Aino-Maija Lakaniemi

Biogenic hydrogen and methane production from Chlorella vulgaris and Dunaliella tertiolecta biomass

BackgroundMicroalgae are a promising feedstock for biofuel and bioenergy production due to their high photosynthetic efficiencies, high growth rates and no need for external organic carbon supply. In this study, utilization of Chlorella vulgaris (a fresh water microalga) and Dunaliella tertiolecta (a marine microalga) biomass was tested as a feedstock for anaerobic H2 and CH4 production.ResultsAnaerobic serum bottle assays were conducted at 37°C with enrichment cultures derived from municipal anaerobic digester sludge. Low levels of H2 were produced by anaerobic enrichment cultures, but H2 was subsequently consumed even in the presence of 2-bromoethanesulfonic acid, an inhibitor of methanogens. Without inoculation, algal biomass still produced H2 due to the activities of satellite bacteria associated with algal cultures. CH4 was produced from both types of biomass with anaerobic enrichments. Polymerase chain reaction-denaturing gradient gel electrophoresis profiling indicated the presence of H2-producing and H2-consuming bacteria in the anaerobic enrichment cultures and the presence of H2-producing bacteria among the satellite bacteria in both sources of algal biomass.ConclusionsH2 production by the satellite bacteria was comparable from D. tertiolecta (12.6 ml H2/g volatile solids (VS)) and from C. vulgaris (10.8 ml H2/g VS), whereas CH4 production was significantly higher from C. vulgaris (286 ml/g VS) than from D. tertiolecta (24 ml/g VS). The high salinity of the D. tertiolecta slurry, prohibitive to methanogens, was the probable reason for lower CH4 production.

Anaerobic conversion of microalgal biomass to sustainable energy carriers – A review

This review discusses anaerobic production of methane, hydrogen, ethanol, butanol and electricity from microalgal biomass. The amenability of microalgal biomass to these bioenergy conversion processes is compared with other aquatic and terrestrial biomass sources. The highest energy yields (kJ g(-1) dry wt. microalgal biomass) reported in the literature have been 14.8 as ethanol, 14.4 as methane, 6.6 as butanol and 1.2 as hydrogen. The highest power density reported from microalgal biomass in microbial fuel cells has been 980 mW m(-2). Sequential production of different energy carriers increases attainable energy yields, but also increases investment and maintenance costs. Microalgal biomass is a promising feedstock for anaerobic energy conversion processes, especially for methanogenic digestion and ethanol fermentation. The reviewed studies have mainly been based on laboratory scale experiments and thus scale-up of anaerobic utilization of microalgal biomass for production of energy carriers is now timely and required for cost-effectiveness comparisons.

Growth of Chlorella vulgaris and associated bacteria in photobioreactors.

The aim of this study was to test three flat plate photobioreactor configurations for growth of Chlorella vulgaris under non‐axenic conditions and to characterize and quantify associated bacterial communities. The photobioreactor cultivations were conducted using tap water‐based media to introduce background bacterial population. Growth of algae was monitored over time with three independent methods. Additionally, the quantity and quality of eukaryotes and bacteria were analysed using culture‐independent molecular tools based on denaturing gradient gel electrophoresis (PCR‐DGGE) and quantitative polymerase chain reaction (QPCR). Static mixers used in the flat plate photobioreactors did not generally enhance the growth at the low light intensities used. The maximum biomass concentration and maximum specific growth rate were 1.0 g l−1 and 2.0 day−1 respectively. Bacterial growth as determined by QPCR was associated with the growth of C. vulgaris. Based on PCR‐DGGE, bacteria in the cultures mainly originated from the tap water. Bacterial community profiles were diverse but reproducible in all flat plate cultures. Most prominent bacteria in the C. vulgaris cultures belonged to the class Alphaproteobacteria and especially to the genus Sphingomonas. Analysis of the diversity of non‐photosynthetic microorganisms in algal mass cultures can provide useful information on the public health aspects and unravel community interactions.

Electricity generation from tetrathionate in microbial fuel cells by acidophiles

Inorganic sulfur compounds, such as tetrathionate, are often present in mining process and waste waters. The biodegradation of tetrathionate was studied under acidic conditions in aerobic batch cultivations and in anaerobic anodes of two-chamber flow-through microbial fuel cells (MFCs). All four cultures originating from biohydrometallurgical process waters from multimetal ore heap bioleaching oxidized tetrathionate aerobically at pH below 3 with sulfate as the main soluble metabolite. In addition, all cultures generated electricity from tetrathionate in MFCs at pH below 2.5 with ferric iron as the terminal cathodic electron acceptor. The maximum current and power densities during MFC operation and in the performance analysis were 79.6 mA m(-2) and 13.9 mW m(-2) and 433 mA m(-2) and 17.6 mW m(-2), respectively. However, the low coulombic efficiency (below 5%) indicates that most of the electrons were directed to other processes, such as aerobic oxidation of tetrathionate and unmeasured intermediates. The microbial community analysis revealed that the dominant species both in the anolyte and on the anode electrode surface of the MFCs were Acidithiobacillus spp. and Ferroplasma spp. This study provides a proof of concept that tetrathionate serves as electron donor for biological electricity production in the pH range of 1.2-2.5.

Eukaryotic and prokaryotic microbial communities during microalgal biomass production.

Eukaryotic and bacterial communities were characterized and quantified in microalgal photobioreactor cultures of freshwater Chlorella vulgaris and marine Dunaliella tertiolecta. The microalgae exhibited good growth, whilst both cultures contained diverse bacterial communities. Both cultures included Proteobacteria and Bacteroidetes, while C. vulgaris cultures also contained Actinobacteria. The bacterial genera present in the cultures were different due to different growth medium salinities and possibly different extracellular products. Bacterial community profiles were relatively stable in D. tertiolecta cultures but not in C. vulgaris cultures likely due to presence of ciliates (Colpoda sp.) in the latter. The presence of ciliates did not, however, cause decrease in total number of C. vulgaris or bacteria during 14 days of cultivation. Quantitative PCR (qPCR) reliably showed relative microalgal and bacterial cell numbers in the batch cultures with stable microbial communities, but was not effective when bacterial communities varied. Raw culture samples were successfully used as qPCR templates.

Selecting an indigenous microalgal strain for lipid production in anaerobically treated piggery wastewater

The aim of this study was to select a potential microalgal strain for lipid production and to examine the suitability of anaerobically treated piggery wastewater as a nutrient source for production of lipid-rich biomass with the selected microalga. Biomass and lipid productivity of three microalgal strains (Chlorella sorokiniana CY1, Chlorella vulgaris CY5 and Chlamydomonas sp. JSC-04) were compared by using different media, nitrogen sources, and nitrogen concentrations. The highest lipid content and productivity (62.5 wt%, 162 mg/L/d) were obtained with C. vulgaris with BG-11 with 62 mg N/L. Secondly, C. vulgaris was cultivated in sterilized, diluted (1-20×), anaerobically treated piggery wastewater. Biomass production decreased and lipid content increased, when wastewater was more diluted. The highest lipid content of 54.7 wt% was obtained with 20× dilution, while the highest lipid productivity of 100.7 mg/L/d with 5× dilution. Piggery wastewater is a promising resource for mass production of oleaginous microalgal biomass.

Mine wastewater treatment using Phalaris arundinacea plant material hydrolyzate as substrate for sulfate-reducing bioreactor.

A low-cost substrate, Phalaris arundinacea was acid hydrolyzed (Reed Canary Grass hydrolyzate, RCGH) and used to support sulfate reduction. The experiments included batch bottle assays (35 degrees C) and a fluidized-bed bioreactor (FBR) experiment (35 degrees C) treating synthetic mine wastewater. Dry plant material was also tested as substrate in batch bottle assays. The batch assays showed sulfate reduction with the studied substrates, producing 540 and 350mgL(-1) dissolved sulfide with RCGH and dry plant material, respectively. The soluble sugars of the RCGH presumably fermented into volatile fatty acids and hydrogen, which served as electron donors for sulfate reducing bacteria. A sulfate reduction rate of 2.2-3.3gL(-1)d(-1) was obtained in the FBR experiment. The acidic influent was neutralized and the highest metal precipitation rates were 0.84g FeL(-1)d(-1) and 15mg ZnL(-1)d(-1). The sulfate reduction rate in the FBR was limited by the acetate oxidation rate of the sulfate-reducing bacteria.

Long-term stability of bioelectricity generation coupled with tetrathionate disproportionation.

To prevent uncontrolled acidification of the environment, reduced inorganic sulfur compounds (RISCs) can be bioelectrochemically removed from water streams. The long-term stability of bioelectricity production from tetrathionate (S4O6(2-)) was studied in highly acidic conditions (pH<2.5) in two-chamber fed-batch microbial fuel cells (MFCs). The maximum current density was improved from previously reported 80mAm(-2) to 225mAm(-2) by optimizing the external resistance. The observed reaction products of tetrathionate disproportionation were sulfate and elemental sulfur. In long-term run, stable electricity production was obtained for over 700days with the average current density of 150mAm(-2). The internal resistance of the MFC decreased over time and no biofouling was observed. This study shows that tetrathionate is an efficient substrate also for long-term bioelectricity production.

Effects of different nickel species on autotrophic denitrification driven by thiosulfate in batch tests and a fluidized-bed reactor

Nickel is a common heavy metal and often occurs with nitrate (NO3-) in effluents from mining and metal-finishing industry. The present study investigates the effects of increasing concentrations (5-200mgNi/L) of NiEDTA2- and NiCl2 on autotrophic denitrification with thiosulfate (S2O32-) in batch tests and a fluidized-bed reactor (FBR). In batch bioassays, 50 and 100mgNi/L of NiEDTA2- only increased the transient accumulation of NO2-, whereas 25-100mgNi/L of NiCl2 inhibited denitrification by 9-19%. NO3- and NO2- were completely removed in the FBR at feed NiEDTA2- and NiCl2 concentrations as high as 100 and 200mgNi/L, respectively. PCR-DGGE revealed the dominance of Thiobacillus denitrificans and the presence of the sulfate-reducing bacterium Desulfovibrio putealis in the FBR microbial community at all feed nickel concentrations investigated. Nickel mass balance, thermodynamic modeling and solid phase characterization indicated that nickel sulfide, phosphate and oxide precipitated in the FBR during NiCl2 injection.

Impact of heavy metals on denitrification of simulated mining wastewaters

Thespreading of nitrogenous compounds into the environment is a common challenge duringmining industries. Typical explosives used in mining are N-based compoundswhich lead to nitrogen contamination of groundwater and water bodies. In goldextraction, cyanide used as lixiviant is also another source of nitrogen pollution.The present work aims to investigate the effect of heavy metals ondenitrification using batch bioassays. Cu, Ni, Co and Fe influence ondenitrification process was studied at pH 7.0. Below the soluble concentrationof 62 mg/L, Ni did not inhibit denitrification, whereas denitrification wasrepressed at soluble Ni concentration above 62 mg/L. At 122 mg/L of soluble Ni,50% inhibition of denitrification was observed. Below soluble concentration of86 mg/L, Co exerted no inhibitory effect on nitrate removal but moderatelydecreased the denitrification rate. Cu slowed denitrification down resulting in40% of nitrate removal averagely at the soluble concentration below 1 mg/L. Onthe contrary, Fe supplementationresulted in iron oxidation and soluble Fe concentrations ranging from 0.4-1.6mg/L that stimulated denitrification. Thepresent work indicates that denitrification can tolerate heavy metals and canbe suitable for acid mine drainage remediation.