Cylindrospermopsin Production Research Articles

Multiplex PCR assay for Cylindrospermopsis raciborskii and cylindrospermopsin-producing cyanobacteria.

Water bodies are routinely monitored for the presence of potentially toxic cyanobacteria; however, the methodology for confirming toxicity is currently complex and expensive. Here we describe the application of gene-based technology to rapidly identify cylindrospermopsin-producing cyanobacteria, specifically, Cylindrospermopsis raciborskii. A multiplex polymerase chain reaction (PCR) test was developed that simultaneously identified polyketide synthase (pks) and peptide synthetase (ps) determinants associated with cylindrospermopsin production and distinguished C. raciborskii from other cylindrospermopsin-producing cyanobacteria of the species Anabaena bergii and Aphanizomenon ovalisporum, by targeting the rpoC1 gene. Twenty-one C. raciborskii, 5 A. bergii, 10 Aph. ovalisporum isolates and 3 environmental samples all yielded PCR results consistent with their toxicological status, as assessed by high-performance liquid chromatography coupled to mass spectrometry or matrix-assisted laser desorption ionization-time-of-flight mass spectrometry, and C. raciborskii was always correctly identified. The PCR test is a rapid, reliable, and economical way of assessing the toxic potential of cyanobacterial blooms formed by these organisms.

The Palm Island mystery disease 20 years on: a review of research on the cyanotoxin cylindrospermopsin.

Poisoning of humans resulting from consumption of water affected by the toxic cyanobacterium Cylindrospermopsis raciborskii was first reported almost 20 years ago from Palm Island, northern Queensland, Australia. Since that time a great deal has been learned about this organism and cylindrospermopsin (CYN), the toxin it produces. This article reviews the information now available to us. It summarizes aspects of the chemistry of the toxin-now known to be produced by some cyanobacterial species other than C. raciborskii-and its biosynthesis and chemical synthesis in vitro, as well as its detection and measurement by chemical and biological assay. Some of the factors affecting toxin production by cultured isolates of C. raciborskii are reviewed and the conditions that cause its release from the cells described. The occurrence of CYN in water bodies and the management strategies used to minimize the harmful effects of the toxin are outlined. These include a range of water-treatment practices now in place to remove CYN-producing organisms and/or to neutralize the toxin together with some management procedures that have been tried, with varying degrees of success, to prevent buildup of blooms of the offending organisms. Some of the public-health considerations arising from exposure to water supplies affected by CYN are summarized along with the risk factors and guidance values as they are currently applied. Among the more recent developments described are those that come from the application of molecular techniques for characterizing toxic and nontoxic strains and for exploring the genetic aspects of CYN production.

Phenotypical variation in a toxic strain of the phytoplankter, Cylindrospermopsis raciborskii (nostocales, cyanophyceae) during batch culture

A nonaxenic strain of Cylindrospermopsis raciborskii Woloszynska (AWT 205) was grown in batch culture, with and without nitrate as the primary N source. Rapid log-phase growth with nitrate was 1.0 doubling/day versus 0.3 doubling/day without nitrate. Cylindrospermopsin (CYN) production was measured by HPLC. The rate of intracellular CYN production matched cell division rate for both the diazotrophies at cell densities less than 10(7) cell/ml. At cell density > 10(7) cell/ml, additional resource limitation in batch culture slowed log-phase growth to 0.04 division/day and cell division and CYN production decoupled. Intracellular CYN concentration increased at a rate of 0.08 doubling/day, twice the cell division rate. Extracellular CYN as a proportion of the total CYN increased from 20% during the rapid growth phase, to 50% during the slow growth phase. The total CYN yield from cultures grown out to stationary phase (55 days) exceeded 2 mg CYN/I. C. raciborskii cells in log-phase growth, exposed to 1 ppm copper (as copper sulphate), lysed within 24 hours. After copper treatment, all CYN was in the filterable fraction. These findings imply that in naturally occurring blooms of C. raciborskii, the movement of intracellular CYN into solution will be the greatest during stationary phase, when intracellular concentrations are highest and cell lysis is more frequent. The application of algicides that promote cell lysis will exacerbate this effect.

The effect of temperature on growth and cylindrospermopsin content of seven isolates of Cylindrospermopsis raciborskii (Nostocales, Cyanophyceae) from water bodies in northern Australia

This study investigates the effect of temperature (range, 20–35°C) on the growth and cylindrospermopsin (CYL) content of seven isolates of Cylindrospermopsis raciborskii grown in batch culture. The concentration of CYL, measured late in the exponential growth phase for cultures grown at 25°C, ranged from below detection (< 2 × 10−5% freeze-dried weight) for isolates from Lake Julius (20°08′S; 139°44′E) and an aquaculture pond in Townsville (19°16′S; 146°49′E) up to 0.46% for an isolate from Solomon Dam, Palm Island (18°45′S; 146°35′E). Maximum growth rates for all isolates occurred at temperatures between 25 and 30°C. Only one isolate failed to grow at 20°C. For isolates containing undetectable concentrations of CYL, the production of this compound could not be induced by an increase in temperature. For strains producing detectable concentrations, there was a strong negative correlation between CYL content and temperature. While all the isolates grew well at 35°C, none produced any detectable amount of CYL. When cultures of isolates that normally produce CYL were transferred from 35°C to a lower temperature, production of CYL was restored. In the cultured isolates, less than 10% of the total CYL was released from exponentially growing cells. In one CYL-producing isolate, extracellular CYL accounted for c. 50% of the total in stationary phase (20 day old) cultures.