Mercedes Ballesteros

Microalgae autoflocculation: an alternative to high-energy consuming harvesting methods

With increasing concerns regarding energy and environment, algae biofuel is generating considerable interest around the world. Nevertheless, the harvesting step required before downstream biomass processing is a major bottleneck. Commonly employed methods include addition of chemicals or use of mechanical equipment that increase dramatically the biofuel production cost. This review deals with naturally occurring processes that can help offset those costs by causing microalgae flocculation. Interaction theories are briefly reviewed. In addition, operational parameters such as pH, irradiance, nutrients, dissolved oxygen, and temperature effect on microalgae flocculation are evaluated. Finally, microalgae flocculation is also considered from an ecological point of view by taking advantage of their interaction with other microorganisms.

Algaculture integration in conventional wastewater treatment plants: Anaerobic digestion comparison of primary and secondary sludge with microalgae biomass

This study evaluated the feasibility of using microalgae biomass as feedstock for anaerobic digestion together with other biomasses (primary and secondary sludge) normally generated in WWTP. Raw microalgae biomass anaerobic biodegradability (33%) was higher than that of secondary sludge (23%). Thermal pretreatment enhanced 62% and 16% methane yield for Chlorellavulgaris and secondary sludge, respectively. When both substrates were codigested, methane yields remained low. On the other hand, primary sludge supported the highest anaerobic biodegradability (97%) and when combined with thermally pretreated C. vulgaris, methane yields were higher (13-17%) than the ones expected theoretically. Despite the high protein content of those substrates and the high nitrogen mineralization, no ammonia inhibition was detected. Thereby, this study showed that algae biomass is a potential cosubstrate for biogas production together with municipal wastewater sludge.

Protease cell wall degradation of Chlorella vulgaris: Effect on methane production

In order to optimize the enzymatic dosage and microalgae biomass loads subjected to enzymatic hydrolysis prior anaerobic digestion of Chlorella vulgaris, organic matter solubilisation and methane production were investigated. Experimental data using protease dosage of 0.585 AU g DW(-1) showed that increasing biomass loads up to 65 g L(-1) did not affect markedly the hydrolysis efficiency (51%). Enzymatically pretreated biomasses subjected to anaerobic digestion enhanced methane production by 50-70%. The attempt of decreasing the enzymatic dosages revealed diminished hydrolysis efficiency concomitantly with a decreased methane production enhancement. In agreement with the good results observed for organic matter conversion into biogas, total nitrogen mineralization was attained for enzymatically pretreated biomass. Despite the high protein content of the biomass and the biocatalyst used in the present study no ammonia inhibition was detected.

Enzymatic pretreatment of Chlorella vulgaris for biogas production: influence of urban wastewater as a sole nutrient source on macromolecular profile and biocatalyst efficiency.

Two biocatalysts, namely carbohydrases and proteases, were assessed for organic matter solubilisation and methane yield enhancement of microalgae biomass. This study evidenced Chlorella vulgaris carbohydrate accumulation (40% on VSS basis) when grown in urban wastewater. Despite of the carbohydrate prevailing fraction, protease pretreatment showed higher organic matter hydrolysis efficiency (54%). Microscopic observation revealed that carbohydrases affected slightly the cell wall while protease was not selective to wall constituents. Raw and pretreated biomass was digested at 1.5 kg tCOD m(-3) day(-1) organic loading rate (OLR1) and 20 days hydraulic retention time (HRT). The highest methane yield (137 mL CH4 g COD in(-1)) was achieved in the reactor fed with protease pretreated C. vulgaris. Additionally, anaerobic digestion was conducted at OLR2 (3 kg tCOD m(-3) day(-1)) and HRT (15 days). When compared to raw biomass, methane yield increased 5- and 6.3-fold at OLR1 and OLR2, respectively. No inhibitors were detected during the anaerobic digestion.

Ammonia tolerant inocula provide a good base for anaerobic digestion of microalgae in third generation biogas process.

This study investigated the ability of an ammonia-acclimatized inoculum to digest efficiently protein-rich microalgae for continuous 3rd generation biogas production. Moreover, we investigated whether increased C/N ratio could alleviate ammonia toxicity. The biochemical methane potential (BMP) of five different algae (Chlorella vulgaris)/manure (cattle) mixtures showed that the mixture of 80/20 (on VS basis) resulted in the highest BMP value (431mLCH4 gVS-1), while the BMP of microalgae alone (100/0) was 415mLCH4 gVS-1. Subsequently, anaerobic digestion of those two substrates was tested in continuous stirred tank reactors (CSTR). Despite of the high ammonium levels (3.7-4.2g NH4+-NL-1), CSTR reactors using ammonia tolerant inoculum resulted in relatively high methane yields (i.e. 77.5% and 84% of the maximum expected, respectively). These results demonstrated that ammonia tolerant inocula could be a promising approach to successfully digest protein-rich microalgae and achieve a 3rd generation biogas production.

Acclimation to extremely high ammonia levels in continuous biomethanation process and the associated microbial community dynamics

Acclimatized anaerobic communities to high ammonia levels can offer a solution to the ammonia toxicity problem in biogas reactors. In the current study, a stepwise acclimation strategy up to 10g NH4+-N L-1, was performed in mesophilic (37±1°C) continuously stirred tank reactors. The reactors were co-digesting (20/80 based on volatile solid) cattle slurry and microalgae, a protein-rich, 3rd generation biomass. Throughout the acclimation period, methane production was stable with more than 95% of the uninhibited yield. Next generation 16S rRNA gene sequencing revealed a dramatic microbiome change throughout the ammonia acclimation process. Clostridium ultunense, a syntrophic acetate oxidizing bacteria, increased significantly alongside with hydrogenotrophic methanogen Methanoculleus spp., indicating strong hydrogenotrophic methanogenic activity at extreme ammonia levels (>7g NH4+-N L-1). Overall, this study demonstrated for the first time that acclimation of methanogenic communities to extreme ammonia levels in continuous AD process is possible, by developing a specialised acclimation AD microbiome.

Comparison of Chlorella vulgaris and cyanobacterial biomass: cultivation in urban wastewater and methane production

Anaerobic digestion of microalgae is hampered by its complex cell wall. Against this background, cyanobacteria cell walls render this biomass as an ideal substrate for overcoming this drawback. The aim of the present study was to compare the growth of two cyanobacteria (Aphanizomenon ovalisporum and Anabaena planctonica) and a microalga (Chlorella vulgaris) in urban wastewater when varying the temperature (22, 27 and 32xa0°C). Cyanobacterial optimal growth for both strains was attained at 22xa0°C, while C. vulgaris did not show remarkable differences among temperatures. For all the microorganisms, ammonium removal was higher than phosphate. Biomass collected was subjected to anaerobic digestion. Methane yield of C. vulgaris was 184.8xa0mL CH4 g COD in−1 while with A. ovalisporum and A. planctonica the methane production was 1.2- and 1.4-fold higher. This study showed that cyanobacteria growth rates could be comparable to microalgae while presenting the additional benefit of an increased anaerobic digestibility.

Effect of microalgae storage conditions on methane yields

During the last decade, a lot of research has been focused on identifying the methane yields achievable when using microalgae biomass (fresh and pretreated) as a substrate in anaerobic digestion. Encountered differences are frequently attributed to the different microalgae strains (cell walls and macromolecular profiles) or the different metabolic activities of anaerobic sludge used as inoculum. Nevertheless, under the hypothesis that the state of microalgae upon biomass storage may also play a significant role, this study was designed to evaluate the effect of biomass processing and storage on methane yields and hydrolysis kinetics in batch mode assays. Slight changes in the macromolecular profile distribution of the different tested biomass were observed. Regardless of the time that the biomass was stored, results revealed that frozen biomass doubled the hydrolysis constant and enhanced methane yield by 1.56-fold compared to fresh microalgae biomass (82.4xa0mLxa0CH4xa0gxa0CODxa0in−1). Similar enhancement was obtained with the freeze-dried biomass, and slightly lower values were obtained (1.34-fold) for the biomass kept at 4xa0°C longer than a week. Likewise, the semi-continuously operated reactor fed with microalgae biomass stored for 28xa0days at 4xa0°C did not show any effect in terms of methane production, although nitrogen mineralization was higher than expected. Remarkably, the initial stage of the biomass should be carefully considered for comparison purposes with the available literature on batch mode assays. This study highlights the importance of considering how the biomass is stored before the anaerobic digestion process to avoid misleading conclusions.

Biochemical methane potential of microalgae biomass using different microbial inocula

BackgroundMicroalgae biomass is regarded as a potential feedstock for bioenergy purposes through anaerobic digestion (AD). Even though AD is a well-proven technology, the use of new feedstocks requires in-depth studies. A lot of research has been conducted assessing methane yield without paying attention to the anaerobic microbiome and their activities. For such a goal, the present investigation was designed to link methane yield to those two later sludge characteristics. In this sense, different anaerobic sources were tested, namely adapted to microalgae biomass and adapted to sewage sludge.ResultsDespite the registered differences for the anaerobic microbiome analysis and specific methane activities towards model substrates, sludge adapted to digest sewage sludge did not affect the methane yield of Chlorella sorokiniana and Scenedesmus sp. Opposite to that, sludge samples adapted to digest microalgae exhibited a concomitant increase in methane yield together with increasing digestion temperatures. More specifically, the values attained were 63.4u2009±u20091.5, 79.2u2009±u20093.1 and 108.2u2009±u20091.9xa0mL CH4 g COD in−1 for psychrophilic, mesophilic and thermophilic digestions, respectively. While psycro- and mesophilic digestion supported similar yields (most probably linked to their anaerobic microbiome resemblance), the values attained for thermophilic digestion evidenced the usefulness of having a highly specific microbiome. The relative abundance of Firmicutes, particularly Clostridia, and Proteobacteria together with an important abundance of hydrogenotrophic methanogens was highlighted in this inoculum.ConclusionOverall, this study showed that working with tailored anaerobic microbiome could help avoiding pretreatments devoted to methane yield enhancement.

Volatile fatty acids production from protease pretreated Chlorella biomass via anaerobic digestion: Volatile fatty acids production from protease pretreated Chlorella biomass via anaerobic digestion

Volatile fatty acids (VFAs) produced via anaerobic digestion (AD) are regarded as a low cost production process of building blocks of interest for the chemical industry. In this study, VFAs and methane production were assessed in batch reactors at different temperature ranges (psychrophilic 25°C, mesophilic 35°C, thermophilic 50°C) and different pH values (5.5 and 7.5) using protease pretreated Chlorella sp. biomass as substrate. Acetic acid and propionic acid were the most abundant products (up to 73% of the total VFAs) during the first days independently of the conditions. VFAs concentration decreased over time as methane was produced after a lag phase of 7–10 days. Results showed that best conditions for VFAs production were mesophilic temperature ranges (35°C) at neutral initial pH values (7.5), and psychrophilic temperature ranges (25°C) at low initial pH values (5.5) which resulted in a conversion of the initial COD into VFAs of 48%, respectively.