Water-saving analysis on an effective water reuse system in biodiesel feedstock production based on Chlorella zofingiensis fed-batch cultivation.

The micralgae-based biofuel obtained from dairy wastewater (DWW) is considered a promising source of energy. However, this process consumes water due to the concentration of wastewater being normally too high for some micoralgae cultivation, and dilution is always needed. In this work, the cultivation of microalgae has been examined in non-recirculated water (NR) and recirculated water systems (R). The growth of Chlorella zofingiensis and the nutrient removal of DWW have been recorded. The comparison indicates the R had a little more advantage in biomass and lipid output (1.55, 0.22 g, respectively) than the NR (1.51, 0.20 g, respectively). However, the total chemical oxygen demand (COD), total Kjeldahl nitrogen (TKN), and total phosphorus (TP) removals of the R were lower than those of the NR system during the culture. The highest removal of total COD, TKN, and TP were 85.05%, 93.64%, and 98.45%, respectively. Furthermore, no significant difference has been observed in the higher heating value and lipid content of the biomass of the R and NR. The results show the R can save 30% of the total water input during the culture. All above results indicate the R system has great potential in industry.

Treatment of mixed wastewater by vertical rotating microalgae-bacteria symbiotic biofilm reactor.

A novel vertical rotating microalgae-bacteria symbiotic biofilm reactor was built to treat the mixed wastewater containing municipal and soybean soaking wastewater. The reactor was operated in both sequential batch and semi-continuous modes. Under the sequential batch operation mode, the maximum removal rates for Chemical Oxygen Demand (COD), Total Nitrogen (TN), Total Phosphorus (TP), and Ammonia Nitrogen (NH4+-N) of the mixed wastewater were 95.6 %, 96.1 %, 97.6 %, and 100 %, respectively. During the semi-continuous operation, the water discharge indices decreased gradually and eventually stabilized. At stabilization, the removal rates of COD, TN, and NH4+-N achieved 98 %, 95 %, and 99.9 %, respectively. The maximum biomass productivity of the biofilm was 2.69 g·m-2·d-1. Additionally, the carbohydrate, protein and lipid comprised approximately 22 %, 51 % and 10 % of the dry weight of Chlorella. This study demonstrates the great potential of the microalgae-bacteria symbiotic biofilm system to treat food and domestic wastewater while harvesting microalgal biomass.

Harmony in detoxification: Microalgae unleashing the potential of lignocellulosic pretreatment wastewater for resource utilization.

Lignocellulosic biomass is a pivotal renewable resource in biorefinery process, requiring pretreatment, primarily chemical pretreatment, for effective depolymerization and subsequent transformation. This process yields solid residue for saccharification and lignocellulosic pretreatment wastewater (LPW), which comprises sugars and inhibitors such as phenols and furans. This study explored the microalgal capacity to treat LPW, focusing on two key hydrolysate inhibitors: furfural and vanillin, which impact the growth of six green microalgae. Chlorella sorokiniana exhibited higher tolerance to furfural and vanillin. However, both inhibitors hindered the growth of C. sorokiniana and disrupted algal photosynthetic system, with vanillin displaying superior inhibition. A synergistic inhibitory effect (Q < 0.85) was observed with furfural and vanillin on algal growth. Furfural transformation to low-toxic furfuryl alcohol was rapid, yet the addition of vanillin hindered this process. Vanillin stimulated carbohydrate accumulation, with 50.48 % observed in the 0.1 g/L furfural + 0.1 g/L vanillin group. Additionally, vanillin enhanced the accumulation of C16: 0 and C18: 2, reaching 21.71 % and 40.36 %, respectively, with 0.1 g/L vanillin. This study proposed a microalgae-based detoxification and resource utilization approach for LPW, enhancing the comprehensive utilization of lignocellulosic components. The observed biomass modifications also suggested potential applications for biofuel production, contributing to the evolving landscape of sustainable biorefinery processes.

Discovery of two novel bioactive algicidal substances from Brevibacillus sp. via metabolomics profiling and back-validation.

Identifying potent bacterial algicidal agents is essential for the development of effective, safe, and economically viable algaecides. Challenges in isolating and purifying these substances from complex secretions have impeded progress in this field. Metabolomics profiling, an efficient strategy for identifying metabolites, was pioneered in identifying bacterial algicidal substances in this study. Extracellular secretions from different generations of the algicidal bacterium Brevibacillus sp. were isolated for comprehensive analysis. Specifically, a higher algicidal efficacy was observed in the secretion from Generation 3 (G3) of Brevibacillus sp. compared to Generation 1 (G1). Subsequent metabolomics profiling comparing G3 and 1 revealed 83 significantly up-regulated metabolites, of which 9 were identified as potential algicidal candidates. Back-validation highlighted the potency of 4-acetamidobutanoic acid (4-ABC) and 8-hydroxyquinoline (8-HQL), which exhibited robust algicidal activity with 3d-EC50 values of 6.40 mg/L and 92.90 µg/L, respectively. These substances disrupted photosynthetic activity in M. aeruginosa by ceasing electron transfer in PSⅡ, like the impact exerted by Brevibacillus sp. secretion. These findings confirmed that 4-ABC and 8-HQL were the main algicidal components derived from Brevibacillus sp.. Thus, this study presents a streamlined strategy for identifying bacterial algicidal substances and unveils two novel and highly active algicidal substances. ENVIRONMENTAL IMPLICATION: Harmful cyanobacterial blooms (HCBs) pose significant environmental problems and health effects to humans and other organisms. The increasing frequency of HCBs has emerged as a pressing global concern. Bacterial-derived algicidal substances are expected to serve as effective, safe, and economically viable algaecides against HCBs. This study presents a streamlined strategy for identifying bacterial algicidal substances and unveils two novel substances (4-ABC and 8-HQL). These two substances demonstrate remarkable algicidal activity and disrupt the photosynthetic system in M. aeruginosa. They hold potential as prospective algaecides for addressing HCBs.

Effects of three surfactants on co-conversion of endogenous carbon and nitrogen of dairy wastewater in mesophilic hydrolytic acidification coupled microalgae culture system.

To determine the feasibility of enhancing co-conversion of endogenous C and N in dairy wastewater by surfactants, the effects of varying dosages of sodium lignosulfonate (LS), sodium dodecyl sulfate (SDS), and Tween-80 on mesophilic hydrolytic acidification coupled microalgae culture system were investigated. Tween-80 had a positive effect on hydrolytic acidification, while LS had no clear effect and SDS had a negative effect. Tween-80 significantly increased the C/N ratio in hydrolytic acidification liquor (HAL) (maximum HAc/NH4+-N rate reached 7.90 in 2.9% Tween-80 test). Pyrosequencing analysis demonstrated that community richness and diversity decreased and the proportion of acidobacteria increased with increasing Tween-80 dosage. Furthermore, the effect of Tween-80-enhanced HAL on microalgae (Chlorella pyrenoidosa) growth and nitrogen removal and the assimilation of C and N on the microalgae culture system were investigated. The biomass concentration and a C and N assimilation yield of 4.8% Tween-80 test (1.75 g/L, 825.01 mg/L, 126.68 mg/L) were significantly higher than those of both the low-concentration Tween-80 tests and control. The yield of protein and carbohydrates and higher heating value in the microalgae culture system were also promoted by Tween-80.

Effects of different nitrogen sources and light paths of flat plate photobioreactors on the growth and lipid accumulation of Chlorella sp. GN1 outdoors.

To assess the potential of Chlorella sp. GN1 for producing biodiesel raw materials in flat plate photobioreactors (FPPs) outdoors, we optimized the nitrogen sources and concentrations for the growth of the algae. The effects of different light paths of FPPs on the growth, lipid accumulation, and fatty acids of Chlorella sp. GN1 were also studied. As the light path of the FPPs was reduced, the alga could accumulate lipids rapidly, achieving high lipid content and lipid productivity outdoors. The highest lipid content obtained was 53.5%, when the light path was 5 cm. In addition, the lipid productivity was 66.7 mg L-1 day-1. The main fatty acids were C16/C18, accounting more than 90% of the total fatty acids. Results showed that Chlorella sp. GN1 had the ability to accumulate large quantities of lipids in FPPs outdoors and was a promising microalgal species for biofuel production.

Performance of a microalgal-bacterial consortium system for the treatment of dairy-derived liquid digestate and biomass production.

To enhance the treatment performance of dairy-derived liquid digestate (DLD) using microalgal-bacterial consortium system composed of Chlorella vulgaris and indigenous bacteria (CV), activated sludge was introduced to form a new microalgal-bacterial consortium system (Co-culture). The activated sludge shortened the lag phase and increased the specific growth rate of C. vulgaris (0.56 d-1). The biomass yield in the Co-culture was 2.72 g L-1, which was lower than that in the CV (3.24 g L-1), but the Co-culture had an improved COD (chemical oxygen demand) removal (25.26%) compared to the CV (13.59%). Quantitative PCR and metagenomic analyses demonstrated that microalgae also promoted bacterial growth, but influenced differently on the bacterial communities of indigenous bacteria and activated sludge. Compared with indigenous bacteria, activated sludge was more prone to forming a favorable symbiosis with C. vulgaris. These findings contribute to the construction of efficient microalgal-bacterial consortium system in wastewater treatment.

Treatment of low C/N ratio wastewater and biomass production using co-culture of Chlorella vulgaris and activated sludge in a batch photobioreactor.

The aim of this work was to study the performance of pollutants removal and biomass production by co-culture of Chlorella vulgaris and activated sludge in a batch photobioreactor (PBR), compared with their single system to treat a low C/N ratio (COD/N = 4.3) wastewater. The co-culture system surpassed activated sludge system in terms of nutrients removal and outperformed microalgae alone system in regard to COD removal. Biomass productivity of the co-culture system was 343.3 mg L-1 d-1, and the harvested biomass could be developed as biofuels, animal feeds or soil conditioners due to the improved calorific value and cellular composition compared with activated sludge. The low C/N ratio wastewater enabled bacteria to maintain a relatively low level, hence in favor of microalgae enrichment and nutrient recovery.

Cultivation of Chlorella vulgaris on unsterilized dairy-derived liquid digestate for simultaneous biofuels feedstock production and pollutant removal.

In order to assess viability of microalgae cultivation using unsterilized dairy-derived liquid digestate (DLD) for simultaneous biofuels feedstock production and contaminant removal, four DLD concentrations (25%, 50%, 75% and 100%) were used to grow Chlorella vulgaris in batch photobioreactors (PBRs). The 25% DLD was an ideal alternative medium in that high growth rate (0.69 d-1), high lipid productivity (112.9 mg L-1 d-1) as well as high nutrient removal were attained. The high DLD concentration caused inhibition of microalgal growth, where COD was more inhibitive than ammonium. The presence of bacteria did not influence microalgae production because of limited growth. Microalgal growth reduced the richness and diversity of bacterial community. Furthermore, the species of Bacteroidetes, Candidatus Saccharibacteria, and Chlamydiae rather than Proteobacteria benefited microalgal-bacterial symbiosis. These findings contribute to better application of microalgal-bacterial system for large-scale microalgae cultivation as well as environmental sustainability.

The roles of starch and lipid in Chlorella sp. during cell recovery from nitrogen starvation.

The influence of N-replenishment on cell growth, chlorophyll content, nitrogen uptake, and accumulation of starch and lipid was studied in Chlorella sp. N-starved algal cells were fully recovered within 2days after N-replenishment. Stored starch in N-deficient culture was degraded immediately during recovery process. However, lipid response had a period of delay when suffered from long starvation. During the recovery process, neutral lipid was reduced accompanied by an increase in membrane lipid. It was speculated that starch served as a source of carbon and energy catabolized to support quick recovery of the culture, while lipid preferred to play a structural role that specific fatty acid species were released from storage lipid and as building blocks for quick synthesis of membrane lipid. In light of rapid growth recovery and no net degradation in total fatty acids, a semi-continuous cultivation process might be a potential way to enhance lipid productivity.

Microalgae consortia cultivation in dairy wastewater to improve the potential of nutrient removal and biodiesel feedstock production.

The potential of microalgae consortia used in dairy wastewater treatment combined with microalgae biodiesel feedstock production was evaluated by comparing the nutrient removal of dairy wastewater, the growth of cells, and the lipid content and composition of biomass between monoalgae and microalgae consortia cultivation system. Our results showed that higher chemical oxygen demand (COD) removal (maximum, 57.01-62.86 %) and total phosphorus (TP) removal (maximum, 91.16-95.96 %) were achieved in almost microalgae consortia cultivation system than those in Chlorella sp. monoalgae cultivation system (maximum, 44.76 and 86.74 %, respectively). In addition, microalgae consortia cultivation except the mixture of Chlorella sp. and Scenedesmus spp. reached higher biomass concentration (5.11-5.41 g L(-1)), biomass productivity (730.4-773.2 mg L(-1) day(-1)), and lipid productivity (143.7-150.6 mg L(-1) day(-1)) than those of monoalgae cultivation (4.72 g L(-1), 674.3, and 142.2 mg L(-1) day(-1), respectively) on the seventh day. Furthermore, the fatty acid methyl ester (FAME) profiles indicated the lipids produced from microalgae consortia cultivation system were more suitable for biodiesel production. The microalgae consortia display superiority in dairy wastewater treatment and the getting feedstock for biodiesel production.

Characterization of Chlorococcum pamirum as a potential biodiesel feedstock.

To evaluate the potential of Chlorococcum pamirum for producing biodiesel, the effects of nitrogen, phosphate, initial cell concentrations and NaCl on lipid accumulation and growth were studied. The highest specific growth rate (µ(max)), biomass productivity and lipid content achieved was 1.888 d(-1), 350.1 mg L(-1) day(-1), and 64.9%, respectively. Under nitrogen-deficient condition, the cells accumulated lipids faster at low initial cell concentration. Additional NaCl to nitrogen-deficient media accelerated the lipid accumulation. When adding 10 g L(-1) NaCl to nitrogen-deficient media, the lipid content and productivity of cells cultured outdoors with high initial cell concentration increased from 38.2% and 153 mg L(-1) day(-1) to 54.3% and 192 mg L(-1) day(-1) respectively. Moreover, NaCl enhanced the saturated fatty acids content from 56.40% to 73.41% of total fatty acids. The results show that C. pamirum is a promising organism for biofuel production.

Growth and lipid accumulation characteristics of Scenedesmus obliquus in semi-continuous cultivation outdoors for biodiesel feedstock production.

In an effort to identify suitable microalgal species for biodiesel production, seven species were isolated from various habitats and their growth characteristics were compared. The results demonstrated that a green alga Scenedesmus obliquus could grow more rapidly and synthesize more lipids than other six microalgal strains. S. obliquus grew well both indoors and outdoors, and reached higher µmax indoors than that outdoors. However, the cells achieved higher dry weight (4.36 g L(-1)), lipid content (49.6%) and productivity (183 mg L(-1) day(-1)) outdoors than in indoor cultures. During the 61 days semi-continuous cultivation outdoors, high biomass productivities (450-550 mg L(-1) day(-1)) and µmax (1.05-1.44 day(-1)) were obtained. The cells could also achieve high lipid productivities (151-193 mg L(-1) day(-1)). These results indicated that S. obliquus was promising for lipids production in semi-continuous cultivation outdoors.

Nutrient removal and biodiesel production by integration of freshwater algae cultivation with piggery wastewater treatment.

An integrated approach, which combined freshwater microalgae Chlorella zofingiensis cultivation with piggery wastewater treatment, was investigated in the present study. The characteristics of algal growth, lipid and biodiesel production, and nutrient removal were examined by using tubular bubble column photobioreactors to cultivate C. zofingiensis in piggery wastewater with six different concentrations. Pollutants in piggery wastewater were efficiently removed among all the treatments. The specific growth rate and biomass productivity were different among all the cultures. As the initial nutrient concentration increased, the lipid content of C. zofingiensis decreased. The differences in lipid and biodiesel productivity of C. zofingiensis among all the treatments mainly resulted from the differences in biomass productivity. It is worthy of note that the diluted piggery wastewater with 1900 mg L(-1) COD provided an optimal nutrient concentration for C. zofingiensis cultivation, where the advantageous nutrient removal and the highest productivities of biomass, lipid and biodiesel were presented.

Lipid accumulation and growth characteristics of Chlorella zofingiensis under different nitrate and phosphate concentrations.

To evaluate the potential of the green agla Chlorella zofingiensis as a feedstock for biodiesel production, the effects of nitrogen and phosphate on lipid accumulation and growth of C. zofingiensis were studied. The maximum specific growth rate (µ(max)) reached 2.15 day⁻¹ when the concentration of NaNO3 and K2HPO4·3H2O was 1.0 g L⁻¹ and 0.01 g L⁻¹, respectively. The lipid contents of C. zofingiensis grown in media deficient of nitrogen (65.1%) or phosphate (44.7%) were both higher than that obtained from cells grown in full medium (33.5%). The highest lipid productivity (87.1 mg L⁻¹ day⁻¹) was also obtained from cells grown in nitrogen deficient media, indicating nitrogen deficiency was more effective than phosphate deficiency for inducing lipid accumulation in C. zofingiensis. In addition, the feasibility of cultivating the alga in 60 L flat plate photobioreactors and 10 L bottles outdoors for biodiesel was also tested. It was found that C. zofingiensis could adapt to fluctuating temperatures and irradiance of outdoors and the highest µ(max) and lipid productivity could reach 0.362 day⁻¹ and 26.6 mg L⁻¹ day⁻¹ outdoors, respectively. The lipid production potential of C. zofingiensis is projected to be 31.1 kg ha⁻¹ day⁻¹ in outdoor culture. These results suggested that C. zofingiensis is a promising organism for feedstock production of biofuel and can be used in scaled up culture outdoors.

Lipid accumulation and growth of Chlorella zofingiensis in flat plate photobioreactors outdoors.

Culturing microalgae using natural sunlight is an effective way to reduce the cost of microalgae-based biodiesel production. In order to evaluate the feasibility of culturing Chlorella zofingiensis outdoors for biodiesel production, effects of nitrogen limitation and initial cell concentration on growth and lipid accumulation of this alga were investigated in 60 L flat plate photobioreactors outdoors. The highest µmax and biomass productivity obtained was 0.994 day(-1) and 58.4 mg L(-1)day(-1), respectively. The lipid content was much higher (54.5% of dry weight) under nitrogen limiting condition than under nitrogen sufficient condition (27.3%). With the increasing initial cell concentrations, the lipid contents declined, while lipid concentrations and productivities increased. The highest lipid content, lipid concentration, and lipid productivity obtained was 54.5%, 536 mg L(-1) and 22.3 mg L(-1)day(-1), respectively. This study demonstrated that it was possible to culture C. zofingiensis under outdoor conditions for producing biodiesel feedstock.