Design of a biodegradable carrier for the application of controller bacteria on air-water interfaces.

BACKGROUND: The formulation of a biodegradable carrier which effectively concentrates microorganisms on air-water interfaces is proposed. This avoids the dispersion of bacteria into the bulk liquid phase and at the same time prevents their sedimentation. This formulation can be used in biocontrol and bioremediation treatments where the target is at the position of the air-water interface, as in the case of the treatment of rice diseases caused by Sclerotium oryzae and Rhizoctonia complex. The carrier is an oil-in-water (O/W) emulsion which contains lecithin and chitosan in both phases at different proportions. In a stable formulation, bacteria that are adsorbed onto the surface of oil droplets are carried with them and flowed upward to the air-water interface, due to buoyancy forces. RESULTS: When using the biodegradable carrier, it is possible to recover at least 15-fold more bacteria from the air-water interface than in the case of using the aqueous formulation. CONCLUSION: The emulsion O/W is applied to the surface by dripping, resulting in a homogeneous two-dimensional film distribution. With this application device, the number of bacteria at the air-water interface is significantly increased. © 2019 Society of Chemical Industry.

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.

Kinetic Model of Photoautotrophic Growth of Chlorella sp. Microalga, Isolated from the Setúbal Lagoon.

In this work, a kinetic expression relating light availability in the culture medium with the rate of microalgal growth is obtained. This expression, which is valid for low illumination conditions, was derived from the reactions that take part in the light-dependent stage of photosynthesis. The kinetic expression obtained is a function of the biomass concentration in the culture, as well as of the local volumetric rate of absorption of photons, and only includes two adjustable parameters. To determine the value of these parameters and to test the validity of the hypotheses made, autotrophic cultures of the Chlorella sp. strain were carried out in a modified BBM medium at three CO2 concentrations in the gas stream, namely 0.034%, 0.34% and 3.4%. Moreover, the local volumetric rate of photon absorption was predicted based on a physical model of the interaction of the radiant energy with the suspended biomass, together with a Monte Carlo simulation algorithm. The proposed intrinsic expression of the biomass growth rate, together with the Monte Carlo radiation field simulator, are key to scale up photobioreactors when operating under low irradiation conditions, independently of the configuration of the reactor and of its light source.

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.

The local and observed photochemical reaction rates revisited.

In a broad sense, photochemical reactions proceed through pathways involving several reaction steps. The initiation step is the absorption of energy both by the reactant or sensitizer molecules and in some cases, by the catalyst, leading to intermediate products that ultimately give rise to stable end products. Preferably, the reaction rate expression is derived from a proposed mechanism together with sound simplifying assumptions; otherwise, it may be adopted on an empirical basis. Under a kinetic control regime, the rate expression thus obtained depends on the local rate of photon absorption according to a power law whose exponent very often ranges from one half to unity. The kinetic expression should be valid at every point of the reactor volume. However, due to radiation attenuation in an absorbing and/or scattering medium, the value of the photon absorption rate is always a function of the spatial position. Therefore, the overall photochemical reaction rate will not be uniform throughout the entire reaction zone, and the distinction between local and volume average photochemical reaction rates becomes mandatory. Experimental values of reaction rates obtained from concentration measurements performed in well-mixed reaction cells are, necessarily, average values. Consequently, for validation purposes, experimental results from these cells must be compared with volume averages of the mechanistically or empirically derived local reaction rate expressions. In this work it is shown that unless the rate is first order with respect to the photon absorption rate or the attenuation in the absorbing and/or scattering medium is kept very low, when the averaging operation is not performed, significant errors may be expected.