Respirometry as a tool to quantify kinetic parameters of microalgal mixotrophic growth.

Modeling microalgal mixotrophy is challenging, as the regulation of algal metabolism is affected by many environmental factors. A reliable tool to simulate microalgal behavior in complex systems, such as wastewaters, may help in setting the proper values of operative variables, provided that model parameters have been properly evaluated. In this work, a new respirometric protocol is proposed to quickly obtain the half-saturation constant values for several nutrients. The protocol was first verified for autotrophic exploitation of ammonium and phosphorus (Monod kinetics), as well as of light intensity (Haldane model), further elaborated on specific light supply basis. It was then applied to measure the kinetic parameters of heterotrophic growth. The half-saturation constants for nitrogen and phosphorus resulted comparable with autotrophic ones. The dependence on acetate and dissolved oxygen concentration was assessed. Mixotrophy was modeled as the combination of autotrophic/heterotrophic reactions, implemented in AQUASIM, and validated on batch curves with/without bubbling, under nutrient limitation, and different light intensities. It was shown that the reliability of the proposed respirometric protocol is useful to measure kinetic parameters for nutrients, and therefore to perform bioprocess simulation.

Exploiting symbiotic interactions between Chlorella protothecoides and Brevundimonas diminuta for an efficient single-step urban wastewater treatment.

The application of microalgal bacteria consortia to the treatment of wastewater is receiving increasing attention, meeting the demand for new green and efficient technologies for water remediation. The specificity of the consortium, however, may strongly affect the performance of the treatment. In fact, even though a general exploitation of the O2/CO2 exchange between microalgae and bacteria is effective, some specific interactions may increase the pollutant removal. With this aim, the co-cultivation of Chlorella protothecoides and Brevundimonas diminuta was tested, with particular attention to the removal capability of nitrogen, phosphorus and chemical oxygen demand (COD) from wastewater. Batch experiments were carried out both for the consortium and, separately, for the bacteria and microalgae alone, in order to compare their performances. B. diminuta showed a remarkable capability for removing organic substances and transforming organic nitrogen to ammonium. C. protothecoides efficiently removed nitrogen and phosphorus. As the specific growth rates of the two organisms are different, the co-cultivation was also carried out also in a continuous system, and the effect of hydraulic retention time (HRT) on the steady-state biomass concentration and nutrient removal efficiency was verified. Residence time was found as the main operating variable for obtaining a significant reduction of pollutants from wastewater.

Light intensity affects the mixotrophic carbon exploitation in Chlorella protothecoides: consequences on microalgae-bacteria based wastewater treatment.

Microalgal-bacteria consortia application on wastewater treatment has been widely studied, but a deeper comprehension of consortium interactions is still lacking. In particular, mixotrophic exploitation of organic compounds by microalgae affects gas (CO2 and O2) exchange between microalgae and bacteria, but it is not clear how environmental conditions may regulate algal metabolism. Using a respirometric-based protocol, we evaluated the combined effect of organic carbon and light intensity on oxygen production and consumption by C. protothecoides, and found that the chemical oxygen demand (COD) was not consumed when incident light increased. Batch experiments under different incident lights, with C. protothecoides alone and in consortium with activated sludge bacteria, confirmed the results obtained by respirometry. Continuous system experiments testing the combined effects of light intensity and residence time confirmed that, under limiting light, mixotrophy is preferred by C. protothecoides, and the nutrient (COD, N, P) removal capability of the consortium is enhanced.

Modeling the oxygen inhibition in microalgae: An experimental approach based on photorespirometry.

Microalgae cultivation has been the object of relevant interest for many industrial applications. Where high purity of the biomass/product is required, closed photobioreactors (PBRs) appear to be the best technological solution. However, as well as cost, the major drawback of closed systems is oxygen accumulation, which is well known to be responsible for growth inhibition. Only a few quantitative approaches have attempted to describe and model oxygen inhibition, which is the result of different biological mechanisms. Here, we have applied a photorespirometric protocol to assess and quantify the effect of high oxygen concentration on photosynthetic production rate. In particular, the effects of light intensity and biomass concentration were assessed, resulting in different maximum inhibitory oxygen concentrations. Literature models available were found not to fully represent experimental data as a function of concentration and light. Accordingly, a new formulation was proposed and validated to describe the photosynthetic rate as a function of external oxygen concentration.

An Artificial Cough Maneuver to Remove Secretions From Below the Endotracheal Tube Cuff.

BACKGROUND: Endotracheal suctioning is mandatory to prevent complications caused by the retention of tracheal secretions. Endotracheal suctioning is often performed late, when patients show signs of respiratory and hemodynamic alterations. We conceived a prototype device that, when synchronized with the ventilator, automatically removes secretions collected below the endotracheal tube (ETT) cuff, thus avoiding endotracheal suctioning. The aim of our investigation was to assess the performance of this novel prototype in vitro. METHODS: Three studies were performed to examine the characteristics of the prototype. We tested device's ability to generate an effective artificial cough flow (artificial cough maneuver) > 1 L/s by rapidly deflating the ETT cuff within the time of a sustained inflation (at 30 and at 40 cm H2O) (cough flow study). We also tested the prototype's ability to remove the fluid positioned below the ETT cuff using saline dye (fluid removal study), and to prevent the aspiration of saline dye from above the ETT cuff during the deflation phase of the ETT cuff (aspiration study). The trachea model was positioned at 45° in the aspiration study, and horizontally in the other two studies. RESULTS: In the cough flow study, the prototype provided an effective artificial cough maneuver, with a mean ± SD of 1.78 ± 0.19 L/s (range, 1.42-2.14 L/s). The tracheal pressure after ETT cuff deflation never decreased below the PEEP level. In the fluid removal study, the prototype cleared the fluid from below the ETT cuff and the experimental trachea. No fluid was aspirated from the area above the ETT cuff toward the lower airways. CONCLUSIONS: We conceived an system capable of automatically expelling fluid from below the ETT cuff outside an experimental trachea by generating an artificial cough maneuver. This system may decrease the use of endotracheal suctioning and its complications. Future in vivo studies are needed to confirm this first in vitro evaluation.

Microalgae-bacteria gas exchange in wastewater: how mixotrophy may reduce the oxygen supply for bacteria.

Microalgae-bacteria consortia application to wastewater treatment is considered as a potential and cheap strategy towards a self-sustaining oxygen-carbon dioxide gas exchange. However, microalgae can also carry out mixotrophy, thus reducing the net oxygen production, due to consumption of organic substrates. In this work, respirometric tests were used to quantify the oxygen reduction in the presence of biodegradable COD (chemical oxygen demand), which resulted up to 70%, depending on the biodegradability of the carbon substrate. The implication of mixotrophic metabolism on nutrient removal in urban wastewater was also measured by co-cultivating C. protothecoides with bacteria from activated sludge. To better understand the contribution of different populations, ad hoc experiments under controlled conditions were designed to quantify the nutrient consumption of bacteria and microalgae. Microalgae and bacteria were cultivated together and separately, with and without external bubbling, so to better ascertain the specific role of gas production and nutrient removal. Results showed that microalgae can remove up to 100 and 85% of P and N respectively, but the contribution on COD consumption may affect the net O2 supply to heterotrophic bacteria. However, a mutual COD consumption by microalgae and bacteria was proved by both experimental growth curves and mass balance application, based on stoichiometry experimentally adjusted.