Biomedical subjects

René H Wijffels

Publications and source records attributed to René H Wijffels.

17 recordsLinked to original sources

PubMed2007

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.

Bioreactors↗

PubMed2006

Controlling light-use by Rhodobacter capsulatus continuous cultures in a flat-panel photobioreactor.

The main bottleneck in scale-up of phototrophic fermentation is the low efficiency of light energy conversion to the desired product, which is caused by an excessive dissipation of light energy to heat. The photoheterotrophic formation of hydrogen from acetate and light energy by the microorganism Rhodobacter capsulatus NCIMB 11773 was chosen as a case study in this work. A light energy balance was set up, in which the total bacterial light energy absorption is split up and attributed to its destinations. These are biomass growth and maintenance, generation of hydrogen and photosynthetic heat dissipation. The constants defined in the light energy balance were determined experimentally using a flat-panel photobioreactor with a 3-cm optical path. An experimental method called D-stat was applied. Continuous cultures were kept in a so-called pseudo steady state, while the dilution rate was reduced slowly and smoothly. The biomass yield and maintenance coefficients of Rhodobacter capsulatus biomass on light energy were determined at 12.4 W/m(2) (400-950 nm) and amounted to 2.58 x 10(-8) +/- 0.04 x 10(-8) kg/J and 102 +/- 3.5 W/kg, respectively. The fraction of the absorbed light energy that was dissipated to heat at 473 W/m(2) depended on the biomass concentration in the reactor and varied between 0.80 and 0.88, as the biomass concentration was increased from 2.0 to 8.0 kg/m(3). The process conditions were estimated at which a 3.7% conversion efficiency of absorbed light energy to produced hydrogen energy should be attainable at 473 W/m(2).

Biomass↗

PubMed2006

Hypothesized kinetic models for describing the growth of globular and encrusting demosponges.

The marine sponges Dysidea avara and Chondrosia reniformis (globular forms) were cultured in the laboratory on a diet of viable Phaeodactylum tricornutum cells and dissolved nutrients (algae and fish powders). Our growth data were combined with literature data for Pseudosuberites andrewsi (a globular sponge) and for the encrusting sponges Oscarella lobularis, Hemimycale columella, and Crambe crambe. The suitability of three growth models-linear, exponential, and radial accretive-for describing the growth of globular and encrusting sponges was assessed. Radial accretive growth was determined to be the best model to describe growth of both encrusting and globular sponges. Average growth rates of 0.051+/-0.016 and 0.019+/-0.003 mm/day (calculated as the increase of the radius of the sponge per day) were obtained experimentally for D. avara and C. reniformis, respectively.

Analysis of Variance↗

PubMed2005

Large-scale production of pharmaceuticals by marine sponges: sea, cell, or synthesis?

Marine sponges are known to produce an overwhelming array of secondary metabolites with pharmaceutical potential. The technical and economical potential of using marine sponges for large-scale production of these compounds was assessed for two cases: the anticancer molecule halichondrin B from a Lissodendoryx sp., and avarol from Dysidea avara for its antipsoriasis activity. An economic and technical analysis was done for three potential production methods: mariculture, ex situ culture (in tanks), and cell culture. We concluded that avarol produced by mariculture or ex situ culture could become a viable alternative to currently used pharmaceuticals for the treatment of psoriasis. Production of halichondrin B from sponge biomass was found to not be a feasible process, mainly due to the extremely low concentration of the compound in the sponge. Technical feasibility was also analyzed for five alternatives: chemical synthesis, wild harvest, primmorph culture, genetic modification and semi-synthesis. It was concluded that the latter two approaches could prove to be valuable methods for the production of pharmaceuticals, based on chemical structures of secondary metabolites present in trace amounts in marine sponges.

Animals↗

PubMed2005

Marine sponges as pharmacy.

Marine sponges have been considered as a gold mine during the past 50 years, with respect to the diversity of their secondary metabolites. The biological effects of new metabolites from sponges have been reported in hundreds of scientific papers, and they are reviewed here. Sponges have the potential to provide future drugs against important diseases, such as cancer, a range of viral diseases, malaria, and inflammations. Although the molecular mode of action of most metabolites is still unclear, for a substantial number of compounds the mechanisms by which they interfere with the pathogenesis of a wide range of diseases have been reported. This knowledge is one of the key factors necessary to transform bioactive compounds into medicines. Sponges produce a plethora of chemical compounds with widely varying carbon skeletons, which have been found to interfere with pathogenesis at many different points. The fact that a particular disease can be fought at different points increases the chance of developing selective drugs for specific targets.

Animals↗

PubMed2005

Optimization of biomass, vitamins, and carotenoid yield on light energy in a flat-panel reactor using the A-stat technique.

Acceleration-stat (A-stat) cultivations in which the dilution rate is continuously changed at a constant acceleration rate, leading to different average light intensities inside the photobioreactor, can supply more information and reduce experimental time compared with chemostat cultivations. The A-stat was used to optimize the biomass and product yield of continuous cultures of the microalgae D. tertiolecta in a flat-panel reactor. In this study, four different accelerations were studied, a pseudo steady state was maintained at acceleration rates of 0.00016 and 0.00029 h(-2) and results were similar to those of the chemostat. An increase in the acceleration rate led to an increase in the deviation between results obtained in the A-stat and in the chemostats. We concluded that it is advantageous to use the A-stat instead of chemostats to determine culture characteristics and optimize a specific photobioreactor. The effect of average light intensity inside the photobioreactor on the production of vitamins C and E, lutein, and beta-carotene was studied using the A-stat. The highest concentrations of these products were 3.48 +/- 0.46, 0.33 +/- 0.06, 5.65 +/- 0.24, and 2.36 +/- 0.38 mg g(-1), respectively. These results were obtained at different average light intensities, showing the importance of optimizing each product on light intensity.

Ascorbic Acid↗

PubMed2004

The life and death of sponge cells.

Cell viability is an essential touchstone in the study of the effect of medium components on cell physiology. We developed a flow-cytometric assay to determine sponge-cell viability, based on the combined use of fluorescein diacetate (FDA) and propidium iodide (PI). Cell fluorescence measurements based on incubation of cells with FDA or PI resulted in a useful and reproducible estimate of the viability of primary sponge-cell cultures. We studied the effects of temperature, ammonium, and the fungicide amphotericin B on the viability of a primary-cell culture from the marine sponge Suberites domuncula using the aforementioned flow-cytometric assay. S. domuncula cells die rapidly at a temperature of >or=22 degrees C, but they are insensitive to ammonium concentrations of up to 25 mM. Amphotericin B, which is frequently used in sponge-cell culture media, was found to be toxic to S. domuncula cells.

Amphotericin B↗

PubMed2004

Overcoming shear stress of microalgae cultures in sparged photobioreactors.

In the present work we identified and quantified the effect of hydrodynamic stress on two different microalgae strains, Dunaliella tertiolecta and D. salina, cultivated in bench-scale bubble columns. The cell death rate constant increased with increasing gas-entrance velocity at the sparger. Dunaliella salina was slightly more sensitive than D. tertiolecta. The critical gas-entrance velocities were approximately 50 and 30 m s(-1) for D. tertiolecta and D. salina, respectively. The effects of gas-flow rate, culture height, and nozzle diameter on the death rate constant were also studied. From these results it was concluded that bubble rising and bubble bursting are not responsible for cell death. Regarding nozzle diameter, small nozzles were more detrimental to cells. The bubble formation at the sparger was found to be the main event leading to cell death.

Air Pressure↗

PubMed2003

Hydrodynamic stress and lethal events in sparged microalgae cultures.

The effect of high superficial gas velocities in continuous and batch cultures of the strains Dunaliella tertiolecta, Chlamydomonas reinhardtii wild-type and cell wall-lacking mutant was studied in bubble columns. No cell damage was found for D. tertiolecta and C. reinhardtii (wild-type) up to superficial gas velocities of 0.076 and 0.085 m s(-1), respectively, suggesting that high superficial gas velocities alone cannot be responsible for cell death and, consequently, bubble bursting cannot be the sole cause for cell injury. A death rate of 0.46 +/- 0.08 h(-1) was found for C. reinhardtii (cell wall-lacking mutant) at a superficial gas velocity of 0.076 m s(-1), and increased to 1.01 +/- 0.29 h(-1) on increasing superficial gas velocity to 0.085 m s(-1). Shear sensitivity is thus strain-dependent and to some extent the cell wall plays a role in the protection against hydrodynamic shear. When studying the effect of bubble formation at the sparger in batch cultures of D. tertiolecta by varying the number of nozzles, a death rate of 0.047 +/- 0.016 h(-1) was obtained at high gas entrance velocities. D. tertiolecta was cultivated in a pilot-plant reactor under different superficial gas velocities of up to 0.026 m s(-1), with relatively low gas entrance velocities and no cell damage was observed. There is some indication that the main parameter causing cell death and damage was the gas entrance velocity at the sparger.

Animals↗

PubMed2003

Microalgae cultivation in air-lift reactors: modeling biomass yield and growth rate as a function of mixing frequency.

The slow development of microalgal biotechnology stems from the failure in the design of large-scale photobioreactors where light energy is efficiently utilized. Due to the light gradient inside the reactor and depending on the mixing properties, algae are subjected to certain light/dark cycles where the light period is characterized by a light gradient. These light/dark cycles will determine productivity and biomass yield on light energy. Air-lift reactors can be used for microalgae cultivation and medium-frequency light/dark cycles will be found in these systems. Light/dark cycles are associated with two basic parameters: first, the light fraction, i.e., the ratio between the light period and the cycle time and second, the frequency of the light/dark cycle. In the present work, light/dark cycles found in air-lift reactors were simulated taking into account the light gradient during the light period. The effect of medium-frequency cycle time (10-100 s) and light fraction (0.1-1) on growth rate and biomass yield on light energy of the microalgae Dunaliella tertiolecta was studied. The biomass yield and growth rates were mainly affected by the light fraction, while cycle time had little influence. Response surface methodology was used and a statistical model describing the effect of light fraction and cycle time on growth rate and biomass yield on light energy was developed. The use of the model as a reactor design criterion is discussed.

Biomass↗

PubMed2003

Progress towards a controlled culture of the marine sponge Pseudosuberites andrewsi in a bioreactor.

Explants of the tropical sponge Pseudosuberites andrewsi were fed with the marine diatom Phaeodactylum tricornotum. The food was supplied either as intact algae or as a filtered crude extract. Growth (measured as an increase in underwater weight) was found in both experiments. The explants fed with intact algae increased to an average underwater weight of 255% of the initial weight in 45-60 days. The explants fed with crude extract increased to an average of 200% of the initial weight in 30 days. These results show that it is possible to grow a sponge using a single microorganism species as a food source. In addition, it was demonstrated that sponges are also capable of growing on non-particulate food. Therefore, this study is an important step forward towards the development of controlled, in vivo sponge cultures.

Animal Feed↗

PubMed2003

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.

Biomass↗

PubMed2003

Optimisation of cultivation parameters in photobioreactors for microalgae cultivation using the A-stat technique.

Light availability inside the reactor is often the bottleneck in microalgal cultivation and for this reason much attention is being given to light limited growth kinetics of microalgae, aiming at the increase of productivity in photobioreactors. Steady-state culture characteristics are commonly used for productivity optimisation and for cell physiology studies in continuous cultures, and are normally achieved using chemostat cultivations. In the present study, we investigated the applicability of a new and dynamic cultivation method called acceleration-stat (A-stat) to microalgae cultivations where light is the limiting substrate. In the A-stat, the dilution rate is increased at a constant rate. This acceleration rate should be a compromise between a short cultivation time, in order to make it a fast process, and the metabolic adaptation rate of the microorganism to changes in the environment. Simulations of the A-stat were done with different acceleration rates to have an indication of the best rate to use. An A-stat was performed in a pilot plant bubble column (65 l) with Dunaliella tertiolecta as a model organism, and results showed that a pseudo steady state was maintained throughout the experiment. From this work, it was concluded that the A-stat can be used as a fast and accurate tool to determine kinetic parameters and to optimise any specific type of photobioreactor.

Bioreactors↗

PubMed2003

Marine biotechnology in education: a competitive approach.

This paper describes the development of a practical, which is taught to third year biotechnology students. We wanted to motivate the students by making them responsible for a research project. Competition was added as a stimulus for interaction between the students. A virtual company called CaroTech employed the students for 2 weeks. They worked in groups of two persons and each group was responsible for a 0.8 l flat panel photobioreactor. They had to produce as much beta-carotene as possible using the marine alga strain Dunaliella salina in this photobioreactor. On the first day, students developed a strategy to obtain optimal algal growth rate. They putted this plan into practice the second day and while cultivating the organism, they developed a second strategy how and when to stress the alga to initiate beta-carotene production. At the end of the ninth day, the total amount of beta-carotene was measured. To stimulate competition, the group that produced the most beta-carotene obtained half a point bonus on the final practical mark. On the tenth day, each group presented their results and an evaluation of their chosen strategies to the CaroTech board. Most groups were successful in growing algae. In the second phase some groups failed to stress the alga. The best group produced more than two times beta-carotene than the runner-up. The students were motivated by being responsible for their own results and the competitive approach. All students liked the practical and indicated that they learned a lot by following this practical.

Bioreactors↗

PubMed2003

Vitamin E (alpha-tocopherol) production by the marine microalgae Dunaliella tertiolecta and Tetraselmis suecica in batch cultivation.

Batch experiments with Dunaliella tertiolecta and Tetraselmis suecica were performed to investigate alpha-tocopherol (alpha-T) production in time, in order to assess the effect of light availability per cell on the production of this antioxidant. In D. tertiolecta alpha-T content increased during growth, in other words, as the cell density increased and the light availability per cell decreased. In T. suecica the pattern was different: alpha-T content was highest during the exponential phase, decreased significantly during the linear phase and increased again towards the end of the cultivation. Chlorophyll (chl-a) content of T. suecica cells decreased after the exponential phase, instead of the expected increase typically observed in shade-adapted cells, suggesting that the culture was nutrient limited. An experiment with extra nutrients showed that chl-a and alpha-T content increased significantly during both the linear and the stationary phase when compared with values in nutrient-deficient conditions. No correlation between alpha-T and chl-a was observed. Our results indicate that diminished light availability does not limit alpha-T production in D. tertiolecta and T. suecica, opening up the possibility of using high cell density, light-limited cultures for the production of this commercially interesting compound.

Adaptation, Physiological↗

PubMed2003

Realizing the promises of marine biotechnology.

High-quality research in the field of marine biotechnology is one of the key-factors for successful innovation in exploiting the vast diversity of marine life. However, fascinating scientific research with promising results and claims on promising potential applications (e.g. for pharmaceuticals, nutritional supplements, (feed-)products for aquaculture and bioremediation solutions) is not the only factor to realise the commercial applications of marine biotechnology. What else is needed to exploit the promising potential of marine biotechnology and to create new industrial possibilities? In the study project 'Ocean Farming-Sustainable exploitation of marine organisms', we explore the possibilities of marine organisms to fulfill needs, such as safe and healthy food, industrial (raw) materials and renewable energy in a sustainable way. One of the three design groups is envisioning the future of strong land-based 'marine' market chains. Marine biotechnology is one of the foci of attention in this design group. This article provides a model of future-oriented thinking in which a variety of experts actively participate.

Aquaculture↗

PubMedDate not supplied

Sponge-cell culture? A molecular identification method for sponge cells.

Dissociated sponge cells are easily confused with unicellular organisms. This has been an obstacle in the development of sponge-cell lines. We developed a molecular detection method to identify cells of the sponge Dysidea avara in dissociated cell cultures. The 18S ribosomal RNA gene from a Dysidea avara specimen was sequenced and compared to eukaryotic 18S rDNA sequences picked up from a proliferating cell culture that originated from a dissociated Dysidea avara specimen. Our method proved unambiguously that this was not a sponge-cell culture. Therefore, it provides a valuable tool for further research on sponge-cell cultures.

Animals↗