Presence of the neurotoxin ß-N-methylamino-L-alanine in irrigation water and accumulation in cereal grains with human exposure risk.

The present study demonstrates the presence of the neurotoxin ß-N-methylamino-L-alanine and its cyanobacterial producers in irrigation water and grains of some cereal plants from farmlands irrigated with Nile River water in Egypt. BMAA detected by LC-MS/MS in phytoplankton samples was found at higher concentrations of free form (0.84-11.4 µg L-1) than of protein-bound form (0.16-1.6 µg L-1), in association with the dominance of cyanobacteria in irrigation water canals. Dominant cyanobacterial species isolated from these irrigation waters including Aphanocapsa planctonica, Chroococcus minutus, Dolichospermum lemmermanni, Nostoc commune, and Oscillatoria tenuis were found to produce different concentrations of free (4.8-71.1 µg g-1 dry weight) and protein-bound (0.1-11.4 µg g-1 dry weight) BMAA. In the meantime, BMAA was also detected in a protein-bound form only in grains of corn (3.87-4.51 µg g-1 fresh weight) and sorghum (5.1-7.1 µg g-1 fresh weight) plants, but not in wheat grains. The amounts of BMAA accumulated in these grains correlated with BMAA concentrations detected in relevant irrigation water canals. The presence of BMAA in cereal grains would constitute a risk to human and animal health upon consumption of contaminated grains. The study, therefore, suggests continuous monitoring of BMAA and other cyanotoxins in irrigation waters and edible plants to protect the public against exposure to such potent toxins.

Biotransformation and detoxification of saxitoxin by Bacillus flexus in batch experiments.

Saxitoxins (STXs) are carbamate alkaloid neurotoxins produced by some species of cyanobacteria. They are water soluble and relatively stable in the natural environment, and thereby represent a risk to animal and human health through a long-time exposure. STXs cannot be sufficiently removed by conventional water treatment methods. Therefore, this study investigates the potential STX biodegradation and detoxification by bacteria as a promising method for toxin removal. STX biodegradation experiments were conducted using Bacillus flexus SSZ01 strain in batch cultures. The results revealed that SSZ01 strain grew well and rapidly detoxified STX, with no lag phase observed. STX detoxification by SSZ01 strain was initial-toxin-concentration-dependent. The highest biotransformation rate (10 µg STX L-1 day-1) the pseudo-first-order kinetic constant (0.58 d-1) were obtained at the highest initial toxin concentration (50 µg L-1) and the lowest ones (0.06 µg STX L-1 day-1 and 0.14 d-1, respectively) were recorded at the lowest initial concentration (0.5 µg L-1). STX biotransformation rate increased with temperature, with highest occurred at 30 ºC. This rate was also influenced by pH, with highest obtained at pH8 and lowest at higher and lower pH values. HPLC chromatograms showed that STX biotransformation peak is corresponding to the least toxic STX analog (disulfated sulfocarbamoyl-C1 variant). The Artemia-based toxicity assay revealed that this biotransformation byproduct was nontoxic. This suggests the potential application of this bacterial strain in slow sand filters for cyanotoxin removal in water treatment plants. Being nontoxic, this byproduct needs to be assayed for its therapeutic effects toward neurodegenerative diseases.

Performance evaluation of Moringa oleifera seeds aqueous extract for removing Microcystis aeruginosa and microcystins from municipal treated-water.

Introduction: Toxic microcystins (MCs) produced by cyanoprokaryotes -particularly by the cosmopolitan cyanobacterium Microcystis aeruginosa- pose adverse effects on aquatic organisms and their ecosystem and may also cause serious impacts on human health. These harmful monocyclic heptapeptides are the most prevalent cyanotoxins reported in freshwaters and must be eliminated for avoiding MCs release in receiving water bodies. Hence, this work aimed to test the efficacy of Moringa oleifera seeds water-based extract (MO) as a natural coagulant for removing cyanobacteria (especially M. aeruginosa), microalgae, and its associated MCs from pre-treated municipal wastewaters. Methodology: Four different MO coagulant doses (25, 50, 75 and 100 mg L-1) were investigated for cyanobacteria and microalgae removal by conventional coagulation assays and morphology-based taxonomy studies. Additionally, water turbidity and chlorophyll a (Chl a) content were also determined. Further, the presence and concentration of MCs soluble in water, remaining in the particulate fraction, and flocculated within the residual sludge were assessed using high-performance liquid chromatography coupled with diode array detection (HPLC-DAD). Results: The treatment with MO at 100 mg L-1 substantially reduced the number of cyanobacterial and microalgal species in the treated samples (average removal rate of 93.8% and 86.9%, respectively). These results agreed with a ∼44% concomitant reduction in Chl a and ∼97% reduction in water turbidity (a surrogate marker for suspended solids content). Notably, MCs concentrations in the treated water were significantly lowered to 0.6 ± 0.1 µg L-1 after addition of 100 mg L-1 MO. This value is below the WHO recommended limits for MCs presence in drinking water (<1.0 µg L-1). Discussion: The present study provides promising insights into the applicability of MO as a cost-effective, reliable, and sustainable natural coagulant, particularly for using in developing countries, to eliminate harmful cyanobacteria and cyanotoxins in municipal water treatment facilities.

The link between microcystin levels in groundwater and surface Nile water, and assessing their potential risk to human health.

Although groundwater is an important source for drinking and irrigation water worldwide, particularly in arid countries, they have been paid little attention to their contamination with microcystins (MCs) compared to surface water. Our study is the fourth one reporting existence of MCs in groundwater due to surface-water and groundwater interaction. Dissolved MCs in groundwater were found with higher concentrations in summer (0.1 to 0.84 µg L-1) than in winter (0-0.06 µg L-1), in association with MCs detected in nearby surface Nile water. The chronic daily intake (CDI) of MCs for both adults and children (0-0.003 µg kg-1 body weight d-1) in groundwater were lower than the chronic reference dose (RfD, 0.003 µg kg-1 body weight d-1) during winter, with hazard quotient less than 1. Conversely, CDI values exceeded the reference dose during summer for both adults (0.005-0.024 µg kg-1 body weight d-1) and children (0.012-0.05 µg kg-1 body weight d-1), with hazard quotient greater than 1. This indicates that MCs concentrations in these groundwater wells might pose adverse health effects to both adults and children during summer, but not during winter. The study provides evidence for the risk of cyanotoxins in groundwater close to cyanobacteria-contaminated surface water. Therefore, regular monitoring for cyanotoxins in groundwater supplies used for drinking-water should be undertaken when cyanobacteria bloom events are noted in nearby surface waters.

Fungal biodegradation and removal of cyanobacteria and microcystins: potential applications and research needs.

Harmful cyanobacterial blooms (HCB) have severe impacts on marine and freshwater systems worldwide. They cause oxygen depletion and produce potent cyanotoxins that have detrimental effects on human and environmental health and deteriorate the water quality. Biological treatment of the water for control of cyanobacterial blooms and removal of cyanotoxins can be a more economical and environment-friendly way, as they do not result in production of undesirable by-products. Most biological treatments of cyanobacteria and cyanotoxins have concentrated largely on bacteria, with little attention paid to algicidal fungi. Therefore, this review aims to provide an overview of the current status and the main progresses achieved in fungal biodegradation of HCB and cyanotoxin research. The available data revealed that 15 fungal species had high lytic activity against cyanobacteria, and 6 species were capable of degrading microcystins (MCs). Some fungal species (e.g., Aurobasidium pullulans and Trichoderma citrinoviride) have been identified to selectively inhibit the growth of cyanobacteria rather than beneficial species of other algal groups. Interestingly, some fungal strains (Trichaptum abietinum, Trichoderma citrinoviride) exhibited di-functional trait, being efficient in lysing cyanobacteria and degrading MCs released from the cells after decay. Beyond a comprehensive review of algicidal and toxin-degrading activities of fungi, this paper also identifies and prioritizes research gaps in algicidal fungi. The review also gives insights to the potential applications of algicidal fungi for removal of cyanobacterial blooms and their cyanotoxins from the aquatic environment.

Impacts of Microcystins on Morphological and Physiological Parameters of Agricultural Plants: A Review.

Cyanobacteria are a group of photosynthetic prokaryotes that pose a great concern in the aquatic environments related to contamination and poisoning of wild life and humans. Some species of cyanobacteria produce potent toxins such as microcystins (MCs), which are extremely aggressive to several organisms, including animals and humans. In order to protect human health and prevent human exposure to this type of organisms and toxins, regulatory limits for MCs in drinking water have been established in most countries. In this regard, the World Health Organization (WHO) proposed 1 µg MCs/L as the highest acceptable concentration in drinking water. However, regulatory limits were not defined in waters used in other applications/activities, constituting a potential threat to the environment and to human health. Indeed, water contaminated with MCs or other cyanotoxins is recurrently used in agriculture and for crop and food production. Several deleterious effects of MCs including a decrease in growth, tissue necrosis, inhibition of photosynthesis and metabolic changes have been reported in plants leading to the impairment of crop productivity and economic loss. Studies have also revealed significant accumulation of MCs in edible tissues and plant organs, which raise concerns related to food safety. This work aims to systematize and analyze the information generated by previous scientific studies, namely on the phytotoxicity and the impact of MCs especially on growth, photosynthesis and productivity of agricultural plants. Morphological and physiological parameters of agronomic interest are overviewed in detail in this work, with the aim to evaluate the putative impact of MCs under field conditions. Finally, concentration-dependent effects are highlighted, as these can assist in future guidelines for irrigation waters and establish regulatory limits for MCs.

Growth inhibition of Microcystis aeruginosa and adsorption of microcystin toxin by the yeast Aureobasidium pullulans, with no effect on microalgae.

This study evaluates the inhibitory effect of a yeast strain, Aureobasidium pullulans KKUY0701, isolated from decayed cyanobacterial bloom against harmful cyanobacterium Microcystis aeruginosa and determines the ability of this strain to remove microcystin (MC) toxin from the water. The antialgal activity of this yeast strain was assayed by co-cultivation with M. aeruginosa, diatom, and green algal species. The MC adsorption experiment was conducted in the presence of living and heat-inactivated yeast cells. Both yeast cells and filtrates caused a rapid reduction in the growth of M. aeruginosa, with complete death and cell lysis occurring after 3 days. The yeast strain did not exhibit any inhibitory effect on either green algae or diatoms. Both living and heat-inactivated yeast cells were capable of adsorption of MC on their surfaces. Inactivated yeast exhibited higher adsorption capacity and lower intensity than living yeast for the adsorption of MC toxin. The results of this study suggest that this yeast strain could be employed to selectively reduce cyanobacterial blooms in freshwaters. Moreover, the application of heat-inactivated yeast's biomass for toxin adsorption gives new possibilities in drinking water treatment plants.

Thermostable Cellulase Biosynthesis from Paenibacillus alvei and its Utilization in Lactic Acid Production by Simultaneous Saccharification and Fermentation.

Cellulosic date palm wastes may have beneficial biotechnological applications for eco-friendly utilization. This study reports the isolation of thermophilic cellulase-producing bacteria and their application in lactic acid production using date palm leaves. The promising isolate was identified as Paenibacillus alvei by 16S rRNA gene sequencing. Maximum cellulase production was acquired using alkaline treated date palm leaves (ATDPL) at 48 h and yielded 4.50 U.mL-1 FPase, 8.11 U.mL-1 CMCase, and 2.74 U.mL-1 ß-glucosidase. The cellulase activity was optimal at pH 5.0 and 50°C with good stability at a wide temperature (40-70°C) and pH (4.0-7.0) range, demonstrating its suitability in simultaneous saccharification and fermentation. Lactic acid fermentation was optimized at 4 days, pH 5.0, 50°C, 6.0% cellulose of ATDPL, 30 FPU/ g cellulose, 1.0 g. L-1 Tween 80, and 5.0 g. L-l yeast extract using Lactobacillus delbrueckii. The conversion efficiency of lactic acid from the cellulose of ATDPL was 98.71%, and the lactic acid productivity was 0.719 g. L-1 h-1. Alkaline treatment exhibited a valuable effect on the production of cellulases and lactic acid by reducing the lignin content and cellulose crystallinity. The results of this study offer a credible procedure for using date palm leaves for microbial industrial applications.

Erratum to "Thermostable cellulase biosynthesis from Paenibacillus alvei and its utilization in lactic acid production by simultaneous saccharification and fermentation".

[This corrects the article DOI: 10.1515/biol-2020-0019.].

Concentrations of cylindrospermopsin toxin in water and tilapia fish of tropical fishponds in Egypt, and assessing their potential risk to human health.

Unlike microcystin, cylindrospermospin (CYN) concentrations in fishpond water and their accumulation in fish tissues have been largely unexplored. This study determined CYN levels in water and tilapia fish organs from three tropical fishponds in southern Egypt. Water and fish samples were collected monthly from fishponds for 12 months (Oct 2012 to Sep 2013). The results revealed that six CYN-producing species of cyanobacteria dominated phytoplankton populations and formed blooms in these fishponds during warm months. Among these species, Anabaena affinis, Planktothrix agardhii, Cylindrospermopsis catemaco, and C. philippinensis were assigned as CYN producers for the first time in the present study. The highest cell densities of CYN-producing species in fishponds were recorded in August and September 2013, correlating with high temperature, pH and nutrient concentrations. Dissolved CYN was found in fishpond waters at levels (0.3-2.76 µg L-1) very close to those of particulate CYN (0.4-2.37 µg L-1). CYN was also estimated in tilapia fish organs at levels up to 417 ng g-1 in the intestines, 1500 ng g-1 in the livers, and 280 ng g-1in edible muscles. Compared to the recommended guideline (0.03 µg kg-1 day-1), the estimated daily intake (EDI) of CYN in our samples of edible muscles exceeded this limit by a factor of 1.3-14 during summer and autumn. This might represent a risk to human health upon consumption of such contaminated fish muscles. Therefore, fishponds worldwide should be monitored for the presence toxic cyanobacteria to protect humans from their potent toxins.

Potentially harmful microalgae and algal blooms in the Red Sea: Current knowledge and research needs.

Harmful algal blooms (HABs) have increased throughout the world's coastal oceans during the last century mostly due to water eutrophication and climate change. These blooms are often accompanied by extreme extensive negative impacts to fisheries, coastal resources, public health and local economies. However, limited studies have reported HAB events in Red Sea coastal waters. This article reviews potentially harmful microalgae in the Red Sea, based on available published information during the last 3 decades. Five harmful algal blooms were recorded in the Red Sea; of which 3 blooms are formed by dinoflagellates (Noctiluca scintillans, Pyrodinuium bahamense, Protoperidinium quinquecorne), one by raphidophytes (Heterosigma akashiwo) and one by cyanobacteria (Trichodesmium erythraeum). Additionally, mangrove swamps in the Red Sea were occupied by cyanobacterial mats, which contain microcystin and saxitoxin-producing species. The existing data in this review could be a catalyst for the establishment of monitoring and management program for HABs and their toxins in Red Sea coastal waters. This review also identifies current research gaps and suggests future research directions.

Bioavailability of bound microcystins in mice orally fed with contaminated tilapia edible tissues: Implications to human health.

This study investigates for the first time in vivo potential release of bound microcystins (MCs) from edible fish tissues. Bound MCs were released in the mouse digestive system and reached the liver (2.1-14.3 ng g-1) and blood (1-5 ng ml-1). The released MCs were active and negatively affected liver functions of treated mice. The estimated daily intake (EDI) for bound MCs (93.5 µg kg-1 day-1) is about 2300 times higher than the WHO safe EDI (0.04 µg kg-1 day-1), compared to lower EDI for free MCs (0.002 µg kg-1 day-1). Therefore, bound MCs in fish tissues should be considered when fish are monitored for human consumption.

Occurrence of cyanobacteria and microcystin toxins in raw and treated waters of the Nile River, Egypt: implication for water treatment and human health.

Monitoring of cyanobacteria and their associated toxins has intensified in raw water sources of drinking water treatment plants (WTPs) in most countries of the world. However, it is not explored yet for Egyptian WTPs. Therefore, this study was undertaken to investigate the occurrence of cyanobacteria and their microcystin (MC) toxins in the Nile River source water of Damietta WTP during warm months (April-September 2013) and to evaluate the removal efficiency of both cyanobacterial cells and MCs by conventional methods used in this plant as a representative of Egyptian drinking WTPs. The results showed that the source water at the intake of Damietta WTP contained dense cyanobacterial population (1.1-6.6 × 107 cells L(-1)) dominated by Microcystis aeruginosa. This bloom was found to produce MC-RR and MC-LR. Both cyanobacterial cell density and intracellular MCs in the intake source water increased with the increase in temperature and nutrients during the study period, with maximum values obtained in August. During treatment processes, cyanobacterial cells were incompletely removed by coagulation/flocculation/sedimentation (C/F/S; 91-96.8%) or sand filtration (93.3-98.9%). Coagulation/flocculation induced the release of MCs into the ambient water, and the toxins were not completely removed or degraded during further treatment stages (filtration and chlorination). MCs in outflow tank water were detected in high concentrations (1.1-3.6 µg L - 1), exceeding WHO provisional guideline value of 1 µg L - 1 for MC-LR in drinking water. Based on this study, regular monitoring of cyanobacteria and their cyanotoxins in the intake source water and at different stages at all WTPs is necessary to provide safe drinking water to consumers or to prevent exposure of consumers to hazardous cyanobacterial metabolites.

Growth inhibition of the cyanobacterium Microcystis aeruginosa and degradation of its microcystin toxins by the fungus Trichoderma citrinoviride.

Harmful cyanobacterial blooms are recognized as a rapidly expanding global problem that threatens human and ecosystem health. Many bacterial strains have been reported as possible agents for inhibiting and controlling these blooms. However, such algicidal activity is largely unexplored for fungi. In this study, a fungal strain kkuf-0955, isolated from decayed cyanobacterial bloom was tested for its capability to inhibit phytoplankton species in batch cultures. The strain was identified as Trichoderma citrinoviride Based on its morphological characteristics and DNA sequence. Microcystis aeruginosa co-cultivated with living fungal mycelia rapidly decreased after one day of incubation, and all cells completely died and lysed after 2 days. The fungal filtrate of 5-day culture also exhibited an inhibitory effect on M. aeruginosa, and this inhibition increased with the amount of filtrate and incubation time. Conversely, green algae and diatoms have not been influenced by either living fungal mycelia or culture filtrate. Interestingly, the fungus was not only able to inhibit Microcystis growth but also degraded microcystin produced by this cyanobacterium. The toxins were completely degraded within 5 days of incubation with living fungal mycelia, but not significantly changed with fungal filtrate. This fungus could be a potential bioagent to selectively control Microcystis blooms and degrade microcystin toxins.

Selective inhibition of toxic cyanobacteria by ß-carboline-containing bacterium Bacillus flexus isolated from Saudi freshwaters.

A bacterial strain SSZ01 isolated from a eutrophic lake in Saudi Arabia dominated by cyanobacterial blooms, showed an antialgal activity against cyanobacteria species. Based on the analysis of the 16S rDNA gene sequence, the isolated strain (SSZ01) most likely belonged to the genus Bacillus with a 99% similarity to Bacillus flexus strain EMGA5. The thin layer chromatography (TLC) analysis of the ethyl acetate extract of this bacterium revealed that this strain can produce harmine and norharmane compared to different ß-carboline analog standards. Harmine and norharmane were also detected in considerable amounts in bacterial growth medium, indicating a potential excretion of these compounds into the aquatic environment. The crude extract of Bacillus flexus as well as pure materials of harmine and norharmane inhibited the growth of tested species of cyanobacteria. However, the bacterial crude extract has a higher toxicity against tested species of cyanobacteria than harmine and norharmane. In addition, harmine was more toxic to cyanobacteria than norharmane. On the other hand, neither pure compounds of harmine and norharmane nor crude bacterial extract showed any antialgal activity against tested species of green algae. The results of the present study suggest that B. flexus SSZ01 or its crude extract containing harmine and norharmane could be a candidate for the selective control of cyanobacterial blooms without affecting other algal species.

Grazing on Microcystis aeruginosa and degradation of microcystins by the heterotrophic flagellate Diphylleia rotans.

Cyanobacterial toxins can cause damage in aquatic ecosystems worldwide, as well as the poisoning of livestock, plants and humans when ingested in large amounts. Although many studies investigated grazing of harmful cyanobacteria by metazoan plankton, grazing of cyanobacteria by hetertophic flagellates is largely unexplored. This laboratory study investigated grazing of toxic Microcystis aeruginosa by the heterotrophic flagellate Diphylleia rotans isolated from a Saudi hypertrophic lake. D. rotans was able to feed on M. aeruginosa with estimated ingestion (10.2 to 16.5 prey flagellate (-1)d(-1)) and specific growth rates (0.71 to 0.99d(-1)) differed with the increase in the initial density of the flagellate. Grazing increased microcystin production within Microcystis cells during first two days of incubation, and afterwards induced the release of these toxins into the medium. The concentrations of releasing microcystin were strongly reduced in grazing-treated cultures compared to controls, indicating the degradation of microcystins by D. rotans growing under axenic conditions. Taken these results, D. rotans can play an important role in the reduction of Microcystis biomass and microcystin toxins, and thus could be used as a safe bioagent for the biocontrol of harmful algal blooms in aquatic environments.

Assessment of cylindrospermopsin toxin in an arid Saudi lake containing dense cyanobacterial bloom.

This study reports the presence of the cyanobacterial toxin cylindrospermopisn (CYN) and its producer Cylindrospermopsis raciborskii for the first time in Saudi freshwater sources. C. raciborskii was found in Gazan Dam Lake water with two morphotypes (coiled and straight). The appearance and cell density of this species was significantly positively related to high temperature and high ammonium concentrations, and negatively with nitrate and phosphate concentrations in the lake. Intracellular concentrations of CYN (4-173 µg L(-1)) were associated with C. raciborskii rather than other cyanobacteria with a maximal value obtained in June 2011, coinciding with the highest bloom of this species (19 × 10(7) trichome L(-1)). CYN cell quotas (0.6-14.6 pg cell(-1)) varied significantly along the study period and correlated with most environmental factors. The results of ELISA and liquid chromatography-mass spectrometry proved that the CYN production by strains of this species was isolated from this lake during the present study, with an amount reaching 568 µg g(-1). Extracellular CYN was also detected in cell-free lake water at concentrations 0.03-23.3 µg L(-1), exceeding the drinking water guideline value of 1 µg L(-1) during the Apr-Jul period. As this lake is an important source for drinking and irrigation waters, CYN monitoring should be included in the environmental and health risk assessment plans of these water bodies.

Biodegradation of cylindrospermopsin toxin by microcystin-degrading bacteria isolated from cyanobacterial blooms.

Cylindrospermopsin (CYN) is highly water soluble and stable cyanotoxin which can persist in water sources and pose health hazards to human, animals and plants. Consequently, there is a need to fully understand the fate of CYN in the aquatic environment, in particular, its biodegradation by endemic organisms. Although there is evidence of CYN degradation by microbial population, bacterial strains responsible for the toxin degradation are not yet identified. This study reports for the first time CYN degradation by Bacillus strain (AMRI-03) isolated from cyanobacterial blooms. In degradation batch experiment, the strain grew well in the presence of CYN without showing any lag period, and this growth increased with the initial CYN concentration. CYN degradation occurred rapidly, and the complete degradation was dependent on the initial CYN concentration. It occurred after 6 days at the highest concentration (300 µg L(-1)) compared to 7 and 8 days at lower concentrations (10 & 100 µg L(-1)). Also, the degradation rate correlated positively with the initial CYN concentration with maximum value (50 µg L(-1) day(-1)) obtained at the highest CYN concentration. Furthermore, the biodegradation rate of CYN by this strain depended remarkably on temperature and pH. The highest biodegradation rates were obtained at 25 and 30 °C, and at pH 7 and pH 8. Taken that such microcystin-degrading strain can also degrade CYN, bacterial strains reported worldwide as microcystin degraders could be tested for their capability of CYN degradation.

Microcystin production in epiphytic cyanobacteria on submerged macrophytes.

Cyanotoxins have been largely studied in planktonic and benthic cyanobacteria, but microcystin (MCYST) production in epiphytic cyanobacteria has not been reported yet. The present study reports for the first time the MCYST production in epiphytic cyanobacteria on submerged macrophytes. During this study, four common submerged macrophytes in eutrophic pond in Saudi Arabia were surveyed for the presence of toxic epiphytic cyanobacteria. The results showed that chlorophyll-a and total biovolume of epiphytic cyanobacteria differed significantly among submerged plants with highest values obtained in Stratiotes aloides and lowest in Elodea canadensis. Epiphytic materials collected from Ceratophyllum demersum and S. aloides had higher species diversities than materials collected from E. canadensis and Myriophyllum verticillatum. The cyanobacteria, Merismopedia tenuissima and Leptolyngbya boryana were recorded with a high abundance in epiphytic materials collected from all submerged macrohpytes. Based on Enzyme-linked immunosorbent assay (ELISA), these two species were found to produce MCYSTs (MCYSTs) with concentrations of 1438 and 630 microg g(-1) dry weight, respectively. HPLC analysis of the methanolic extracts of the two species showed that M. tenuissima extract contained MCYST-RR and -LR/demethyl LR plus 3 minor unidentified MCYSTs, while L. boryana extract contained MCYST-YR, -LR/demethyl LR, and 2 minor unidentified MCYSTs. This study suggests that epiphytic species should be considered during monitoring of toxic cyanobacteria in water sources.