Anaerobic digestion of microalgae as a necessary step to make microalgal biodiesel sustainable

The potential of microalgae as a source of biofuels and as a technological solution for CO₂ fixation is subject to intense academic and industrial research. In the perspective of setting up massive cultures, the management of large quantities of residual biomass and the high amounts of fertilizers must be considered. Anaerobic digestion is a key process that can solve this waste issue as well as the economical and energetic balance of such a promising technology. Indeed, the conversion of algal biomass after lipid extraction into methane is a process that can recover more energy than the energy from the cell lipids. Three main bottlenecks are identified to digest microalgae. First, the biodegradability of microalgae can be low depending on both the biochemical composition and the nature of the cell wall. Then, the high cellular protein content results in ammonia release which can lead to potential toxicity. Finally, the presence of sodium for marine species can also affect the digester performance. Physico-chemical pretreatment, co-digestion, or control of gross composition are strategies that can significantly and efficiently increase the conversion yield of the algal organic matter into methane. When the cell lipid content does not exceed 40%, anaerobic digestion of the whole biomass appears to be the optimal strategy on an energy balance basis, for the energetic recovery of cell biomass. Lastly, the ability of these CO₂ consuming microalgae to purify biogas and concentrate methane is discussed.     

Life-cycle assessment of microalgae culture coupled to biogas production

Due to resource depletion and climate change, lipid-based algal biofuel has been pointed out as an interesting alternative because of the high productivity of algae per hectare and per year and its ability to recycle CO₂ from flue gas. Another option for taking advantage of the energy content of the microalgae is to directly carry out anaerobic digestion of raw algae in order to produce methane and recycle nutrients (N, P and K). In this study, a life-cycle assessment (LCA) of biogas production from the microalgae Chlorella vulgaris is performed and the results are compared to algal biodiesel and to first generation biodiesels. These results suggest that the impacts generated by the production of methane from microalgae are strongly correlated with the electric consumption. Progresses can be achieved by decreasing the mixing costs and circulation between different production steps, or by improving the efficiency of the anaerobic process under controlled conditions. This new bioenergy generating process strongly competes with others biofuel productions.     

Improving carbohydrate production of Chlorella sorokiniana NIES‐2168 through semi‐continuous process coupled with mixotrophic cultivation

Biofuels from microalgae is now a hot issue of great potential. However, achieving high starch productivity with photoautotrophic microalgae is still challenging. A feasible approach to enhance the growth and target product of microalgae is to conduct mixotrophic cultivation. The appropriate acetate addition combined with CO₂ supply as dual carbon sources (i.e., mixotrophic cultivation) could enhance the cell growth of some microalgae species, but the effect of acetate‐mediated mixotrophic culture mode on carbohydrate accumulation in microalgae remains unclear. Moreover, there is still lack of the information concerning how to increase the productivity of carbohydrates from microalgae under acetate‐amended mixotrophic cultivation and how to optimize the engineering strategies to achieve the goal. This study was undertaken to develop an optimal acetate‐contained mixotrophic cultivation system coupled with effective operation strategies to markedly improve the carbohydrate productivity of Chlorella sorokiniana NIES‐2168. The optimal carbohydrate productivity of 695 mg/L/d was obtained, which is the highest value ever reported. The monosaccharide in the accumulated carbohydrates is mainly glucose (i.e., 85–90%), which is very suitable for bio‐alcohols fermentation. Hence, by applying the optimal process developed in this study, C. sorokiniana NIES‐2168 has a high potential to serve as a feedstock for subsequent biofuels conversion.     

Enhancement of acid resistance of Scenedesmus dimorphus by acid adaptation

When using flue gas as carbon source for microalgae cultivation, the resulting acidic environment caused by SO _X and NO _X can inhibit microalgal growth. In this study, Scenedesmus dimorphus acquired increased acid resistance by prior exposure to sublethal acid stress; a process defined as acid adaptation. Among the five algal species tested, S. dimorphus showed the highest level of acid tolerance to extreme acid challenge (exposure to pH 3.0). Non-adapted and acid-adapted exponential algal cells were used as inocula for tubular photobioreactors aerated with 2 % CO₂. Previously adapted at pH 4.0 for 1 h, S. dimorphus developed highest growth rate under extreme acidic condition, and the maximum biomass concentration and specific growth rate at pH 3.0 (3.638?±?0.074 g?L^?1 and 1.037?±?0.008 d^?1, respectively) were respectively 14.22 and 10.79 % higher than those of non-adapted cells. Moreover, acid-adapted cells could tolerate lower pH of 2.5, at which the growth of non-adapted cells was totally inhibited. All the results indicated that acid adaptation was an effective approach for the acid resistance enhancement of microalgae.     

Feasibility study of biogas upgrading coupled with nutrient removal from anaerobic effluents using microalgae-based processes

The present research was conducted to simultaneously optimize biogas upgrading and carbon and nutrient removal from centrates in a 180-L high-rate algal pond interconnected to an external CO₂ absorption unit. Different biogas and centrate supply strategies were assessed to increase biomass lipid content. Results showed 99 % CO₂ removal efficiencies from simulated biogas at liquid recirculation rates in the absorption column of 9.9 m³ m^?2 h^?1, concomitant with nitrogen and phosphorus removal efficiencies of 100 and 82 %, respectively, using a 1:70 diluted centrate at a hydraulic retention time of 7 days. The lipid content of the harvested algal–bacterial biomass remained low (2.9–11.2 %) regardless of the operational conditions, with no particular trend over time. The good settling characteristics of the algal–bacterial flocs resulted in harvesting efficiencies over 95 %, which represents a cost-effective alternative for algal biomass reutilization compared to conventional physical–chemical techniques. Finally, high microalgae biodiversity was found regardless of the operational conditions.     

Integrated biogas upgrading and hydrogen utilization in an anaerobic reactor containing enriched hydrogenotrophic methanogenic culture

Biogas produced by anaerobic digestion, is mainly used in a gas motor for heat and electricity production. However, after removal of CO₂, biogas can be upgraded to natural gas quality, giving more utilization possibilities, such as utilization as autogas, or distant utilization by using the existing natural gas grid. The current study presents a new biological method for biogas upgrading in a separate biogas reactor, containing enriched hydrogenotrophic methanogens and fed with biogas and hydrogen. Both mesophilic‐ and thermophilic anaerobic cultures were enriched to convert CO₂ to CH₄ by addition of H₂. Enrichment at thermophilic temperature (55°C) resulted in CO₂ and H₂ bioconversion rate of 320 mL CH₄/(gVSS h), which was more than 60% higher than that under mesophilic temperature (37°C). Different dominant species were found at mesophilic‐ and thermophilic‐enriched cultures, as revealed by PCR–DGGE. Nonetheless, they all belonged to the order Methanobacteriales, which can mediate hydrogenotrophic methanogenesis. Biogas upgrading was then tested in a thermophilic anaerobic reactor under various operation conditions. By continuous addition of hydrogen in the biogas reactor, high degree of biogas upgrading was achieved. The produced biogas had a CH₄ content, around 95% at steady‐state, at gas (mixture of biogas and hydrogen) injection rate of 6 L/(L day). The increase of gas injection rate to 12 L/(L day) resulted in the decrease of CH₄ content to around 90%. Further study showed that by decreasing the gas–liquid mass transfer by increasing the stirring speed of the mixture the CH₄ content was increased to around 95%. Finally, the CH₄ content around 90% was achieved in this study with the gas injection rate as high as 24 L/(L day). Biotechnol. Bioeng. 2012; 109: 2729–2736. © 2012 Wiley Periodicals, Inc.     

The effect of gradual increase in salinity on the biomass productivity and biochemical composition of several marine,halotolerant<Emphasis Type="Bold">,</Emphasis> and halophilic microalgae

Open ponds are the preferred cultivation system for large-scale microalgal biomass production. To be more sustainable, commercial scale biomass production should rely on seawater, as freshwater is a limiting resource, especially in places with high irradiance. If seawater is used for both pond fill and evaporative volume makeup, salinity of the growth media will rise over time. It is not possible for any species to achieve optimum growth over the whole saline spectrum (from seawater salinity level up to salt saturation state). In this study, we investigated the effects of gradual salinity increase (between 35 and 233 ppt) on biomass productivity and biochemical composition (lipid and carbohydrate) of six marine, two halotolerant, and a halophilic microalgae. A gradual and slow stepped salinity increase was found to expand the salinity tolerance range of tested species. A gradual reduction in biomass productivity and maximum photochemical efficiency was observed as a consequence of increased salinity in all tested species. Among the marine microalgae, Tetraselmis showed highest biomass productivity (32 mg L^?1 day^?1) with widest salinity tolerance range (35 to 109 ppt). Halotolerant Amphora and Navicula were able to grow from 35 ppt to 129 ppt salinity. Halophilic Dunaliella was the only species capable of growing between 35 and 233 ppt and showed highest lipid content (56.2%) among all tested species. This study showed that it should be possible to maintain high biomass in open outdoor cultivation utilizing seawater by growing Tetraselmis, Amphora, and Dunaliella one after another as salinity increases in the cultivation system.     

Mixed Wastewater Coupled with CO2 for Microalgae Culturing and Nutrient Removal

Biomass, nutrient removal capacity, lipid productivity and morphological changes of Chlorella sorokiniana and Desmodesmus communis were investigated in mixed wastewaters with different CO₂ concentrations. Under optimal condition, which was 1:3 ratio of swine wastewater to second treated municipal wastewater with 5% CO₂, the maximum biomass concentrations were 1.22 g L^-1 and 0.84 g L^-1 for C. sorokiniana and D. communis, respectively. Almost all of the ammonia and phosphorus were removed, the removal rates of total nitrogen were 88.05% for C. sorokiniana and 83.18% for D. communis. Lipid content reached 17.04% for C. sorokiniana and 20.37% for D. communis after 10 days culture. CO₂ aeration increased intracellular particle numbers of both microalgae and made D. communis tend to be solitary. The research suggested the aeration of CO₂ improve the tolerance of microalgae to high concentration of NH₄-N, and nutrient excess stress could induce lipid accumulation of microalgae.     

High biomass production and CO<Subscript>2</Subscript> fixation from biogas by <Emphasis Type="Italic">Chlorella</Emphasis> and <Emphasis Type="Italic">Scenedesmus</Emphasis> microalgae using tequila vinasses as culture medium

Tequila vinasses (TVs) generated during Tequila production are brown liquid residues rich in nutrients. The nutrient content of agro-industrial effluents represents an excellent resource to support low-cost biomass production of microalgae; nonetheless, it is crucial to select the suitable microalgal strain to attain the highest biomass production in each residue used. In this study, biomass production, CO₂ fixation from biogas, and cell compound accumulation by Chlorella vulgaris U162, Chlorella sp., Scenedesmus obliquus U169, and Scenedesmus sp. using biodigested and filtered TVs as culture medium were evaluated and compared with the conventional microalgal culture media, C30, BG-11, Bold 3N, and Bristol. The four microalgae evaluated attained the highest biomass production and CO₂ fixation rate cultured in both residues, accumulating mainly carbohydrates and proteins although the most appropriate microalga to be cultured in TVs was Chlorella sp., recording 2.30 g L^?1. Moreover, the nutrient ratio of filtered TVs was ideal to support biomass production while biodigested TVs need to be supplemented with nitrogen. Overall, these results demonstrated that tequila vinasses are an excellent resource to support high and quick biomass production of microalgae, which can be used to obtain biofuels as ethanol, biogas, and supplement food depicting an extra benefit during the appropriate disposal of this residue.     

Landscape genetic structure of natural populations of Acacia caven in Argentina

Acacia caven is a South American species which shows remarkable climate tolerance and ecological adaptability; as such, this species is suitable for colonizing anthropogenically degraded sites. This species is widely distributed, and six varieties have been described based on both morphological traits and molecular markers. Moreover, Aronson (1992) suggests that, for this species, geographical separation could be associated with ecological differentiation. In this study, amplified fragment length polymorphisms were used to study genetic variation within and among 15 populations of A. caven from five eco-regions of Argentina and to investigate (1) whether the varieties are genetically coherent, (2) whether the varieties correspond consistently to a single eco-region, (3) the proportion of the species diversity explained within and among varieties and eco-regions. Eight of the 225 bands appear to be under positive selection. The remaining 217 neutral loci showed a high percentage of polymorphism (99.1%). The estimates of genetic diversity H _j were generally high. The F _ST (0.315) was highly significant, providing evidence for genetic structure among populations. Hierarchical analysis of molecular variance indicated that variation among eco-regions was 8.2% and highly significant. The higher component of variance was found within populations (67.5%). STRUCTURE analysis suggested that the optimal number of K?=?11. The results showed that, in most cases, geographic separation is associated with ecological differentiation. Since differentiation of A. caven populations studied here in eco-regions was highly significant, sampling should include a large number of trees within populations as well as covering the wide ecological diversity of the species.     

Tolerance of the photosynthetic apparatus to acidification of the growth medium as a possible determinant of CO<Subscript>2</Subscript>-tolerance of the symbiotic microalga <Emphasis Type="Italic">Desmodesmus</Emphasis> sp. IPPAS-2014

The symbiotic unicellular chlorophyte Desmodesmus sp. IPPAS-2014 capable of growth at extremely high CO₂ levels prohibitive for most other microalgae is an interesting model for studies of CO₂ tolerance mechanisms and a promising organism for CO₂ biocapture. We studied the initial (0-60 min) phase of acclimation of this microalga to an abrupt decrease in pH of the medium sparged with air/20% CO₂ mixture. Acclimation of the culture to these conditions was accompanied by a sharp decrease in photochemical activity of the chloroplast followed by its recovery with a characteristic time of 10-50 min. We hypothesize that acidification of the cultivation medium by dissolving CO₂ plays a key role in the observed decrease in the photochemical activity. The possible role of photosynthetic apparatus tolerance to abrupt acidification in overall high tolerance of symbiotic microalgae to extremely high CO₂ levels is discussed.     

High-CO2 tolerance in microalgae: possible mechanisms and implications for biotechnology and bioremediation

Recent developments in the field of microalgal biotechnology, including CO₂ biomitigation and the discovery of new species of microalgae that are tolerant to extremely high CO₂ levels (40–100 vol%), have renewed interest in the physiological effects and mechanisms of high-CO₂ tolerance in photoautotrophs. Photosynthetic apparatus state transitions that increase ATP generation, upregulation of H⁺-ATPases pumping protons out of the cell, rapid shutdown of CO₂-concentrating mechanisms, and adjustment of membranes’ fatty acid composition are currently believed to be the key mechanisms governing cellular pH homeostasis and hence microalgae’s tolerance to high CO₂ levels, which is especially characteristic of extremophile and symbiotic species. The mechanisms governing acclimation to high CO₂ comprise the subject of this review and are discussed in view of the use of CO₂ enrichment to increase the productivity of microalgal cultures, as well as the practice of carbon capture from flue gases.     

Comparative life cycle assessment of autotrophic cultivation of Scenedesmus dimorphus in raceway pond coupled to biodiesel and biogas production

Life cycle assessment (LCA) of indigenous freshwater microalgae, Scenedesmus dimorphus, cultivation in open raceway pond and its conversion to biodiesel and biogas were carried out. The LCA inventory inputs for the biogas scenario was entirely based on primary data obtained from algal cultivation (in pilot scale raceway pond), harvesting, and biogas production; while only the downstream processing involved in biodiesel production namely drying, reaction and purification were based on secondary data. Overall, eight scenarios were modeled for the integrated process involving: algae-based CO₂ capture and downstream processing scenarios for biodiesel and biogas along with impact assessment of nutrient addition and extent of recycling in a life cycle perspective. The LCA results indicated a huge energy deficit and net CO₂ negative in terms of CO₂ capture for both the biodiesel and biogas scenarios, majorly due to lower algal biomass productivity and higher energy requirements for culture mixing. The sensitivity analysis indicated that variability in the biomass productivity has predominant effect on the primary energy demand and global warming potential (GWP, kg CO₂ eq.) followed by specific energy consumption for mixing algal culture. Furthermore, the LCA results indicated that biogas conversion route from microalgae was more energy efficient and sustainable than the biodiesel route. The overall findings of the study suggested that microalgae-mediated CO₂ capture and conversion to biodiesel and biogas production can be energy efficient at higher biomass productivity (> 10 g m⁻² day⁻¹) and via employing energy-efficient systems for culture mixing (< 2 W m⁻³).

     

A comparison of desiccation tolerance among 12 species of chironomid larvae

Tolerance to desiccation was compared among 12 Japanese species of chironomid larvae under the condition of 60% in relative humidity at 25.5?°C. Three parameters were assessed: time to 50% survival (T ₅₀), water loss at 50% survival (WL₅₀) and water loss rate (WLR). T ₅₀, WL₅₀ and WLR were determined as measures of desiccation tolerance, dehydration tolerance, and dehydration resistance, respectively. T ₅₀ was 64.4–142 min for most species, except Propsilocerus akamusi (Tokunaga) which took 872 min. WL₅₀ was 60.6–82.4% for all species. WLR was only 0.0664% per minute for Pr. akamusi, while it was 0.629–1.50% for the other species. These results showed that Pr. akamusi had a high desiccation tolerance due to a high preventive ability of evaporation from body surface. T ₅₀ showed no significant relationships to WL₅₀ or WLR among the 12 species, while there was a significant positive relationship between WL₅₀ and WLR. These results suggest that chironomid species have a trade-off tendency that a species has a high tolerance – low resistance or a high resistance – low tolerance for dehydration.     

Perspectives on microalgal CO2-emission mitigation systems — A review

The problem of climate change arising mainly from CO₂ emission is currently a critical environmental issue. Biofixation using microalgae has recently become an attractive approach to CO₂ capture and recycling with additional benefits of downstream utilization and applications of the resulting microalgal biomass. This review summarizes the history and strategies of microalgal mitigation of CO₂ emissions, photobioreactor systems used to cultivate microalgae for CO₂ fixation, current microalgae harvesting methods, as well as applications of valuable by-products. It is of importance to select appropriate microalgal species to achieve an efficient and economically feasible CO₂-emission mitigation process. The desired microalgae species should have a high growth rate, high CO₂ fixation ability, low contamination risk, low operation cost, be easy to harvest and rich in valuable components in their biomass.     

Co-digestion of manure and whey for in situ biogas upgrading by the addition of H2: process performance and microbial insights

In situ biogas upgrading was conducted by introducing H₂ directly to the anaerobic reactor. As H₂ addition is associated with consumption of the CO₂ in the biogas reactor, pH increased to higher than 8.0 when manure alone was used as substrate. By co-digestion of manure with acidic whey, the pH in the anaerobic reactor with the addition of hydrogen could be maintained below 8.0, which did not have inhibition to the anaerobic process. The H₂ distribution systems (diffusers with different pore sizes) and liquid mixing intensities were demonstrated to affect the gas-liquid mass transfer of H₂ and the biogas composition. The best biogas composition (75:6.6:18.4) was obtained at stirring speed 150 rpm and using ceramic diffuser, while the biogas in the control reactor consisted of CH₄ and CO₂ at a ratio of 55:45. The consumed hydrogen was almost completely converted to CH₄, and there was no significant accumulation of VFA in the effluent. The study showed that addition of hydrogen had positive effect on the methanogenesis, but had no obvious effect on the acetogenesis. Both hydrogenotrophic methanogenic activity and the concentration of coenzyme F₄₂₀ involved in methanogenesis were increased. The archaeal community was also altered with the addition of hydrogen, and a Methanothermobacter thermautotrophicus related band appeared in a denaturing gradient gel electrophoresis gel from the sample of the reactor with hydrogen addition. Though the addition of hydrogen increased the dissolved hydrogen concentration, the degradation of propionate was still thermodynamically feasible at the reactor conditions.     

Cryopreservation of some useful microalgae species for biotechnological exploitation

Culture collections of microalgae represent a biological resource for scientific research and biotechnological applications. When compared to the current methods of maintenance and sub-culturing, cryopreservation minimizes labor costs and is an effective method for maintaining a large range of species over long periods with high stability. In order to determine the best cryopreservation method for microalgae species with great biotechnological potential, three freezing protocols were employed using different cryoprotectants (dimethyl sulfoxide—Me₂SO; methanol—MeOH). Three marine microalgae species (Thalassiosira weissflogii; Nannochloropsis oculata, and Skeletonema sp.) were cooled by directly plunging into liquid nitrogen (?196°C) and with two-step controlled cooling protocols (?18°C and ?80°C pre-treatments). After storage periods ranging from 10 to 120 days, viability was determined by the ability of cells to actively grow again. Results obtained for T. weissflogii showed that this species could be preserved at ultra-low temperature (?196°C) for 10 and 30 days with 10?% Me₂SO and 5?% MeOH when employed a controlled cooling protocol (?80°C). N. oculata was successfully cryopreserved either by direct freezing or with controlled cooling protocols. N. oculata samples presented good responses when treated with 5?% Me₂SO, 10?% Me₂SO, 5?% MeOH and even without any cryoprotectant. Skeletonema sp. did not survive cryopreservation in any of the tested conditions. The results indicate the difficulty in establishing common protocols for different microalgae species, being necessary further studies for a better understanding of cell damages during freezing and thawing conditions for each species.     

Mineral nutrient requirements of cucumber seedlings

Mineral nutrient requirements for maximum growth rate of cucumber (Cucumis sativus L.) seedlings are estimated on the basis of three criteria. 1. The optimal weight proportions among the nutrients present in the seedlings. In relation to N = 100, close to 75 K, 13 P, 9 Ca, and 9 Mg are required. 2. The optimal ratio between the nitrogen sources NH₄ and NO₃ in the solution. The best growth is recorded with about equivalent amounts. Nitrate alone is also highly productive. Cucumber prefers nitrate and is sensitive to high ammonium concentrations. 3. The optimal total concentration in the solution corresponds to 200 to 300 milligrams of nitrogen per liter, with the proportions of the nutrients according to criterion 1. Simple growth methods are suggested in which the three criteria are fulfilled.     

Flue gas compounds and microalgae: (Bio-)chemical interactions leading to biotechnological opportunities

Flue gases are a resource yet to be fully utilised in microalgal biotechnology, not only to moderate the anthropogenic effects on our climate, but also to steer microalgal resource management towards innovative applications of microalgal biomass compounds. These gases, both untreated and treated into current discharge standards, contain CO₂, N₂, H₂O, O₂, NO_x, SO_x, C_xH_y, CO, particulate matter, halogen acids and heavy metals. To better steer and engineer flue gas-fed microalgal cultures, all these compounds need to be considered. Therefore, here, we review (i) the chemical composition and treatment technologies of flue gas, (ii) the uptake pathways and removal of the different compounds in microalgae reactors, and (iii) the tolerance and effects on microalgae of all flue gas compounds. By emphasising the interactions between microalgae and flue gas compounds, we envisage new pathways for microalgal biomass valorisation such as enzyme production for environmental technology, novel biogas production and biosequestration of minerals. Furthermore, we highlight fundamental and applied research niches that merit further investigation.