Luuc R. Mur

ON THE DOMINANCE OF FILAMENTOUS CYANOBACTERIA IN SHALLOW, TURBID LAKES

The phytoplankton community of eutrophic shallow lakes is often dominated by filamentous cyanobacteria of the family Oscillatoriaceae. In this paper we follow two independent approaches to show that this situation is likely to be one of two alternative stable states of the algal community. First we analyze patterns of cyanobacterial dominance observed in the field, and show that these patterns imply that the algal community is a hysteretic system with two alternative equilibria. Then, we construct a simple competition model to show that hysteresis should in fact be expected from differences in physiology between cyanobacteria and algae. The basic mechanism is that cyanobacteria are the superior competitors under conditions of low light, but also promote such conditions, as they can cause a higher turbidity per unit of phosphorus than other algae. This mechanism of hys- teresis offers an explanation for the resistance of cyanobacteria dominance in shallow lakes to restoration efforts by means of nutrient reduction.

Effects of Light on the Microcystin Content of Microcystis Strain PCC 7806

ABSTRACT Many cyanobacteria produce microcystins, hepatotoxic cyclic heptapeptides that can affect animals and humans. The effects of photosynthetically active radiation (PAR) on microcystin production by Microcystis strain PCC 7806 were studied in continuous cultures. Microcystis strain PCC 7806 was grown under PAR intensities between 10 and 403 μmol of photons m−2 s−1 on a light-dark rhythm of 12 h -12 h. The microcystin concentration per cell, per unit biovolume and protein, was estimated under steady-state and transient-state conditions and on a diurnal timescale. The cellular microcystin content varied between 34.5 and 81.4 fg cell−1 and was significantly positively correlated with growth rate under PAR-limited growth but not under PAR-saturated growth. Microcystin production and PAR showed a significant positive correlation under PAR-limited growth and a significant negative correlation under PAR-saturated growth. The microcystin concentration, as a ratio with respect to biovolume and protein, correlated neither with growth rate nor with PAR. Adaptation of microcystin production to a higher irradiance during transient states lasted for 5 days. During the period of illumination at a PAR of 10 and 40 μmol of photons m−2 s−1, the intracellular microcystin content increased to values 10 to 20% higher than those at the end of the dark period. Extracellular (dissolved) microcystin concentrations were 20 times higher at 40 μmol of photons m−2 s−1 than at 10 μmol of photons m−2 s−1 and did not change significantly during the light-dark cycles at both irradiances. In summary, our results showed a positive effect of PAR on microcystin production and content of Microcystis strain PCC 7806 up to the point where the maximum growth rate is reached, while at higher irradiances the microcystin production is inhibited.

Isolation and characterization of extracellular polysaccharides from the epipelic diatoms Cylindrotheca closterium and Navicula salinarum

The production and composition of extracellular polymeric substances (EPS) in axenic batch cultures of the benthic marine epipelic diatoms Navicula salinarum and Cylindrotheca closterium were investigated. EPS was secreted into the medium and the bulk was loosely associated with the cells. Neither N. salinarum nor C. closterium formed a well-defined polysaccharide capsule. EPS of both N. salinarum and C. closterium consisted predominantly of polysaccharide but small quantities of protein were present as well. EPS also contained uronic acids and SO4 −2 groups. Analysis of monosaccharides using gas chromatography showed that for both species glucose and xylose were the main constituents, but several other monosaccharides were present in smaller quantities. Two fractions of EPS were distinguished: a small amount was secreted into the medium and a second fraction was extracted in water at 30 °C. For both species the two fractions differed somewhat in composition, indicating that they represented two different...

Application of light‐emitting diodes in bioreactors: Flashing light effects and energy economy in algal culture (Chlorella pyrenoidosa)

Light‐emitting diodes (LEDs) were used as the sole light source in continuous culture of the green alga Chlorella pyrenoidosa. The LEDs applied show a peak emission at 659 nm with a half‐power bandwidth of 30 nm. Selection of this wavelength range, which is optimal for excitation of chlorophylls a and b in their “red” absorption bands makes all photons emitted potentially suitable for photosynthesis. No need for additional supply of blue light was found. A standardized panel with 2 LEDs cm−2 fully covered one side of the culture vessel. At standard voltage in continuous operation the light output of the diode panel appeared more than sufficient to reach maximal growth. Flash operation (5‐μs pulse duration) enables potential use of higher operating voltages which may render up to three times more light output. Flat airlift fermentor‐type continuous culture devices were used to estimate steady state growth rates of Chlorella pyrenoidosa as a function of the light flux (μmol photons · m−2 · s−1) and the flashing frequency of the light‐emitting diodes (which determines the duration of the dark “off” time between the 5‐μs “on” pulses). At the fixed voltage and turbidostat setting applied a 20‐kHz frequency, which equals dark periods of 45 μs, still permitted the maximum growth rate to become nearly reached. Lower frequencies fell short of sustaining the maximal growth rate. However, the light flux decrease resulting from lowering of the flash frequency appeared to reduce the observed growth rates less than in the case of a similar flux decrease with light originating from LEDs in continuous operation. Flash application also showed reduction of the quantum requirement for oxygen evolution at defined frequencies. The frequency domain of interest was between 2 and 14 kHz. LEDs may open interesting new perspectives for studies on optimization of mixing in mass algal culture via the possibility of separation of interests in the role of modulation on light energy conversion and saturation of nutrient supply. Use of flashing LEDs in indoor algal culture yielded a major gain in energy economy in comparison to luminescent light sources.

Exopolysaccharide production by the epipelic diatom Cylindrotheca closterium: effects of nutrient conditions

During the stationary phase of a batch culture of the epipelic diatom Cylindrotheca closterium, accumulation of exopolysaccharides and intracellular carbohydrates was observed. When nitrogen was added to the culture in the stationary phase, growth was resumed and the accumulation of exopolysaccharides was delayed. This indicated that nitrogen depletion caused cessation of growth, and stimulated exopolysaccharide accumulation. Exopolysaccharide accumulation was also stimulated when cells were either resuspended in medium lacking N or P, or when they were inoculated in medium with low concentrations of N or P. Growth was not immediately affected by low N or P concentrations. S depletion only resulted in exopolysaccharide accumulation when growth was affected. Si or Fe depletion did not stimulate exopolysaccharide accumulation, even when growth rates were lowered. Apparently, stimulation of exopolysaccharide accumulation is dependent on the type of nutrient depletion. Intracellular storage carbohydrates did not accumulate when cells were incubated at low N or P concentrations. Cells grown with ammonium as nitrogen source produced more carbohydrates (both extracellular and intracellular) than cells grown with nitrate as nitrogen source, indicating that both exopolysaccharides and intracellular carbohydrates accumulated as a result of overflow metabolism.

The Microcystin Composition of the Cyanobacterium Planktothrix agardhii Changes toward a More Toxic Variant with Increasing Light Intensity

ABSTRACT The cyanobacterium Planktothrix agardhii, which is dominant in many shallow eutrophic lakes, can produce hepatotoxic microcystins. Currently, more than 70 different microcystin variants have been described, which differ in toxicity. In this study, the effect of photon irradiance on the production of different microcystin variants by P. agardhii was investigated using light-limited turbidostats. Both the amount of the mRNA transcript of the mcyA gene and the total microcystin production rate increased with photon irradiance up to 60 μmol m−2 s−1, but they started to decrease with irradiance greater than 100 μmol m−2 s−1. The cellular content of total microcystin remained constant, independent of the irradiance. However, of the two main microcystin variants detected in P. agardhii, the microcystin-DeRR content decreased twofold with increased photon irradiance, whereas the microcystin-DeLR content increased threefold. Since microcystin-DeLR is considerably more toxic than microcystin-DeRR, this implies that P. agardhii becomes more toxic at high light intensities.

Efficiency of light utilization of Chlamydomonas reinhardtii under medium-duration light/dark cycles

The light regime inside a photobioreactor is characterized by a light gradient with full (sun)light at the light-exposed surface and darkness in the interior of the bioreactor. Consequently, depending on the mixing characteristics, algae will be exposed to certain light/dark cycles. In this study the green alga Chlamydomonas reinhardtii was cultivated under five different light regimes: (1) continuous illumination; (2) a square-wave light/dark cycle with a light fraction (epsilon) of 0.5 and a duration (t(c)) of 6.1 s; (3) epsilon=0.5, t(c)=14.5 s; (4) epsilon=0.5, t(c)=24.3 s and (5) epsilon=0.8, t(c)=15.2 s. The biomass yield on light energy, protein per photons, decreased under light/dark cycles (epsilon=0. 5) in comparison to continuous light (CL), from 0.207 (CL) to 0.117-0.153 g mol(-1) (epsilon=0.5). Concomitantly, the maximal specific photosynthetic activity, oxygen production per protein, decreased from 0.94 (CL) to 0.64-0.66 g g(-1) h(-1) (epsilon=0.5). Also the quantum yield of photochemistry, yield of the conversion of light energy into chemical energy, decreased from 0.47 (CL) to 0. 23 (epsilon=0.5, t(c)=24.3 s). Apparently, C. reinhardtii is not able to maintain a high photosynthetic capacity under medium-duration light/dark cycles and since specific light absorption did not change, light utilization efficiency decreased in comparison to continuous illumination.

Photosynthetic efficiency of Dunaliella tertiolecta under short light/dark cycles

Abstract Inside photobioreactors algae are exposed to light/dark fluctuations. In this study the marine green alga Dunaliella tertiolecta was cultivated under short light/dark cycles of 3/3 s, 94/94 ms and 31/156 ms as found in medium and short light-path reactors. The photon flux density (PFD) during the light period was 440–455 μmol m −2 s −1 and, under the 31/156 ms cycle only, 1025 μmol m −2 s −1 . The photosynthetic efficiency was determined and expressed as the biomass yield on light energy in gram protein produced per mol of photons absorbed. The yield under the 94/94 ms cycle was higher than the yield under continuous light of 440–455 μmol m −2 s −1 . Apparently a light integration effect occurs under the 94/94 ms cycle together with an increase of the photosynthetic efficiency. On the contrary, at the 3/3 s cycle the efficiency decreased. Also under the 31/156 ms cycle, with 1025 μmol m −2 s −1 in the light period, the yield was lower than under continuous light. The results obtained under the 31/156 ms light/dark cycle are discussed with respect to the performance of short light-path flat panel photobioreactors operated at high biomass densities.

Modelling vertical migration of the cyanobacterium Microcystis

Computer models can be helpful tools to provide abetter understanding of the mechanisms responsible forthe complex movements of cyanobacteria resulting fromchanges in buoyancy and mixing of the water column ina lake. Kromkamp & Walsby (1990) developed a verticalmigration model for Oscillatoria, that wasbased on the experimentally determinedrelationship between the rates of density change andphoton irradiance in this cyanobacterium. To adaptthis model to Microcystis, we determinedrelated changes in carbohydrate content in cultures ofMicrocystis. Samples were incubated at variousconstant values of photon irradiance and then placedin the dark. The changes in carbohydrate content ofthe cells during these incubations were investigated.The relationship between the ratio of carbohydrate toprotein and cell density in Microcystis wasestablished to permit conversion of the rates ofcarbohydrate change to rates of density change. Byplotting the calculated rates of density changeagainst the values of photon irradiance experiencedduring the incubations, an irradiance-response curveof density change was established. The curve showed adistinct maximum at 278 µmol photons m-2s-1. At higher values of photon irradiance, therate of density change was strongly inhibited. Apositive linear correlation was found between celldensity and the rates of density decrease in the dark.The validity of the use of rate equations of densitychange, which are based on short-term incubations atconstant values of photon irradiance, to predictdensity changes in Microcystis in fluctuatinglight regimes was tested. This was accomplished bymeasuring the time course of change in carbohydratecontent of two continuous cultures of Microcystis, which were submitted to fluctuatinglight regimes, and comparing the results with thechanges in the carbohydrate contents of these culturespredicted by the rate equations of carbohydratechange. The results showed good agreement: the rateequations of density change were therefore introducedinto the model to simulate vertical migration of Microcystis. The model predicts that the maximummigration depth of Microcystis will increasewith colony size up to a maximum of 200 µm radius.The effect of colony size on the net increase in celldensity during the light period was also investigatedwith the model. It predicts that small colonies havea higher net increase in cell density than largecolonies, but are inhibited at high photon irradiancesat the surface.

Growth Kinetics of Oscillatoria agardhii Gomont in Continuous Culture, Limited in its Growth by the Light Energy Supply

Summary: Growth efficiency (c) and specific maintenance rate constant (μe) were determined in continuous cultures of the cyanobacterium Oscillatoria agardhii Gomont, in light energy-limiting conditions, according to the formula μ = cq E – μe in which q E is the specific light energy uptake rate. Values of the efficiency factor c varied with irradiance from 0.23 at 0.5 W m−2 to 0.05 at 40 W m−2. The specific maintenance rate constant was 0.001 h−1. The culture pH value influenced c and μe . A diurnal light/dark cycle with a 16 h photoperiod did not affect either of the two parameters, μe and c. The relationship between growth rate and light energy uptake rate, embodied in the above formula, was also valid for nitrogen (nitrate)-limited cultures.