ERIC-PCR technique applied to monitoring and quantification of DNA damage during water disinfection process.

In this work, we propose a novel application of ERIC-PCR technique to study DNA damage after ultraviolet radiation (UV) and peracetic acid (PAA) treatment for water disinfection purpose. The efficacy of both treatments on E. coli suspension was evaluated by two approaches: through monitoring of inactivation by conventional culture technique, and by analyzing DNA damage with ERIC-PCR. All the experiments were carried out in a batch reactor, using three intensities of UV-C radiation (10.5, 4.2 and 2.1 mW/cm2) and different PAA concentrations (4 to 16 ppm). Both treatments produced bacterial inactivation in a dose-response fashion. Based on the results of bacterial count we obtained an index of inactivation (INACI). For each sample, DNA extraction was performed and evaluated by ERIC-PCR. Qualitative modifications were observed in ERIC-PCR band patterns for all the UV-C radiation intensities used, but no changes were detected at any of the PAA concentrations. The banding pattern modifications observed are consequence of the interruption of Taq polymerase enzyme amplification-activity, caused by the presence of alterations on the DNA structure (dimer and hydrates formation). Furthermore, an index was proposed to measure DNA damage (DNADI) regarding the changes in the relative optical density values of the amplification products. A linear correlation was obtained with a high correspondence between the inactivation index (INACI) and the DNA damage index (DNADI), that was expressed as DNADI = 0.05881×INACI. This approach proves that ERIC-PCR is a feasible and valuable tool for detecting and quantifying DNA damage and it may provide a useful strategy for bacterial identification, tracking changes in DNA and providing reliable and reproducible data.

Kinetic model of water disinfection using peracetic acid including synergistic effects.

The disinfection efficiencies of a commercial mixture of peracetic acid against Escherichia coli were studied in laboratory scale experiments. The joint and separate action of two disinfectant agents, hydrogen peroxide and peracetic acid, were evaluated in order to observe synergistic effects. A kinetic model for each component of the mixture and for the commercial mixture was proposed. Through simple mathematical equations, the model describes different stages of attack by disinfectants during the inactivation process. Based on the experiments and the kinetic parameters obtained, it could be established that the efficiency of hydrogen peroxide was much lower than that of peracetic acid alone. However, the contribution of hydrogen peroxide was very important in the commercial mixture. It should be noted that this improvement occurred only after peracetic acid had initiated the attack on the cell. This synergistic effect was successfully explained by the proposed scheme and was verified by experimental results. Besides providing a clearer mechanistic understanding of water disinfection, such models may improve our ability to design reactors.

A novel approach to explain the inactivation mechanism of Escherichia coli employing a commercially available peracetic acid.

The chemical inactivation of Escherichia coli employing a commercial mixture of peracetic acid (PAA) was studied. For this purpose, experiments were carried out using dilutions of the unmodified mixture, and also the same mixture but altered with hydrogen peroxide (HP) previously inhibited. Also, these results were compared to those obtained before employing HP alone. It was found that the mixture is much more efficient than HP and PAA acting separately. Furthermore, it was found that PAA without HP is much more efficient than HP alone. A plausible explanation is presented. The homolysis of PAA would give rise to a chain reaction that generates a significant number of highly oxidizing radicals. An attacking scheme to bacteria in two stages is proposed, where the initial step, mainly caused by PAA, is very fast and eliminates some specific components of the bacteria that would otherwise inhibit the parallel action of HP. Thereafter, the emergence of a potentiating synergetic action of the second oxidant seems to be immediately unveiled.