Constitutive cylindrospermopsin pool size in Cylindrospermopsis raciborskii under different light and CO2 partial pressure conditions.

Cylindrospermopsin (CYN) and 7-deoxy-cylindrospermopsin (dCYN) are potent hepatotoxic alkaloids produced by numerous species of cyanobacteria, including the freshwater Cylindrospermopsis raciborskii. C. raciborskii is an invasive cyanobacterium, and the study of how environmental parameters drive CYN production has received significant interest from water managers and health authorities. Light and CO2 affect cell growth and physiology in photoautotrophs, and these are potential regulators of cyanotoxin biosynthesis. In this study, we investigated how light and CO2 affect CYN and dCYN pool size as well as the expression of the key genes, cyrA and cyrK, involved in CYN biosynthesis in a toxic C. raciborskii strain. For cells growing at different light intensities (10 and 100 µmol photons m(-2) s(-1)), we observed that the rate of CYN pool size production (µCYN) was coupled to the cell division rate (µc) during batch culture. This indicated that CYN pool size under our experimental conditions is constant and cell quotas of CYN (QCYN) and dCYN (QdCYN) are fixed. Moreover, a lack of correlation between expression of cyrA and total CYN cell quotas (QCYNs) suggests that the CYN biosynthesis is regulated posttranscriptionally. Under elevated CO2 (1,300 ppm), we observed minor effects on QCYN and no effects on expression of cyrA and cyrK. We conclude that the CYN pool size is constitutive and not affected by light and CO2 conditions. Thus, C. raciborskii bloom toxicity is determined by the absolute abundance of C. raciborskii cells within the water column and the relative abundance of toxic and nontoxic strains.

Light and phosphate competition between Cylindrospermopsis raciborskii and Microcystis aeruginosa is strain dependent.

The hypothesis that outcomes of phosphorus and light competition between Cylindrospermopsis raciborskii and Microcystis aeruginosa are strain dependent was tested experimentally. Critical requirements of phosphorus (P*) and of light (I*) of two strains of each species were determined through monoculture experiments, which indicated a trade-off between species and also between Microcystis strains. Competition experiments between species were performed using the weakest predicted competitors (with the highest values of P* and of I*) and with the strongest predicted competitors (with the lowest values of P* and of I*). Under light limitation, competition between the weakest competitors led C. raciborskii to dominate. Between the strongest competitors, the opposite was observed, M. aeruginosa displaced C. raciborskii, but both strains co-existed in equilibrium. Under phosphate limitation, competition between the weakest competitors led C. raciborskii to exclude M. aeruginosa, and between the strongest competitors, the opposite was observed, M. aeruginosa displaced C. raciborskii, but the system did not reach an equilibrium and both strains were washed out. Hence, outcomes of the competition depended on the pair of competing strains and not only on species or on type of limitation. We concluded that existence of different trade-offs among strains and between species underlie our results showing that C. raciborskii can either dominate or be displaced by M. aeruginosa when exposed to different conditions of light or phosphate limitation.

Effects of oxidative and physical treatments on inactivation of Cylindrospermopsis raciborskii and removal of cylindrospermopsin.

The presence of toxic cyanobacterial blooms (or blue-green algae) in water bodies used either as drinking water or for recreational purposes may present serious health risks for the human population. In this study, the removal of the chemical toxin, cylindrospermopsin, via free chlorine, chlorine dioxide, monochloramine, permanganate, ozone, and UV irradiation was studied. Ozone and free chlorine were found to be highly effective for cylindrospermopsion removal while the other disinfectants were ineffective. Ozone and free chlorine were also determined to be highly effective for the inactivation of the cyanobacteria, Cylindrospermopsis raciborskii, at typical water treatment exposures, chlorine dioxide, monochloramine, and permanganate were only marginally effective at inactivation of Cylindrospermopsis raciborskii.

Application of N-TiO2 for visible-light photocatalytic degradation of Cylindrospermopsis raciborskii - More difficult than that for photodegradation of Microcystis aeruginosa ?

Nowadays Cylindrospermopsis raciborskii (C. raciborskii) and the metabolites produced, such as cylindrospermopsin (CYN), pose a serious threat to the ecosystem. Advanced oxidation technologies have been verified as constituting a very promising means to eliminate the risk from harmful algae. But so far little research has focused on the visible-light photocatalytic destruction of C. raciborskii cells and the degradation of their metabolites. In our study, N-doped TiO2 (N-TiO2) was used to degrade C. raciborskii and the degradation was compared with that of the Microcystis aeruginosa (M. aeruginosa). Results showed that although the photodegradation of C. raciborskii was more difficult than that of M. aeruginosa, the treatment with N-TiO2 was still satisfactory. After adding 200 mg/L N-TiO2, C. raciborskii cells (5 × 106 cells/mL) were completely destroyed within 20 h under visible light irradiation, and nearly 90% of the organic matter and CYN in the suspensions were also degraded, thereby markedly improving the water quality. The photocatalytic process starts with damage to the cell membrane resulting in the leakage of internal components. Subsequently, the leaked metabolites were oxidised by the reactive oxidizing species produced by N-TiO2. Thus, the application of N-TiO2 is a promising method for the treatment of C. raciborskii.

Combined Effect of Light and Temperature on the Production of Saxitoxins in Cylindrospermopsis raciborskii Strains.

Cylindrospermopsis raciborskii is a potentially toxic freshwater cyanobacterium that can tolerate a wide range of light and temperature. Due to climatic changes, the interaction between light and temperature is studied in aquatic systems, but no study has addressed the effect of both variables on the saxitoxins production. This study evaluated the combined effect of light and temperature on saxitoxins production and cellular quota in C. raciborskii. Experiments were performed with three C. raciborskii strains in batch cultures under six light intensities (10, 40, 60, 100, 150, and 500 µmol of photons m-2 s-1) and four temperatures (15, 20, 25, and 30 °C). The growth of C. raciborskii strains was limited at lower temperatures and the maximum growth rates were obtained under higher light combined with temperatures equal or above 20 °C, depending on the strain. In general, growth was highest at 30 °C at the lower light intensities and equally high at 25 °C and 30 °C under higher light. Highest saxitoxins concentration and cell-quota occurred at 25 °C under high light intensities, but were much lower at 30 °C. Hence, increased temperatures combined with sufficient light will lead to higher C. raciborskii biomass, but blooms could become less toxic in tropical regions.

The impacts of a high CO2 environment on a bicarbonate user: the cyanobacterium Cylindrospermopsis raciborskii.

The potentially toxic cyanobacterium Cylindrospermopsis raciborskii (Woloszynska) Seenya et Subba Raju, originally described as a tropical-subtropical species, is increasingly found in temperate regions and its range is expanding. Climate change is hypothesised to be a factor in this expansion. We studied the effects of dissolved inorganic carbon (DIC) and pH on growth and photosynthesis of this species. We prepared six treatments in a continuous culture (turbidostat) grown at high light, two at low light, and eight in batch cultures grown under low light, by manipulating pH, HCO(3)(-) and CO(2) to assess the effect, if any, of these parameters on the growth rate, inorganic carbon acquisition and photosynthetic parameters of C. raciborskii. When the turbidostats were grown at 100 µmol photons (PAR) m(-2) s(-1), HCO(3)(-) concentration and pH had a positive effect on growth rate; the specific growth rate in 6 mM HCO(3)(-), for example, was twice what it was in 0.6 mM HCO(3)(-) (0.84 ± 0.10 and 0.44 ± 0.04 d(-1) respectively). Growth was lower in turbidostats grown at 20 µmol photons (PAR) m(-2) s(-1). Isotope disequilibrium experiments showed that the contribution of HCO(3)(-) to DIC acquisition is proportionately greater at the higher light. The maximum relative electron transport rate (rETR(max)) was significantly higher at the higher light, while the slope of the linear portion of the rETR(max) versus irradiance curve (α) was unchanged. In low light batch cultures, increasing HCO(3)(-) concentration and pH had a negative effect on growth, while CO(2) concentration had a small, positive effect. This species of cyanobacterium has an efficient CCM and under ideal growing conditions gets most of its carbon from HCO(3)(-). It may, therefore, be at a competitive disadvantage in a high CO(2) world.

Morphology, genetic diversity, temperature tolerance and toxicity of Cylindrospermopsis raciborskii (Nostocales, Cyanobacteria) strains from Thailand and Japan.

Cylindrospermopsis raciborskii is a planktonic, nostocalean cyanobacterium, which produces an alkaloid heptatoxin, cylindrospermopsin. We performed morphological observations, 16S rDNA sequence analysis, PCR fingerprint analysis of short tandemly repeated repetitive (STRR) sequences, temperature tolerances and toxin analysis to characterize 24 strains of this toxic cyanobacterium isolated from Thailand and Japan. All strains shared common morphological traits characteristic of C. raciborskii and showed high 16S rDNA sequence similarity, forming a defined cluster together with the reference strains from Australia. In particular, some of the Thai strains shared 99.9% to 100% similarity with the Australian strains. Various combinations of STRR primers revealed different and unique DNA band patterns among strains of C. raciborskii. The phylogenetic tree revealed two main clusters of C. raciborskii strains, the Thai/Japan-Shinobazugaike cluster (cluster I) and the Japan-Gonoike cluster (cluster II). Cluster I was further divided into two subclusters, A (only Thai strains) and B (one Thai strain and the Japan-Shinobazugaike strains). Thus, the results from 16S rDNA and STRR analyses showed no clear geographical distinction between Japanese and Thai strains and between Thai and Australian strains. Thai strains were separated into adaptive and non-adaptive groups to low temperature (15 and 17.5 degrees C) and Japanese strains were composed of only low-temperature-adaptive ones. The toxin cylindrospermopsin was detected in some strains of cluster I-A and in one strain of cluster II. We conclude that C. raciborskii is a species that has recently begun to invade, and a species with different physiological strains or ecotypes in temperature tolerance; the toxin is synthesized without any relation to phylogenetic or genetic clusters and to geography.