A primary-structure analysis of the 16s rRNA gene was performed with 10 strains representing five described and one unidentified species of the genus Microcystis. The phylogenies determined illustrate the evolutionary affiliations among Microcystis strains, other cyanobacteria, and related plastids and bacteria. A cluster of 10 strains that included hepatotoxic isolates identified as Microcystis aeruginosa formed a monophyletic group. However, the genus Microcystis appeared to be polyphyletic and contained two strains that clustered with unicellular cyanobacteria belonging to the genus Synechococcus. The clustering of related Microcystis strains, including strains involved in the production of the cyclic peptide toxin microcystin, was consistent with cell morphology, gas vacuolation, and the low G+C contents of the genomes. The Microcystis lineage was also distinct from the lineage containing the unicellular genus Synechocystis and the filamentous, heterocystforming genus Nostoc. The secondary structure of a Microcystis 16s rRNA molecule was determined, and genus-specific sequence signatures were used to design primers that permitted identification of the potentially toxic cyanobacteria belonging to the genus Microcystis via DNA amplification.The Cyanobacteria is a diverse bacterial phylum with respect to form, function, and habitat. On the basis of microfossil and geochemical evidence the origin of cyanobacterium-like organisms has been dated to the late Precambrian era. On the basis of the results of phylogenetic studies workers have inferred that the cyanobacterial phylum is 1 of the 11 bacterial phyla (32,42). The possibility that eukaryotic chloroplasts arose from a cyanobacterial ancestor by a symbiotic event has also been inferred on the basis of molecular data (8, 38).Members of the genus Microcystis are a major cause of freshwater noxious cyanobacterial blooms, which have a broad geographical distribution. The microcystins, a family of cyclic heptapeptide toxins that are not synthesized ribosomally (3) and are produced by most members of this genus, cause acute hepatotoxicity in agricultural livestock. Recent epidemiological data have also linked chronic subacute consumption of microcystins to human liver tumor promotion (5). The increasing occurrence of Microcystis blooms in major sources of human drinking water makes identification and prediction of these toxic blooms very important.Due to the variably expressed and minor morphological and developmental characteristics used for identification, classification of cyanobacterial strains at the genus or species level may be ambiguous (29), particularly when laboratory cultures and environmental isolates are compared (7). The current cyanobacterial taxonomy does not provide an unequivocal system for identification of the toxigenic and bloom-forming genus Microcystis (20). Depending on the taxonomic treatises used for classification, which differ in their emphasis on the cell size, shape, buoyancy, and toxicity of the planktonic, freshwater cyanobacteria, d...
Cylindrospermopsis raciborskii, a tropical blooming species of cyanobacterium (blue-green alga), was isolated from the domestic water supply reservoir on Palm Island, a continental island off the tropical northeast coast of Australia. This species, not previously known to be toxic, was shown to be severely hepatotoxic for mice. The 50% lethal dose at 24 h after injection was found to be 64 ± 5 mg of freeze-dried culture per kg of mouse. The principal lesion produced was centrilobular to massive hepatocyte necrosis, but various degrees of injury were also seen in the kidneys, adrenal glands, lungs, and intestine. The possible implication of this finding in relation to an incident of hepatoenteritis in humans living on the island is discussed.
We mapped the distribution of mat forming cyanobacteria along the thermal gradient from 30 to 80 degrees C, in nine hot spring districts in northern Thailand. Nineteen genera and 36 species were identified by morphometric analysis. Water temperature was the predominant determinant of community structure in the springs. The diversity of cyanobacterial morphotypes fell as temperature increased. Water chemistry (pH, alkalinity and ammonia concentration) was a much weaker descriptor of the floral similarity between the springs. The morphotypes which dominated all springs were Synechococcus lividus and Synechococcus sp. (>40 and <80 degrees C) and Phormidium boryanum (>30 and <60 degrees C). The occurrence of Synechococcus lividus and Synechococcus sp. in every spring at 70 degrees C or more, implied there was no regional barrier to the distribution of these highly thermophilic taxa. Conversely, there were regional differences in the diversity of mat communities growing below 60 degrees C. The most depauperate flora were in the northernmost springs (SKP, TPN, PD, JS) and the springs further south around Chiang Mai had more diverse flora, suggestive of barriers to the dispersal of some taxa. More discriminating analyses using molecular tools will be required to determine whether the ubiquitous distribution Synechoccoccus morphotypes above 60 degrees C masks a genotypic diversity, comparable to the morphotype diversity observed below 60 degrees C.
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.
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