Growth inhibition of Microcystis aeruginosa by a glycolipid-type compound from Bacillus subtilis C1.

We attempted to identify the compound responsible for the growth inhibition of Microcystis aeruginosa occurring when a culture broth of Bacillus subtilis C1 was added to the medium. The active compound was purified from B. subtilis C1 culture broth by adsorption chromatography and HPLC, and was identified as a type of glycolipid based on 1H NMR and MS analyses. The purified active compound completely inhibited the growth of M. aeruginosa at a concentration of 10 microgram/ml. This is the first report of a glycolipid produced by a Bacillus strain that has potential as an agent for the selective control of bloom-forming M. aeruginosa.

Alternative alert system for cyanobacterial bloom, using phycocyanin as a level determinant.

Chlorophyll a concentration and cyanobacterial cell density are regularly employed as dual criteria for determinations of the alert level for cyanobacterial bloom. However, chlorophyll a is not confined only to the cyanobacteria, but is found universally in eukaryotic algae. Furthermore, the determination of cyanobacterial cell counts is notoriously difficult, and is unduly dependent on individual variation and trained skill. A cyanobacteria-specific parameter other than the cell count or chlorophyll a concentration is, accordingly, required in order to improve the present cyanobacterial bloom alert system. Phycocyanin has been shown to exhibit a strong correlation with a variety of bloom-related factors. This may allow for the current alert system criteria to be replaced by a three-stage alert system based on phycocyanin concentrations of 0.1, 30, and 700 microg/L. This would also be advantageous in that it would become far more simple to conduct measurements without the need for expensive equipment, thereby enabling the monitoring of entire lakes more precisely and frequently. Thus, an alert system with superior predictive ability based on high-throughput phycocyanin measurements appears feasible.

Simple method for a cell count of the colonial Cyanobacterium, Microcystis sp.

The cell counting of colonial Microcystis spp. is a rather difficult and error-prone proposition, as this genus forms irregularly-shaped and irregularly-sized colonies, which are packed with cells. Thus, in order to facilitate a cell count, four methods of dividing the colonies into single cells were compared, including vortexing, sonication, TiO2 treatment, and boiling. As a result, the boiling method was determined to generate the greatest number of single cells from a colony, and all colonies were found to have divided completely after only 6 min of treatment. Furthermore, no significant cell destruction, which might alter the actual cell density, was detected in conjunction with the boiling method (P = 0.158). In order to compute the cell number more simply, the relationship between the colony size and the cell number was determined, via the boiling method. The colony volume, rather than the area or diameter was correlated more closely with the cell number (r2 = 0.727), thereby suggesting that the cell numbers of colonial Microcystis sp. can also be estimated effectively from their volumes.

Dynamic variation of toxic and non-toxic Microcystis proportion in the eutrophic Daechung Reservoir in Korea.

This study was conducted to determine the environmental factors affecting the level of potentially toxic Microcystis. The long-term tendencies of temperature, precipitation, and water quality factors were analyzed to determine the environmental characteristics of the Daechung Reservoir in Korea, and water samples were directly collected to analyze the dynamics of toxic and non-toxic Microcystis at weekly intervals from May to October 2012. Microcystis was the dominant genus during the study period, and it was composed of potentially toxic and non-toxic Microcystis. The fraction of potentially toxic Microcystis ranged from 6.0% to 61.1%. The amount of toxic Microcystis was highly related to the intracellular microcystin concentration (r = 0.760, P < 0.01). Therefore, the fraction of potentially toxic Microcystis is an important concern in Microcystis blooming because the intracellular microcystin concentration may reflect microcystin levels in the water. The prevalence of potentially toxic Microcystis was highly related to water temperature in Daechung Reservoir (r = 0.585, P < 0.01). Thus, temperature increase during Microcystis blooming may lead to more frequent toxic Microcystis blooms in eutrophic water bodies.

Annual variation of Microcystis genotypes and their potential toxicity in water and sediment from a eutrophic reservoir.

The relative genetic diversity of microcystin-producing Microcystis in the water and sediment of the Daechung Reservoir, Korea, was investigated over an entire year, including the cyanobacterial bloom season. The cells of potentially toxic Microcystis strains containing mcyJ genotypes and cells containing the genus-specific cpcBA gene were quantified by a real-time PCR. The ratio of cells with mcyJ genotypes to the total Microcystis population in the water body was the highest (68.3%) in August when the cyanobacterial bloom reached its peak and the microcystin concentration in the water began to increase. A denaturing gradient gel electrophoresis profile analysis of the mcyJ genotypes performed to monitor any changes in the toxic Microcystis population showed the appearance of new genotypes and the disappearance of existing genotypes in the reservoir water collected during the summer months, when compared with the profile for the samples collected in spring and autumn. However, very little change was observed over the course of the year as regards the population diversity of the sediment samples.

Selective control of cyanobacteria by surfactin-containing culture broth of Bacillus subtilis C1.

Of several types of chemical surfactants and biosurfactants, only the culture broth of Bacillus subtilis C1 containing surfactin at 10 mg l(-1) completely inhibited the growth of Microcystis aeruginosa, a bloom-forming cyanobacterium in highly eutrophic lakes. The broth with 10 mg surfactin l(-1) also removed 85% of the maximally grown M. aeruginosa (chlorophyll-a concentration, 1000 microg l(-1)) within 2 d, and the removal efficiency was enhanced by Ca2+ over 1 mM. The growth of Anabaena affinis, another bloom-forming cyanobacterium, was also inhibited about 70% with surfactin at 10 mg l(-1) broth. However, the effect of the broth was delayed over 3 d in the green algae, Chlorella vulgaris and Scenedesmus sp., and was negligible in a diatom, Navicula sp., indicating the potential for the selective control of cyanobacterial blooms.

Growth inhibition of Cyanobacteria by ultrasonic radiation: laboratory and enclosure studies.

The growth of Microcystis aeruginosa UTEX 2388 was repressed by ultrasonic radiation and resulted in an increased chlorophyll a content and cell size, suggesting the inhibition of cell division. However, growth was recovered immediately after the interruption of ultrasonication. In addition to the disruption of gas vesicles, other mechanisms of growth inhibition were also investigated. Although free radicals were produced by ultrasonication and hydrogen peroxide, the resulting lipid peroxidation in the cells was not comparable, indicating minimal damage by the free radicals. Ultrasonic radiation late in the day was found to be most effective in reducing the growth rate of M. aeruginosa, and this timing also corresponded to the phase of daily cell division. In an enclosure experiment, ultrasonic radiation reduced the pH, DO, total nitrogen, and total phosphorus, whereas it increased the water temperature, conductivity, and orthophosphate concentration. The algal cell density and chlorophyll a concentration drastically decreased after 3 d of ultrasonication, plus the cyanobacterial proportion was selectively reduced as compared to other algal species. Accordingly, ultrasonic radiation would appear to have considerable potential as an effective control method for cyanobacterial blooms.