Determining the probability of cyanobacterial blooms: the application of Bayesian networks in multiple lake systems.

A Bayesian network model was developed to assess the combined influence of nutrient conditions and climate on the occurrence of cyanobacterial blooms within lakes of diverse hydrology and nutrient supply. Physicochemical, biological, and meteorological observations were collated from 20 lakes located at different latitudes and characterized by a range of sizes and trophic states. Using these data, we built a Bayesian network to (1) analyze the sensitivity of cyanobacterial bloom development to different environmental factors and (2) determine the probability that cyanobacterial blooms would occur. Blooms were classified in three categories of hazard (low, moderate, and high) based on cell abundances. The most important factors determining cyanobacterial bloom occurrence were water temperature, nutrient availability, and the ratio of mixing depth to euphotic depth. The probability of cyanobacterial blooms was evaluated under different combinations of total phosphorus and water temperature. The Bayesian network was then applied to quantify the probability of blooms under a future climate warming scenario. The probability of the "high hazardous" category of cyanobacterial blooms increased 5% in response to either an increase in water temperature of 0.8°C (initial water temperature above 24°C) or an increase in total phosphorus from 0.01 mg/L to 0.02 mg/L. Mesotrophic lakes were particularly vulnerable to warming. Reducing nutrient concentrations counteracts the increased cyanobacterial risk associated with higher temperatures.

Current approaches to cyanotoxin risk assessment and risk management around the globe.

Toxic cyanobacteria became more widely recognized as a potential health hazard in the 1990s, and in 1998 the World Health Organization (WHO) first published a provisional Guideline Value of 1 µg L-1 for microcystin-LR in drinking-water. In this publication we compare risk assessment and risk management of toxic cyanobacteria in 17 countries across all five continents. We focus on the three main (oral) exposure vehicles to cyanotoxins: drinking-water, water related recreational and freshwater seafood. Most countries have implemented the provisional WHO Guideline Value, some as legally binding standard, to ensure the distribution of safe drinking-water with respect to microcystins. Regulation, however, also needs to address the possible presence of a wide range of other cyanotoxins and bioactive compounds, for which no guideline values can be derived due to insufficient toxicological data. The presence of microcystins (commonly expressed as microcystin-LR equivalents) may be used as proxy for overall guidance on risk management, but this simplification may miss certain risks, for instance from dissolved fractions of cylindrospermopsin and cyanobacterial neurotoxins. An alternative approach, often taken for risk assessment and management in recreational waters, is to regulate cyanobacterial presence - as cell numbers or biomass - rather than individual toxins. Here, many countries have implemented a two or three tier alert level system with incremental severity. These systems define the levels where responses are switched from Surveillance to Alert and finally to Action Mode and they specify the short-term actions that follow. Surface bloom formation is commonly judged to be a significant risk because of the elevated concentration of microcystins in a scum. Countries have based their derivations of legally binding standards, guideline values, maximally allowed concentrations (or limits named otherwise) on very similar scientific methodology, but underlying assumptions such as bloom duration, average body size and the amount of water consumed while swimming vary according to local circumstances. Furthermore, for toxins with incomplete toxicological data elements of expert judgment become more relevant and this also leads to a larger degree of variation between countries' thresholds triggering certain actions. Cyanobacterial blooms and their cyanotoxin content are a highly variable phenomenon, largely depending on local conditions, and likely concentrations can be assessed and managed best if the specific conditions of the locality are known and their impact on bloom occurrence are understood. Risk Management Frameworks, such as for example the Water Safety Plan concept of the WHO and the 'bathing water profile' of the European Union are suggested to be effective approaches for preventing human exposure by managing toxic cyanobacteria from catchment to consumer for drinking water and at recreational sites.

Attachment, re-mobilization, and inactivation of bacteriophage MS2 during bank filtration following simulation of a high virus load and an extreme rain event.

Viruses, including human pathogenic viruses, can persist in water. For producing drinking water from surface water via bank filtration, natural attenuation capacities and the fate of viruses during the passage of aquatic sediments are of particular interest. Moreover, the increasing frequency of extreme hydrological events necessitate re-evaluation of the sustainability and efficacy of processes removing viruses. For this purpose, we performed bank sediment filtration experiments using a mesocosm in a technical-scale experimental facility that simulates a field situation under more tightly controlled conditions. We used the bacteriophage MS2 as a surrogate for enteric viruses to study the transport of different viral loads through the bank sediment. Additionally, we simulated a heavy rain event to investigate the re-mobilization of initially attached virus particles. We quantified the abundance of infectious MS2 phages by plaque assay and the total number of MS2 particles by qPCR. Also, we differentiated pore water concentrations by depths of the sediment column and investigated attachment to the sediment matrix at the end of the individual experimental phases. Bank filtration over a vertical distance of 80 cm through sandy sediment revealed a virus removal efficiency of 0.8 log10 for total MS2 particles and 1.7 log10 for infectious MS2 particles, with an initial phage concentration of 1.84 × 108 gene copies mL-1. A low load of infectious MS2 (1.9 × 106 plaque forming units mL-1) resulted in a greater removal efficiency (3.0 log10). The proportion of infectious MS2 phages of the total MS2 particle mass steadily decreased over time, i.e., in the course of individual breakthrough curves and with sediment depth. The simulated pulse of rainwater caused a front of low ionic strength water which resulted in pronounced phage remobilization. The high proportion of infectious MS2 among the detached phages indicated that attachment to the sediment matrix may substantially conserve virus infectivity. Therefore, the re-mobilization of previously attached viruses owing to hydrological extremes should be considered in water quality assessment and monitoring schemes.

Comment on "Models predict planned phosphorus load reduction will make Lake Erie more toxic".

Hellweger et al. (Reports, 27 May 2022, pp. 1001) predict that phosphorus limitation will increase concentrations of cyanobacterial toxins in lakes. However, several molecular, physiological, and ecological mechanisms assumed in their models are poorly supported or contradicted by other studies. We conclude that their take-home message that phosphorus load reduction will make Lake Erie more toxic is seriously flawed.

Water and phosphorus mass balance of Lake Tegel and Schlachtensee - a modelling approach.

Management models for aquatic systems can be used to determine which measures in the watershed or in the water body have been effective and/or which one should be used in future. The newly developed management models presented in the following for Lake Tegel and Schlachtensee are empirical and lake specific. The values for the unknown factors are estimated by an iterative process using optimisation routines and sensitivity analysis methods. The resulting models describe the water and phosphorus balance of each lake. The Lake Tegel water balance model calculates the unknown water inflow from the River Havel depending on the other main in- and outflows with very good validation results. The phosphorus models of both lakes quantify mixing of the upper and lower water body as well as sedimentation and release from the sediment as functions of measured variables. For Lake Tegel, management scenarios were run indicating effective management interventions. For Lake Schlachtensee, the phosphorus model captured the variations in the hypolimnion well but produced poorer results for the epilimnion because of unknown external phosphorus loads. For these the model indicated possible sources and magnitudes.

Seasonal dynamics of cylindrospermopsin and cyanobacteria in two German lakes.

Cylindrospermopsin (CYN) is a potent hepatotoxin produced by different cyanobacteria of the order Nostocales. Questions of major concern are: which species produce CYN, which are the seasonal patterns of CYN dynamics and how are they regulated? Therefore, we studied for the first time the seasonal dynamics of particulate and dissolved CYN concentrations, cyanobacterial abundance and environmental factors in two German lakes over 2 years. Total CYN reached maximum concentrations of 0.34 and 1.80 microg L(-1) in Melangsee and Langer See, respectively. In both lakes, maxima of the dissolved CYN fraction occurred later in the season than those of the particulate fraction, and it reached higher concentrations. This indicates that CYN is poorly decomposed and accumulates in the water. The cyanobacterial community in both lakes included several potentially CYN-producing species that did not correlate with CYN concentrations. Significant correlations between the particulate CYN concentrations and species biovolume were only found for Aphanizomenon gracile (r(s)=0.803) in Langer See indicating that this species is a CYN producer. Different correlations of CYN with abiotic factors in the two lakes indicate the presence of further undetected CYN producers as well as different regulation mechanisms of their dynamics and the variability of CYN.

Fatal Neurotoxicosis in Dogs Associated with Tychoplanktic, Anatoxin-a Producing Tychonema sp. in Mesotrophic Lake Tegel, Berlin.

In May 2017, at least 12 dogs showed signs of acute neurotoxicosis after swimming in or drinking from Lake Tegel, a mesotrophic lake in Berlin, Germany, and several of the affected dogs died shortly afterwards despite intensive veterinary treatment. Cyanobacterial blooms were not visible at the water surface or the shorelines. However, detached and floating water moss (Fontinalis antipyretica) with high amounts of Tychonema sp., a potential anatoxin-a (ATX) producing cyanobacterium, was found near the beaches where the dogs had been swimming and playing. Necropsies of two of the dogs revealed no specific lesions beside the anamnestic neurotoxicosis. ATX was detected in concentrations up to 8700 µg L-1 in the stomach contents, while other (neuro)toxic substances were not found. In the aqueous fraction of Fontinalis/Tychonema clumps sampled after the casualties, ATX was found in concentrations up to 1870 µg L-1. This is the first report of a dense population of Tychonema sp. in stands of Fontinalis resulting in high ATX contents. This case emphasizes the need for further investigation of potentially toxic, non-bloom forming cyanobacteria in less eutrophic water bodies and underlines the novel challenge of developing appropriate surveillance schemes for respective bathing sites.

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.

Accumulation of cyanobacterial toxins in freshwater "seafood" and its consequences for public health: a review.

This review summarizes and discusses the current understanding of human exposure to cyanobacterial toxins in "seafood" collected from freshwater and coastal areas. The review consists of three parts: (a) the existing literature on concentrations of cyanobacterial toxins in seafood is reviewed, and the likelihood of bioaccumulation discussed; (b) we derive cyanotoxin doses likely to occur through seafood consumption and propose guideline values for seafood and compare these to guidelines for drinking water; and (c) we discuss means to assess, control or mitigate the risks of exposure to cyanotoxins through seafood consumption. This is discussed in the context of two specific procedures, the food specific HACCP-approach and the water-specific Water Safety Plan approach by the WHO. Risks of exposure to cyanotoxins in food are sometimes underestimated. Risk assessments should acknowledge this and investigate the partitioning of exposure between drinking-water and food, which may vary depending on local circumstances.

First report on cylindrospermopsin producing Aphanizomenon flos-aquae (Cyanobacteria) isolated from two German lakes.

Three single-filament isolates of Aphanizomenon flos-aquae from two German lakes were found to produce remarkable amounts of the cyanobacterial hepatotoxin cylindrospermopsin (CYN). CYN-synthesis of the strains were evidenced both by LC-MS/MS analysis and detection of PCR products of gene fragments which are implicated in the biosynthesis of the toxin. The strains contain CYN in the range of 2.3-6.6 mg g(-1) of cellular dry weight. To our knowledge this is the first report of CYN in A. flos-aquae.

Microcystins (cyanobacterial toxins) in surface waters of rural Bangladesh: pilot study.

In Bangladesh the exposure of millions of inhabitants to water from (shallow) tube wells contaminated with high geogenic loads of arsenic is a major concern. As an alternative to the costly drilling of deep wells, the return to the use of surface water as a source of drinking water is considered. In addition to the well-known hazards of water borne infectious diseases associated with the use of surface water, recently the potential public health implications of toxic cyanobacteria have been recognized. As a first step towards a risk assessment for cyanotoxins in Bangladesh surface waters, seston samples of 79 ponds were analysed in late summer 2002 for the presence of cyanobacteria and microcystins (MCYST), the most frequently detected cyanobacterial toxins worldwide. Microcystins could be detected in 39 ponds, mostly together with varying abundance of potentially microcystin-producing genera such as Microcystis, Planktothrix and Anabaena. Total microcystin concentrations ranged between <0.1 and > 1,000 microg l(-1), and more than half of the positive samples contained high concentrations of more than 10 microg l(-1). The results clearly show that concentrations of microcystins well above the provisional WHO guideline value of 1 microg l(-1) MCYST-LR can be frequently detected in Bangladesh ponds. Thus, an increasing use of surface water for human consumption introduces a risk of replacing one health hazard by another and therefore needs to be accompanied by cyanotoxin hazard assessments.

Attenuation and colloidal mobilization of bacteriophages in natural sediments under anoxic as compared to oxic conditions.

Redox conditions are known to affect the fate of viruses in porous media. Several studies report the relevance of colloid-facilitated virus transport in the subsurface, but detailed studies on the effect of anoxic conditions on virus retention in natural sediments are still missing. Therefore, we investigated the fate of viruses in natural flood plain sediments with different sesquioxide contents under anoxic conditions by considering sorption to the solid phase, sorption to mobilized colloids, and inactivation in the aqueous phase. Batch experiments were conducted under oxic and anoxic conditions at pH values between 5.1 and 7.6, using bacteriophages MS2 and PhiX174 as model viruses. In addition to free and colloid-associated bacteriophages, dissolved and colloidal concentrations of Fe, Al and organic C as well as dissolved Ca were determined. Results showed that regardless of redox conditions, bacteriophages did not adsorb to mobilized colloids, even under favourable charge conditions. Under anoxic conditions, attenuation of bacteriophages was dominated by sorption over inactivation, with MS2 showing a higher degree of sorption than PhiX174. Inactivation in water was low under anoxic conditions for both bacteriophages with about one log10 decrease in concentration during 16 h. Increased Fe/Al concentrations and a low organic carbon content of the sediment led to enhanced bacteriophage removal under anoxic conditions. However, even in the presence of sufficient Fe/A-(hydr)oxides on the solid phase, bacteriophage sorption was low. We presume that organic matter may limit the potential retention of sesquioxides in anoxic sediments and should thus be considered for the risk assessment of virus breakthrough in the subsurface.

Distribution of microcystin-producing and non-microcystin-producing Microcystis sp. in European freshwater bodies: detection of microcystins and microcystin genes in individual colonies.

Microcystis is a well-known cyanobacterial genus frequently producing hepatotoxins named microcystins. Toxin production is encoded by microcystin genes (mcy). This study aims (i) to relate the mcy occurrence in individual colonies to the presence of microcystin, (ii) to assess whether morphological characteristics (morphospecies) are related to the occurrence of mcy genes, and (iii) to test whether there are geographical variations in morphospecies specificity and abundance of mcy genes. Individual colonies of nine different European countries were analysed by (1) morphological characteristics, (2) PCR to amplify a gene region within mcyA and mcyB indicative for microcystin biosynthesis, (3) matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF MS) to detect microcystins. Almost one hundred percent of the colonies predicted to produce microcystins by PCR analysis were found to contain microcystins. A high similarity in microcystin variants in the different colonies selected from lakes across Europe was demonstrated. The different morphospecies varied in the frequency with which they contained mcy genes. Most colonies (>75%) of M. aeruginosa and M. botrys contained the mcy genes, whereas < or = 20% of the colonies identified as M. ichthyoblabe and M. viridis gave a PCR product of the mcy genes. No colonies of M. wesenbergii gave a PCR product of either mcy gene. In addition, a positive relationship was found between the size of the colony and the frequency of those containing the mcy genes. It is concluded that the analysis of morphospecies is indicative for microcystin production, although the quantitative analysis of microcystin concentrations in water remains indispensable for hazard control.

Nitrogen limitation promotes accumulation and suppresses release of cylindrospermopsins in cells of Aphanizomenon sp.

As the biosynthesis of cylindrospermopsin (CYN) is assumed to depend on nitrogen availability, this study investigated the impact of nitrogen availability on intra- and extracellular CYN and deoxy-CYN (D-CYN) contents in three Aphanizomenon strains from temperate waters. Nitrogen deficient (-N) cultures showed a prolonged growth phase and intracellular toxin accumulation by a factor of 2-6. In contrast, cultures with additional nitrate supply (+N) did not accumulate CYN within the cells. Instead, the maximum conceivable CYN release estimated for dead cells (identified by SYTOX Green staining) was much lower than the concentrations of dissolved CYN actually observed, suggesting these cultures actively release CYN from intact cells. Furthermore, we found remarkably altered proportions of CYN to D-CYN: as batch cultures grew, the proportion of D-CYN increased by up to 40% in +N medium, whereas D-CYN remained constant or decreased slightly in -N medium. Since +N cultures showed similar toxin patterns as -P cultures with increased extracellular CYNs and higher proportion of D-CYN we conclude that nitrogen limitation may affect the way the cells economize resources, especially the yield from phosphorus pools, and that this has an impact on CYN production and release. For water management, these result imply that nutrient availability not only determines the abundance of potentially CYN-producing cyanobacteria, but also the amount of extracellular CYNs (challenging drinking-water treatment) as well as the ratio of D-CYN to CYN (affecting toxicity).

Transport and removal of viruses in saturated sand columns under oxic and anoxic conditions--Potential implications for groundwater protection.

To protect groundwater as a drinking water resource from microbiological contamination, protection zones are installed. While travelling through these zones, concentrations of potential pathogens should decline to levels that pose no risks to human health. Removal of viruses during subsurface passage is influenced by physicochemical conditions, such as oxygen concentration, which also affects virus survival. The aim of our study was to evaluate the effect of redox conditions on the removal of viruses during sand filtration. Experiments in glass columns filled with medium-grained sand were conducted to investigate virus removal in the presence and absence of dissolved oxygen. Bacteriophages MS2 and PhiX174, as surrogates for human enteric viruses were spiked in pulsed or in continuous mode and pumped through the columns at a filter velocity of about 1m/d. Virus breakthrough curves were analyzed by calculating total viral elimination and fitted using one-dimensional transport models (CXTFIT and HYDRUS-1D). While short-term experiments with pulsed virus application showed only small differences with regard to virus removal under oxic and anoxic conditions, a long-term experiment with continuous dosing revealed a clearly lower elimination of viruses under anoxic conditions. These findings suggest that less inactivation and less adsorption of viruses in anoxic environments affect their removal. Therefore, in risk assessment studies aimed to secure drinking water resources from viral contamination and optimization of protection zones, the oxic and anoxic conditions in the subsurface should also be considered.

Microcystin elimination during sediment contact.

Microcystins (MCYSTs) are a group of structurally similar toxic peptides produced by cyanobacteria ("blue-green algae") which occur frequently in surface waters worldwide. Reliable elimination is necessary when using these waters as drinking water sources. Bank filtration and artificial groundwater recharge utilize adsorption and degradation processes in the subsurface, commonly through sand and gravel aquifers, for the elimination of a wide range of substances during drinking water (pre-) treatment. To obtain parameters for estimating whether MCYST breakthrough is likely in field settings, we tested MCYST elimination in laboratory experiments (batch experiments, column experiments) under a range of conditions. Adsorption coefficients (k(d)-values) obtained from batch studies ranged from 0.2 mL/g for filter sand to 11.6 mL/g for fine grained aquifer materials with 2% fine grains (<63 microm) and 0.8% organic matter. First order degradation rates in column studies reached 1.87 d(-1) under aerobic conditions and showed high variations under anoxic conditions (<0.01-1.35 d(-1)). These results show that, next to sediment texture, redox conditions play an important role for MCYST elimination during sediment passage. Biodegradation was identified as the dominating process for MCYST elimination in sandy aquifer material.

Retention and degradation of the cyanobacterial toxin cylindrospermopsin in sediments - the role of sediment preconditioning and DOM composition.

Recent results show that cylindrospermopsin is more frequent and widespread in surface waters than previously assumed. Studies on the fate of CYN in sediments are lacking, but this is important if these resources are used for drinking-water production via sediment passage. Therefore, the aim of our study was to determine a) CYN retention in two sandy sediments as a function of flow rate, CYN concentration, the presence of DOM and the content of fines (1% and 4%, respectively) and b) the influence of sediment preconditioning and DOM composition of the water (aquatic DOM versus DOM released from lysed cells) on CYN degradation. Retention of CYN proved negligible under the investigated conditions. Degradation in virgin sediments showed the highest lag phases (20 days). Preconditioned sediments showed no lag phase. The presence of aquatic DOM yielded highest degradation rates (kappa(1)=0.46 and 0.49 day(-1)) without a lag phase. Readily available organic carbon sources were preferentially metabolized and hence induced a lag phase. Thus, the presence and composition of DOM in the water proved important for both CYN degradation rates in preconditioned sediments and for the lag phase. Cylindrospermopsin degradation took place solely in the sediment and not in the water body.

Toxic cyanobacterial blooms in reservoirs under a semiarid mediterranean climate: the magnification of a problem.

Sicilian reservoirs constitute the most important water resources available on the island. During summer 2001, the intense water utilization of Lake Arancio reservoir reduced the water level significantly, which coincided with the formation of intense blooms formed by the microcystin (MC)-producing cyanobacterium Microcystis aeruginosa. During summer 2003, Lake Arancio was continuously filled and the vertical stratification of the water column was maintained resulting in five to sixfold lower cell numbers of M. aeruginosa. For both years, a significant relationship between MC net production and Microcysytis cell growth was observed, implying that Microcystis cell numbers can be used to infer MC concentrations in water. Unexpectedly, dense blooms of the MC-producing cyanobacterium Planktothrix rubescens occurred during winter 2005/2006 in the reservoirs Lake Pozzillo, Prizzi, Nicoletti, and Garcia but have not been reported earlier. In this season, MC concentrations higher than those recorded in summer were measured, implying that monitoring of Mediterranean drinking water reservoirs needs to be intensified during winter, a season usually considered to be less prone to the formation of cyanobacterial blooms.

Occurrence of the cyanobacterial toxin cylindrospermopsin in northeast Germany.

The frequent occurrence of the cyanobacterial toxin cylindrospermopsin (CYN) in the (sub)tropics has been largely associated with cyanobacteria of the order Nostocales of tropical origin, in particular Cylindrospermopsis raciborskii. C. raciborskii is currently observed to spread northwards into temperate climatic zones. In addition, further cyanobacteria of the order Nostocales typically inhabiting water bodies in temperate regions are being identified as CYN-producers. Therefore, data on the distribution of CYN in temperate regions are necessary for a first assessment of potential risks due to CYN in water used for drinking and recreation. A total of 127 lakes situated in the north-eastern part of Germany were investigated in 2004 for the presence of the toxin CYN and the phytoplankton composition. The toxin could be detected in half of the lakes (n = 63) and in half of 165 samples (n = 88). Concentrations reached up to 73.2 microg CYN/g DW. CYN thus proved more widely distributed than previously demonstrated. The analyses of phytoplankton data suggest Aphanizomenon sp. and Anabaena sp. as important CYN producers in Germany, and confirm recent findings of Aphanizomenon flos-aquae as CYN-producing species frequently inhabiting water bodies in temperate climatic regions. The data shown here suggest that CYN may be an important cyanobacterial toxin in German water bodies and that further data are needed to assess this.