Nutrient removal and biomass production of marine microalgae cultured in recirculating aquaculture systems (RAS) water with low phosphate concentration.

This study was conducted to investigate the feasibility of microalgal biomass production and nutrient removal from recirculating aquaculture systems (RAS) water (RASW) with low phosphate concentration. For this purpose, Nannochloropsis oculata, Pavlova gyrans, Tetraselmis suecica, Phaeodactylum tricornutum, and their consortium were cultivated in RASW and RASW supplemented with vitamins (+V). Among them, N. oculata showed the maximum biomass production of 0.4 g/L in RASW. Vitamins supplementation significantly increased the growth of T. suecica from 0.16 g/L in RASW to 0.33 g/L in RASW + V. Additionally, T. suecica showed the highest nitrate (NO3-N) removal efficiency of 80.88 ± 2.08 % in RASW and 83.82 ± 2.08 % in RASW + V. Accordingly, T. suecica was selected for scaling up study of microalgal cultivation in RASW and RASW supplemented with nitrate (RASW + N) in 4-L airlift photobioreactors. Nitrate supplementation enhanced the growth of T. suecica up to 2.2-fold (day 15). The fatty acid nutritional indices in T. suecica cultivated in RASW and RASW + N showed optimal polyunsaturated fatty acids (PUFAs)/saturated fatty acid (SFAs), omega-6 fatty acid (n-6)/omega-3 fatty acid (n-3), indices of atherogenicity (IA), and thrombogenicity (IT)). Overall, the findings of this study revealed that despite low phosphate concentration, marine microalgae can grow in RASW and relatively reduce the concentration of nitrate. Furthermore, the microalgal biomass cultivated in RASW consisting of pigments and optimal fatty acid nutritional profile can be used as fish feed, thus contributing to a circular bioeconomy.

Strategies for livestock wastewater treatment and optimised nutrient recovery using microalgal-based technologies.

Global sustainable development faces several challenges in addressing the needs of a growing population. Regarding food industries, the heightening pressure to meet these needs has resulted in increased waste generation. Thus, recognising these wastes as valuable resources is crucial to integrating sustainable models into current production systems. For instance, the current 24 billion tons of nutrient-rich livestock wastewater (LW) generated yearly could be recovered and valorised via biological uptake through microalgal biomass. Microalgae-based livestock wastewater treatment (MbLWT) has emerged as an effective technology for nutrient recovery, specifically targeting carbon, nitrogen, and phosphorus. However, the viability and efficacy of these systems rely on the characteristics of LW, including organic matter and ammonium concentration, content of suspended solids, and microbial load. Thus, this systematic literature review aims to provide guidance towards implementing an integral MbLWT system for nutrient control and recovery, discussing several pre-treatments used in literature to overcome the challenges regarding LW as a suitable media for microalgae cultivation.

Coelastrella terrestris for Adonixanthin Production: Physiological Characterization and Evaluation of Secondary Carotenoid Productivity.

A novel strain of Coelastrella terrestris (Chlorophyta) was collected from red mucilage in a glacier foreland in Iceland. Its morphology showed characteristic single, ellipsoidal cells with apical wart-like wall thickenings. Physiological characterization revealed the presence of the rare keto-carotenoid adonixanthin, as well as high levels of unsaturated fatty acids of up to 85%. Initial screening experiments with different carbon sources for accelerated mixotrophic biomass growth were done. Consequently, a scale up to 1.25 L stirred photobioreactor cultivations yielded a maximum of 1.96 mg·L-1 adonixanthin in free and esterified forms. It could be shown that supplementing acetate to the medium increased the volumetric productivity after entering the nitrogen limitation phase compared to autotrophic control cultures. This study describes a promising way of biotechnological adonixanthin production using Coelastrella terrestris.

Crypthecodinium cohnii Growth and Omega Fatty Acid Production in Mediums Supplemented with Extract from Recycled Biomass.

Crypthecodinium cohnii is a marine heterotrophic dinoflagellate that can accumulate high amounts of omega-3 polyunsaturated fatty acids (PUFAs), and thus has the potential to replace conventional PUFAs production with eco-friendlier technology. So far, C. cohnii cultivation has been mainly carried out with the use of yeast extract (YE) as a nitrogen source. In the present study, alternative carbon and nitrogen sources were studied: the extraction ethanol (EE), remaining after lipid extraction, as a carbon source, and dinoflagellate extract (DE) from recycled algae biomass C. cohnii as a source of carbon, nitrogen, and vitamins. In mediums with glucose and DE, the highest specific biomass growth rate reached a maximum of 1.012 h-1, while the biomass yield from substrate reached 0.601 g·g-1. EE as the carbon source, in comparison to pure ethanol, showed good results in terms of stimulating the biomass growth rate (an 18.5% increase in specific biomass growth rate was observed). DE supplement to the EE-based mediums promoted both the biomass growth (the specific growth rate reached 0.701 h-1) and yield from the substrate (0.234 g·g-1). The FTIR spectroscopy data showed that mediums supplemented with EE or DE promoted the accumulation of PUFAs/docosahexaenoic acid (DHA), when compared to mediums containing glucose and commercial YE.

Photoautotrophic and Mixotrophic Cultivation of Polyhydroxyalkanoate-Accumulating Microalgae Consortia Selected under Nitrogen and Phosphate Limitation.

Microalgae consortia were photoautotrophically cultivated in sequencing batch photobioreactors (SBPRs) with an alteration of the normal growth and starvation (nutrient limitation) phases to select consortia capable of polyhydroxyalkanoate (PHA) accumulation. At the steady state of SBPR operation, the obtained microalgae consortia, selected under nitrogen and phosphate limitation, accumulated up to 11.38% and 10.24% of PHA in their biomass, which was identified as poly(3-hydroxybutyrate) (P3HB). Photoautotrophic and mixotrophic batch cultivation of the selected microalgae consortia was conducted to investigate the potential of biomass and PHA production. Sugar source supplementation enhanced the biomass and PHA production, with the highest PHA contents of 10.94 and 6.2%, and cumulative PHA productions of 100 and 130 mg/L, with this being achieved with sugarcane juice under nitrogen and phosphate limitation, respectively. The analysis of other macromolecules during batch cultivation indicated a high content of carbohydrates and lipids under nitrogen limitation, while higher protein contents were detected under phosphate limitation. These results recommended the selected microalgae consortia as potential tools for PHA and bioresource production. The mixed-culture non-sterile cultivation system developed in this study provides valuable information for large-scale microalgal PHA production process development following the biorefinery concept.

Highly Valuable Polyunsaturated Fatty Acids from Microalgae: Strategies to Improve Their Yields and Their Potential Exploitation in Aquaculture.

Microalgae have a great potential for the production of healthy food and feed supplements. Their ability to convert carbon into high-value compounds and to be cultured in large scale without interfering with crop cultivation makes these photosynthetic microorganisms promising for the sustainable production of lipids. In particular, microalgae represent an alternative source of polyunsaturated fatty acids (PUFAs), whose consumption is related to various health benefits for humans and animals. In recent years, several strategies to improve PUFAs' production in microalgae have been investigated. Such strategies include selecting the best performing species and strains and the optimization of culturing conditions, with special emphasis on the different cultivation systems and the effect of different abiotic factors on PUFAs' accumulation in microalgae. Moreover, developments and results obtained through the most modern genetic and metabolic engineering techniques are described, focusing on the strategies that lead to an increased lipid production or an altered PUFAs' profile. Additionally, we provide an overview of biotechnological applications of PUFAs derived from microalgae as safe and sustainable organisms, such as aquafeed and food ingredients, and of the main techniques (and their related issues) for PUFAs' extraction and purification from microalgal biomass.

Contribution of vitamin B12 to biogas upgrading and nutrient removal by different microalgae-based technology.

The algae-based technology has a positive effect on the treatment of biogas slurry and the purification of biogas, while vitamin B12 (VB12) is one of the important regulatory substances in the algae-based cultivation system. In this study, different concentrations of VB12 were used in three microalgal treatment technologies to assess their effect on simultaneous removal of nutrients from biogas slurry and removal of CO2 from raw biogas. Results showed that Chlorella vulgaris exhibited higher growth rate, mean daily productivity, chlorophyll a content, carbonic anhydrase activity and better photosynthetic properties when co-cultivated with Ganoderma lucidum, rather than when co-cultivated with activated sludge or under mono-cultivation. Maximum mean chemical oxygen demand, total nitrogen, total phosphorus and CO2 removal efficiencies were found to be 84.29 ± 8.28%, 83.27 ± 8.14%, 85.27 ± 8.46% and 65.71 ± 6.35%, respectively when microalgae were co-cultivated with Ganoderma lucidum under 100 ng L-1 of VB12. This study shows the potential of microalgae and fungi co-cultivation supplemented with VB12 for the simultaneous upgradation of biogas production as well as for the purification of biogas slurry.

Feedback between bottom-up and top-down control of stream biofilm mediated through eutrophication effects on grazer growth.

Algal biofilms in streams are simultaneously controlled by light and nutrient availability (bottom-up control) and by grazing activity (top-down control). In addition to promoting algal growth, light and nutrients also determine the nutritional quality of algae for grazers. While short-term experiments have shown that grazers increase consumption rates of nutrient-poor algae due to compensatory feeding, nutrient limitation in the long run can constrain grazer growth and hence limit the strength of grazing activity. In this study, we tested the effects of light and phosphorus availability on grazer growth and thus on the long-term control of algal biomass. At the end of the experiment, algal biomass was significantly affected by light, phosphorus and grazing, but the interactive effects of the three factors significantly changed over time. At both high light and phosphorus supply, grazing did not initially reduce algal biomass, but the effect of grazing became stronger in the final three weeks of the experiment. Snail growth was enhanced by light, rather than phosphorus, suggesting that algal quantity rather than quality was the main limiting factor for grazer growth. Our results highlight the role of feedback effects and the importance of long-term experiments in the study of foodweb interactions.

Secretomics: a biochemical footprinting tool for developing microalgal cultivation strategies.

Microalgae offer a promising source of biofuel and a wide array of high-value biomolecules. Large-scale cultivation of microalgae at low density poses a significant challenge in terms of water management. High-density microalgae cultivation, however, can be challenging due to biochemical changes associated with growth dynamics. Therefore, there is a need for a biomarker that can predict the optimum density for high biomass cultivation. A locally isolated microalga Cyanobacterium aponinum CCC734 was grown with optimized nitrogen and phosphorus in the ratio of 12:1 for sustained high biomass productivity. To understand density-associated bottlenecks secretome dynamics were monitored at biomass densities from 0.6 ± 0.1 to 7 ± 0.1 g/L (2 to 22 OD) in batch mode. Liquid chromatography coupled with mass spectrometry identified 880 exometabolites in the supernatant of C. aponinum CCC734. The PCA analysis showed similarity between exometabolite profiles at low (4 and 8 OD) and mid (12 and 16 OD), whereas distinctly separate at high biomass concentrations (20 and 22 OD). Ten exometabolites were selected based on their role in influencing growth and are specifically present at low, mid, and high biomass concentrations. Taking cues from secretome dynamics, 5.0 ± 0.5 g/L biomass concentration (16 OD) was optimal for C. aponinum CCC734 cultivation. Further validation was performed with a semi-turbidostat mode of cultivation for 29 days with a volumetric productivity of 1.0 ± 0.2 g/L/day. The secretomes-based footprinting tool is the first comprehensive growth study of exometabolite at the molecular level at variable biomass densities. This tool may be utilized in analyzing and directing microalgal cultivation strategies and reduction in overall operating costs.

Application of uniform design to evaluate the different conditions on the growth of algae Prymnesium parvum.

Prymnesium parvum is an environmentally harmful algae and well known for its toxic effects to the fish culture. However, there is a dearth of studies on the growth behavior of P. parvum and information on how the availability of nutrients and environmental factors affect their growth rate. To address this knowledge gap, we used a uniform design approach to quantify the effects of major nutrients (N, P, Si and Fe) and environmental factors (water temperature, pH and salinity) on the biomass density of P. parvum. We also generated the growth model for P. parvum as affected by each of these nutrients and environmental factors to estimate optimum conditions of growth. Results showed that P. parvum can reach its maximum growth rate of 0.789, when the water temperature, pH and salinity is 18.11 °C, 8.39, and 1.23‰, respectively. Moreover, maximum growth rate (0.895-0.896) of P. parvum reached when the concentration of nitrogen, phosphorous, silicon and iron reach 3.41, 1.05, 0.69 and 0.53 mg/l, respectively. The order of the effects of the environmental factors impacting the biomass density of P. parvum was pH > salinity > water temperature, while the order of the effects of nutrients impacting the biomass density of P. parvum was nitrogen > phosphorous > iron > silicon. These findings may assist to implement control measures of the population of P. parvum where this harmful alga threatens aquaculture industry in the waterbodies such as Ningxia region in China.

Insights into the physiology of Chlorella vulgaris cultivated in sweet sorghum bagasse hydrolysate for sustainable algal biomass and lipid production.

Supplementing cultivation media with exogenous carbon sources enhances biomass and lipid production in microalgae. Utilization of renewable organic carbon from agricultural residues can potentially reduce the cost of algae cultivation, while enhancing sustainability. In the present investigation a medium was developed from sweet sorghum bagasse for cultivation of Chlorella under mixotrophic conditions. Using response surface methodology, the optimal values of critical process parameters were determined, namely inoculum cell density (O.D.750) of 0.786, SSB hydrolysate content of the medium 25% v/v, and zero medium salinity, to achieve maximum lipid productivity of 120 mg/L/d. Enhanced biomass (3.44 g/L) and lipid content (40% of dry cell weight) were observed when the alga was cultivated in SSB hydrolysate under mixotrophic conditions compared to heterotrophic and photoautotrophic conditions. A time course investigation revealed distinct physiological responses in terms of cellular growth and biochemical composition of C. vulgaris cultivated in the various trophic modes. The determined carbohydrate and lipid profiles indicate that sugar addition to the cultivation medium boosts neutral lipid synthesis compared to structural lipids, suggesting that carbon flux is channeled towards triacylglycerol synthesis in the cells. Furthermore, the fatty acid profile of lipids extracted from mixotrophically grown cultures contained more saturated and monosaturated fatty acids, which are suitable for biofuel manufacturing. Scale-up studies in a photobioreactor using SSB hydrolysate achieved a biomass concentration of 2.83 g/L consisting of 34% lipids and 26% carbohydrates. These results confirmed that SSB hydrolysate is a promising feedstock for mixotrophic cultivation of Chlorella and synthesis of algal bioproducts and biofuels.

Influence of Nitrogen and Phosphorus on Microalgal Growth, Biomass, Lipid, and Fatty Acid Production: An Overview.

Microalgae can be used as a source of alternative food, animal feed, biofuel, fertilizer, cosmetics, nutraceuticals and for pharmaceutical purposes. The extraction of organic constituents from microalgae cultivated in the different nutrient compositions is influenced by microalgal growth rates, biomass yield and nutritional content in terms of lipid and fatty acid production. In this context, nutrient composition plays an important role in microalgae cultivation, and depletion and excessive sources of this nutrient might affect the quality of biomass. Investigation on the role of nitrogen and phosphorus, which are crucial for the growth of algae, has been addressed. However, there are challenges for enhancing nutrient utilization efficiently for large scale microalgae cultivation. Hence, this study aims to highlight the level of nitrogen and phosphorus required for microalgae cultivation and focuses on the benefits of nitrogen and phosphorus for increasing biomass productivity of microalgae for improved lipid and fatty acid quantities. Furthermore, the suitable extraction methods that can be used to utilize lipid and fatty acids from microalgae for biofuel have also been reviewed.

Kinetics Growth and Recovery of Valuable Nutrients from Selangor Peat Swamp and Pristine Forest Soils Using Different Extraction Methods as Potential Microalgae Growth Enhancers.

Soil extracts are useful nutrients to enhance the growth of microalgae. Therefore, the present study attempts for the use of virgin soils from Peninsular Malaysia as growth enhancer. Soils collected from Raja Musa Forest Reserve (RMFR) and Ayer Hitam Forest Reserve (AHFR) were treated using different extraction methods. The total dissolved nitrogen (TDN), total dissolved phosphorus (TDP), and dissolved organic carbon (DOC) concentrations in the autoclave methods were relatively higher than natural extraction with up to 132.0 mg N/L, 10.7 mg P/L, and 2629 mg C/L, respectively for RMFR. The results of TDN, TDP, and DOC suggested that the best extraction methods are autoclaved at 121 °C twice with increasing 87%, 84%, and 95%, respectively. Chlorella vulgaris TRG 4C dominated the growth at 121 °C twice extraction method in the RMRF and AHRF samples, with increasing 54.3% and 14%, respectively. The specific growth rate (µ) of both microalgae were relatively higher, 0.23 d-1 in the Ayer Hitam Soil. This extract served well as a microalgal growth promoter, reducing the cost and the needs for synthetic medium. Mass production of microalgae as aquatic feed will be attempted eventually. The high recovery rate of nutrients has a huge potential to serve as a growth promoter for microalgae.

Mineral phosphorus drives glacier algal blooms on the Greenland Ice Sheet.

Melting of the Greenland Ice Sheet is a leading cause of land-ice mass loss and cryosphere-attributed sea level rise. Blooms of pigmented glacier ice algae lower ice albedo and accelerate surface melting in the ice sheet's southwest sector. Although glacier ice algae cause up to 13% of the surface melting in this region, the controls on bloom development remain poorly understood. Here we show a direct link between mineral phosphorus in surface ice and glacier ice algae biomass through the quantification of solid and fluid phase phosphorus reservoirs in surface habitats across the southwest ablation zone of the ice sheet. We demonstrate that nutrients from mineral dust likely drive glacier ice algal growth, and thereby identify mineral dust as a secondary control on ice sheet melting.

Effect of iron and phosphorus on the microalgae growth in co-culture.

Iron and phosphorus (P) are the important micro- and macro-nutrient for microalgae growth, respectively. However, the effect of iron and P on microalgae growth in co-culture associating with the formation of dominate algae has not been investigated before. In the current study, Anabaene flos-aquae, Chlorella vulgaris and Melosira sp. were co-cultivated under the addition of different initial iron and P to reveal the effect of iron and phosphorus on the growth of microalgae. The results showed that the mean growth rate of A. flos-aquae, C. vulgaris and Melosira was 0.270, 0.261 and 0.062, respectively, indicating that the A. flos-aquae and C. vulgaris algae are liable to be the dominant algae while the growth of Melosira was restrained when co-cultured. The ratio of Fe to P has a significant impact on the growth of microalgae and could be regarded as an indicator of algae growth. Microalgae showed a much more obvious uptake of iron compared to that of P. The information obtained in the current study was useful for the forecast of water quality and the control of microalgae bloom.

Effects of different phosphorus concentrations on growth and biochemical composition of Desmodesmus communis (E.Hegewald) E.Hegewald.

The members of the family Scenedesmaceae has the most widely used microalgae species in algal biotechnology studies because of their fast growth rate, quality of nutrition content and lipid accumulation under nutrient-limiting conditions. However, the biochemical responses of the species under phosphorus (P) limiting conditions are still unknown. The growth and biochemical composition of Desmodesmus communis in response to different phosphorus concentrations were investigated in this research. Five different phosphorus conditions were used: control (BG11); excess treatments (50% P+, 75% P+) and limited treatments (50% P-, 75% P-The highest cell concentration was observed in 75% P+ (725.6 × 104 cells/mL), whereas the highest dry weight concentration (1.81 mg/L) was found in 50% P- medium. The highest total lipid (4.94%) accumulation was found in the 50% P + medium and the maximum protein (49.5%) content was detected in 50% P- medium. Fatty acid and amino acid compositions change according to P concentration. PUFAs concentrations are higher than SFAs and MUFAs. Therefore the microalgae biomass obtained from this study cannot be used for biodiesel production although it is more suitable for nutritional supplement productions.

Using microalgae for remediation of crude petroleum oil-water emulsions.

Crude petroleum oil spills are among the most important organic contaminations. While the separated oils accumulated on the surface water are relatively easily removed, the emulsified portions are more difficult to remove and pose significant threats to the environment. Bioremediation using bacteria has proven to be an effective method, but the biomass produced in this case does not have any significant remunerative value. In this work, microalgae were proposed to combine emulsified oil remediation process with the potential of microalgae as a biofuel feedstock, thus enhancing the economic and environmental benefits of the process. A freshwater strain of Chlorella vulgaris was grown in water containing different concentrations of emulsified crude oil at different temperatures. The specific growth rate (µmax ) of the microalgae for each initial oil concentration was determined and was found to increase with the increase in initial oil concentration. For example, at 30°C, the specific growth rate, µ increased from 0.477 to 0.784 per day as the oil concentration increased from 57 to 222 mg/L. At 30°C, the effect of substrate concentration agreed with that of the microalgae growth, whereas at 40°C, the drop in oil concentration decreased with the increase in concentration. The results were fitted to a modified Monod kinetics model that used specific interfacial area as the influential substrate instead of the actual concentration. The results of this study clearly show the potential of using microalgae for emulsified oil remediation at relatively high concentrations.

Novel Insights into Phosphorus Deprivation Boosted Lipid Synthesis in the Marine Alga Nannochloropsis oceanica without Compromising Biomass Production.

Nannochloropsis oceanica represents a preferred oleaginous alga for producing lipids. Here we found that phosphorus deprivation (PD) caused a severe decrease in protein and a considerable increase in lipids including triacylglycerol (TAG), yet it had little effect on the carbohydrate level and biomass production of N. oceanica. The combinatorial analysis by integrating physiological, biochemical, and transcriptomic data unraveled the molecular mechanisms underlying PD-induced lipid accumulation. Albeit attenuating the Calvin-Benson cycle, PD stimulated the C4-like pathway to maintain CO2 fixation for biomass production. PD attenuated nitrogen utilization and enhanced protein catabolism thus leading to protein decrease, from which the carbon was likely salvaged into the stimulated tricarboxylic acid cycle for supplying lipid synthesis with carbon precursors. The impairment of TAG catabolism by downregulating certain lipases rather than the stimulation of TAG assembly pathways contributed to PD-boosted TAG increase. These findings provide novel insights into PD-induced lipogenesis without compromising biomass production by N. oceanica.

Effects of Nitrogen Supplementation Status on CO2 Biofixation and Biofuel Production of the Promising Microalga Chlorella sp. ABC-001.

The use of microalgal biomass as feedstock for biofuels has been discussed for decades as it provides a sustainable approach to producing fuels for the future. Nonetheless, its feasibility has not been established yet and various aspects of biomass applications such as CO2 biofixation should also be explored. Therefore, in this study, the CO2 biofixation and lipid/carbohydrate production potential of Chlorella sp. ABC-001 were examined under various nitrogen concentrations. The highest biomass productivity and CO2 biofixation rate of 0.422 g/l/d and 0.683 g/l/d, respectively, were achieved under a nitrogen-rich condition (15 mM nitrate). Carbohydrate content was generally proportional to initial nitrate concentration and showed the highest value of 41.5% with 15 mM. However, lipid content showed an inverse relationship with nitrogen supplementation and showed the highest value of 47.4% with 2.5 mM. In consideration as feedstock for biofuels (bioethanol, biodiesel, and biogas), the sum of carbohydrate and lipid contents were examined and the highest value of 79.6% was achieved under low nitrogen condition (2.5 mM). For lipid-based biofuel production, low nitrogen supplementation should be pursued. However, considering the lower feasibility of biodiesel, pursuing CO2 biofixation and the production of carbohydrate-based fuels under nitrogenrich condition might be more rational. Thus, nitrogen status as a cultivation strategy must be optimized according to the objective, and this was confirmed with the promising alga Chlorella sp. ABC-001.

New insights on improved growth and biogas production potential of Chlorella pyrenoidosa through intermittent iron oxide nanoparticle supplementation.

In the present work, the effect of α-Fe2O3-nanoparticles (IONPs) supplementation at varying doses (0, 10, 20 and, 30 mg L-1) at the intermittent stage (after 12th day of growth period) was studied on the growth and biogas production potential of Chlorella pyrenoidosa. Significant enhancements in microalgae growth were observed with all the tested IONPs doses, the highest (2.94 ± 0.01 g L-1) being at 20 mg L-1. Consequently, the composition of the biomass was also improved. Based on the precedent determinations, theoretical chemical oxygen demand (CODth) as well as theoretical and stoichiometric methane potential (TMP, and SMP) were also estimated. The CODth, TMP, SMP values indicated IONPs efficacy for improving biogas productivity. Further, the biochemical methane potential (BMP) test was done for IONPs supplemented biomass. The BMP test revealed up to a 25.14% rise in biogas yield (605 mL g-1 VSfed) with 22.4% enhanced methane content for 30 mg L-1 IONPs supplemented biomass over control. Overall, at 30 mg L-1 IONPs supplementation, the cumulative enhancements in biomass, biogas, and methane content proffered a net rise of 98.63% in biomethane potential (≈ 2.86 × 104 m3 ha-1 year-1) compared to control. These findings reveal the potential of IONPs in improving microalgal biogas production.