Economic cost of cyanobacterial blooms.

Cyanobacterial blooms impact upon the water quality, environmental and ecological status of water bodies and affect most of the uses we make of water. The extent of the impact depends upon the type, size and frequency of the blooms, the size of the water body affected, the uses made of the water and the treatment options available to respond to the blooms. The impacts therefore vary considerably from place to place. Overall costs should also account for the planning and remedial actions taken to prevent future blooms.

Risk Assessment Workgroup report.

The Risk Assessment Work Group focused on six charge questions related to CHABS, cyanobacteria and their toxins. The charge questions covered the following topics: Research needed to reduce uncertainty in establishing health based guidelines. Research that minimize the cost and maximize the benefits of various regulatory approaches. Exposure pathways for receptors of concern. Data available to support the derivation of health-based guideline values for harmful cyanobacterial algal blooms. Ecological services that guidelines or regulations should protect? A framework for making risk management determinations that incorporates consideration of the characteristics of CHABs, the risk for human health, ecosystem viability, and the costs and benefits of CHABs detection and management? The Work Group concluded that there is a considerable amount of human case-study data and information from animal studies to demonstrate that cyanobacterial toxins pose a hazard to humans, domestic animals, wildlife, and the ecosystem. However, the data on dose-response are limited and confounded by a lack of sufficient pure toxin to conduct most of the toxicological studies that will be needed in order to answer remaining questions on risk, and to provide the data for quantitative dose-response analysis. The Work Group recommended that research on purification or synthesis of pure toxin must be accomplished before the large scale studies to establish dose-response relationships will be possible. As the necessary-pure toxins become available, the Work Group recommended that studies be prioritized by the impact that they will have on reducing the uncertainty in the risk assessment in order to minimize the research costs and maximize the risk assessment benefits. Use of quantitative structure activity relationships (QSAR) and toxicity equivalency factor studies are also recommended as approaches for filling dose-response data gaps. The Work Group recognized that CHABs rarely introduce single toxins into the water supply. Under CHAB conditions, affected water is likely to contain a variety of toxins in varying concentrations that may change over the duration of the bloom. Accordingly, research on cyanotoxin interactions is needed, along with the development of risk assessment approaches for CHAB mixtures. The development of simple, accurate analytical methods that can be utilized by most analytical laboratories or used in the field was recognized as a major data need for establishing exposure potential and monitoring bloom conditions. Most currently available methods are time-consuming and/or costly. Human exposure to cyanobacterial toxins can occur through ingestion of contaminated drinking water, plus dermal contact and/or inhalation of aerosols while bathing and showering in tap water. Treatment can reduce the concentrations of both the toxins and the bacteria in the treated water but there is still much to be learned about the effectiveness of most treatment technologies on cyanobacteria and toxin removal. Human exposure to cyanobacteria and their toxins also occurs through incidental ingestion, dermal contact, and inhalation of aerosols during recreational use of surface waters, ingestion of contaminated fish and other foods of aquatic origin, and/or BGAS supplements. Establishing intakes and duration parameters for these exposure scenarios will facilitate the application of risk assessment approaches to these situations.

Application of the neuroblastoma assay for paralytic shellfish poisons to neurotoxic freshwater cyanobacteria: interlaboratory calibration and comparison with other methods of analysis.

Paralytic shellfish poisons (PSPs) are produced by freshwater cyanobacteria and pose a threat to human and animal drinking-water supplies. The wide range of toxin analogues (and the likelihood that further analogues remain to be discovered) means that chromatographic methods are not always reliable indicators of toxicity. Although the mouse bioassay remains the method of choice in the seafood industry, its use is increasingly being questioned on ethical grounds. The cell-based Neuro-2A neuroblastoma toxicity assay is an alternative bioassay validated for testing shellfish extracts, so it was of interest to determine its applicability with the different suite of toxin analogues produced by cyanobacteria. Cyanobacterial bloom samples from Australia, Brazil, and France were assayed using the neuroblastoma assay, liquid chromatography-tandem mass spectrometry (LC-MS/MS), high-performance liquid chromatography with postcolumn derivatization and fluorescence detection, and the Jellett Rapid Test for PSP. To assess interlaboratory variability, the neuroblastoma assay was set up in laboratories in Paris (France) and Adelaide (Australia). Neuroblastoma and chromatographic methods gave comparable results except in the case of the neurotoxic Brazilian samples: LC-MS/MS did not detect the putative new PSPs contained in these samples. Inter- and intralaboratory variability of the neuroblastoma assay was typical of biological assays but no greater than that found for interassay variability between different chromatographic determinations. The batch of Jellett Rapid Tests for PSP used did not yield quantitative results. Overall, the neuroblastoma assay was useful as a screening assay for determination of toxicity caused by saxitoxin neurotoxins in freshwater cyanobacteria, having the advantage of being sensitive to unidentified toxins that currently cannot be quantified by chromatographic means.

Extraction of cyanobacterial endotoxin.

To simplify our efforts in acquiring toxicological information on endotoxins produced by cyanobacteria, a method development study was undertaken to identify relatively hazard-free and efficient procedures for their extraction. One article sourced and two novel methods were evaluated for their ability to extract lipopolysaccharides (LPSs) or endotoxins from cyanobacteria. The Limulus polyphemus amoebocyte lysate (LAL) assay was employed to compare the performance of a novel method utilizing a 1-butanol-water (HBW) solvent system to that of Westphal's (1965) phenol-water system (HPW) for the extraction of endotoxin from various cyanobacteria. The traditional HPW method extracted from 3- to 12-fold more endotoxin from six different cyanobacterial blooms and culture materials than did the novel HBW method. In direct contrast, the novel HBW method extracted ninefold more endotoxin from a non-microcystin producing Microcystis aeruginosa culture as compared to the HPW method. A solvent system utilizing N,N'-dimethylformamide-water (HDW) was compared to both the HPW and HBW methods for the extraction of endotoxin from natural samples of Anabaena circinalis, Microcystis flos-aquae, and a 1:1 mixture of Microcystis aeruginosa/Microcystisflos-aquae. The LAL activities of these extracts showed that the novel HDW method extracted two- and threefold more endotoxin from the Anabaena sample that did the HBW and HPW methods, respectively. The HDW method also extracted approximately 1.5-fold more endotoxin from the Microcystis flos-aquae sample as compared to both the HBW and HPW methods. On the other hand, the HBW method extracted 2- and 14-fold more endotoxin from the Microcystis flos-aquae/Microcystis aeruginosa mixture than did the HPW and HDW methods, respectively. Results of this study demonstrate that significant disparities exist between the physicochemical properties of the cell wall constituents not only of different cyanobacterial species but also of different strains of the same cyanobacterial species, as showing by the varying effectiveness of the solvent systems investigated. Therefore, a sole method cannot be regarded as universal and superior for the extraction of endotoxins from cyanobacteria. Nevertheless, the ability of the novel HBW and HDW methods to utilize easily handled organic solvents that are less hazardous than phenol render them attractive alternatives to the standard HPW method.