Microcystin Levels Research Articles

The effects of temperature and nutrients on the growth and dynamics of toxic and non-toxic strains of Microcystis during cyanobacteria blooms

In temperate latitudes, toxic cyanobacteria blooms often occur in eutrophied ecosystems during warm months. Many common bloom-forming cyanobacteria have toxic and non-toxic strains which co-occur and are visually indistinguishable but can be quantified molecularly. Toxic Microcystis cells possess a suite of microcystin synthesis genes ( mcyA– mcyJ), while non-toxic strains do not. For this study, we assessed the temporal dynamics of toxic and non-toxic strains of Microcystis by quantifying the microcystin synthetase gene ( mcyD) and the small subunit ribosomal RNA gene, 16S (an indicator of total Microcystis), from samples collected from four lakes across the Northeast US over a two-year period. Nutrient concentrations and water quality were measured and experiments were conducted which examined the effects of elevated levels of temperatures (+4 °C), nitrogen, and phosphorus on the growth rates of toxic and non-toxic strains of Microcystis. During the study, toxic Microcystis cells comprised between 12% and 100% of the total Microcystis population in Lake Ronkonkoma, NY, and between 0.01% and 6% in three other systems. In all lakes, molecular quantification of toxic ( mcyD-possessing) Microcystis was a better predictor of in situ microcystin levels than total cyanobacteria, total Microcystis, chlorophyll a, or other factors, being significantly correlated with the toxin in every lake studied. Experimentally enhanced temperatures yielded significantly increased growth rates of toxic Microcystis in 83% of experiments conducted, but did so for non-toxic Microcystis in only 33% of experiments, suggesting that elevated temperatures yield more toxic Microcystis cells and/or cells with more mcyD copies per cell, with either scenario potentially yielding more toxic blooms. Furthermore, concurrent increases in temperature and P concentrations yielded the highest growth rates of toxic Microcystis cells in most experiments suggesting that future eutrophication and climatic warming may additively promote the growth of toxic, rather than non-toxic, populations of Microcystis, leading to blooms with higher microcystin content.

Simultaneous determination of microcystin contaminations in various vertebrates (fish, turtle, duck and water bird) from a large eutrophic Chinese lake, Lake Taihu, with toxic Microcystis blooms

This is the first to conduct simultaneous determination of microcystin (MC) contaminations in multi-groups of vertebrates (fish, turtle, duck and water bird) from Lake Taihu with Microcystis blooms. MCs (-RR, -YR, -LR) in Microcystis scum was 328 μg g − 1 DW. MCs reached 235 μg g − 1 DW in intestinal contents of phytoplanktivorous silver carp, but never exceeded 0.1 μg g − 1 DW in intestinal contents of other animals. The highest MC content in liver of fish was in Carassius auratus (150 ng g − 1 DW), followed by silver carp and Culter ilishaeformis, whereas the lowest was in common carp (3 ng g − 1 DW). In livers of turtle, duck and water bird, MC content ranged from 18 to 30 ng g − 1 DW. High MC level was found in the gonad, egg yolk and egg white of Nycticorax nycticorax and Anas platyrhynchos, suggesting the potential effect of MCs on water bird and duck embryos. High MC contents were identified for the first time in the spleens of N. nycticorax and A. platyrhynchos (6.850 and 9.462 ng g − 1 DW, respectively), indicating a different organotropism of MCs in birds. Lakes with deaths of turtles or water birds in the literatures had a considerably higher MC content in both cyanobacteria and wildlife than Lake Taihu, indicating that toxicity of cyanobacteria may determine accumulation level of MCs and consequently fates of aquatic wildlife.

Identification of toxigenic Microcystis strains after incidents of wild animal mortalities in the Kruger National Park, South Africa

The eutrophic process potentially caused by a high urine and faecal load resulting from an unusually high hippopotamus ( Hippopotamus amphibious) density in the Nhlanganzwane Dam, Kruger National Park, South Africa, triggered a chain of events characterised by an increase in the growth of primary producers ( Microcystis aeruginosa). This increase in M. aeruginosa biomass was followed by bio-intoxication incidents in wild animals. In this study, we determine if a M. aeruginosa bloom with a total microcystin level of 23,718 μg l −1 have been responsible for mortalities of megaherbivores in the Nhlanganzwane Dam. We further use microcystin molecular markers derived from the mcy gene cluster to identify potentially toxigenic environmental Microcystis strains in the dam during the occurrence of animal intoxications. The estimated total microcystin-LR daily intake by an adult male white rhinoceros ( Ceratotherium simum) from cyanobacterial-contaminated water of the dam during the toxic event was an order of magnitude higher (754.29 μg kg −1 bw) in comparison with the lowest observed adverse effecting level (LOAEL) value measured for pigs in a previous study by other authors. In this study the presence of toxic cyanobacterial strains was confirmed with the use of molecular markers that detected the presence of the mcy gene cluster responsible for the production of toxin by M. aeruginosa.

Method validation of microcystins in water and tissue by enhanced liquid chromatography tandem mass spectrometry

A liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI–MS/MS) method has been developed and validated to identify and quantify trace levels of cyanotoxins or microcystins (MC) in water, bivalves and fish tissue with enhanced sensitivity and specificity. The method enables confirmation and quantification of six MCs (MC-LA, LF, LR, LW, RR and YR) with a single chromatographic run. The applied chromatography also allows determination of certain MC metabolites (Desmethyl-LR and -RR). By using LC-ESI–MS/MS in multiple reaction monitoring (MRM) mode, the limit of detection and quantitation for the microcystins studied, were determined to be between 0.2 and 1 pg on column (5:1 S/N ratio). These values are below the 2 pg detection limits found in the available literature.

Changes in the GST activity of the mussel Mytilus galloprovincialis during exposure and depuration of microcystins

Mussels are quite resistant to cyanotoxins and their resistance may be because of an efficient metabolization of cyanotoxins by glutathione-S-transferases (GST) activity. Nevertheless, other secondary metabolites may interfere with the detoxication efficiency. The accumulation and depuration of hepatotoxins produced by the freshwater cyanobacterium Microcystis aeruginosa in the mussel Mytilus galloprovincialis were studied. Mussels were fed twice a day 1.5x10(5) cells/mL of the toxic cyanobacterium, which produces microcystins (MCs) -FR, -LR and -WR, for 4 days. After that period, the animals were placed in toxin-free water and were fed the green algae Ankistrodesmus sp. During 2 weeks, the concentration of the toxin in the mussels was monitored using an ELISA assay. Mussels showed a maximum detectable level of MCs of 0.38 microg/g mussels dry weight (DW) during the accumulation period and 0.37 microg MC/g mussel DW by day 4 of the depuration period. Then there was a decrease trend with peaks of toxin at days 8 and 12 of the depuration period. The activity of the detoxication enzymes GST was studied and the results showed that the peaks of toxin in the mussels coincide with an increase in the activity of GST. These results support the hypothesis that the rise of the toxin level on days 4, 8 and 12 of the depuration period in the mussels may be related to the renewal of protein phosphatases and subsequent release of unbound toxins.

Necessity of Screening Water Chestnuts for Microcystins After Cyanobacterial Blooms Break Out

Water chestnut is one of the most popular vegetables in Asian countries that grows in shallow water. Eighteen water chestnut samples were collected from Lake Tai and six samples were bought at markets in Wuxi, China, in October 2007. Extraction solution of water chestnut was cleaned up with a solid phase extraction column and immunoaffinity chromatography cartridges, then the microcystin (MC) level was detected by indirect competitive enzyme-linked immunosorbent assay (ELISA) and liquid chromatography-mass spectrometry (LC-MS). The results of ELISA showed that there were six samples collected from Lake Tai which contained MCs; the highest level of total MCs was 7.02 ng/g. The results of LC-MS confirmed that MC-LR and MC-RR were present in five samples. The highest level of MC-LR was 1.02 ng/g and that of MC-RR was 4.44 ng/g. Heavy cyanobacterial blooms had occurred, and MCs were detected in water at the points in Lake Tai where MCs occurred in water chestnuts collected in 2007. MCs were not detected in the six samples bought at Wuxi markets. The results suggest that MCs can accumulate in water chestnuts, which is a potential hazard for human health.

Widespread Distribution and Identification of Eight Novel Microcystins in Antarctic Cyanobacterial Mats

The microcystin (MC) content and cyanobacterial community structure of Antarctic microbial mat samples collected from 40 ponds, lakes, and hydroterrestrial environments were investigated. Samples were collected from Bratina Island and four of the Dry Valleys, Wright, Victoria, Miers, and Marshall. Enzyme-linked immunosorbent assays (ELISAs), liquid chromatography-mass spectrometry (LC-MS), and protein phosphatase 2A (PP-2A) inhibition assays resulted in the identification of low levels (1 to 16 mg/kg [dry weight]) of MCs in all samples. A plot of indicative potencies of MCs (PP-2A inhibition assay/ELISA ratio) versus total MCs (ELISA) showed a general decrease in potency, as total MC levels increased, and a clustering of values from discrete geographic locations. LC-tandem MS analysis on selected samples identified eight novel MC congeners. The low-energy collisional activation spectra were consistent with variants of [D-Asp(3)] MC-RR and [D-Asp(3)] MC-LR containing glycine [Gly(1)] rather than alanine and combinations of homoarginine [hAr(2)] or acetyldemethyl 3-amino-9-methoxy-2,6,8-trimethyl-10-phenyl-4,6-decadienoic acid (acetyldemethyl ADDA) [ADMAdda(5)] substitutions. Nostoc sp. was identified as a MC producer using PCR amplification of a region of the 16S rRNA gene and the aminotransferase domain of the mcyE gene. Automated ribosomal intergenic spacer analysis (ARISA) was undertaken to enable a comparison of cyanobacterial mat community structure from distant geographical locations. Two-dimensional multidimensional scaling ordination analysis of the ARISA data showed that in general, samples from the same geographic location tended to cluster together. ARISA also enabled the putative identification of the MC-producing Nostoc sp. from multiple samples.

Cyanotoxins in Tidal Waters of Chesapeake Bay

Abstract Cyanobacteria blooms have long been described for Chesapeake Bay nontidal and tidal waters, but measurable toxin has only been recently recorded. During September 2000, the earliest tidal-water records of cyanotoxins in the Bay identified microcystin from a Microcystis-dominated bloom on the Sassafras River. Between 2000 and 2006, opportunistic samples collected from cyanobacteria blooms were analyzed for toxin concentration to better inform natural resource, agriculture, and human-health management agencies on potential bloom-related health risks. The hepatotoxin microcystin was detected most frequently and over a range of concentrations from 2.9 × 10−2 to 6.58 × 102 μg L−1. Microcystin levels exceeded literature-based chronic drinking-water guidance values of 1 μg L−1 and recreational safety guidance for children of 10 μg L−1 in 71% and 31% of samples, respectively. Samples from tidal fresh and oligohaline habitats showed a log-normal distribution of toxin concentrations, and microcystin had po...

Abundance and toxicity of Planktothrix rubescens in the pre-alpine Lake Ammersee, Germany

Regular occurrences of the cyanobacterium Planktothrix rubescens have been observed in several lakes that have undergone recent re-oligotrophication, e.g. Lake Ammersee. Planktothrix species are known to produce microcystins, potent phosphatase inhibitors that have been associated with morbidities and mortalities in humans and animals. The aim of this study was to characterise the temporal and spatial abundance and toxicity of P. rubescens in Lake Ammersee. P. rubescens cell densities and biovolumes were calculated via fluorescence image analyses. P. rubescens was present during the entire observation period from 1999 to 2004, albeit at different cell densities. Maximum biovolumes of 45 cm 3 m −2 were observed in May 2001. Filaments were regularly distributed over the entire water column during winter and stratified in distinct metalimnic layers during summer, reaching maximum cell densities of ≤15,000 (winter) and ≤77,000 cells ml −1 (summer). The results demonstrate that P. rubescens abundance is strongly influenced by water transparency, i.e. illumination in the metalimnion. Moreover, the P. rubescens abundance appears to result from regular phosphate depletion in the epilimnion, possibly additionally benefiting from high nitrogen loads. Microcystin (MC) was detectable in 27 and 38 of 54 seston samples via HPLC and Adda-ELISA measurements, respectively. The main microcystin congeners in the seston samples were [Asp 3]-MC-RR and [Asp 3,Dhb 7]-MC-RR. Microcystin concentrations correlated significantly with the respective phycoerythrin (PE)-concentrations. The variation in the MC/PE-ratios was low suggesting that the microcystin production of P. rubescens in Lake Ammersee is consistent and indicates that the appearance of P. rubescens coincides with measurable microcystin levels. Moreover, the observation of pronounced metalimnic oxygen depletions appears to be causally related to recurring high P. rubescens abundance. In conclusion the results suggest that aquatic organisms such as indigenous fish populations (e.g. coregonids) are regularly confronted with potentially adverse P. rubescens densities, which might provide a possible explanation for the often observed impaired health and growth retardation of coregonid populations in P. rubescens containing pre-alpine lakes.

Harmful phytoplankton blooms and fish mortality in a eutrophicated reservoir of northeast Brazil

The aim of this work was to study the eutrophication in the tropical freshwater ecosystems and the consequent cyanobacterial bloom formation and economical damage to fisheries and harmful effects to public health. Mass fish mortality due to toxin producing cyanobacterial blooms was registered during December 2003 in Marechal Dutra Reservoir, Acari/RN, Northeast Brazil. Phytoplankton and fish samplings were carried out on alternate days during the episode of fish mortality and monthly during January to June 2004. The cyanobacterial toxin was identified and quantified from the seston samples and liver of the dead fishes using the standard HPLC method. The results indicated that the toxic blooms of Cylindrospermopsis raciborskii and Microcystis aeruginosa were persistent for two weeks and represented 90% of the phytoplankton species assemblages. The lethally affected fishes were Oreochromis niloticus, Plagioscion squamosissimus, Cichla monoculus, Prochilodus brevis, Hoplias malabaricus and Leporinus friderici. The microcystin levels varied from 0.07 to 8.73µg L-1 the seston samples and from 0.01 to 2.59µg g-1in the liver samples of the fishes during the bloom period.

Evaluation of the human health threat associated with the hepatotoxin microcystin in the muscle and liver tissues of yellow perch (Perca flavescens)

During the summer of 2006, the western basin of Lake Erie experienced a bloom of the toxigenic cyanobacterium Microcystis aeruginosa . Across 11 sites, intracellular, particulate-bound microcystin levels in the seston increased to levels that exceeded World Health Organization guidelines for drinking water exposure (1 µg toxin·L–1). In contrast, toxin concentrations in yellow perch ( Perca flavescens ) muscle tissue (n = 68) declined from June to August, were negatively related to algal toxin levels, and never exceeded a conservative chronic exposure concentration estimated using proposed United States Environmental Protection Agency (US EPA) guidelines. Microcystin concentrations in yellow perch liver exceeded US EPA chronic exposure guidelines, were on average 125 times higher than muscle toxin concentrations per unit dry weight, and varied little throughout the summer. With current guidelines, humans do not appear to be at risk when consuming the muscle tissue of Lake Erie yellow perch collected during large-scale cyanobacterial blooms. However, this study highlights the need for a better understanding of the trophic transfer of cyanobacterial toxins through aquatic food webs in diverse ecosystems with an emphasis on understanding if these compounds could accumulate sufficiently to affect human health.

Recreational Exposure to Low Concentrations of Microcystins During an Algal Bloom in a Small Lake

We measured microcystins in blood from people at risk for swallowing water or inhaling spray while swimming, water skiing, jet skiing, or boating during an algal bloom. We monitored water samples from a small lake as a Microcystis aeruginosa bloom developed. We recruited 97 people planning recreational activities in that lake and seven others who volunteered to recreate in a nearby bloom-free lake. We conducted our field study within a week of finding a 10-μg/L microcystin concentration. We analyzed water, air, and human blood samples for water quality, potential human pathogens, algal taxonomy, and microcystin concentrations. We interviewed study participants for demographic and current health symptom information. Water samples were assayed for potential respiratory viruses (adenoviruses and enteroviruses), but none were detected. We did find low concentrations of Escherichia coli, indicating fecal contamination. We found low levels of microcystins (2 μg/L to 5 μg/L) in the water and (

Recreational Exposure to Low Concentrations of Microcystins During an Algal Bloom in a Small Lake

We measured microcystins in blood from people at risk for swallowing water or inhaling spray while swimming, water skiing, jet skiing, or boating during an algal bloom. We monitored water samples from a small lake as a Microcystis aeruginosa bloom developed. We recruited 97 people planning recreational activities in that lake and seven others who volunteered to recreate in a nearby bloom-free lake. We conducted our field study within a week of finding a 10-μg/L microcystin concentration. We analyzed water, air, and human blood samples for water quality, potential human pathogens, algal taxonomy, and microcystin concentrations. We interviewed study participants for demographic and current health symptom information. Water samples were assayed for potential respiratory viruses (adenoviruses and enteroviruses), but none were detected. We did find low concentrations of Escherichia coli, indicating fecal contamination. We found low levels of microcystins (2 μg/L to 5 μg/L) in the water and (<0.1 ng/m3) in the aerosol samples. Blood levels of microcystins for all participants were below the limit of detection (0.147μg/L). Given this low exposure level, study participants reported no symptom increases following recreational exposure to microcystins. This is the first study to report that water-based recreational activities can expose people to very low concentrations of aerosol-borne microcystins; we recently conducted another field study to assess exposures to higher concentrations of these algal toxins.

Monolithic solid-phase extraction for the rapid on-line monitoring of microcystins in surface waters

We describe the development and optimization of a sensitive and selective screening method for the measurement of trace levels of microcystins in surface waters. Several sample preparation techniques were compared, including solid-phase microextraction (SPME), particle-based solid-phase extraction (SPE), and monolith-based SPE. A flow-injection (FI) based approach employing a reversed-phase monolithic SPE column was found to be optimal. Quantification was performed by directly interfacing the FI-based SPE system to an electrospray ionization-mass spectrometer (ESI-MS). To more safely simulate peptidyl toxins such as the microcystins, a model peptide (i.e., angiotensin II) was used for method optimization. Sample loading flow rate and volume, eluent composition, and elution flow rate were optimized. Sample throughput was six samples per hour, a detection limit of 1.31 ng angiotensin II was demonstrated for a linear dynamic range from 1–1,000 ng and 3.4% relative standard deviation (n = 4, 100 ng sample). Sample volumes up to 1,000 ml of surface water could be loaded onto the monolithic SPE disk without exceeding the sorbent’s capacity. Unlike conventional particle-based SPE methods, the monolithic SPE disk does not need to be replaced between samples and could be used indefinitely. The FI-based SPE-ESI-MS method was successfully applied to the determination of microcystin-LR, the most common of the microcystins, in environmental samples and was demonstrated for the direct monitoring of chlorinated drinking water, with trends tracked over a period of eight months.

Toxicity of microcystin‐LR, a cyanobacterial toxin, to multiple life stages of the burrowing mayfly, Hexagenia, and possible implications for recruitment

Burrowing mayflies, genus Hexagenia, were extirpated from the major water bodies of North America in the early 1950s, paralleling an increase in eutrophication and organic pollution, and a decrease in dissolved oxygen concentrations. Burrowing mayflies recolonized the western basin of Lake Erie, but remain absent in other former habitats such as Oneida Lake, New York. Eutrophication is commonly associated with a shift in the phytoplankton community toward dominance by cyanobacteria, and therefore, one class of cyanobacterial toxins, microcystins, were investigated as a contributing factor to Hexagenia's eradication or as an impediment to recolonization. Laboratory experiments were conducted to determine if microcystin-LR (MC-LR) produced negative effects on Hexagenia at three points within its life cycle: egg, hatchling nymph (<24-h old, <1 mm total length), and pre-emergence nymph (>17 mm). Treatment concentrations ranged from the guideline set by the World Health Organization for drinking water (0.001 microg mL(-1)) to 0.1 microg mL(-1) for the egg experiment and 10 microg mL(-1) for the nymph trials. Eggs showed a delay in hatching and an altered distribution of hatching over the study period when submerged in 0.1 microg mL(-1) MC-LR (an elevated concentration representative of bloom scum). The 72-h (1.1 microg mL(-1)) and 96-h (0.049 microg mL(-1)) LC(50) values for hatchling nymphs exceeded typical bloom concentrations of North American lakes, (0.01 microg mL(-1)). Large nymphs were more tolerant of the toxin, as indicated by 100% survival over seven days exposure to 10 microg mL(-1), suggesting older larvae can withstand brief encounters with high microcystin levels for at least short periods of time. The sensitivity of young nymphs and eggs to MC-LR may have implications for the recruitment of the genus in water bodies with persistent summer cyanobacterial blooms.

First reported case of turtle deaths during a toxic Microcystis spp. bloom in Lake Oubeira, Algeria

Microcystins analysis was conducted in field cyanobacterial bloom samples and dead terrapin tissues from Lake Oubeira (Algeria) with an aim of studying the cause of the mortality of the freshwater terrapin species Emys orbicularis and Mauremys leprosa during October 2005. The deaths of these two terrapin species were observed during a bloom of Microcystis spp. The total microcystin content per phytoplankton biomass evaluated with the methanol extraction-protein phosphatase methodology was 1.12 mg MCYST-LR equivalents/g dried bloom material. The analysis of this bloom extract by the LC/MS technique demonstrated the presence of three microcystin variants: microcystin-LR (MCYST-LR), microcystin-YR (MCYST-YR), and microcystin-RR (MCYST-RR). Microcystins were also detected in fresh carcasses of terrapin liver, viscera and muscle tissues using the GC/MS after Lemieux oxidation method and the PP2A inhibition assay. The high level of microcystins detected using the Lemieux oxidation-GC/MS method in the liver tissue (1192.8 μg MCYST-LR equivalent/g dw) and in the viscera tissue (37.19 μg MCYST-LR equivalent/g dw) of the species M. leprosa and E. orbicularis, respectively, and the liver crumbling observed after the necropsy examination of the fresh carcass of M. leprosa support the possibility that cyanobacterial microcystins contribute to the turtle mortalities.

Reduction of trihalomethane formation and detoxification of microcystins in tap water by ozonation

The efficiency of ozonation in comparison to chlorination for removal of microcystins and production of trihalomethanes (THMs) in water was investigated. One hundred and ninety water samples of ozone and chlorine treated water were collected at a water treatment plant between August 2004 and March 2005. The level of THMs, total organic carbon and residual chlorine were determined. Protein phosphatase 2A inhibition assay was used to detect microcystins and the presence of microcystins was confirmed by HPLC. The results show that 91.5% of the THM species in treated water was chloroform and 8.5% was bromodichloromethane. The mean THM level± standard error of mean in chlorinated water (CW) (45.1±3.0 μg/L) was higher than the mean of THM level in ozonated water (OW) (18.6±2.2 μg/L). In addition, no OW sample exceeded the first stage U.S. EPA maximum THM contaminant level for drinking water (80 μg/L) and only 8% of these samples exceeded the second stage level (40 μg/L). On the other hand, 3% of CW samples exceeded 80 μg/L and 68% exceeded the 40 μg/L level. The microcystin level in all water samples was below the WHO guideline value (1 μg/L) for drinking water.

Tissue Distribution and Depuration of the Extracted Hepatotoxic Cyanotoxin Microcystins in Crucian Carp (Carassius carassius) Intraperitoneally Injected at a Sublethal Dose

An acute toxicity experiment was conducted by intraperitoneal injection with a sublethal dose of extracted microcystins (MCs), 50 μg MC-LR (where L = leucine and R = arginine) equivalent/kg body weight (BW), to examine tissue distribution and depuration of MCs in crucian carp (Carassius carassius). Liver to body weight ratio increased at 3, 12, 24, and 48 h postinjection compared with that at 0 h (p < 0.05). MC concentrations in various tissues and aquaria water were analyzed at 1, 3, 12, 24, 48, and 168 h postinjection using liquid chromatography coupled with mass spectrometry (LC-MS). The highest concentration of MCs (MC-RR + MC-LR) was found in blood, 2–270 ng/g dry weight (DW), followed by heart (3–100 ng/g DW) and kidney (13–88 ng/g DW). MC levels were relatively low in liver, gonad, intestine, spleen, and brain. MC contents in gills, gallbladder, and muscle were below the limit of detection. Significant negative correlation was present between MC-RR concentration in blood and that in kidney, confirming that blood was important in the transportation of MC-RR to kidney for excretion. Rapid accumulation and slow degradation of MCs were observed in gonad, liver, intestine, spleen, and brain. Only 0.07% of injected MCs were detected in liver. The recovery of MCs in liver of crucian carp seemed to be dose dependent.

Microcystin accumulation in liver and muscle of tilapia in two large Brazilian hydroelectric reservoirs

The objective of this study was to measure levels of the toxin microcystin in different tissues of fish known to feed on cyanobacteria during toxic bloom events. Wild Nile and redbreast tilapia ( Oreochromis niloticus and Tilapia rendalli) were sampled from the catch of artisanal fishermen at eutrophic stations of Funil and Furnas reservoirs in southeastern Brazil. Phytoplankton communities in the two reservoirs were quite different taxonomically, but not dissimilar in microcystin content (200 μg g dry weight (DW) seston −1 at Funil, 800 μg gDW seston −1 at Furnas). All of the 27 fish sampled contained microcystin, ranging from 0.8 to 32.1 μg g liver −1 and from 0.9 to 12.0 ng g muscle −1. Most microcystin variants found in seston were also found in fish liver . T. rendalli had the lowest concentration in both tissues when compared to O. niloticus. In both reservoirs, one of every four fish sampled, always O. niloticus, had a level of microcystins beyond the World Health Organization tolerable daily intake (8 ng g tissue −1) and represented a risk for consumers. It is possible that closer study of inter-species variability in toxin burden in cyanobacteria-impacted water bodies will permit the development of guidelines for fish consumption that will better protect public health.

First Observation of Microcystins in Tunisian inland waters: a threat to river mouths and lagoon ecosystems

1 - Microcystin pollution is known to affect different types of inland water bodies: river mouths and coastal lagoons may be affected by local production as well as by transportation through the freshwater network. Physicochemical and biological water quality, including the total microcystin concentration, was investigated from July to December 2003 in the reservoir Hjar, Tunisia. 2 - Microcystin levels and characterization of the different microcystin variants present were measured by protein phosphatase inhibition assays (PP2A) and by high performance liquid chromatography coupled to diode array detector and tandem mass spectrometry, respectively. The microscopic examination of the phytoplankton samples showed the dominance of the Oscillatoria and Pseudoanabaena genera. The total (particulate and dissolved) microcystin concentrations in the reservoir water varied between 23.4 and 7455.2 ng/l microcystin-LR equivalent per liter. 3 - The highest MCYST concentration was observed in November 2003. The analysis of the field bloom extract from this month by HPLC coupled to photodiode-array detector revealed the presence of five peaks having characteristic spectra of microcystins with a maximum of absorbance at 238 nm. HPLC/MS/MS analysis of this sample demonstrated the presence of three variants of microcystins: microcystin-LR (MCYST-LR), microcystin- (MCYST-RR), microcystin- (MCYST-YR). Therefore, estuaries of rivers contaminated by cyanobacteria toxins may play an important role on the transfer of these cyanotoxins through food chains.