Double-stranded pBS plasmid DNA was irradiated with gamma rays at doses ranging from 1 to 12 kGy and electron beams from 1 to 10 kGy. Fragment-size distributions were determined by direct visualization, using atomic force microscopy with nanometer-resolution operating in non-tapping mode, combined with an improved methodology. The fragment distributions from irradiation with gamma rays revealed discrete-like patterns at all doses, suggesting that these patterns are modulated by the base pair composition of the plasmid. Irradiation with electron beams, at very high dose rates, generated continuous distributions of highly shattered DNA fragments, similar to results at much lower dose rates found in the literature. Altogether, these results indicate that AFM could supplement traditional methods for high-resolution measurements of radiation damage to DNA, while providing new and relevant information.
Persistent harmful scenarios associated with disposal of radioactive waste, high-background radiation areas and severe nuclear accidents are of great concern regarding consequences to both human health and the environment. Of particular concern is the extracellular DNA in aquatic environments contaminated by radiological substances. Strand breaks induced by radiation promote decrease in the transformation efficiency for extracellular DNA. The focus of this study is the quantification of DNA damage following long-term exposure (over one year) to low doses of natural uranium (an alpha particle emitter) to simulate natural conditions, since nothing is known about alpha radiation induced damage to extracellular DNA. A high-resolution Atomic Force Microscope was used to evaluate DNA fragments. Double-stranded plasmid pBS as a model for extracellular DNA was exposed to different amounts of natural uranium. It was demonstrated that low concentrations of U in water (50 to 150 ppm) produce appreciable numbers of double strand breaks, scaling with the square of the average doses. The importance of these findings for environment monitoring of radiological pollution is addressed.
Microcystins (MCs) are toxins profusely synthesized by cyanobacteria, causing livestock poisonings and endangering human health. We design and execute an experiment to investigate the attenuation (degradation) of microcystins by exposing them to gamma radiation and electron beams at doses of 0 (control), 3, 5, 10 and 15 kGy. The experimental conditions simulate microcystin contamination of aquatic environments; we thus consider (1) microcystins inside whole cells and extracellular dissolved in water, simulated by non-sonicated and sonicated cells, respectively, and (2) two acute microcystin concentrations within water. Toxicity tests of Microcystis aeruginosa detected immobilization (i.e., paralysis) of Ceriodaphniasilvestrii exposed to aqueous crude extracts of irradiated and non-irradiated M. aeruginosa (NPLJ-4 strain) at concentrations of 45 and 90 mg.L-1 (mg dry weight of freeze-dried material), and the results were analyzed using the Trimmed Spearman-Karber statistical program to obtain 48-h EC50, the average effective concentration causing immobility in 50% of organisms after 48 hours.
We conclude that electron beams are effective physical agents for toxin attenuation (degradation) and reach 100% effectiveness at 5 kGy and above; their efficiency is two orders of magnitude greater than that of gamma radiation.
This new body of information contributes to (1) remediating environmental water sources; (2) designing water/wastewater treatment facilities; (3) combatting chronic microcystin environmental contamination; and (4) inspiring further studies to promote the use of biomonitors (e.g., Cladocerans) to detect and evaluate microalgae contamination.
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