Cylindrospermopsin Concentrations Research Articles

Toxic and non-toxic cyanobacterial biomass as a resource for sustainable agriculture: A lettuce cultivation experiment

Cyanobacteria represent a promising resource for sustainable agriculture, as they have demonstrated the ability to restore soil fertility even after death and decay. However, several cyanobacteria can also release secondary metabolites, such as cyanotoxins, which may compromise the quality of agricultural products and pose a potential risk to human health. Depending on the concentration of exposure, few studies reported deleterious effects on plant species when irrigated with cylindrospermopsin (CYN) contaminated water, impairing plant growth and leading to food product contamination, while other studies show promoting effects on plant yield. To evaluate the potential of cyanobacterial biomass (cyanotoxin-containing or not) as a sustainable resource for soil amendment, biostimulants or fertilizers for lettuce cultivation, a study was carried out that consisted of the culture of lettuce plants under controlled conditions, in soil: (1) with no extra nutrient addition (control) and supplemented with 0.6 g of freeze-dried Raphidiopsis raciborskii biomass of (2) a non-CYN-producing strain, (3) a CYN-producing strain, and (4) the same CYN-producing strain pasteurized. Results showed no significant differences in photosystem II efficiency with the amendment addition. On the contrary, shoot fresh weight significantly increased in lettuce plants grown with the cyanobacterial biomass addition, especially in condition (3). In addition, there were significant differences in mineral concentrations in lettuce leaves after the cyanobacterial biomass addition, such as K, Na, Ca, P, Mg, Mn, Zn, Cu, Mo, and Co. CYN accumulation was detected under conditions (3) and (4), with concentrations observed in descending order from roots > soil > shoot. Nevertheless, the CYN concentration in edible tissues did not exceed the WHO-proposed tolerable daily intake of 0.03 μg/kg/day. These findings suggest that incorporating cyanobacterial biomass as a soil amendment, biostimulant or fertilizer for lettuce cultivation, even with trace amounts of CYN (1–40 μg/g), may enhance plant yield without leading to cyanotoxin accumulation in edible tissues above the WHO-recommended tolerable daily intake.

Nephrotoxicity of Cylindrospermopsin (CYN) and Microcystin-LR (MC-LR) on Mammalian Kidney: Wistar Rat as a Model Assessment

Naturally derived cyanotoxins, cylindrospermopsin (CYN), and microcystin-LR (MC-LR) have shown hepatotoxic and nephrotoxic effects in several studies. The present study aimed to determine the possible nephrotoxicity of MC-LR and CYN on mammalian kidneys using male Wistar rats as an animal model. Potential nephrotoxicity was evaluated at different doses of CYN (0.175 μg.kg-1, 0.140 μg.kg-1, 0.105 μg.kg-1) and MC-LR (0.105 μg.kg-1, 0.070 μg.kg-1, 0.035 μg.kg-1) was observed. Water samples from dug wells contaminated with CYN (0.161 μg.kg-1) and MC-LR (0.091 μg.kg-1) from the Padaviya area in Anuradhapura, Sri Lanka were used as environmental samples. The control groups were treated with distilled water. The exposure time of rats to the toxin was 90 days. Evaluation of urinary creatinine, serum creatinine, and Kidney Injury Molecule-1 (KIM-1) were estimated using standard protocols. A significant increase in serum creatinine levels was observed in all CYN and MC-LR treated groups (p<0.05) after 7 and 42 days of exposure, respectively, compared to control. It was found a decrease of urine creatinine when rats were treated with different concentrations of CYN and MC-LR (p<0.05) after 7 days compared to the control. The highest KIM-1 concentrations were recorded at 0.175 μg.kg-1 of CYN and 0.105 μg.kg-1 of MC-LR. The concentrations of KIM-1 in the control groups for CYN-treated and MC-LR-treated were not detected. Luminal protein, nuclear pyknosis, mild tubular epithelial swelling, vascular congestion, and interstitial inflammation in CYN and MC-LR treated groups were common. No predominant changes were observed in the control groups treated with CYN and MC-LR. The results of the present study confirm that the consumption of CYN and MC-LR-contaminated water may lead to kidney injury in Wistar rats.

Environmentally Friendly and Cost-Effective Approaches to Reduce Toxin Content in Toxic Cyanobacterial Biomasses

Cyanobacterial outgrowths are naturally occurring processes in eutrophic aquatic ecosystems. Furthermore, as a result of climate change and anthropogenic pollution, cyanobacteria harmful algal blooms (CyanoHABs) are expanding worldwide. CyanoHABs are considered a threat to human health and environment due to the production of potent toxic substances, but at the same time, valuable products can be obtained from these microorganisms. The main objective of this study was to test straightforward and cost-effective methods to reduce the toxin content of cyanobacterial biomass for the exploitation of this important biological resource. To carry out this study, lyophilized or hydrated biomass from microcystin-LR (MC-LR) producing Microcystis aeruginosa and cylindrospermopsin (CYN) producing Chrysosporum ovalisporum strains were subjected to the following treatments: (1) thermal (50 °C); (2) ultraviolet (UV) radiation; (3) ozone; and (4) sunlight, for periods varying between 2 and 12 h. MC-LR and CYN concentrations were quantified by LC-MS and compared between experimental groups. The results show a significant reduction in the amount of MC-LR in M. aeruginosa biomass (lyophilized and hydrated) exposed to sunlight. Since no other treatment reduced MC-LR in M. aeruginosa biomass, this molecule was demonstrated to be very stable. Regarding CYN, the concentration of this toxin in C. ovalisporum biomass was significantly reduced with the exposure to UV radiation, to approximately 51% of the initial concentration after 2 h of exposure; 86% reduction after 5 h of exposure; and 77% reduction after 12 h of exposure. Overall, this study demonstrates that the toxicity of cyanobacterial biomass can be reduced by employing environmentally friendly and cost-effective treatments with sunlight and UV radiation.

Consumption of toxic benthic cyanobacteria by two common demersal fish: Growth, antioxidant and liver histopathology responses

Benthic freshwater cyanobacteria have the potential to produce toxins. Compared with more extensively studied plankton species, little is known about the impact of harmful benthic cyanobacteria on aquatic organisms. As demersal fish are usually in direct contact with benthic cyanobacteria, it is important to understand their interactive effects. This study investigated the physio-chemical responses of two demersal fish (Xenocypris davidi and Crucian carp) after exposure to benthic Oscillatoria (producing cylindrospermopsin, 2 × 106 cells/mL) for 7 days. Interestingly, benthic Oscillatoria had less adverse effects on X. davidi than C. carp. The two demersal fish effectively ingested Oscillatoria, but Oscillatoria cell sheathes could not be fully digested in C. carp intestines and led to growth inhibition. Oscillatoria consumption induced oxidative stress and triggered alterations in detoxification enzyme activities in the X. davidi liver. Superoxide dismutase (SOD) and glutathione reductase (GR) activities significantly increased in the C. carp liver, but catalase (CAT) and detoxification enzymes glutathione S-transferase (GST) and glutathione (GSH) activities were insignificantly changed. This suggested that C. carp may have a relatively weak detoxification capacity for toxic Oscillatoria. Oscillatoria ingestion led to more pronounced liver pathological changes in C. carp, including swelling, deformation, and loss of cytoskeleton structure. Simultaneously, fish consumption of Oscillatoria increased extracellular cylindrospermopsin concentration. These results provide valuable insights into the ecological risks associated with benthic cyanobacteria in aquatic ecosystems.

Biodegradation of the cyanotoxin cylindrospermopsin by Bacillus cereus, Micrococcus luteus and Alcaligenes faecalis 

Cylindrospermopsin (CYN) is a cyanotoxin found in natural waters, with potential risk to human health through the inhibition of protein synthesis. Despite the implementation of conventional water treatment procedures, complete removal of CYN remains a question due to its heat-stable nature. Hence, contamination of water sources with CYN is a challenge in providing safe drinking water throughout the world. The present study was conducted to test the ability to degrade CYN at 280C and pH 7, of four bacterial strains: Bacillus cereus-Y, Bacillus cereus-S (B. cereus-S), Micrococcus luteus, and Alcaligenes faecalis, which were previously isolated from different water sources as different hydrocarbon degraders. 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 280C 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.

Coupling of cylindrospermopsin and pho-harboring Verrucomicrobia supports the formation of Raphidiopsis blooms in low-phosphorus waters

Cylindrospermopsin (CYN) can induce phytoplankton community to secrete alkaline phosphatase (ALP), which is one of the important strategies for the bloom-forming cyanobacterium Raphidiopsis to thrive in extremely low-phosphorus (P) waters. However, how bacterioplankton community, another major contributor to ALPs in waters, couples to Raphidiopsis through CYN, and the role of this coupling in supporting the dominance of Raphidiopsis in nature remain largely unknown. Here, we conducted microcosm experiments to address this knowledge gap, using a combination of differential filtration-based and metagenomics-based methods to identify the sources of ALPs. We found that, compared with algal-derived ALPs, bacteria-derived ALPs exhibited a more pronounced and sensitive response to CYN. This response to CYN was enhanced under low-P conditions. Interestingly, we found that Verrucomicrobia made the largest contribution to the total abundance of pho genes, which encode ALPs. Having high gene abundance of the CYN-sensing PI3K-AKT signaling pathway, Verrucomicrobia's proportion increased with higher concentrations of CYN under low-P conditions, thereby explaining the observed increase in pho gene abundance. Compared with other cyanobacterial genera, Raphidiopsis had a higher abundance of the pst gene. This suggests that Raphidiopsis exhibited a greater capacity to uptake the inorganic P generated by ALPs secreted by other organisms. Overall, our results reveal the mechanism of CYN-induced ALP secretion and its impact on planktonic P-cycling, and provide valuable insights into the role of CYN in supporting the formation of Raphidiopsis blooms.

Cylindrospermopsin exposure promotes redox unbalance and tissue damage in the liver of Poecilia reticulata, a neotropical fish species

ABSTRACT 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.

Cyanotoxins Increase Cytotoxicity and Promote Nonalcoholic Fatty Liver Disease Progression by Enhancing Cell Steatosis.

Freshwater prokaryotic cyanobacteria within harmful algal blooms produce cyanotoxins which are considered major pollutants in the aquatic system. Direct exposure to cyanotoxins through inhalation, skin contact, or ingestion of contaminated drinking water can target the liver and may cause hepatotoxicity. In the current study, we investigated the effect of low concentrations of cyanotoxins on cytotoxicity, inflammation, modulation of unfolded protein response (UPR), steatosis, and fibrosis signaling in human hepatocytes and liver cell models. Exposure to low concentrations of microcystin-LR (MC-LR), microcystin-RR (MC-RR), nodularin (NOD), and cylindrospermopsin (CYN) in human bipotent progenitor cell line HepaRG and hepatocellular carcinoma (HCC) cell lines HepG2 and SK-Hep1 resulted in increased cell toxicity. MC-LR, NOD, and CYN differentially regulated inflammatory signaling, activated UPR signaling and lipogenic gene expression, and induced cellular steatosis and fibrotic signaling in HCC cells. MC-LR, NOD, and CYN also regulated AKT/mTOR signaling and inhibited autophagy. Chronic exposure to MC-LR, NOD, and CYN upregulated the expression of lipogenic and fibrosis biomarkers. Moreover, RNA sequencing (RNA seq) data suggested that exposure of human hepatocytes, HepaRG, and HCC HepG2 cells to MC-LR and CYN modulated expression levels of several genes that regulate non-alcoholic fatty liver disease (NAFLD). Our data suggest that low concentrations of cyanotoxins can cause hepatotoxicity and cell steatosis and promote NAFLD progression.

Cytotoxic Effects and Oxidative Stress Produced by a Cyanobacterial Cylindrospermopsin Producer Extract versus a Cylindrospermopsin Non-Producing Extract on the Neuroblastoma SH-SY5Y Cell Line.

The incidence and interest of cyanobacteria are increasing nowadays because they are able to produce some toxic secondary metabolites known as cyanotoxins. Among them, the presence of cylindrospermopsin (CYN) is especially relevant, as it seems to cause damage at different levels in the organisms: the nervous system being the one most recently reported. Usually, the effects of the cyanotoxins are studied, but not those exerted by cyanobacterial biomass. The aim of the present study was to assess the cytotoxicity and oxidative stress generation of one cyanobacterial extract of R. raciborskii non-containing CYN (CYN-), and compare its effects with those exerted by a cyanobacterial extract of C. ovalisporum containing CYN (CYN+) in the human neuroblastoma SH-SY5Y cell line. Moreover, the analytical characterization of potential cyanotoxins and their metabolites that are present in both extracts of these cultures was also carried out using Ultrahigh Performance Liquid Chromatography-Mass Spectrometry, in tandem (UHPLC-MS/MS). The results show a reduction of cell viability concentration- and time-dependently after 24 and 48 h of exposure with CYN+ being five times more toxic than CYN-. Furthermore, the reactive oxygen species (ROS) increased with time (0-24 h) and CYN concentration (0-1.11 µg/mL). However, this rise was only obtained after the highest concentrations and times of exposure to CYN-, while this extract also caused a decrease in reduced glutathione (GSH) levels, which might be an indication of the compensation of the oxidative stress response. This study is the first one performed in vitro comparing the effects of CYN+ and CYN-, which highlights the importance of studying toxic features in their natural scenario.

Paramecium jenningsi effectively grazes on toxic Raphidiopsis raciborskii and degrades cylindrospermopsin: Implications for control harmful cyanobacterial blooms

Harmful cyanobacterial blooms (HCB) represent a serious threat to aquatic ecosystems due to their production for potent toxins that have negative effects on plant, animal and human health. Although grazing process can be exploited to reduce harmful cyanobacteria in eutrophic waters, the feeding capacity and detoxification capability of the grazer should be considered as key traits of a safe biocontrol agent. In this study, laboratory experiments were conducted to investigate the grazing rate of the ciliate Paramecium jenningsi on toxic Raphidiopsis raciborskii and potential cylindrospermopsin (CYN) degradation during feeding process. P. jenningsi efficiently ingested R. raciborskii cells with high growth (0.1- 0.26 d−1) and grazing rates (0.1-1.4 × 104 cyanobacterial cells ciliate −1 d−1), varying with the initial ciliate density. CYN concentrations within Raphidiopsis cells displayed an elevation under grazing conditions (5.3-7.2 pg cell−1) compared to control cultures (4.4 pg cell−1). CYN concentrations within the ciliate cells showed a reduction (degradation rate =121-276 pg ciliate−1 d−1) and became undetectable after all Raphidiopsis cells were grazed out, indicating that this ciliate could not accumulate CYN within its body during feeding process. Therefore, P. jenningsi could be used as a safe bioagent for the biocontrol of HCBs in water sources. However, a further in situ study is needed for a possible application of Paramecium species for mitigation of toxic cyanobacteria and their cyanotoxins in the natural environment.

Ferrate(VI) mediated degradation of the potent cyanotoxin, cylindrospermopsin: Kinetics, products, and toxicity

The presence of cylindrospermopsin (CYN), a potent cyanotoxin, in drinking water sources poses a tremendous risk to humans and the environment. Detailed kinetic studies herein demonstrate ferrate(VI) (FeVIO42−, Fe(VI)) mediated oxidation of CYN and the model compound 6-hydroxymethyl uracil (6-HOMU) lead to their effective degradation under neutral and alkaline solution pH. A transformation product analysis indicated oxidation of the uracil ring, which has functionality critical to the toxicity of CYN. The oxidative cleavage of the C5=C6 double bond resulted in fragmentation of the uracil ring. Amide hydrolysis is a contributing pathway leading to the fragmentation of the uracil ring. Under extended treatment, hydrolysis, and extensive oxidation lead to complete destruction of the uracil ring skeleton, resulting in the generation of a variety of products including nontoxic cylindrospermopsic acid. The ELISA biological activity of the CYN product mixtures produced during Fe(VI) treatment parallels the concentration of CYN. These results suggest the products do not possess ELISA biological activity at the concentrations produced during treatment. The Fe(VI) mediated degradation was also effective in the presence of humic acid and unaffected by the presence of common inorganic ions under our experimental conditions. The Fe(VI) remediation of CYN and uracil based toxins appears a promising drinking water treatment process.

Photo-Fenton oxidation of cylindrospermopsin at neutral pH with LEDs

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.

Water quality, microcystin-LR, and Cylindrospermopsin contamination status of spring and dug well water in CKDu high, low, and non-prevalent areas of Sri Lanka

Cyanotoxins, Microcystin LR (MC-LR) and Cylindrospermopsin (CYN) are hepato and nephro toxins and considered as one of the hypotheses for Chronic Kidney Disease of Unknown Etiology (CKDu) in Sri Lanka. Recent studies have revealed that significant number of dug wells in the North Central and Uva provinces where CKDu is prevalent are contaminated with cyanotoxins MC-LR and CYN. In the present study, cyanotoxin; MC-LR, CYN, and potential toxin-producing cyanobacteria were studied in 330 dug wells and 9 spring water samples collected from CKDu high prevalent areas in several Divisional Secretariats (DS) and Grama Niladhari (GN) divisions of Anuradhapura (Padaviya DS, Medawachchiya DS, Kebithigollewa DS), Polonnaruwa (Medirigiriya DS), Badulla (Girandurukotte GN) and Ampara (Dehiaththakandiya DS) districts, low prevalent areas in Anuradhapura district (Galnewa DS, Rajanganaya DS), non-prevalent areas in Angunakolapelessa DS of the Hambanthota district. General water quality parameters: temperature, pH, conductivity, Dissolved Oxygen (DO), Nitrate (N-NO3-), Nitrite (N-NO2-), Ammonia (N-NH3), Total Phosphorus (TP), and Total Hardness (TH) were determined using standard spectrophotometric and titrimetric methods. BACON test kits were used following manufactures instructions to quantify the CYN, and MC-LR concentrations by using the ELISA plate reader. Cyanobacteria density and species composition were determined under light microscopy following the standard algae and cyanobacteria keys. With the exception of Electrical Conductivity, all of the water quality parameters tested remained within the SLSI drinking water quality standard range. Microcystis spp. cell densities ranged from 14±3 to 1590±256 cells mL-1, where MC-LR concentration ranging from 0.04±0.01 to 3.89±0.02 µg L-1. At different CYN concentrations (0.04±0.01 to 3.59±0.05 µg L-1), Cylindrospermopsis spp. cell densities varied from 15 ±4 to 615±112 cells mL-1. Well water collected from Hambanthota district and spring water was collected Kebithigollewa DS, in Anuradhapura did not record cyanotoxins and cyanobacteria. The factor analysis classified well waters into three clusters of high, low, and non-prevalence areas with presence and absence of cyanobacteria and cyanotoxins. In contrast, springs were classified to a separate cluster. High concentrations of MC-LR and CYN were found in wells where CKDu high prevalent 85% of Padaviya DS and 80% of Medirigiriya DS, respectively. High Nitrate-N (3.01±0.56 mg L-1) and Nitrite-N (0.69±0.09 mg L-1) concentrations were recorded in well water collected from high CKDu prevalence areas. Thus, the results of the present study showed a relationship between cyanobacteria, cyanotoxins and CKDu records in the study area. Accordingly, further comprehensive studies are needed to be carried out to confirm the relationship between cyanotoxins and CKDu in Sri Lanka as cyanotoxin were listed in the WHO report as one of the possible reasons for CKDu.

Trace determination of multiple hydrophilic cyanotoxins in freshwater by off- and on-line solid phase extraction coupled to liquid chromatography-tandem mass spectrometry

Hydrophilic cyanotoxins (HCTs), such as paralytic shellfish toxins (PSTs), anatoxin-a (ATX-a), and cylindrospermopsin (CYN) are highly toxic and toxin-producing algae are widely distributed worldwide. However, HCTs, especially PSTs, are rarely reported in freshwater due to analytical limitations. This may result in an underestimation of the ecological risks and health risks. This study developed a new method to detect ATX-a, CYN, and thirteen common PSTs in freshwater simultaneously by using off-line solid phase extraction (SPE) and liquid chromatography-tandem mass spectrometry (LC-MS/MS). The limits of detection (LODs) of all targets were lower than 0.05 μg/L, which could meet the regulatory requirements for monitoring of HCTs in drinking water in different countries and regions. To improve the detection sensitivities for trace PSTs, a method based on off-line SPE and on-line SPE-LC-MS/MS was established with LOD around 0.001 μg/L. GTX1&4, GTX2&3, and GTX5 were detected in freshwater in China for the first time, highlighting that overall communities are facing potential risks of exposure to various PSTs in China. High concentrations of ATX-a and CYN were also detected in freshwater from Northern China. The proposed method helps to understand the pollution status of HCT in water bodies, especially during the non-algal bloom period.

Bioremoval of Cylindrospermopsis raciborskii cells and cylindrospermopsin toxin in batch culture by the yeast Aureobasidium pullulans.

This study describes the ability of a yeast strain, Aureobasidium pullulans KKUY0701 isolated from eutrophic lake to eliminate Cylindrospermopsis raciborskii and cylindrospermopsin (CYN) toxin. The anti-cyanobacterial activity of this yeast strain was evaluated by growing with living cells and filtrate of C. raciborskii. CYN bioremoval was assayed using living and heat-inactivated yeast cells. Both living cells and filtrate of this yeast strain were able to suppress the growth of C. raciborskii, with total cell death occurring at day 2 and day 3, respectively. Living and inactivated yeast cells, but not yeast filtrate, reduced CYN concentrations released into cyanobacterial cultures, indicating that this toxin might be removed from the culture medium via absorption onto yeast surface rather than enzymatic biodegradation. The adsorption experiments also confirmed the elimination of CYN by living and heat-inactivated yeast. Nevertheless, inactivated yeast exhibited higher capacity (K = 3.3) and intensity (n = 1.4) than living yeast (K = 1.9, n = 1) for CYN adsorption. The study suggests that this yeast strain could be employed for bioremediation of Cylindrospermopsis blooms in freshwaters. Additionally, heat-inactivated yeast biomass could be used in slow sand filters for elimination of CYN in drinking water treatment plants.

Co-occurrence of microcystin and cylindrospermopsin in hypereutrophic Mahakanadarawa and Nachchaduwa reservoirs

Microcystins (MCs) and cylindrospermopsin (CYN) are the most abundant toxins produced by cyanobacteria in tropical freshwaters. We studied the spatial distribution of MC and CYN in two multipurpose reservoirs, Mahakanadarawa and Nachchaduwa in Anuradhapura district in Sri Lanka in September 2020. Fourteen water quality parameters, phytoplankton composition, chlorophyll-a, MC and CYN concentrations were analyzed in triplicate in 25 sampling sites from each reservoir. Both reservoirs were at hypereutrophic status. Microcystis was the dominant cyanobacteria with 0-3.75 x 103 cell/mL in Mahakanadarawa and 1-7 x 103 cell/mL in Nachchaduwa. Besides Microcystis, no other potential MC-producing cyanobacteria were observed. In Mahakanadarawa, MC was detected in the range of 0.11-1.63 µg/L which was above the WHO permissible level (1.0 µg/L) for drinking water. Although comparatively high Microcystis cell density was present in Nachchaduwa, its MC concentration was low (0.06-0.17 µg/L). The CYN concentration in Nachchaduwa was above the WHO permissible level (0.7 µg/L) for drinking water. It was 0.20-1.02 µg/L in Nachchaduwa and 0.03-0.08 µg/L in Mahakanadarawa. We did not observe any potential CYN-producing cyanobacteria in either of the reservoirs. There was no relationship between the spatial distribution pattern of MC and Microcystis cell density in both reservoirs. Although the majority of physico-chemical properties of water indicated suitability for drinking, co-occurrence of high concentrations of MC and CYN indicated their unsuitability for drinking. Hence, this study highlights the necessity for routing detection of cyanotoxins in both reservoirs. Further, our findings alarm potential health risks for the local community that relies on Mahakanadarawa and Nachchaduwa reservoirs for drinking, irrigation and fisheries.

Determination of Cylindrospermopsin in Surface and Treated Water using Liquid Chromatography-Tandem Mass Spectrometry

Cylindrospermopsin (CYN) is an emerging freshwater cyanobacterial toxin, and its reports on toxicity toward the human liver and kidney tissues has drawn a lot of attention. An appropriate analytical method is necessary to determine the presence of this emerging cyanobacterial toxin in water resources including drinking water; therefore, it is necessary to develop a sensitive analytical method for CYN detection. In this study, we developed a simple and sensitive analytical method for CYN detection using liquid chromatography-tandem mass spectrometry (LC-MS/MS) using direct injection. The method was validated for linearity of calibration, limit of detection, limit of quantitation, accuracy, and precision. The limit of detection and quantitation were in the range of 0.029 μg/L and 0.091 μg/L, respectively. Accuracy and precision were also obtained within an acceptable range. The optimized method was used to measure the concentrations of CYN in the surface water from each weir areas of the Geum River, Nakdong River. Additionally, this method was applied to samples of drinking water obtained from the treatment plants of the Geum River, Nakdong River for each process.

Cyanobacterium Raphidiopsis raciborskii and its toxin in Buon Phong reservoir, Dak Lak province, Vietnam

This study investigated the variation of the cyanobacterium Raphidiopsis raciborskii population under the influence of physicochemical parameters from May 2019 to April 2020 in the Buon Phong reservoir. The correlations between these parameters were explored by using Principal Component Analysis (PCA) and Pearson correlation analysis. The cylindrospermopsin (CYN) toxin in isolated strains from the reservoir was confirmed by using High-performance liquid chromatography (HPLC) analyses. R. raciborskii presented throughout the year in the reservoir with a biovolume from 0.12 to 9.14 mm3 L-1. Four R. raciborskii strains (CBP2, CBP3, CBP4, and CBP5) were successfully isolated and confirmed to produce CYN by the HPLC results. The highest concentration in isolated strains was 0.345 μg g-1 DW in the CBP4 strain. The PCR results of genes responsible for CYN biosynthesis showed that the PCR amplicons of cyrB and CyrC were amplified in two toxic strains (CBP2 and CBP3), while the amplicons of both cyrB and cyrC were not observed in two remaining toxic strains. In addition, such abiotic factors as temperature and nutrients played major roles in the abundance of R. raciborskii. Moreover, the biovolume of R. raciborskii positively correlated with the CYN concentrations in the Buon Phong reservoir.

The cyanotoxin cylindrospermopsin slows down cell cycle progression and extends metaphase duration in immortalised human airway epithelial cells

Cylindrospermopsin (CYN) is a cyanobacterial toxin that occurs worldwide in aquatic environments. It is known for its delayed effects upon oral uptake in animals and humans. A less well studied route of CYN internalisation is the inhalation of CYN-contaminated aerosols. We analyzed potential effects of different CYN concentrations (1, 2.5 and 5 μmol/l) on cultures of immortalised human bronchial epithelial cells (16HBE14o−). Impedance, a proxi for cell attachment to the culture support, cell spreading, cell growth and cell proliferation, was measured using an Acea iCELLigence device. Cell division rate and metaphase duration were determined using time lapse movies (Nikon Biostation II) of CYN-exposed cell cultures. Western blot studies were used to determine expression levels of cell cycle regulator proteins, the cyclins B1, D1 and A2. Our investigations revealed that exposure of cells to CYN concentrations of 1 μmol/l or higher led to a concentration- and time-dependent attenuation of impedance development as well as cell proliferation rate, and an extension of the metaphase of the cell cycle. CYN-mediated downregulation of cyclins B1 and D1 may be part of the underlying cell physiological mechanism. These results indicate that exposure of airways in humans and animals to aerosolised CYN over longer periods may be harmful.

Cylindrospermopsin impairs zebrafish (Danio rerio) embryo development

Cyanotoxins are among common contaminants that can impair human, animal, and environmental health. Cylindrospermopsin (CYN) is an abundant form of cyanotoxins elevated following algal bloom in the water worldwide. Previous studies have described CYN effects on several organs in mammals. However, little is known about its toxicity mechanisms in other vertebrates. This study aims to characterize the developmental effects of CYN using zebrafish larvae as an aquatic model organism. A wide range of CYN concentrations (0–2000 μg/L) was tested using a morphometric approach for survival, hatching, various growth and developmental abnormalities. We also investigated the expression of genes related to oxidative stress, osmoregulation, and thyroid function. Exposure to CYN resulted in decreased growth, increased developmental abnormalities such as pericardial and yolk sac edema as well as swim bladder absence. In addition, CYN increased tr1a, and decreased dio1 and dio3 transcript levels which are involved in thyroid-mediated function. It also increased transcript levels related to oxidative stress, including hsp70, ahr1a, cyp1a, gpx and cat. Lastly, CYN exposure increased aqp3a and decreased dab2, which are involved in osmoregulation with a threshold of 10 μg/L. The present study demonstrates multiple effects of exposure to environmentally relevant CYN concentrations in zebrafish embryos.