freshwater algal toxin produced during the proliferation of harmful cyanobacteria, known as cyanobacterial algal blooms (cyano-HABs). Recently, the effects of global warming have facilitated the growth of cyano-HABs, leading to their worldwide occurrence and an increase in toxin-related damage to aquatic ecosystems. CYN is known to exhibit strong cell toxicity upon ingestion, inhibiting protein synthesis and glutathione production and, ultimately, leading to cell death. In addition to cell toxicity, CYN exhibits skin toxicity, genotoxicity, and hepatotoxicity. It can also affect other organs, such as the kidneys (causing tubular necrosis), thymus (causing atrophy), and heart (causing pericardial and myocardial hemorrhage). The standard method used for CYN detection to date, enzyme-linked immunosorbent assay (ELISA), has several drawbacks: it is complex, time-consuming, and requires trained researchers. Recently, biosensors have been shown to offer numerous advantages, such as their simplicity, portability, and rapidity, making them suitable for onsite applications. Consequently, recent studies have actively explored the latest biosensor-based technologies for CYN detection. This review discusses the recent advances in CYN detection platforms that utilize several types of biosensors. [ABSTRACT FROM AUTHOR]

Introduction: Cyanobacterial blooms produce toxins that may become aerosolized, increasing health risks through inhalation exposures. Health related effects on the lower respiratory tract caused by these toxins are becoming better understood. However, nasal exposures to cyanotoxins remain understudied, especially for those with neurotoxic potential. Here, we present a case series study evaluating exposure to β-N-methylamino-l-alanine (BMAA), a cyanobacterial toxin linked to neurodegenerative disease, in postmortem olfactory tissues of individuals with varying stages of Alzheimer’s disease (AD). Methods: Olfactory bulb (Ob) tissues were collected during autopsies performed between 2014 and 2017 from six South Florida brain donors (ages 47–78) with residences less than 140 m from a freshwater body. A triple quadrupole tandem mass spectrometry (UHPLC-MS/MS) method validated according to peer AOAC International guidelines was used to detect BMAA and two BMAA isomers: 2,4-diaminobutyric acid (2,4-DAB) and N-(2-aminoethyl)glycine (AEG). Quantitative PCR was performed on the contralateral Ob to evaluate the relative expression of genes related to proinflammatory cytokines (IL-6 & IL-18), apoptotic pathways (CASP1 & BCL2), and mitochondrial stress (IRF1 & PINK1). Immunohistochemistry was also performed on the adjacent olfactory tract (Ot) to evaluate co-occurring neuropathology with BMAA tissue concentration. Results: BMAA was detected in the Ob of all cases at a median concentration of 30.4 ng/g (Range

With the rapid advancement of nanotechnology and its widespread applications, increasing amounts of manufactured and natural nanoparticles (NPs) have been tested for their potential utilization in treating harmful cyanobacterial blooms (HCBs). NPs can be used as a photocatalyst, algaecide, adsorbent, flocculant, or coagulant. The primary mechanisms explored for NPs to mitigate HCBs include photocatalysis, metal ion-induced cytotoxicity, physical disruption of the cell membrane, light-shielding, flocculation/coagulation/sedimentation of cyanobacterial cells, and the removal of phosphorus (P) and cyanotoxins from bloom water by adsorption. As an emerging and promising chemical/physical approach for HCB mitigation, versatile NP-based technologies offer great advantages, such as being environmentally benign, cost-effective, highly efficient, recyclable, and adaptable. The challenges we face include cost reduction, scalability, and impacts on non-target species co-inhabiting in the same environment. Further efforts are required to scale up to real-world operations through developing more efficient, recoverable, reusable, and deployable NP-based lattices or materials that are adaptable to bloom events in different water bodies of different sizes, such as reservoirs, lakes, rivers, and marine environments.

True branching is a facultative characteristic only known from two cyanobacteria in the Aphanizomenonaceae, Umezakia natans and Dolichospermum brachiatum. In both cases, its expression has been associated with environmental stress, and its practical use as a diacritical feature has been previously evaluated. In this study, we undertook further evaluation of the phylogeny of Umezakia natans and its relationship to Chrysosporum ovalisporum as a previous study suggested the two were potentially congeneric. We used combined morphological, phylogenetic, and phylogenomic approaches to determine their relatedness using new strains available from a broad geographic range. Phylogenetic analysis based on 16S rRNA gene sequences showed that Australian C. ovalisporum and Japanese U. natans strains clustered together with accessions of C. ovalisporum originating from Australia, Israel, and Spain, with high p‐distance similarity values (99.5%–99.9%). Additionally, differences between the two species in the 16S–23S ITS region was low (0%–2.5%). The average nucleotide identity of the U. natans and C. ovalisporum strains was also high (ANI of > 99.5 and AF > 0.9) and supported a genus‐level separation from Chrysosporum bergii (83 ANI between clusters). Furthermore, in culture, strains of both species grown in vitamin‐free media showed facultative true branching, a feature not previously known in C. ovalisporum. Collectively, the results support unification of C. ovalisporum and U. natans according to the principle of priority as Umezakia ovalisporum. [ABSTRACT FROM AUTHOR]

Water resources are critically important, but also pose risks of exposure to toxic and pathogenic microbes. Increasingly, a concern is toxic cyanobacteria, which have been linked to the death and disease of humans, domesticated animals, and wildlife in freshwater systems worldwide. Management approaches successful at reducing cyanobacterial abundance and toxin production have tended to be short-term solutions applied on small scales (e.g., algaecide application) or solutions that entail difficult multifaceted investments (e.g., modification of landscape and land use to reduce nutrient inputs). However, implementation of these approaches can be undermined by microbial species interactions that (a) provide toxic cyanobacteria with protection against the method of control or (b) permit toxic cyanobacteria to be replaced by other significant microbial threats. Understanding these interactions is necessary to avoid such scenarios and can provide a framework for novel strategies to enhance freshwater resource management via systems science (e.g., pairing existing physical and chemical approaches against cyanobacteria with ecological strategies such as manipulation of natural enemies, targeting of facilitators, and reduction of benthic occupancy and recruitment). Here, we review pertinent examples of the interactions and highlight potential applications of what is known.

An immunosensor for the assay of toxic biological warfare agents is a biosensor suitable for detecting hazardous substances such as aflatoxin, botulinum toxin, ricin, Shiga toxin, and others. The application of immunosensors is used in outdoor assays, point-of-care tests, as a spare method for more expensive devices, and even in the laboratory as a standard analytical method. Some immunosensors, such as automated flow-through analyzers or lateral flow tests, have been successfully commercialized as tools for toxins assay, but the research is ongoing. New devices are being developed, and the use of advanced materials and assay techniques make immunosensors highly competitive analytical devices in the field of toxic biological warfare agents assay. This review summarizes facts about current applications and new trends of immunosensors regarding recent papers in this area. [ABSTRACT FROM AUTHOR]

Cylindrospermopsin (CYN) is a potent cyanobacterial toxin found in freshwaters worldwide. In this work, the feasibility of the photo-Fenton process under neutral pH using light emitting diodes as irradiation source for the removal of this hazardous cyanotoxin from freshwater was investigated. The impact of the kind of iron chelating agent (ethylenediamine-N, N′-disuccinic acid vs. ethylenedinitrilotetraacetic acid) as well as the effect of the main operating conditions viz. H2O2 dose, Fe(III) load, initial CYN concentration, and Fe(III):EDDS molar ratio on the performance of the process was systematically evaluated. EDDS was selected as the most appropriate iron chelating agent considering the kinetics of the process and the environmental impact (Vibrio fischeri and Artemia salina). Under optimized conditions ([H2O2] = 30 mg L−1; [Fe(III)] = 5 mg L−1; Fe(III):ligand = 1:0.5 (molar ratio)), complete removal of CYN was achieved in 15-min reaction time. Furthermore, the catalytic system showed to be effective in real water matrices (river and reservoir waters) spiked with CYN. Although the presence of inorganic ions (mainly HCO3−/CO32−) and dissolved organic carbon decreased the oxidation rate of CYN due to scavenging reactions and iron coordination, respectively, complete elimination of the cyanotoxin was achieved in all cases. The fate of EDDS along the process was also evaluated to demonstrate that the catalytic system investigated, apart from its effectiveness, warrants the complete absence of residues after reaction. Therefore, the proposed system constitutes a promising method for cyanotoxin treatment either as a drinking water treatment step in conventional plants or as a potential remediation strategy in the natural environment. [ABSTRACT FROM AUTHOR]

A new microcystin-producing mangrove cyanobacterium, Aliinostoc bakau sp. nov., was isolated from a tropical mangrove in Penang, Malaysia, and characterized using combined morphological and phylogenetic approaches. Cultures were established in liquid media of different salinities (0, 7, 14, 21, 28, and 35 ppt). Optimal growth observed at both 7 and 14 ppt was consistent with the origin of the strain from an estuarine mangrove environment. Phylogenetic analysis based on the 16S rRNA gene strongly indicated that the strain is a member of the genus Aliinostoc and is distinct from other currently sequenced species in the genus. The sequences and secondary structure of the 16S–23S ITS region D1–D1’ and Box–B helices provided further confirmation that the new species is clearly distinct from previously described Aliinostoc species. Amplification of the mcyE gene fragment associated with the production of microcystin in A. bakau revealed that it is identical to that in other known microcystin-producing cyanobacteria. Analysis of the extracts obtained from this strain by HPLC-MS/MS confirmed the presence of microcystin variants (MC-LR and -YR) at concentrations of 0.60 μg/L and MC-RR at a concentration of 0.30 μg/L. This is the first record of microcystin production from Aliinostoc species in tropical mangrove habitats.

Although cyanobacteria are commonly associated with eutrophic lakes, they are the basic compo-nents of phytoplankton communities in lakes that have different trophic statuses. In inland waters, both nutrient loading from watersheds and warmer conditions promote phytoplankton growth and cause extensive cyanobacterial blooms. Certain bloom-forming cyanobacterial species can pose a healthrisktohumansandaquaticecosystemsthroughcyanotoxinproduction.Theaimofthisstudy was to evaluate the cyanobacterial composition and toxins in five reservoirs and two natural lakes in the Küçük Menderes River Basin, all with varying trophic statuses. Within this scope, sam-pleswerecollectedinautumn2017andspring2018.CyanobacterialspecieswereenumeratedaccordingtotheUtermöhlmethod.CyanotoxinsampleswereanalyzedusingHPLC.Tofindthetrophic status of the water bodies, the Trophic State Index (TSI) developed by Carlson (1977) was used and Total Phosphorus (TP), Secchi Depth (SD), and Chlorophyll-ɑ (chl-ɑ) measurements were performed. Cyanobacterial abundance, species composition, and cyanotoxin production differed significantly between the lakes and reservoirs. A total of 13 cyanobacteria species were identified including potential cyanotoxin producers such as Microcystis, Aphanizomenon, and Dolichospermum. According to the TSI, three reservoirs were mesotrophic and the other four waterbodies had eutro-phic-hypereutrophic conditions. Microcystis is the most common bloom-forming freshwater cyano-bacteria in the Küçük Menderes River Basin. However, microcystin concentrations were relatively low and the highest microcystin concentration was detected in the Tahtalı Reservoir at 9 μg/L. The Küçük Menderes River Basin is under water-stressed conditions and the cyanobacteria blooms in the region might pose another threat for wildlife and humans.

Fish mortality is associated with harmful algal blooms, although whether toxicity is related directly to the presence of cyanotoxins or the prevailing water chemistry remains unclear. Similarly, while planktivorous fish may be exposed to toxin through the diet, the hazard posed by waterborne extracellular toxin to carnivorous fish is less well understood. In this study rainbow trout (Oncorhynchus mykiss) were exposed for up to 28 d to waterborne microcystin-LR at nominal concentrations of 1.5 and 50 µg L−1 (measured values 2 and 49 µg L−1, respectively). The former represents the Canadian drinking water guideline, and the latter an elevated environmental level. This study hypothesised that waterborne toxin exposure would specifically impact gill function, and given the importance of this tissue in freshwater fish ion regulation, effects on plasma ions and branchial ion transporter activity would be observed. Microcystin-LR exposure resulted in a significant and persistent hypocalcaemia at the higher exposure concentration, but plasma sodium and branchial activities of the sodium/potassium ATPase, proton ATPase and calcium ATPase enzymes remained unaffected. An in vitro assessment failed to show any effect of microcystin-LR on branchial calcium ATPase activity even at exposure concentrations as high as 1000 µg L−1. A transient increase in hepatic alkaline phosphatase activity was also observed at 49 µg L−1, but there were no effects of toxin exposure on branchial or hepatic lactate dehydrogenase activity. These results suggest that microcystin-LR exposure does not have a general effect on ion regulation, but instead produces a novel and specific impact on calcium metabolism in rainbow trout, although the mechanism underlying this effect remains unknown.

Nodularin (NOD) is a potent toxin produced by Nodularia spumigena cyanobacteria. Usually, NOD co-exists with other microcystins in environmental waters, a class of cyanotoxins secreted by certain cyanobacteria species, which makes identification difficult in the case of mixed toxins. Herein we report a complete theoretical DFT-vibrational Raman characterization of NOD along with the experimental drop-coating deposition Raman (DCDR) technique. In addition, we used the vibrational characterization to probe SERS analysis of NOD using colloidal silver nanoparticles (AgNPs), commercial nanopatterned substrates with periodic inverted pyramids (KlariteTM substrate), hydrophobic Tienta® SpecTrimTM slides, and in-house fabricated periodic nanotrenches by nanoimprint lithography (NIL). The 532 nm excitation source provided more well-defined bands even at LOD levels, as well as the best performance in terms of SERS intensity. This was reflected by the results obtained with the KlariteTM substrate and the silver-based colloidal system, which were the most promising detection approaches, providing the lowest limits of detection. A detection limit of 8.4 × 10−8 M was achieved for NOD in solution by using AgNPs. Theoretical computation of the complex vibrational modes of NOD was used for the first time to unambiguously assign all the specific vibrational Raman bands. [ABSTRACT FROM AUTHOR]

Microcystins are produced by multifaceted organisms called cyanobacteria, which are integral to Africa's freshwater environments. The excessive proliferation of cyanobacteria caused by rising temperature and eutrophication leads to the production and release of copious amounts of microcystins, requiring critical management and control approaches to prevent the adverse environmental and public health problems associated with these bioactive metabolites. Despite hypotheses reported to explain the phylogeography and mechanisms responsible for cyanobacterial blooms in aquatic water bodies, many aspects are scarcely understood in Africa due to the paucity of investigations and lack of uniformity of experimental methods. Due to a lack of information and large-scale studies, cyanobacteria occurrence and genetic diversity are seldom reported in African aquatic ecosystems. This review covers the diversity and geographical distribution of potential microcystin-producing and non-microcystin-producing cyanobacterial taxa in Africa. Molecular analyses using housekeeping genes (e.g., 16S rRNA, ITS, rpoC1, etc.) revealed significant sequence divergence across several cyanobacterial strains from East, North, West, and South Africa, but the lack of uniformity in molecular markers employed made continent-wise phylogenetic comparisons impossible. Planktothrix agardhii, Microcystis aeruginosa, and Cylindrospermopsis raciborskii (presently known as Raphidiopsis raciborskii) were the most commonly reported genera. Potential microcystin (MCs)-producing cyanobacteria were detected using mcy genes, and several microcystin congeners were recorded. Studying cyanobacteria species from the African continent is urgent to effectively safeguard public and environmental health because more than 80% of the continent has no data on these important microorganisms and their bioactive secondary metabolites. [ABSTRACT FROM AUTHOR]

Water resources are critically important, but also pose risks of exposure to toxic and pathogenic microbes. Increasingly, a concern is toxic cyanobacteria, which have been linked to the death and disease of humans, domesticated animals, and wildlife in freshwater systems worldwide. Management approaches successful at reducing cyanobacterial abundance and toxin production have tended to be short-term solutions applied on small scales (e.g., algaecide application) or solutions that entail difficult multifaceted investments (e.g., modification of landscape and land use to reduce nutrient inputs). However, implementation of these approaches can be undermined by microbial species interactions that (a) provide toxic cyanobacteria with protection against the method of control or (b) permit toxic cyanobacteria to be replaced by other significant microbial threats. Understanding these interactions is necessary to avoid such scenarios and can provide a framework for novel strategies to enhance freshwater resource management via systems science (e.g., pairing existing physical and chemical approaches against cyanobacteria with ecological strategies such as manipulation of natural enemies, targeting of facilitators, and reduction of benthic occupancy and recruitment). Here, we review pertinent examples of the interactions and highlight potential applications of what is known. [ABSTRACT FROM AUTHOR]

Cylindrospermopsin (CYN) is a cyanobacterial toxin that occurs in aquatic environments worldwide. It is known for its delayed effects in animals and humans such as inhibition of protein synthesis or genotoxicity. The molecular targets and the cell physiological mechanisms of CYN, however, are not well studied. As inhalation of CYN-containing aerosols has been identified as a relevant route of CYN uptake, we analyzed the effects of CYN on protein expression in cultures of immortalized human bronchial epithelial cells (16HBE14o−) using a proteomic approach. Proteins whose expression levels were affected by CYN belonged to several functional clusters, mainly regulation of protein stability, cellular adhesion and integration in the extracellular matrix, cell proliferation, cell cycle regulation, and completion of cytokinesis. With a few exceptions of upregulated proteins (e.g., ITI inhibitor of serine endopeptidases and mRNA stabilizer PABPC1), CYN mediated the downregulation of many proteins. Among these, centrosomal protein 55 (CEP55) and osteonectin (SPARC) were significantly reduced in their abundance. Results of the detailed semi-quantitative Western blot analyses of SPARC, claudin-6, and CEP55 supported the findings from the proteomic study that epithelial cell adhesion, attenuation of cell proliferation, delayed completion of mitosis, as well as induction of genomic instability are major effects of CYN in eukaryotic cells. [ABSTRACT FROM AUTHOR]