Morphological changes during akinete germination in Cylindrospermopsis raciborskii (Nostocales, Cyanobacteria)

The occurrence of the cyanobacterium Cylindrospermopsis raciborskii (Woloszynska) Seenayya and Subba Raju is a global water quality issue. The misidentification of C. raciborskii in the past is a major concern for water quality users, considering the reported cases of human and livestock poisonings associated with the cyanobacterium. Many of the available taxonomic descriptions for this species provide little or no detail of the morphology of early developmental phases that may assist with identification. Therefore, typifying the morphological changes throughout the entire life cycle for such a species requires urgent attention. In this study, five distinct morphological phases identified using a new culturing technique are reported for the process of akinete germination in C. raciborskii. Before the terminal emergence of three to four cell germlings through a ruptured akinete envelope (phase 3), mature akinetes (phase 1) elongated and the endospore separated from the akinete envelope (phase 2). After the association with the envelope was lost, four-cell germlings (phase 4a) matured into young trichomes of more than four cells (phase 4b). Throughout the process of germination, internal granular structures decreased in size and were irregular in shape in germlings and young trichomes. The culturing technique, which used a Sedgwick-Rafter cell, was successful in its application but was limiting in that the development of young trichomes after phase 4b could not be monitored.

Factors affecting akinete differentiation in Cylindrospermopsis raciborskii (Nostocals, Cyanobacteria)

1. Cylindrospermopsis raciborskii is a potentially toxic freshwater cyanobacterium which can produce akinetes (reproductive spores) that on germinating can contribute to future populations. To further understand factors controlling the formation of these specialised cells, the effects of diurnal temperature fluctuations (magnitude and frequency), in combination with different light intensities and phosphorus concentrations were investigated under laboratory conditions. 2. Akinete differentiation was affected by the frequency of temperature fluctuations. Maximum akinete concentrations were observed in cultures that experienced multiple diurnal temperature fluctuations. 3. Akinete concentrations increased with increasing magnitude of temperature fluctuation. A maximum akinete concentration was achieved under multiple diurnal temperature fluctuations with a magnitude of 10degreesC (25degreesC to 15degreesC). 4. A fourfold increase in light intensity (25-100 mumol m(-2) s(-1)) resulted in an approximate 14-fold increase in akinete concentration. 5. High filterable reactive phosphorus (FRP) concentrations (> 70 mug L-1) in the medium, combined with a multiple diurnal temperature fluctuation of 10degreesC, supported the development of the highest akinete concentration.

Chlorination for degrading saxitoxins (paralytic shellfish poisons) in water

Chlorination was investigated as a treatment option for degrading and thus removing saxitoxins (paralytic shellfish poisons, PSPs) produced by cyanobacteria (blue-green algae) from water. It was found to be effective with the order of ease of degradation of the saxitoxins being GTX5 (B1) similar to dcSTX > STX > GTX3 similar to C2 > C1 > GTX2. However the effectiveness of chlorine was pH dependent. Degradation as a function of pH was not linear with the degree of degradation increasing rapidly at around pH 7.5. At pH 9 > 90% removal was possible provided a residual of 0.5 mg l(-1) free chlorine was present after 30 min contact time. The more effective degradation at higher pH was unexpected as chlorine is known to be a weaker oxidant under these conditions. The more effective degradation, then, must be due to the toxins, which are ionisable molecules, being present in a form at higher pH which is more susceptible to oxidation. The feasibility of using chlorine to remove saxitoxins during water treatment will therefore depend strongly on the pH of the water being chlorinated. Degradation may be improved by pH adjustment but may not be a practical solution. Although saxitoxins were degraded in that the parent compounds were not detected by chemical analysis, there is no indication as to the nature of the degradation products. However, acute toxicity as determined by the mouse bioassay was eliminated.

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 mug/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 mug/l were found for MCs, PSPs and CYN, respectively. However, only traces (< 1.0 mug/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. (C) 2004 Elsevier Ltd. All rights reserved.

Genotoxicity investigation of chlorinated degradation products of a cyanobacterial toxin, cylindrospermopsin

Cylindrospermopsin (CYN), a potent cyanobacterial hepatotoxin produced by Cylindrospermopsis raciborskii and other cyanobacteria, is regularly found in water supplies in many parts of the world and has been associated with the intoxication of humans and livestock.Water treatment via chlorination can degrade the toxin effectively but result in the production of several byproducts. In this study, male and female Balb/c mice were injected via the intraperitoneal (IP) route with a single dose of 10 mg/kg 5-chlorouracil and 10 mg/kg 5-chloro-6-hydroxymethyluracil; these two compounds are the predicted chlorinated degradation products of CYN.DNA was isolated from the mouse livers and examined for strand breakage by alkaline gel electrophoresis (pH 12). The median molecular length (MML) of the DNA distributed in the gel was determined by estimating the midpoint of the DNA size distribution by densitometry. The toxicity of 5-chlorouracil (as measured by DNA strand breakage) was significantly influenced by time from dosing. There was no significant difference in MML between mice dosed with 5-chloro-6-hydroxymethyluracil and the controls. In another experiment, mice were dosed with 0, 0.1, 1, 10 and 100 mg/kg body weight 5-chlorouracil and 0, 0.1, 1, 10 and 20 mg/kg 5-chloro-6-hydroxymethyluracil via IP injection. The heart, liver, kidney, lung and spleen were removed, fixed and examined under electron microscopy. Liver was the main target organ. The EM results revealed marked distortion on the nuclear membrane of liver cells in mice dosed with 1.0 mg/kg 5-chlorouracil or 10 mg/kg 5-chloro-6-hydroxymethyluracil, or higher.

Phosphate uptake and growth kinetics of Trichodesmium (Cyanobacteria) isolates from the North Atlantic Ocean and the Great Barrier Reef, Australia

We compared inorganic phosphate (P-i) uptake and growth kinetics of two cultures of the diazotrophic cyanobacterium Trichodesmium isolated from the North Atlantic Ocean (IMS101) and from the Great Barrier Reef, Australia (GBRTRLI101). Phosphate-limited cultures had up to six times higher maximum P-i uptake rates than P-replete cultures in both strains. For strain GBRTRLI101, cell-specific P-i uptake rates were nearly twice as high, due to larger cell size, but P-specific maximum uptake rates were similar for both isolates. Half saturation constants were 0.4 and 0.6 muM for P-i uptake and 0.1 and 0.2 muM for growth in IMS101 and GBRTRLI101, respectively. Phosphate uptake in both strains was correlated to growth rates rather than to light or temperature. The cellular phosphorus quota for both strains increased with increasing P-i up to 1.0 muM. The C:P ratios were 340-390 and N:P ratios were 40-45 for both strains under severely P-limited growth conditions, similar to reported values for natural populations from the tropical Atlantic and Pacific Oceans. The C:P and N:P ratios were near Redfield values in medium with >1.0 muM P-i. The North Atlantic strain IMS101 is better adapted to growing on P-i at low concentrations than is GBRTRLI101 from the more P-i-enriched Great Barrier Reef. However, neither strain can achieve appreciable growth at the very low (nanomolar) P-i concentrations found in most oligotrophic regimes. Phosphate could be an important source of phosphorus for Trichodesmium on the Great Barrier Reef, but populations growing in the oligotrophic open ocean must rely primarily on dissolved organic phosphorus sources.

Epidemiology of recreational exposure to freshwater cyanobacteria - an international prospective cohort study

Background: Case studies and anecdotal reports have documented a range of acute illnesses associated with exposure to cyanobacteria and their toxins in recreational waters. The epidemiological data to date are limited; we sought to improve on the design of some previously conducted studies in order to facilitate revision and refinement of guidelines for exposure to cyanobacteria in recreational waters. Methods: A prospective cohort study was conducted to investigate the incidence of acute symptoms in individuals exposed, through recreational activities, to low ( cell surface area < 2.4 mm(2)/mL), medium ( 2.4 - 12.0 mm(2)/mL) and high (> 12.0 mm(2)/mL) levels of cyanobacteria in lakes and rivers in southeast Queensland, the central coast area of New South Wales, and northeast and central Florida. Multivariable logistic regression analyses were employed; models adjusted for region, age, smoking, prior history of asthma, hay fever or skin disease ( eczema or dermatitis) and clustering by household. Results: Of individuals approached, 3,595 met the eligibility criteria, 3,193 (89%) agreed to participate and 1,331 (37%) completed both the questionnaire and follow-up interview. Respiratory symptoms were 2.1 (95% CI: 1.1 - 4.0) times more likely to be reported by subjects exposed to high levels of cyanobacteria than by those exposed to low levels. Similarly, when grouping all reported symptoms, individuals exposed to high levels of cyanobacteria were 1.7 ( 95% CI: 1.0 - 2.8) times more likely to report symptoms than their low-level cyanobacteria-exposed counterparts. Conclusion: A significant increase in reporting of minor self-limiting symptoms, particularly respiratory symptoms, was associated with exposure to higher levels of cyanobacteria of mixed genera. We suggest that exposure to cyanobacteria based on total cell surface area above 12 mm(2)/mL could result in increased incidence of symptoms. The potential for severe, life-threatening cyanobacteria-related illness is likely to be greater in recreational waters that have significant levels of cyanobacterial toxins, so future epidemiological investigations should be directed towards recreational exposure to cyanotoxins.