Image analysis of light distribution in a photobioreactor

Light intensity is a crucial factor that determines the growth of photosynthetic cells. This study analyzed the light distribution in a photobioreactor by processing images, captured with a digital camera, of a rectangular photobioreactor containing Synechococcus sp. PCC6801 as a model microorganism. The gray-scale images obtained clearly demonstrate the variation of the light-distribution profiles upon changing cell concentrations and external light intensity. Image-processing techniques were also used to predict the cell density in the photobioreactor. By analyzing the digitized image data with a neural network model, we were able to predict the cell concentrations in the photobioreactor with a <5% error.     

Enclosed outdoor photobioreactors: light regime,photosynthetic efficiency,scale-up,and future prospects

Enclosed outdoor photobioreactors need to be developed and designed for large-scale production of phototrophic microorganisms. Both light regime and photosynthetic efficiency were analyzed in characteristic examples of state-of-the-art pilot-scale photobioreactors. In this study it is shown that productivity of photobioreactors is determined by the light regime inside the bioreactors. In addition to light regime, oxygen accumulation and shear stress limit productivity in certain designs. In short light-path systems, high efficiencies, 10% to 20% based on photosynthetic active radiation (PAR 400 to 700 nm), can be reached at high biomass concentrations (>5 kg [dry weight] m(-3)). It is demonstrated, however, that these and other photobioreactor designs are poorly scalable (maximal unit size 0.1 to 10 m(3)), and/or not applicable for cultivation of monocultures. This is why a new photobioreactor design is proposed in which light capture is physically separated from photoautotrophic cultivation. This system can possibly be scaled to larger unit sizes, 10 to >100 m(3), and the reactor liquid as a whole is mixed and aerated. It is deduced that high photosynthetic efficiencies, 15% on a PAR-basis, can be achieved. Future designs from optical engineers should be used to collect, concentrate, and transport sunlight, followed by redistribution in a large-scale photobioreactor.     

An integrated solar and artificial light system for internal illumination of photobioreactors

Exploitation of photosynthetic cells for the production of useful metabolites requires efficient photobioreactors. Many laboratory scale photobioreactors have been reported but most of them are extremely difficult to scale up. Furthermore, the use of open ponds and outdoor tubular photobioreactors is limited by the requirement for large spaces and the difficulty in maintaining sterile conditions. In view of this, we have designed and constructed an internally illuminated stirred tank photobioreactor. The photobioreactor is simple, heat sterilizable and mechanically agitated like the conventional stirred tank bioreactors. Furthermore, it can easily be scaled up while maintaining the light supply coefficient and thus the productivity constant. A device was installed for collecting solar light and distributing it inside the reactor through optical fibers. It was equipped with a light tracking sensor so that the lenses rotate with the position of the sun. This makes it possible to use solar light for photosynthetic cell cultivation in indoor photobioreactors. As a solution to the problems of night biomass loss and low productivity on cloudy days, an artificial light source was coupled with the solar light collecting device. A light intensity sensor monitors the solar light intensity and the artificial light is automatically switched on or off, depending on the solar light intensity. In this way, continuous light supply to the reactor is achieved by using solar light during sunny period, and artificial light at night and on cloudy days.     

Scale-Up of flat plate photobioreactors considering diffuse and direct light characteristics

This study investigates the scaling of photobioreactor productivity based on the growth of Nannochloropsis salina incorporating the effects of direct and diffuse light. The scaling and optimization of photobioreactor geometry was analyzed by determining the growth response of a small-scale system designed to represent a core sample of a large-scale photobioreactor. The small-scale test apparatus was operated at a variety of light intensities on a batch time scale to generate a photosynthetic irradiance (PI) growth dataset, ultimately used to inform a PI growth model. The validation of the scalability of the PI growth model to predict productivity in large-scale systems was done by comparison with experimental growth data collected from two geometrically different large-scale photobioreactors operated at a variety of light intensities. For direct comparison, the small-scale and large-scale experimental systems presented were operated similarly and in such a way to incorporate cultivation relevant time scales, light intensities, mixing, and nutrient loads. Validation of the scalability of the PI growth model enables the critical evaluation of different photobioreactor geometries and design optimization incorporating growth effects from diffuse and direct light. Discussion focuses on the application of the PI growth model to assess the effect of diffuse light growth compared to direct light growth for the evaluation of photobioreactors followed by the use of the model for photobioreactor geometry optimization on the metric of areal productivity.     

Productivity analysis of outdoor chemostat culture in tubular air-lift photobioreactors

Net productivity and biomass night losses in outdoor chemostat cultures ofPhaeodactylum tricornutum were analyzed in two tubular airlift photobioreactors at different dilution rates, photobioreactor surface/volume ratios and incident solar irradiance. In addition, an approximate model for the estimation of light profile and average irradiance inside outdoor tubular photobioreactors was proposed. In both photobioreactors, biomass productivity increased with dilution rate and daily incident solar radiation except at the highest incident solar irradiances and dilution rates, when photoinhibition effect was observed in the middle of the day. Variation of estimated average irradiance vs mean incident irradiance showed two effects: first, the outdoor cultures are adapted to average irradiance, and second, simultaneous photolimitation and photoinhibition took place at all assayed culture conditions, the extent of this phenomena being a function of the (incident)¹ irradiance and light regime inside the culture. Productivity ranged between 0.50 and 2.04 g L^–1 d^–1 in the tubular photobioreactor with the lower surface/volume ratio (S/V = 77.5 m^–1) and between 1.08 and 2.76 g L^–1 d^–1 in the other (S/V = 122.0 m^–1). The optimum dilution rate was 0.040 h^–1 in both reactors. Night-time biomass losses were a function of the average irradiance inside the culture, being lower in TPB0.03 than TPB0.06, due to a better light regime in the first. In both photobioreactors, biomass night losses strongly decreased when the photoinhibition effect was pronounced. However, net biomass productivity also decreased due to lower biomass generation during the day. Thus, optimum culture conditions were obtained when photolimitation and photoinhibition were balanced.     

A theoretical consideration on oxygen production rate in microalgal cultures

Because algal cells are so efficient at absorbing incoming light energy, providing more light energy to photobioreactors would simply decrease energy conversion efficiency. Furthermore, the algal biomass productivity in photobioreactor is always proportional to the total photosynthetic rate. In order to optimize the productivity of algal photobioreactors (PBRs), the oxygen production rate should be estimated. Based on a simple model of light penetration depth and algal photosynthesis, the oxygen production rate in high-density microalgal cultures could be calculated. The estimated values and profiles of oxygen production rate by this model were found to be in accordance with the experimental data. Optimal parameters for PBR operations were also calculated using the model.     

Photobioreactors for microalgal cultures: A Lagrangian model coupling hydrodynamics and kinetics

Closed photobioreactors have to be optimized in terms of light utilization and overall photosynthesis rate. A simple model coupling the hydrodynamics and the photosynthesis kinetics has been proposed to analyze the photosynthesis dynamics due to the continuous shuttle of microalgae between dark and lighted zones of the photobioreactor. Microalgal motion has been described according to a stochastic Lagrangian approach adopting the turbulence model suitable for the photobioreactor configuration (single vs. two‐phase flows). Effects of light path, biomass concentration, turbulence level and irradiance have been reported in terms of overall photosynthesis rate. Different irradiation strategies (internal, lateral and rounding) and several photobioreactor configurations (flat, tubular, bubble column, airlift) have been investigated. Photobioreactor configurations and the operating conditions to maximize the photosynthesis rate have been pointed out. Results confirmed and explained the common experimental observation that high concentrated cultures are not photoinhibited at high irradiance level. © 2015 American Institute of Chemical Engineers Biotechnol. Prog., 31:1259–1272, 2015     

Photosynthetic efficiency of Chlamydomonas reinhardtii in flashing light

Efficient light to biomass conversion in photobioreactors is crucial for economically feasible microalgae production processes. It has been suggested that photosynthesis is enhanced in short light path photobioreactors by mixing‐induced flashing light regimes. In this study, photosynthetic efficiency and growth of the green microalga Chlamydomonas reinhardtii were measured using LED light to simulate light/dark cycles ranging from 5 to 100 Hz at a light‐dark ratio of 0.1 and a flash intensity of 1000 µmol m⁻² s⁻¹. Light flashing at 100 Hz yielded the same photosynthetic efficiency and specific growth rate as cultivation under continuous illumination with the same time‐averaged light intensity (i.e., 100 µmol m⁻² s⁻¹). The efficiency and growth rate decreased with decreasing flash frequency. Even at 5 Hz flashing, the rate of linear electron transport during the flash was still 2.5 times higher than during maximal growth under continuous light, suggesting storage of reducing equivalents during the flash which are available during the dark period. In this way the dark reaction of photosynthesis can continue during the dark time of a light/dark cycle. Understanding photosynthetic growth in dynamic light regimes is crucial for model development to predict microalgal photobioreactor productivities. Biotechnol. Bioeng. 2011;108: 2905–2913. © 2011 Wiley Periodicals, Inc.     

A model for light distribution and average solar irradiance inside outdoor tubular photobioreactors for the microalgal mass culture

A mathematical model to estimate the solar irradiance profile and average light intensity inside a tubular photobioreactor under outdoor conditions is proposed, requiring only geographic, geometric, and solar position parameters. First, the length of the path into the culture traveled by any direct or disperse ray of light was calculated as the function of three variables: day of year, solar hour, and geographic latitude. Then, the phenomenon of light attenuation by biomass was studied considering Lambert-Beer's law (only considering absorption) and the monodimensional model of Cornet et al. (1900) (considering absorption and scattering phenomena). Due to the existence of differential wavelength absorption, none of the literature models are useful for explaining light attenuation by the biomass. Therefore, an empirical hyperbolic expression is proposed. The equations to calculate light path length were substituted in the proposed hyperbolic expression, reproducing light intensity data obtained in the center of the loop tubes. The proposed model was also likely to estimate the irradiance accurately at any point inside the culture. Calculation of the local intensity was thus extended to the full culture volume in order to obtain the average irradiance, showing how the higher biomass productivities in a Phaeodactylum tricornutum UTEX 640 outdoor chemostat culture could be maintained by delaying light limitation. (c) 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 55: 701-714, 1997.     

Cultivation of a cyanobacterium in an internally radiating air-lift photobioreactor

This study proposes a new design of the internally radiatingphotobioreactor, which combines the advantages of an air-lift bioreactorand an internally radiating system, and an efficient way of supplying lightenergy into the photobioreactor during cell cultivation. For a modelphotosynthetic microorganism, Synechococcus PCC 6301 wascultivated in an internally radiating air-lift photobioreactor. The lightcondition inside the photobioreactor was characterized by the average lightintensity which was calculated from the light distribution model. Sinceexcessive light energy induced photoinhibition at the early growth stage, thestrategy of lumostatic operation was developed in order to maintain thelight condition at an appropriate level during cell cultivation. Based on thecalculation results of the light distribution model, the average light intensitywas regulated at 30, 60, or 90 mol m^-2 s^-1 byincreasing the number of light radiators. The model-based control ofirradiating level enabled us to harvest a larger amount of cells withoutshowing the photoinhibited growth. Other favorable results included thereduction of cultivation time and lower consumption of irradiating power.     

In situ monitoring of cell concentration in a photobioreactor using image analysis: comparison of uniform light distribution model and artificial neural networks

Light intensity is a very important factor that determines the growth of photosynthetic cells. In this study, the light distribution in a photobioreactor was analyzed by processing the images captured with a digital camera. The contour images obtained by filtering the original images clearly showed the effects of the cell concentration and external light intensity on the light distribution. Image-processing techniques were then applied to predict the cell density in the photobioreactor. To correlate the cell concentration with the light intensity in the photobioreactor, the captured images were processed using two different approaches. The first method involved the use of an average gray value after deriving a simplified model equation that could be related to the cell density. The second method involved the use of local points instead of a representative value. In this case, an artificial neural network model was adopted to infer the cell density from the information of the local points. By using these two methods, it was possible to relate the image data to the cell concentration. Finally, we compared these two methods with regard to their accuracy, easiness, and effectiveness.     

Prediction of volumetric productivity of an outdoor photobioreactor

Volumetric productivity of Monodus subterraneus cultivated in an outdoor pilot-plant bubble column was predicted with a mathematical model. Two border cases to model the photobioreactor were chosen. Firstly, a model with no light integration in which it is assumed that microalgae can adapt immediately to local light conditions. Secondly, full light integration implicating that microalga can convert all absorbed light with a photosynthetic yield based on average light intensity. Because temperature and light conditions in our photobioreactor changed during the day, photosynthetic yields at any combination of temperature and light intensity were needed. These were determined in repeated-batch lab-scale experiments with an experimental design. The model was evaluated in an outdoor bubble column at different natural light conditions and different temperatures. Volumetric productivities in the bubble column were predicted and compared with experimental volumetric productivities. The light integration model over-estimated productivity, while the model in which we assumed no light integration under-estimated productivity. Light integration occurred partly (47%) during the period investigated. The average observed biomass yield on light was 0.60 g.mol(-1). The model of partly light integration predicted an average biomass yield on light of 0.57 g.mol(-1) and predicted that productivity could have been increased by 19% if culture temperature would have been maintained at 24 degrees C.     

Light energy supply in plate-type and light diffusing optical fiber bioreactors

The light distribution profiles of plate-type photobioreactors were investigated. Light reaching individual channels of a plate module is dependent on the orientation of the module to the sun, the position of the channel within a plate and the position of the plate. The highest incident radiation was measured at the south oriented side of the first channel of the front plate. The light intensity decreased from top to ground channels. Different types of light diffusing optical fibers (LDOF) were characterized with respect to their applicability in photobioreactor systems.     

The oxygen evolution methodology affects photosynthetic rate measurements of microalgae in well‐defined light regimes

Designing photobioreactors correctly is a must for the success of microalgal mass production. Optimal photobioreactor design requires a precise knowledge of photosynthesis dynamics in fluctuating light conditions and hence a method for the measurement of photosynthetic rates in specific light regimes. However, it is not uncommon in literature that experimental protocols used to obtain oxygen generation rates are described ambiguously and the reported rates of photosynthesis vary widely depending on the methodology. Additionally, quite a number of methods overlook certain aspects that can affect the estimated rates significantly, and can therefore affect photobioreactor design. We have developed a method based on oxygen evolution measurements that accurately determines photosynthetic rates under well‐defined light regimes. Our experimental protocol takes into account most of the issues that can affect the rates of oxygen generation, such as depletion of nutrients during the measurements and precision of the measurements. We have focused on the basic applications in photobioreactor design and used a dynamic model of photosynthesis to analyze our results and compare them with available published data. The results suggest that our oxygen evolution method is consistent. Biotechnol. Bioeng. 2010;106: 228–237. © 2010 Wiley Periodicals, Inc.     

Analyzing and modeling of photobioreactors by combining first principles of physiology and hydrodynamics

Mixing in photobioreactors is known to enhance biomass productivity considerably, and flow dynamics play a significant role in the reactor's performance, as they determine the mixing and the cells' movement. In this work we focus on analyzing the effects of mixing and flow dynamics on the photobioreactor performance. Based on hydrodynamic findings from the CARPT(Computer Automated Radioactive Particle Tracking) technique, a possible mechanism for the interaction between the mixing and the physiology of photosynthesis is presented, and the effects of flow dynamics on light availability and light intensity fluctuation are discussed and quantitatively characterized. Furthermore, a dynamic modeling approach is developed for photobioreactor performance evaluation, which integrates first principles of photosynthesis, hydrodynamics, and irradiance distribution within the reactor. The results demonstrate the reliability and the possible applicability of this approach to commercially interesting microalgae/cyanobacteria culture systems.     

Synechococcus sp. (PTCC 6021) cultivation under different light irradiances—Modeling of growth rate-light response

Synechococcus sp. (PTCC 6021), a cyanobacterium species, was cultivated in an internally illuminated photobioreactor. The reactor was designed to achieve a monoseptic cultivation of the species. The goal was to study the growth–irradiance behavior of Synechococcus sp. (PTCC 6021). To accomplish this, different initial light irradiances were implemented inside the photobioreactor and the growth of the cells was monitored. It was observed that cell growth increased with higher light intensity until the photoinhibition occurrence at light irradiance higher than 250?μE?m^?2?s^?1. The maximum OD₆₀₀, maximum growth rate, and biomass productivity increased, and hence the extinction coefficient decreased, with the increase in light irradiance before photoinhibition. The maximum optical density (OD₆₀₀) of 5.91 was obtained with irradiance below 250?μE?m^?2?s^?1 during a growth period of 80 days. The modified Monod function could model the growth–irradiance of cells with satisfactory agreement with the experimental data. The comparison of growth–irradiance of the studied species with other photosynthetic organisms showed the same trend as for cyanobacteria with photoinhibition.     

Photobioreactor for cultivation and real‐time,in‐situ measurement of O2 and CO2 exchange rates,growth dynamics,and of chlorophyll fluorescence emission of photoautotrophic microorganisms

A detailed knowledge about the dynamics of phytoplanktonic photosynthesis and respiration is crucial for the determination of primary productivity in open oceans as well as for biotechnological applications. The dynamics are best studied in photobioreactors that are able to simulate natural conditions in such, that light can be modulated not only diurnally but also mimicking effects of solar elevation angle from sunrise to sunset, variable cloudiness, light modulation in refractory sun flecks due to water waves, or light intermittence due to turbulent flow in dense suspensions. In addition, high performance photobioreactors ought to be able to monitor in real time photosynthetic and respiratory activities as well as culture growth. Here, we demonstrate performance of a newly designed bench‐top laboratory photobioreactor that meets these needs, with a study of green alga Scenedesmus quadricauda. The algal suspension was exposed to simulated daily variations of total photosynthetic active irradiance and spectral profile, with a larger proportion of red photons in the morning and evening hours. The instrument monitored automatically the culture growth by measuring the optical densities at 735 nm and 680 nm and by measuring steady state and maximal chlorophyll fluorescence emission yields. The photochemical yields were estimated from chlorophyll fluorescence data. These widely used but rather indirect yield estimates were confronted with direct measurements of oxygen evolution and consumption quantum yields. The CO₂ fluxes in and out of the culture media as well as the dissolved CO₂ in algal suspension were also recorded. The experiments demonstrated potential of the new photobioreactor to reveal minute modulations in gas exchange rates as well as to yield data for calculation of photon requirement of oxygen evolution in the suspension volume that is key technological parameter for planning of large scale photobioreactors as well as key optimization parameter for strain selection.     

Acclimation of photosynthesis to light and canopy nitrogen distribution: an interpretation

BACKGROUND AND AIMS: Acclimation of photosynthesis to light and its connection with canopy nitrogen (N) distribution are considered. An interpretation of a proportionality between light-saturated photosynthesis and local averaged leaf irradiance is proposed by means of a simple model. MODEL: The model assumes (a) local irradiance drives synthesis of photosynthetic protein from metabolic N; (b) photosynthetic N is slowly degraded over approx. 5-7 d; (c) metabolic N is equally available through the canopy. CONCLUSIONS: The kinetics of acclimation at different light levels may provide a way of parameterizing and testing the model. The model provides a rationale for the proportionality assumption mentioned above, which, while it is consistent with much experimental work, is valuable because it allows canopy photosynthesis to be calculated analytically.