Francesco G. Gentili

A Single-Step Method for Rapid Extraction of Total Lipids from Green Microalgae

Microalgae produce a wide range of lipid compounds of potential commercial interest. Total lipid extraction performed by conventional extraction methods, relying on the chloroform-methanol solvent system are too laborious and time consuming for screening large numbers of samples. In this study, three previous extraction methods devised by Folch et al. (1957), Bligh and Dyer (1959) and Selstam and Öquist (1985) were compared and a faster single-step procedure was developed for extraction of total lipids from green microalgae. In the single-step procedure, 8 ml of a 2∶1 chloroform-methanol (v/v) mixture was added to fresh or frozen microalgal paste or pulverized dry algal biomass contained in a glass centrifuge tube. The biomass was manually suspended by vigorously shaking the tube for a few seconds and 2 ml of a 0.73% NaCl water solution was added. Phase separation was facilitated by 2 min of centrifugation at 350 g and the lower phase was recovered for analysis. An uncharacterized microalgal polyculture and the green microalgae Scenedesmus dimorphus, Selenastrum minutum, and Chlorella protothecoides were subjected to the different extraction methods and various techniques of biomass homogenization. The less labour intensive single-step procedure presented here allowed simultaneous recovery of total lipid extracts from multiple samples of green microalgae with quantitative yields and fatty acid profiles comparable to those of the previous methods. While the single-step procedure is highly correlated in lipid extractability (r2 = 0.985) to the previous method of Folch et al. (1957), it allowed at least five times higher sample throughput.

Microalgal biomass and lipid production in mixed municipal, dairy, pulp and paper wastewater together with added flue gases.

The aim of the study was to grow microalgae on mixed municipal and industrial wastewater to simultaneously treat the wastewater and produce biomass and lipids. All algal strains grew in all wastewater mixtures; however, Selenastrum minutum had the highest biomass and lipids yields, up to 37% of the dry matter. Nitrogen and phosphorus removal were high and followed a similar trend in all three strains. Ammonium was reduced from 96% to 99%; this reduction was due to algal growth and not to stripping to the atmosphere, as confirmed by the amount of nitrogen in the dry algal biomass. Phosphate was reduced from 91% to 99%. In all strains used the lipid content was negatively correlated to the nitrogen concentration in the algal biomass. Mixtures of pulp and paper wastewater with municipal and dairy wastewater have great potential to grow algae for biomass and lipid production together with effective wastewater treatment.

Co-digestion of cultivated microalgae and sewage sludge from municipal waste water treatment

In this study two wet microalgae cultures and one dried microalgae culture were co-digested in different proportions with sewage sludge in mesophilic and thermophilic conditions. The aim was to evaluate if the co-digestion could lead to an increased efficiency of methane production compared to digestion of sewage sludge alone. The results showed that co-digestion with both wet and dried microalgae, in certain proportions, increased the biochemical methane potential (BMP) compared with digestion of sewage sludge alone in mesophilic conditions. The BMP was significantly higher than the calculated BMP in many of the mixtures. This synergetic effect was statistically significant in a mixture containing 63% (w/w VS based) undigested sewage sludge and 37% (w/w VS based) wet algae slurry, which produced 23% more methane than observed with undigested sewage sludge alone. The trend was that thermophilic co-digestion of microalgae and undigested sewage sludge did not give the same synergy.

Quantification and characterisation of fatty acid methyl esters in microalgae: Comparison of pretreatment and purification methods

A systematic qualitative and quantitative analysis of fatty acid methyl esters (FAMEs) is crucial for microalgae species selection for biodiesel production. The aim of this study is to identify the best method to assess microalgae FAMEs composition and content. A single-step method, was tested with and without purification steps-that is, separation of lipid classes by thin-layer chromatography (TLC) or solid-phase extraction (SPE). The efficiency of a direct transesterification method was also evaluated. Additionally, the yield of the FAMEs and the profiles of the microalgae samples with different pretreatments (boiled in isopropanol, freezing, oven-dried and freeze-dried) were compared. The application of a purification step after lipid extraction proved to be essential for an accurate FAMEs characterisation. The purification methods, which included TLC and SPE, provided superior results compared to not purifying the samples. Freeze-dried microalgae produced the lowest FAMEs yield. However, FAMEs profiles were generally equivalent among the pretreatments.

Anaerobic co-digestion of sludge and microalgae grown in municipal wastewater – a feasibility study

In this study a natural mix of microalgae grown in wastewater of municipal character was co-digested with sewage sludge in mesophilic conditions, in both batch and semi-continuous modes. The semi-continuous experiment was divided into two periods with OLR1 (organic loading rate) of 2.4 kg volatile solids (VS) m-3 d-1 and HRT1 (hydraulic retention time) of 15 days, and OLR2 of 3.5 kg VS m-3 d-1 and HRT2 of 10 days, respectively. Results showed stable conditions during both periods. The methane yield was reduced when adding microalgae (from 200 ± 25 NmL CH4 g VSin-1, to 168 ± 22 NmL CH4 g VSin-1) but VS reduction was also decreased by 51%. This low digestibility was confirmed in the anaerobic batch test. However, adding microalgae improved the dewaterability of the digested sludge. The high heavy metals content in the microalgae resulted in a high heavy metals content in the digestate, making it more difficult to reuse the digestate as fertilizer on arable land. The heavy metals are thought to originate from the flue gas used as a CO2 source during the microalgae cultivation. Therefore the implementation of CO2 mitigation via algal cultivation requires careful consideration regarding the source of the CO2-rich gas.