Delayed fluorescence as a direct indicator of diurnal variation in quantum and radiant energy utilization efficiencies of phytoplankton

We compared delayed fluorescence (DF) excitation spectrometry with radiocarbon (¹⁴C) technique using a monoalgal culture of Chlorella vulgaris grown under natural temperature and irradiance. This was done by monitoring the DF, in parallel to quantum efficiency (QE) and index of radiant energy utilization efficiency (Ψ) as calculated on the basis of carbon uptake measurements by radiocarbon technique. During the diurnal cycle, temperature, irradiance, and chlorophyll (Chl) contents were monitored in the algal culture that was kept in an open transparent plastic tank submerged at the surface of Lake Kinneret, Israel. The DF signal correlated with both the QE (r ² = 0.869, p<0.01) and Ψ (r ² = 0.977, p<0.01) during a diurnal cycle. We suggest that, besides the measurement of active Chl and phytoplankton population composition, the DF signal provides additional information on the QE and Ψ in phytoplankton population.     

Modeling of lutein production by heterotrophic <Emphasis Type="Italic">Chlorella</Emphasis> in batch and fed-batch cultures

Chlorella is a promising alternative resource of lutein (xanthophyll) production as it can be cultivated heterotrophically in fermentors. In this paper, a kinetic model for lutein production by heterotrophic Chlorella pyrenoidosa was developed based on batch cultivations in 250-ml flasks and a 19-l fermentor. The model was validated by experimental data from two fed-batch cultivations performed in the same fermentor. The dynamic behavior of lutein production by C. pyrenoidosa with various concentrations of glucose and nitrogen was analyzed based on the kinetic model. Model-based analyses suggested that glucose concentrations between 5 and 24 g/l and nitrogen concentrations between 0.7 and 12 g/l during the cultivation were favorable for lutein production by heterotrophic C. pyrenoidosa. It also showed that fed-batch cultivations are more suitable for efficient production of lutein than batch ones. The results obtained in this study may contribute to commercial lutein production by heterotrophic Chlorella.     

Using biosorption to enrich the biomass of <Emphasis Type="Italic">Chlorella vulgaris </Emphasis>with microelements to be used as mineral feed supplement

The paper discusses biosorption of Cr(III), Cu(II), Mn(II), Zn(II) and Co(II) to the biomass of Chlorella vulgaris, to produce a biologically bound, concentrated form of microelements. The kinetics of biosorption was described with a pseudo-second order equation and equilibrium with the Langmuir isotherm. The mechanism of biosorption was identified as cation-exchange with alkaline metals. Cation-exchange capacity was evaluated as 4.07 meq g⁻¹. The effect of operation conditions, pH and temperature, on biosorption performance was investigated and the best operation conditions for biosorption were selected (pH 5, temperature 25 °C). The maximum sorption capacity of microelements was determined in single-metal system at pH 5 and 25 °C: Zn(II) 3.30 meq g⁻¹, Cu(II) 1.77 meq g⁻¹, Co(II) 1.75 meq g⁻¹, Cr(III) 1.74 meq g⁻¹, Mn(II) 0.764 meq g⁻¹. Biosorption experiments were also carried out in multi-metal system. The biomass of C. vulgaris enriched with microelements via the process of biosorption in both single- and multi-metal system was discussed in terms of preparation of feed supplement for laying hens and piglets. The experiments showed that 1 kg of conventional feed for laying hens can be supplemented with 0.20 g of the biomass enriched with microelements and for piglets with 0.15 g of the preparation.     

Identification and quantification of indole-3-acetic acid in the kelp Laminaria japonica Areschoug and its effect on growth of marine microalgae

A method was established for the identification and quantification of indole-3-acetic acid (IAA) in extracts of the kelp Laminaria japonica. An IAA content of 90–95 μg kg⁻¹ fresh weight in kelp extract was determined by high performance liquid chromatography (HPLC). IAA identification was based on a combination of co-chromatography and comparative chromatography with a standard, analysis of UV spectra, and atmospheric pressure electrospray mass spectrometry (APESI-MS). IAA was isolated by silica gel chromatography and HPLC. The effect on the growth of four marine microalgae of the pure IAA isolated from kelp extract was investigated. Exogenously added IAA from kelp enhanced the growth of Chlorella sp., Dunaliella salina and Porphyridium cruentum, but not that of Chaetoceros muelleri. IAA from kelp significantly inhibited the accumulation of soluble cellular proteins in Chlorella sp. and P. cruentum, and had a very significant effect on chlorophyll biosynthesis in Chlorella sp. However, there was no obvious effect of IAA on the regulation of biosynthesis of cellular polysaccharides in these four marine microalgae.     

Are Freshwater Mixotrophic Ciliates Less Sensitive to Solar Ultraviolet Radiation than Heterotrophic Ones?1

We tested whether mixotrophic ciliates are more resistant to solar ultraviolet radiation (UVR) than heterotrophic ones because symbiotic algae can provide self-shading by cell matter absorption and eventually by direct UV screening from mycosporine-like amino acids (MAAs). Sensitivity of a natural assemblage to solar radiation was tested in experiments in the original lake and in a more UV transparent alpine lake after transplantation of the ciliates. In both lakes, the assemblage was exposed either to full sunlight, to photosynthetically active radiation only, or kept in the dark. In each lake, exposure was for 5 h at the surface and at the depth corresponding to the 10% attenuation depth at 320 nm. Overall, when the assemblage was exposed to surface UVR, only one out of four dominant mixotrophic ciliates, Vorticella chlorellata, was more resistant than heterotrophic species. The higher UV resistance in V. chlorellata was related to the presence of MAAs and the high percentage of ciliate volume occupied by algal symbionts. Our results indicate that effects of UVR were species-specific and depended on efficient screening of these wavelengths, but also on the depth preference of the ciliates and thus, on their previous exposure history to UVR.     

利用市政污水培养Chlorella vulgaris生产生物柴油

为了考察利用南昌市政污水规模化培养富油微藻生产生物柴油,同时达到净化污水的目的,取南昌市青山湖污水处理厂未经任何处理的市政污水作为普通小球藻(Chlorella vulgaris)生长的培养液。监测了C.vulgaris在市政污水中连续培养10 d的特定生长率、生物质产量以及与之相关的市政污水中氨氮(NH4+-N)、总磷(TP)、化学需氧量(COD)、总悬浮固体(TSS)和挥发性悬浮固体(VSS)的清除情况。实验表明:营养物质的水平显著地影响了C.vulgaris的生长。C.vulgaris的生长率在培养8 d后达到最大,OD680为2.856,总的生物质产量日均最大积累速率为0.01 g/L,油脂含量为干质量的18%,油脂的平均日产量为0.001 g/L。培养10 d内NH4+-N、TP和COD的去除率分别为50.0%、32.1%和26.0%,TSS和VSS的日平均去除速率分别为0.01 g/L和0.006 1 g/L。     

31P-NMR STUDIES ON INORGANIC POLYPHOSPHATES IN MICROALGAE1

Using “P nuclear magnetic resonance analysis, total inorganic polyphosphate in algae could be quantitatively estimated, For this purpose the algal suspension, which had been kept in cold trichloroacetic acid, was further treated with 6 mM EDTA, or the cells were kept in 2 N KOH containing 100 mM EDTA for 18 h at 37°C. These simple methods avoid hydrolysis of cellular inorganic polyphosphate and, therefore, are useful for the study of phosphorus metabolism in algae. The effects of these treatments on visualization of the signal for inorganic polyphosphate in nuclear magnetic resonance spectra were discussed in comparison with in vivo, ‘P nuclear magnetic resonance spectra of algae.     

Cultivation factors and population size control the uptake of nitrogen by the microalgae Chlorella vulgaris when interacting with the microalgae growth-promoting bacterium Azospirillum brasilense

Growth of and the capacity to take up nitrogen in the freshwater microalgae Chlorella vulgaris were studied while varying the concentrations of ammonium and nitrate, the pH and the source of carbon in a synthetic wastewater growth medium when co-immobilized in alginate beads with the microalgae growth-promoting bacterium Azospirillum brasilense. Analyses of 29 independent experiments showed that co-immobilization of the microalgae with A. brasilense could result in two independent phenomena directly affected by cultivation factors, such as nitrogen species, pH and presence of a carbon source. First, growth of the microalgal population increased without an increase in the capacity of the single cells to take up nitrogen, or second, the capacity of cells to take up nitrogen increased without an increase of the total microalgal population. These phenomena were dependent on the population density of the microalgae, which was in turn affected by cultivation factors. This supports the conclusion that the size of the microalgal population controls the uptake of nitrogen in C. vulgaris cells - the higher the population (regardless the experimental parameters), the less nitrogen each cell takes up.     

Growth kinetics of microalgae in microfluidic static droplet arrays

We investigated growth kinetics of microalgae, Chlorella vulgaris, in immobilized arrays of nanoliter‐scale microfluidic drops. These static drop arrays enabled simultaneous monitoring of growth of single as well as multiple cells encapsulated in individual droplets. To monitor the growth, individual drop volumes were kept nearly intact for more than a month by controlling the permeation of water in and out of the microfluidic device. The kinetic growth parameters were quantified by counting the increase in the number of cells in each drop over time. In addition to determining the kinetic parameters, the cell‐size distribution of the microalgae was correlated with different stages of the growth. The single‐cell growth kinetics of C. vulgaris showed significant heterogeneity. The specific growth rate ranged from 0.55 to 1.52 day^?1 for different single cells grown in the same microfluidic device. In comparison, the specific growth rate in bulk‐scale experiment was 1.12 day^?1. It was found that the average cell size changes significantly at different stages of the cell growth. The mean cell‐size increased from 5.99 ± 1.08 to 7.33 ± 1.3 µm from exponential to stationary growth phase. In particular, when multiple cells are grown in individual drops, we find that in the stationary growth phase, the cell size increases with the age of cell suggesting enhanced accumulation of fatty acids in older cells. Biotechnol. Bioeng. 2012; 109: 2987–2996. © 2012 Wiley Periodicals, Inc.