Impact of pulsed electric fields and mechanical compressions on the permeability and structure of Chlamydomonas reinhardtii cells.

Current research findings clearly reveal the role of the microalga's cell wall as a key obstacle to an efficient and optimal compound extraction. Such extraction process is therefore closely related to the microalga species used. Effects of electrical or mechanical constraints on C. reinhardtii's structure and particularly its cell wall and membrane, is therefore investigated in this paper using a combination of microscopic tools. Membrane pores with a radius between 0.77 and 1.59 nm were determined for both reversible (5 kV∙cm-1) and irreversible (7 kV∙cm-1) electroporation with a 5 µs pulse duration. Irreversible electroporation with longer pulses (10 µs) lead to the entry of large molecules (at least 5.11 nm). Additionally, for the first time, the effect of pulsed electric fields on the cell wall was observed. The combined electrical and mechanical treatment showed a significant impact on the cell wall structure as observed under Transmission Electron Microscopy. This treatment permits the penetration of larger molecules (at least 5.11 nm) within the cell, shown by tracking the penetration of dextran molecules. For the first time, the size of pores on the cell membrane and the structural changes on the microalgae cell wall induced by electrical and mechanical treatments is reported.

Structural changes of Chlamydomonas reinhardtii cells during lipid enrichment and after solvent exposure.

Data are related to Confocal Laser Scanning Microscopy (CLSM) observations of lipid-enriched Chlamydomonas reinhardtii cells under different conditions. Firstly, the impact of stress conditions (nitrogen starvation) on the cell wall structure is assessed. Secondly is described the effect of solvents, in the context of lipid extraction, on the microalga's cell, particularly its lipid droplets, in the perspective of understanding the mechanisms behind solvent extraction of lipids. Furthermore, the role of the cell wall as a barrier to the solvent extraction action is highlighted.

Understanding the mechanisms of lipid extraction from microalga Chlamydomonas reinhardtii after electrical field solicitations and mechanical stress within a microfluidic device.

One way envisioned to overcome part of the issues biodiesel production encounters today is to develop a simple, economically viable and eco-friendly process for the extraction of lipids from microalgae. This study investigates the lipid extraction efficiency from the microalga Chlamydomonas reinhardtii as well as the underlying mechanisms. We propose a new methodology combining a pulsed electric field (PEF) application and mechanical stresses as a pretreatment to improve lipid extraction with solvents. Cells enriched in lipids are therefore submitted to electric field pulses creating pores on the cell membrane and then subjected to a mechanical stress by applying cyclic pressures on the cell wall (using a microfluidic device). Results showed an increase in lipid extraction when cells were pretreated by the combination of both methods. Microscopic observations showed that both pretreatments affect the cell structure. Finally, the dependency of solvent lipid extraction efficiency with the cell wall structure is discussed.

Understanding the fundamental mechanisms of biofilms development and dispersal: BIAM (Biofilm Intensity and Architecture Measurement), a new tool for studying biofilms as a function of their architecture and fluorescence intensity.

Confocal laser scanning microscopy (CLSM) is one of the most relevant technologies for studying biofilms in situ. Several tools have been developed to investigate and quantify the architecture of biofilms. However, an approach to quantify correctly the evolution of intensity of a fluorescent signal as a function of the structural parameters of a biofilm is still lacking. Here we present a tool developed in the ImageJ open source software that can be used to extract both structural and fluorescence intensity from CLSM data: BIAM (Biofilm Intensity and Architecture Measurement). This is of utmost significance when studying the fundamental mechanisms of biofilm growth, differentiation and development or when aiming to understand the effect of external molecules on biofilm phenotypes. In order to provide an example of the potential of such a tool in this study we focused on biofilm dispersion. cis-2-Decenoic acid (CDA) is a molecule known to induce biofilm dispersion of multiple bacterial species. The mechanisms by which CDA induces dispersion are still poorly understood. To investigate the effects of CDA on biofilms, we used a reporter strain of Escherichia coli (E. coli) that expresses the GFPmut2 protein under control of the rrnBP1 promoter. Experiments were done in flow cells and image acquisition was made with CLSM. Analysis carried out using the new tool, BIAM, indicates that CDA affects the fluorescence intensity of the biofilm structures as well as biofilm architectures. Indeed, our results demonstrate that CDA removes more than 35% of biofilm biovolume and suggest that it results in an increase of the biofilm's mean fluorescence intensity (MFI) by more than 26% compared to the control biofilm in the absence of CDA.

Purification of organic acids by chromatography with strong anionic resins: Investigation of uptake mechanisms.

Bio-based organic acids are promising renewable carbon sources for the chemical industry. However energy-consuming purification processes are used, like distillation or crystallization, to reach high purities required in some applications. That is why preparative chromatography was studied as an alternative separation technique. In a previous work dealing with the purification of lactic, succinic and citric acids, the Langmuir model was insufficient to explain the elution profiles obtained with a strong anionic resin. Consequently the Langmuir model was coupled with a usual ion-exchange model to take into account the retention of their conjugate bases (<2%), which are commonly neglected at low pH (<1.5). Elution simulations with both uptake mechanisms fitted very well with experimental pulse tests. Only two parameters were optimized (equilibrium constant of acid uptake and ion-exchange selectivity coefficient of conjugate base) and their value were coherent with experimental and resin suppliers' data. These results confirmed that the singular tailing and apparent delay observed with succinic and citric acids can be explained by the high affinity of succinate and citrate for resin cationic sites. The model was implemented in a preparative chromatography simulation program in order to optimize operating parameters of our pilot-scale ISMB unit (Improved Simulated Moving Bed). The comparison with experimental ISMB profiles was conclusive.

Nitrogen and phosphate removal from wastewater with a mixed microalgae and bacteria culture.

Microalgae are able to convert nutrients (nitrogen and phosphorus) from wastewater into biomass and bio-products, thus improving the sustainability of wastewater treatment. In High Rate Algal Ponds (HRAP), biomass productivity and water treatment efficiency are highly dependent on environmental parameters such as temperature, light intensity and photoperiod. The influence of temperature and photoperiod on biomass productivity and the removal of dissolved nitrogen and phosphorus from municipal wastewater by a native microalgae-bacteria consortium was assessed in batch cultures in view of the development of an HRAP at a larger scale. Temperature affected the growth rate and microalgae biomass production as well as ammonium and phosphate removal rates. At the temperatures 15 and 25 °C, the average total nitrogen and phosphorus removal extents ranged from 72 to 83% and 100% respectively. Additionally 33.0 ± 0.1% of the total nitrogen was eliminated by stripping at 25 °C, and 50 ± 2% was assimilated by the microorganisms under all conditions tested.

Modeling the continuous lactic acid production process from wheat flour.

A kinetic model of the simultaneous saccharification, protein hydrolysis, and fermentation (SSPHF) process for lactic acid production from wheat flour has been developed. The model describes the bacterial growth, substrate consumption, lactic acid production, and maltose hydrolysis. The model was fitted and validated with data from SSPHF experiments obtained under different dilution rates. The results of the model are in good agreement with the experimental data. Steady state concentrations of biomass, lactic acid, glucose, and maltose as function of the dilution rate were predicted by the model. This steady state analysis is further useful to determine the operating conditions that maximize lactic acid productivity.

Magnesium Uptake by the Green Microalga Chlorella vulgaris in Batch Cultures.

The accumulation (internal and superficial distribution) of magnesium ions (Mg(2+)) by the green freshwater microalga Chlorella vulgaris (C. vulgaris) was investigated under autotrophic culture in a stirred photobioreactor. The concentrations of the three forms of Mg(2+) (dissolved, extracellular, and intracellular) were determined with atomic absorption spectroscopy during the course of C. vulgaris growth. The proportions of adsorbed (extracellular) and absorbed (intracellular) Mg(2+) were quantified. The concentration of the most important pigment in algal cells, chlorophyll a, increased over time in proportion to the increase in the biomass concentration, indicating a constant chlorophyll/biomass ratio during the linear growth phase. The mean-average rate of Mg(2+) uptake by C. vulgaris grown in a culture medium starting with 16 mg/l of Mg(2+) concentration was measured. A clear relationship between the biomass concentration and the proportion of the Mg(2+) removal from the medium was observed. Of the total Mg(2+) present in the culture medium, 18% was adsorbed on the cell wall and 51% was absorbed by the biomass by the end of the experiment (765 h). Overall, 69% of the initial Mg(2+) were found to be removed from the medium. This study supported the kinetic model based on a reversible first-order reaction for Mg(2+) bioaccumulation in C. vulgaris, which was consistent with the experimental data.

Nonlinear predictive control for maximization of CO2 bio-fixation by microalgae in a photobioreactor.

In the framework of environment preservation, microalgae biotechnology appears as a promising alternative for CO2 mitigation. Advanced control strategies can be further developed to maximize biomass productivity, by maintaining these microorganisms in bioreactors at optimal operating conditions. This article proposes the implementation of Nonlinear Predictive Control combined with an on-line estimation of the biomass concentration, using dissolved carbon dioxide concentration measurements. First, optimal culture conditions are determined so that biomass productivity is maximized. To cope with the lack of on-line biomass concentration measurements, an interval observer for biomass concentration estimation is built and described. This estimator provides a stable accurate interval for the state trajectory and is further included in a nonlinear model predictive control framework that regulates the biomass concentration at its optimal value. The proposed methodology is applied to cultures of the microalgae Chlorella vulgaris in a laboratory-scale continuous photobioreactor. Performance and robustness of the proposed control strategy are assessed through experimental results.

Detoxifying emulsion for overdosed aspirin intoxication.

Aspirin overdose could lead to intoxication, with the clinical manifestations of vomit, pulmonary edema and severe dyspnea. Stomach washing, emetics and activated charcoal are the common treatments with a limited efficiency for the intoxication. In this study, an active emulsion for aspirin intoxication was prepared with the detoxifying efficiency of 100% in less than 15 min, with the conditions of dodecane used as the oil phase, 8% Abil EM90 as the surfactant and 0.1 mol/L sodium hydroxide as the inner aqueous phase in a volume ratio of 2 between internal aqueous phase and the oil phase.