Evaluation of UV-C induced changes in Escherichia coli DNA using repetitive extragenic palindromic-polymerase chain reaction (REP-PCR).

Ultraviolet radiation is an efficient inactivation method for a broad range of bacteria, viruses and parasites. Inactivation of microorganisms by UV-B and UV-C radiation is driven through modifications in their genomic DNA, being the most stable DNA-lesions different kinds of pyrimidine dimers (PDs) (e.g., cyclobutane pyrimidine dimers (CPDs) and other photoproducts). Taking into account that these modifications inhibit the DNA polymerization in vivo as well as in vitro, in the present work the usefulness of the REP-PCR assay to detect UV-induced changes in the Escherichia coli DNA was evaluated. In vitro amplification of DNA extracted at different times after UV treatment showed a disappearance of amplicons of higher size as time of treatment increases. When the bacteria were let to progress through their dark repair process, modifications in the electrophoretic patterns by REP-PCR were observed again. Amplified bacterial DNA tended to recover the profile showed at the beginning of treatment. In addition, the reappearance of bands of higher molecular size was associated to an increase in their signal intensity probably due to a higher amplification rate. Results of REP-PCR were correlated to the colony-forming ability of E. coli. It was concluded that REP-PCR appears as a rapid, robust, useful complementary methodology to monitor the impact of UV irradiation--at a molecular level--on the inactivation and the mechanisms of repair, applicable on a broad spectrum of microorganisms.

Stratification of the radiation field inside a photobioreactor during microalgae growth.

Light availability is a main issue in autotrophic growth of photosynthetic microorganisms. The change of the suspended cells concentration and that of their chlorophylls content during microalgal growth alters the optical properties of the aqueous suspension. This brings about changes in the properties of the radiation field inside the reactor. In this work, we have computed the evolution in time of the local volumetric rate of absorption of photons inside a photobioreactor by means of a Monte Carlo simulation algorithm previously developed. From this study, we have computed two operational variables that are useful tools for the analysis, performance comparison, optimization and scaling up of photobioreactors: the average rate of photon absorption and the volumetric distribution function of photons absorption rates. Based on these two variables, it is possible to systematically quantify the degree of stratification of the culture medium, which is a decisive aspect that hampers the reactor efficiency regarding the energy usage for biomass production.

Analysis and design of photobioreactors for microalgae production I: method and parameters for radiation field simulation.

Having capabilities for the simulation of the radiation field in suspensions of microalgae constitutes a great asset for the analysis, optimization and scaling-up of photobioreactors. In this study, a combined experimental and computational procedure is presented, specifically devised for the assessment of the coefficients of absorption and scattering, needed for the simulation of such fields. The experimental procedure consists in measuring the radiant energy transmitted through samples of suspensions of microalgae of different biomass concentrations, as well as the forward and backward scattered light. At a microscopic level, suspensions of microalgae are complex heterogeneous media and due to this complexity, in this study they are modeled as a pseudocontinuum, with centers of absorption and scattering randomly distributed throughout its volume. This model was tested on suspensions of two algal species of dissimilar cell shapes: Chlorella sp. and Scenedesmus quadricauda. The Monte Carlo simulation algorithm developed in this study, when used as a supporting subroutine of a main optimization program based on a genetic algorithm, permits the assessment of the physical parameters of the radiation field model. The Monte Carlo algorithm simulates the experiments, reproducing the events that photons can undergo while they propagate through culture samples or at its physical boundaries.

Analysis and design of photobioreactors for microalgae production II: experimental validation of a radiation field simulator based on a Monte Carlo algorithm.

In a previous study, we developed a methodology to assess the intrinsic optical properties governing the radiation field in algae suspensions. With these properties at our disposal, a Monte Carlo simulation program is developed and used in this study as a predictive autonomous program applied to the simulation of experiments that reproduce the common illumination conditions that are found in processes of large scale production of microalgae, especially when using open ponds such as raceway ponds. The simulation module is validated by comparing the results of experimental measurements made on artificially illuminated algal suspension with those predicted by the Monte Carlo program. This experiment deals with a situation that resembles that of an open pond or that of a raceway pond, except for the fact that for convenience, the experimental arrangement appears as if those reactors were turned upside down. It serves the purpose of assessing to what extent the scattering phenomena are important for the prediction of the spatial distribution of the radiant energy density. The simulation module developed can be applied to compute the local energy density inside photobioreactors with the goal to optimize its design and their operating conditions.