Occurrence and elimination of cyanobacterial toxins in two Australian drinking water treatment plants.

In Australian freshwaters, Anabaena circinalis, Microcystis spp. and Cylindrospermopsis raciborskii are the dominant toxic cyanobacteria. Many of these surface waters are used as drinking water resources. Therefore, the National Health and Medical Research Council of Australia set a guideline for MC-LR toxicity equivalents of 1.3 microg/l drinking water. However, due to lack of adequate data, no guideline values for paralytic shellfish poisons (PSPs) (e.g. saxitoxins) or cylindrospermopsin (CYN) have been set. In this spot check, the concentration of microcystins (MCs), PSPs and CYN were determined by ADDA-ELISA, cPPA, HPLC-DAD and/or HPLC-MS/MS, respectively, in two water treatment plants in Queensland/Australia and compared to phytoplankton data collected by Queensland Health, Brisbane. Depending on the predominant cyanobacterial species in a bloom, concentrations of up to 8.0, 17.0 and 1.3 microg/l were found for MCs, PSPs and CYN, respectively. However, only traces (<1.0 microg/l) of these toxins were detected in final water (final product of the drinking water treatment plant) and tap water (household sample). Despite the low concentrations of toxins detected in drinking water, a further reduction of cyanobacterial toxins is recommended to guarantee public safety.     

The use of Lepidium sativum in a plant bioassay system for the detection of microcystin-LR.

Toxin-producing cyanobacteria pose a worldwide health threat to humans and animals due to their increasing presence in both drinking and recreational waters. Detection of microcystins in water generally relies on specialised equipment and a delay of several days for transport and analysis. Little work has, however, been done on establishing a simple, cost-effective and sensitive plant bioassay for the detection of microcystin-LR (MCLR) in water at the WHO Tolerable Daily Intake guideline level of 1 microg/l. We investigated the effect of a MCLR extract at 1 and 10 microg/l on the growth of Lepidium sativum over 6 days. Exposure to 10 microg/l MCLR resulted in a significant decrease in root and leaf lengths and fresh weights of seedlings when compared to the controls. These results were consistent with seedlings exposed to pure MCLR at 10 microg/l. Seedlings exposed to 1 microg/l MCLR showed a significant decrease in root development from day 2 to day 6. Glutathione S-transferase and glutathione peroxidase activities were also significantly raised in plants from days 5 and 4, respectively, at both toxin levels investigated.     

Identification of microcystins in waters used for daily life by people who live on Tai Lake during a serious cyanobacteria dominated bloom with risk analysis to human health

Tai Lake is the third largest freshwater lake in China with annual cyanobacteria blooms. Microcystins produced by these blooms have serious health risks for populations surrounding the lake, especially for people living on Tai Lake, because they usually drink raw lake water after a simple alum treatment. This study presents data on the detection and identification of microcystins in waters used for daily life by people living on Tai Lake, during the cyanobacterial blooming in July 2007. The health risks from drinking these microcystin-polluted waters were also calculated. The main microcystins detected by high-performance liquid chromatography-electrospray ionization mass spectrometry in the water samples collected from two parts of Tai Lake (Wuli Lake and Meiliang Bay) were MC-LR (4.33-12.27 microg/L), MC-RR (8.36-16.91 microg/L) and MC-YR (1.41-5.57 microg/L). Risk assessment showed that the drinking water simply treated by alum was not safe. The lowest calculated hazards ratios in all water samples was 6.4, which indicated that the risk of microcystins exposure from drinking water was over six times higher than the tolerable daily intake (TDI) recommended by The World Health Organization (WHO). Further studies should be conducted to elucidate the relationships between the epidemiology of people living on Tai Lake and microcystins exposure from drinking water.     

Cyanobacteria and Cyanotoxins Occurrence and Removal from Five High-Risk Conventional Treatment Drinking Water Plants

An environmental protection agency EPA expert workshop prioritized three cyanotoxins, microcystins, anatoxin-a, and cylindrospermopsin (MAC), as being important in freshwaters of the United States. This study evaluated the prevalence of potentially toxin producing cyanobacteria cell numbers relative to the presence and quantity of the MAC toxins in the context of this framework. Total and potential toxin producing cyanobacteria cell counts were conducted on weekly raw and finished water samples from utilities located in five US states. An Enzyme-Linked Immunosorbant Assay (ELISA) was used to screen the raw and finished water samples for microcystins. High-pressure liquid chromatography with a photodiode array detector (HPLC/PDA) verified microcystin concentrations and quantified anatoxin-a and cylindrospermopsin concentrations. Four of the five utilities experienced cyanobacterial blooms in their raw water. Raw water samples from three utilities showed detectable levels of microcystins and a fourth utility had detectable levels of both microcystin and cylindrospermopsin. No utilities had detectable concentrations of anatoxin-a. These conventional plants effectively removed the cyanobacterial cells and all finished water samples showed MAC levels below the detection limit by ELISA and HPLC/PDA.     

Microcystin diversity in a Planktothrix rubescens population from Lake Albano (Central Italy).

The cyanobacterium Planktothrix rubescens Anagnostidis & Komarek (previously Oscillatoria rubescens DC ex Gomont) is present in several Italian lakes and it is known to produce cyanotoxins. The dynamics and toxin production of P. rubescens population in Lake Albano, a volcanic crater lake in Central Italy, has been studied for 5 years (January 2001-April 2005). Winter-spring superficial blooms with frequent scums were observed every year. Total microcystin (MC) levels were measured from April 2004 to October 2005 by liquid chromatography-tandem mass spectrometry. MC levels up to 14.2mug/l were measured, with high concentrations found in summer at a 20-25m depth. The intracellular toxin content varied between 1.5 (surface, January 2004) and 0.21pg/cell (surface, May 2004). Six different MCs were detected, the most abundant being two desmethyl-MC-RR isomers. Of the 13 water wells monitored in the Lake Albano area, two of them showed MC contamination during winter, confirming the ability of these toxins to migrate through groundwater towards public water sources. These results highlight the need for further studies on the mobility and fate of these pervasive cyanobacterial toxins.     

Microcystins in potable surface waters: toxic effects and removal strategies

In freshwater, harmful cyanobacterial blooms threaten to increase with global climate change and eutrophication of surface waters. In addition to the burden and necessity of removal of algal material during water treatment processes, bloom‐forming cyanobacteria can produce a class of remarkably stable toxins, microcystins, difficult to remove from drinking water sources. A number of animal intoxications over the past 20 years have served as sentinels for widespread risk presented by microcystins. Cyanobacterial blooms have the potential to threaten severely both public health and the regional economy of affected communities, particularly those with limited infrastructure or resources. Our main objectives were to assess whether existing water treatment infrastructure provides sufficient protection against microcystin exposure, identify available options feasible to implement in resource‐limited communities in bloom scenarios and to identify strategies for improved solutions. Finally, interventions at the watershed level aimed at bloom prevention and risk reduction for entry into potable water sources were outlined. We evaluated primary studies, reviews and reports for treatment options for microcystins in surface waters, potable water sources and treatment plants. Because of the difficulty of removal of microcystins, prevention is ideal; once in the public water supply, the coarse removal of cyanobacterial cells combined with secondary carbon filtration of dissolved toxins currently provides the greatest potential for protection of public health. Options for point of use filtration must be optimized to provide affordable and adequate protection for affected communities. Copyright © 2013 John Wiley & Sons, Ltd.     

Dose-dependent antioxidant responses and pathological changes in tenca (Tinca tinca) after acute oral exposure to Microcystis under laboratory conditions

The effects of cyanobacterial cells containing microcystins (MCs), toxins from cyanobacteria, on oxidative stress biomarkers from liver and kidney of Tenca fish (Tinca tinca) were investigated under laboratory conditions. Moreover, a histopathological study of liver, kidney, heart and intestine tissues was performed. Fish were orally exposed to cyanobacterial cells dosing 0, 5, 11, 25 and 55 microg MC-LR/fish mixed with the food. Results showed a dose-dependent decrease of superoxide dismutase (SOD) activity, and also of catalase (CAT) in the liver. Glutathione levels and protein oxidation, however, were not altered by the exposure to the cyanobacterial material. The microscopic study revealed tissue alterations even at the lower cyanobacterial cells doses. Onion-like hepatocytes in the liver, glomerulopathy in the kidney, loss of myofibrils in the heart and vacuolated enterocytes in the gastrointestinal tract were the main changes observed. These findings suggest that this fresh water fish can be adversely affected by cyanobacterial blooms in their natural habitats.     

Occurrence of toxic blue-green algae in the Kucukcekmece lagoon (Istanbul, Turkey)

The concentration of microcystin (MC) in the Kucukcekmece Lagoon, Istanbul, Turkey, and the physicochemical and biological parameters of water quality were investigated from October 2000 to June 2003. Water samples were collected from surface waters at three sites. Most bloom samples were dominated by Microcystis aeruginosa. The major microcystin variants detected by HPLC-PDA were microcystin-YR and microcystin-LR. Microcystin concentration increased dramatically from early summer to early autumn and thereafter tended to decrease. The toxin concentration found in the filtered samples from surface waters varied between 0.06 and 24.2 microg L(-1) microcystin-LR equivalents. Each year extensive fish mortality was recorded between mid-June and early October, coinciding with heavy algal blooms. A comparison of the conditions associated with cyanotoxin episodes in 2000, 2001, and 2002 showed that the microcystin increase was related to temperature, high concentration of dissolved nutrients, high light intensity (PAR). The highest MC concentrations were recorded at temperatures between 24 degrees C and 28.5 degrees C. Field data showed that the highest MC concentration (>3 microg L(-1)) and the highest cyanobacterial biomass (>30 mg L(-1)) corresponded to a total nitrogen:total phosphorus ratio greater than 7:1. The highest concentrations of M. aeruginosa biomass (173 mg L(-1)) and MC (24.2 microg L(-1) MC-LR equiv.) and the highest salinity (8.8%) were measured concurrently in the lagoon. To our knowledge, this is the first evidence of cyanobacterial toxins in the Kucukcekmece Lagoon.     

Concentrations of particulate and dissolved cylindrospermopsin in 21 Aphanizomenon-dominated temperate lakes.

The cyanobacterial toxin cylindrospermopsin (CYN) is widely distributed in German lakes, but volumetric data for risk assessment are lacking and it is unclear which cyanobacterial species produce CYN in Europe. We therefore analyzed CYN concentration and cyanobacterial composition of 21 German lakes in 2005. CYN was detected in 19 lakes (102 of 115 samples). In total, 45 samples contained particulate CYN only, and 57 contained both dissolved and particulate CYN. The concentrations were 0.002-0.484 microg L(-1) for particulate CYN and 0.08-11.75 microg L(-1) for dissolved CYN with a maximum of 12.1 microg L(-1) total CYN. A drinking water guideline value of 1 microg L(-1) proposed by Humpage and Falconer [2003. Oral toxicity of the cyanobacterial toxin CYN in male Swiss albino mice: determination of no observed adverse effect level for deriving a drinking water guideline value. Environ. Toxicol. 18, 94-103] was exceeded in 18 samples from eight lakes due to high concentrations of dissolved CYN. CYN occurrence in the German lakes could not be ascribed to the three known CYN-producing species Cylindrospermopsis raciborskii, Anabaena bergii and Aphanizomenon flos-aquae, which were detected in some lakes in low abundances. The highest correlation coefficients were observed between particulate CYN and the native Aphanizomenon gracile. It occurred in 98 CYN-positive samples, was the most abundant Nostocales and was the only Nostocales in five samples. This indicates that A. gracile is a potential CYN producer in German lakes.     

Detection of harmful cyanobacteria and their toxins by both PCR amplification and LC-MS during a bloom event.

We briefly report here the occurrence of toxic blooms in the eutrophic reservoir Billings, S?o Paulo city, Brazil. Water samples were collected in May 2004, during a cyanobacterial bloom. The presence of toxic species was confirmed by using PCR amplifications of a fragment region of genes encoding microcystin synthetase-mcyB. The determination of toxins was performed by liquid chromatography coupled with mass spectrometry (LC-MS). LC-MS analyses of the toxins from the bloom revealed variants of microcystins (MC), such as MC-LR, MC-RR and MC-YR. HPLC-FLD was used to determine the paralytic shellfish poisoning (PSP) saxitoxin (STX), neosaxitoxin (NEO), gonyautoxins 2 (GTX2) and 3 (GTX3). GTX2, GTX3 and NEO were detected for the first time in a natural sample from Billings reservoir. These results are a contribution to the knowledge of the biogeography of toxic cyanobacteria and their toxins, specifically in S?o Paulo.     

Cyanobacteria and their toxins in treated-water storage reservoirs in Abha city, Saudi Arabia.

Occurrence of toxic cyanobacteria in drinking and recreational waters poses human health at risk as they can release potent toxins into the water. In the present study, open and covered treated-water storage reservoirs as well as their relevant tap waters in Abha city, Saudi Arabia, were surveyed for the presence of cyanobacteria and their toxins. The results revealed the contamination of most open reservoir and tap waters by algae and cyanobacteria, with an abundance of toxigenic species of cyanobacteria. Depending on the results of the Limulus amebocyte lysate (LAL) assay and enzyme linked immunosorbent assay (ELISA), endotoxins and microcystins (MCYSTs) were found in most open reservoir and tap waters at concentrations up to 32EUml(-1) and 0.3mugml(-1), respectively. The extracts of axenic cultures of most cyanobacterial species isolated from these reservoirs showed activity to LAL assay, with large endotoxin amounts obtained in Calothrix parietina (490EUg(-1)) and Phormidium tenue (210EUg(-1)). Based on ELISA and HPLC analysis for these extracts, only C. parietina can produce MCYSTs (202mugg(-1)) with a profile consisting of MCYST-RR and -LR. This study suggests that open treated-water storage reservoirs should be covered to prevent the presence of cyanobacteria and their toxins in such drinking and recreational waters.     

Cyanobacterial toxins: risk management for health protection

This paper reviews the occurrence and properties of cyanobacterial toxins, with reference to the recognition and management of the human health risks which they may present. Mass populations of toxin-producing cyanobacteria in natural and controlled waterbodies include blooms and scums of planktonic species, and mats and biofilms of benthic species. Toxic cyanobacterial populations have been reported in freshwaters in over 45 countries, and in numerous brackish, coastal, and marine environments. The principal toxigenic genera are listed. Known sources of the families of cyanobacterial toxins (hepato-, neuro-, and cytotoxins, irritants, and gastrointestinal toxins) are briefly discussed. Key procedures in the risk management of cyanobacterial toxins and cells are reviewed, including derivations (where sufficient data are available) of tolerable daily intakes (TDIs) and guideline values (GVs) with reference to the toxins in drinking water, and guideline levels for toxigenic cyanobacteria in bathing waters. Uncertainties and some gaps in knowledge are also discussed, including the importance of exposure media (animal and plant foods), in addition to potable and recreational waters. Finally, we present an outline of steps to develop and implement risk management strategies for cyanobacterial cells and toxins in waterbodies, with recent applications and the integration of Hazard Assessment Critical Control Point (HACCP) principles.     

Toxic cyanobacteria and microcystin concentrations in a public water supply reservoir in the Brazilian Amazonia region.

Toxic cyanobacteria in public water supply reservoirs represent a serious health risk as they can release potent cyanotoxins into the water. In the present study we analyzed surface water collected from the Utinga Reservoir, the main source of drinking water for the city of Belem-PA, in order to characterize the reservoir's cyanobacterial biota and to determine the toxicity of these organisms and the concentration of microcystins in raw and treated water. The cyanobacterial biota included potentially toxic genera such as Aphanizomenon, Microcystis, Nostoc, Oscillatoria, Planktothrix and Radiocystis. Mouse bioassays revealed a 43.6% frequency of hepatotoxic strains, including 30.8% Microcystis viridis strains and 12.8% Radiocystis fernandoi strains, with the lowest LD(100) (ip) of 45 and 75 mg kg(-1) body weight, respectively. Subacute hepatotoxicity was observed for Aphanizomenon cf. gracile and Coelomoron pusillum strains. HPLC analysis confirmed the production of microcystins at maximum concentrations of 4.22 microg mg(-1) dry weight for M. viridis and 2.47 microg mg(-1) for R. fernandoi. Microcystins at concentrations of up to 1.25 microg L(-1) detected in raw water by ELISA, together with a cyanobacterial density of 20,000 cells mL(-1), represents the first report of a bloom of cyanobacteria for an Amazonian water body.     

Microcystin-producing cyanobacteria in recreational waters in southwestern Germany

Toxin-forming species of freshwater cyanobacteria have been detected in all parts of Germany. Because of the high number of lakes (more than 2000) in Baden-Württemberg, in southwestern Germany, only a representative selection of lakes (155) was intensively monitored for mass development of cyanobacteria and the formation of toxins in 1999. Nutrients, dominant algal species, chlorophyll, and toxins were determined from waters with cyanobacterial blooms. Toxin analyses were performed using HPLC, the PP1 inhibition test, and the microcystin ELISA. In 45% of the monitored lakes, the phytoplankton assemblage consisted mainly of cyanobacteria; in an additional 24% of the lakes cyanobacteria were detected. More than 26% of the lakes contained toxigenic cyanobacteria. The highest amount of microcystins detected outside scum areas was 11-560 microg/L. In relation to biomass, cyanobacteria contained up to 1100 microg/g dry matter and up to 2.6 microg/microg Chl a. The dominant form of microcystin detected was microcystin-LR, one of the most toxic known microcystins. The results of the present study have been used to develop a stepwise monitoring scheme for lakes with permanent cyanobacterial populations to protect human health and livestock.     

Effects of cyanobacterial crude extracts from Planktothrix agardhii on embryo-larval development of medaka fish, Oryzias latipes.

Embryonic toxicity from exposure to microcystins, cyclic hepatotoxic heptapeptides from cyanobacteria, receives increasing attention as a public human health biohazard. Using a microinjection technology, we have introduced cyanobacterial extracts from Planktothrix agardhii directly into the vitellus of late neurula embryos (stage 19) of medaka (Oryzias latipes). Microinjection (2 nL) of P. agardhii PMC 75.02 extract containing microcystins (MC) resulted in a dose-dependent mortality of embryos. Survival rates were reduced up to 81% with extract concentrations of 10 mg mL(-1) (EC(50)=7.8 microg mL(-1)). On the other hand, injection of P. agardhii PMC 87.02 extract in which no microcystin could be detected resulted in much less embryonal toxicity (EC(50)=460 microg mL(-1)). In addition, advanced embryonic hatching processing was limited with PMC 75.02 crude extract and less obvious than had been described with pure MC-LR injections. In agreement with the known hepatotoxic effects of microcystin, embryos injected with PMC 75.02 extract consistently displayed hepatobiliary abnormalities. Loss of glycogen content of the hepatocytes and hepatic haemorrhage were evidenced in surviving post-hatching juveniles. Thus, the methodology presented in this paper should be a valuable tool to analyse the effects of crude extracts of cyanobacterial toxins on the development of aquatic vertebrate embryos.     

Toxic cyanobacteria and cyanotoxins in public hot springs in Saudi Arabia.

Toxic cyanobacteria are well reported in rivers, lakes and even marine environments, but the toxin production of cyanobacteria in hot springs is largely unexplored. Therefore, the present study investigated the presence of toxic cyanobacteria and cyanotoxins in public hot springs in Saudi Arabia. The results of an enzyme-linked immunosorbent assay (ELISA) revealed that Saudi spring cyanobacterial mats contained microcystins (MCYSTs) at concentrations ranging from 468 to 512.5 microg g(-1). The Limulus amebocyte lystae (LAL) assay detected lipopolysaccharide (LPS) endotoxins in these mats at concentrations ranging from 433.3 to 506.8 EU g(-1). MCYSTs and endotoxins were also detected in spring waters at levels of 5.7 microg l(-1) and 640 EU ml(-1), respectively, exceeding WHO's provisional guideline value for MCYST-LR in drinking-water. High-performance liquid chromatography (HPLC) analysis revealed that only Oscillatoria limosa and Synechococcus lividus can produce MCYSTs with a profile consisting of MCYST-RR and -LR. Based on the LAL assay, 12 out of 17 cyanobacterial species contained LPS at concentrations ranging from 0.93 to 21.06 EU g(-1). However, not all LPS of these species were toxic to mice. This study suggests that the hot springs in the world including Saudi Arabia should be screened for toxic cyanobacteria to avoid the exposure of people recreating and bathing in spring waters to cyanobacterial toxins.     

An investigation into the effect of selenium supplementation on microcystin hepatotoxicity.

Toxin-producing cyanobacteria pose a worldwide health threat to humans and animals due to their increasing presence in both drinking and recreational waters. Little work has, however, been done on a preventative therapy for anyone at risk of exposure to cyanobacterial toxins. The potential benefits of dietary supplementation of selenium, an antioxidant, to protect against the mouse liver injury induced by the toxin, microcystin-LR, has been investigated. BALB/c mice were pretreated for two weeks with sodium selenite (1.5 microg/mouse/day) before an intraperitoneal injection of microcystin-LR. Selenium-supplementation was found to provide some protection to the action of the toxin. In addition selenium pretreatment reduced the liver damage caused by lethal and sub-lethal toxin doses as reflected in liver pathology, decreased serum ALT and lipid peroxidation levels as well as prevention of glycogen loss compared to non-selenium supplemented toxin treated mice. The increased level of liver glutathione peroxidase activity following selenium-supplementation may indicate the possible route of selenium protection in the mice.     

CyanoHAB occurrence and water irrigation cyanotoxin contamination: ecological impacts and potential health risks

The world-wide occurrence of harmful cyanobacteria blooms "CyanoHAB" in fresh and brackish waters creates problems for all life forms. During CyanoHAB events, toxic cyanobacteria produce cyanotoxins at high levels that can cause chronic and sub-chronic toxicities to animals, plants and humans. Cyanotoxicity in eukaryotes has been mainly focused on animals, but during these last years, data, related to cyanotoxin (mainly microcystins, MCs) impact on both aquatic and terrestrials crop plants irrigated by water containing these toxins, have become more and more available. This last cited fact is gaining importance since plants could in a direct or indirect manner contribute to cyanotoxin transfer through the food chain, and thus constitute a potent health risk source. The use of this contaminated irrigation water can also have an economical impact which appears by a reduction of the germination rate of seeds, and alteration of the quality and the productivity of crop plants. The main objective of this work was to discuss the eventual phytotoxicity of cyanotoxins (microcystins) as the major agricultural impacts induced by the use of contaminated water for plant irrigation. These investigations confirm the harmful effects (ecological, eco-physiological, socio-economical and sanitary risk) of dissolved MCs on agricultural plants. Thus, cyanotoxin phytotoxicity strongly suggests a need for the surveillance of CyanoHAB and the monitoring of water irrigation quality as well as for drinking water.     

First report of (homo)anatoxin-a and dog neurotoxicosis after ingestion of benthic cyanobacteria in The Netherlands

In April and May 2011, three dogs died and one dog became ill after swimming in Lake IJmeer (The Netherlands). At the time, the lake was infested with the benthic cyanobacterial species Phormidium. A Eurasian Coot (Fulica atra) and a Black-headed Gull (Chroicocephalus ridibundus) also died near Lake IJmeer in the same period. One of the dogs and both birds were subjected to a pathological investigation. Furthermore, the Phormidium mat; algal samples from the dikes; contents of the animals’ digestive systems and organ tissues were analysed for the following cyanobacterial toxins: (homo)anatoxin-a; (7-deoxy-)cylindrospermopsin; saxitoxins and gonyautoxins by LC-MS/MS. Samples were also analysed for the nontoxic (homo)anatoxin-a metabolites dihydro(homo)anatoxin-a and epoxy(homo)anatoxin-a. The dog necropsy results indicated neurotoxicosis and its stomach contained Phormidium filaments. Anatoxin-a was detected in the Phormidium mat (272 μg g⁻¹ dry weight, stdev 65, n = 3) and in the dog’s stomach contents (9.5 μg g⁻¹ dry weight, stdev 2.4, n = 3). Both samples also contained the anatoxin-a metabolite dihydroanatoxin-a, and a trace of homoanatoxin-a was detected in the Phormidium mat. The birds were in bad nutritive condition at the time of necropsy and their stomachs and intestines did not contain any cyanobacterial material. Furthermore, no cyanobacterial toxins were detected in their stomachs, intestines and organs and they both had lesions that are not associated with cyanobacterial intoxication. This is the first report of anatoxin-a and homoanatoxin-a occurrence in The Netherlands, these toxins have likely caused the deaths of three dogs. The birds probably died of other causes. Dutch recreational waters are at this moment only screened for pelagic cyanobacterial species, the current bathing water protocol therefore does not protect humans and animals from negative effects of blooms of benthic cyanobacteria.