Principles, challenges, and optimization of indigenous microalgae-bacteria consortium for sustainable swine wastewater treatment.

Indigenous microalgae-bacteria consortium (IMBC) offers significant advantages for swine wastewater (SW) treatment including enhanced adaptability and resource recovery. In this review, the approaches for enriching IMBC both in situ and ex situ were comprehensively described, followed by symbiotic mechanisms for IMBC which involve metabolic cross-feeding and signal transmission. Strategies for enhancing treatment efficiencies of SW-originated IMBC were then introduced, including improving SW quality, optimizing system operating conditions, and adjusting microbial activities. Recommendations for maximizing treatment efficiencies were particularly proposed using a decision tree approach. Moreover, removal/recovery mechanisms for typical pollutants in SW using IMBC were critically discussed. Ultimately, a technical route termed SW-IMBC-Crop-Pig was proposed, to achieve a closed-loop economy for pig farms by integrating SW treatment with crop cultivation. This review provides a deeper understanding of the mechanism and strategies for IMBC's resource recovery from SW.

Attached indigenous microalgal-bacterial consortium with greater stress-resistance facilitated recovery of integrated fixed-film system after experiencing short-term stagnation inhibition.

Stability of integrated fixed-film indigenous microalgal-bacterial consortium (IF-IMBC) requires further investigation. This study focused on the influence of short-term stagnation (STS), caused by influent variations or equipment maintenance, on IF-IMBC. Results showed that the IF-IMBC system experienced initial inhibition followed by subsequent recovery during STS treatment. Enhanced organics utilization was believed to contribute to system recovery. It is proposed that the attached IMBC possessed greater stress resistance. On the one hand, a higher increase in bacteria potentially participating in organic degradation was observed. Moreover, the dominant eukaryotic species significantly decreased in suspended IMBC while its abundance remained stable in the attached state. On the other hand, increased abundance for most functional enzymes was primarily observed in the attached bacteria. This fundamental research aims to bridge the knowledge gap regarding the response of IMBC to variations in operational conditions.

Physiological responses and molecular mechanism of Chlorella sorokiniana to surgical mask exudates in wastewater.

Microalgae-based bioremediation is likely to be challenged by the microplastics (MPs) in wastewater induced by the widely use of surgical masks (SMs) during COVID-19. However, such toxic impact was generally evaluated under high exposure concentrations of MPs, which was not in agreement with the actual wastewater environments. Therefore, this study investigated the microalgal cellular responses to the surgical mask exudates (SMEs) in wastewater and explored the underlying inhibitory mechanism from the molecular perspective. Specifically, 390 items/L SMEs (including 200 items/L MPs which was the actual MP level in wastewater) significantly inhibited nutrient uptake and photosynthetic activities interrupted peroxisome biogenesis and induced oxidative stress which destroyed the structure of cell membrane. Moreover, the SMEs exposure also affected carbon fixation pathways, suppressed ABC transporters while promoted oxidative phosphorylation processes for the ATP accumulation These comprehensive processes led to an 8.5% reduced microalgae growth and variations of cellular biocomponents including lipid, carbohydrate, and protein. The increased carotenoids and consumed unsaturated fatty acid were considered to alleviate the SMEs-induced stress, and the enhanced EPS secretion facilitated the homogeneous aggregation. These findings will enhance current understandings of the SMEs effects in wastewater on microalgae and further improve the practical relevance of microalgae wastewater bioremediation technology.

Coagulation pretreatment coupled with indigenous microalgal-bacterial consortium system for on-site treatment of rural black wastewater.

Improper treatment of rural black wastewater (RBW) presents substantial challenges, including the wastage of resource, environmental contamination, and economic consequences. This study proposed an integrated process for RBW treatment, consisting of coagulation/flocculation (C/F) pretreatment and subsequent inoculation of indigenous microalgal-bacterial consortium (IMBC) for nitrogen recovery, namely C/F-IMBC process. Specifically, the optimal C/F conditions (polyaluminium chloride of 4 g/l, polyacrylamide of 50 mg/l, and pH of 6) were determined through a series of single-factor experiments, considering CN, turbidity, and dissolved organic matter (DOM) removal, economic cost, and potential influence on the water environment. Compared to the sole IMBC system for RBW treatment, the proposed C/F-IMBC process exhibited a remarkable 1.23-fold increase in microalgal growth and a substantial 17.6-22.6 % boost in nitrogen recovery. The altered RBW characteristic induced by C/F pretreatment was supposed to be responsible for the improved system performance. In particular, the abundance of DOM was decreased and its composition was simplified after C/F pretreatment, based on the analysis for excitation-emission matrices with parallel factor and gas chromatography-mass spectrometry, thus eliminating the potential impacts of toxic DOM components (e.g., Bis(2-ethylhexyl) phthalate) on IMBC activity. It should also be noted that C/F pretreatment modified microbial community structure as well, thereby regulating the expression of nitrogen-related genes and enhancing the system nitrogen recovery capacity. For instance, the functional Cyanobacteria responsible for nutrient recovery was enriched by 1.95-fold and genes involved in the assimilatory nitrate reduction to ammonia pathway were increased by 1.52-fold. These fundamental findings are expected to offer insights into the improvement of DOM removal and nitrogen recovery for IMBC-based wastewater treatment system, and provide valuable guidance for the development of sustainable on-site RBW treatment technologies.

Nitrification-denitrification co-metabolism in an algal-bacterial aggregates system for simultaneous pyridine and nitrogen removal.

Photosynthetic oxygenation in algal-bacterial symbiotic (ABS) system was mainly concerned to enhance contaminant biodegradation by developing an aerobic environment, while the role of nitrification-denitrification involved is often neglected. In this study, an algal-bacterial aggregates (ABA) system was developed with algae and activated sludge (PBR-1) to achieve simultaneous pyridine and nitrogen removal. In PBR-1, as high as 150 mg·L-1 pyridine could be completely removed at hydraulic residence time of 48 h. Besides, total nitrogen (TN) removal efficiency could be maintained above 80%. Nitrification-denitrification was verified as the crucial process for nitrogen removal, accounting for 79.3% of TN removal at 180 µmol·m-2·s-1. Moreover, simultaneous pyridine and nitrogen removal was enhanced through nitrification-denitrification co-metabolism in the ABA system. Integrated bioprocesses in PBR-1 including photosynthesis, pyridine biodegradation, carbon and nitrogen assimilation, and nitrification-denitrification, were revealed at metabolic and transcriptional levels. Fluorescence in situ hybridization analysis indicated that algae and aerobic species were located in the surface layer, while denitrifiers were situated in the inner layer. Microelectrode analysis confirmed the microenvironment of ABA with dissolved oxygen and pH gradients, which was beneficial for simultaneous pyridine and nitrogen removal. Mechanism of nitrification-denitrification involved in pyridine and nitrogen removal was finally elucidated under the scale of ABA.

Unraveling the intracellular and extracellular self-defense of Chlorella sorokiniana toward highly toxic pyridine stress.

A bottleneck of microalgae-based techniques for wastewater bioremediation is activity inhibition of microalgae by toxic pollutants. The defense strategies of Chlorella sorokinana against toxic pyridine were studied. Results indicated that pyridine caused photoinhibition and reactive oxygen species overproduction in a concentration-dependent manner. The 50% inhibitory concentration of pyridine (147 mg L-1) destroyed C/N balance, disrupted multiple metabolic pathways of C. sorokinana. In response to pyridine stress, ascorbate peroxidase and catalase activities increased to scavenge reactive oxygen species under pyridine concentrations lower than 23 mg L-1. At higher pyridine concentrations, the activation of calcium signaling pathways and phytohormones represented the predominant defense response. Extracellular polymeric substances increased 3.6-fold in 147 mg L-1 group than control, which interacted with pyridine through hydrophobic and aromatic stacking to resist pyridine entering algal cells. Unraveling the intracellular and extracellular self-defense mechanisms of microalgae against pyridine stress facilitates the development of microalgal-based technology in wastewater bioremediation.

Start-up and maintenance of indigenous microalgae-bacteria consortium treating toilet wastewater through partial nitrification and nitrite-type denitrification.

Microalgae-bacteria consortium (MBC) provides an alternative to sustainable treatment of human toilet wastewater (TWW) and resource recovery. This study compared the conventional activated sludge system and wastewater indigenous MBC system (IMBC) for nitrogen removal in TWW through the coupled partial nitrification (PN) and nitrite-type denitrification process. PN was firstly established by alternating FA and FNA. Subsequently, the successful PN maintenance with the nitrite accumulation rate ranging between 90.1-95.3% was achieved using two strategies: light irradiation with the appropriate specific light energy density at 0.0188-0.0598 kJ/mg VSS and the timely nitrite-type denitrification with the algae-secreted organics as the carbon source, eventually resulting in the nitrite accumulation rate ranging between 90.1-95.3%. In the IMBC-PN system, bacterial metabolism contributed to 91.5% of nitrogen removal and the rest was through microalgal assimilation. This study offers a sustainable hybrid IMBC-PN process for high NH4+-N strength wastewater treatment (e.g., TWW), which theoretically saves 23.5% aeration and 34.2% carbon source as well as reduces 17.0% sludge production.

Effect of extreme pH conditions on methanogenesis: Methanogen metabolism and community structure.

The control of pH is effective for inhibiting methanogenesis in the chain elongation fermentation (CEF) system. However, obscure conclusions exist especially with regard to the underlying mechanism. This study comprehensively explored the responses of methanogenesis in granular sludge at various pH levels, ranging from 4.0 to 10.0, from multiple aspects including methane production, methanogenesis pathway, microbial community structure, energy metabolism and electron transport. Results demonstrated that compared with that at pH 7.0, pH at 4.0, 5.5, 8.5 and 10.0 triggered a 100%, 71.7%, 23.8% and 92.1% suppression on methanogenesis by the end of 3 cycles lasting 21 days. This might be explained by the remarkably inhibited metabolic pathways and intracellular regulations. To be more specific, extreme pH conditions decreased the abundance of the acetoclastic methanogens. However, obligate hydrogenotrophic and facultative acetolactic/hydrogenotrophic methanogens were significantly enriched by 16.9%-19.5 fold. pH stress reduced the gene abundance and/or activity of most enzymes involved in methanogenesis such as acetate kinase (by 81.1%-93.1%), formylmethanofuran dehydrogenase (by 10.9%-54.0%) and tetrahydromethanopterin S-methyltransferase (by 9.3%-41.5%). Additionally, pH stress suppressed electron transport via improper electron carriers and decreased electron amount as evidenced by 46.3%-70.4% reduced coenzyme F420 content and diminished abundance of CO dehydrogenase (by 15.5%-70.5%) and NADH:ubiquinone reductase (by 20.2%-94.5%). pH stress also regulated energy metabolism with inhibited ATP synthesis (e.g., ATP citrate synthase level reduced by 20.1%-95.3%). Interestingly, the protein and carbohydrate content secreted in EPS failed to show consistent responses to acidic and alkaline conditions. Specifically, when compared with pH 7.0, the acidic condition remarkably reduced the levels of total EPS and EPS protein while both levels were enhanced in the alkaline condition. However, the EPS carbohydrate content at pH 4.0 and 10.0 both decreased. This study is expected to promote the understanding of the pH control-induced methanogenesis inhibition in the CEF system.

Impacts of 2-bromoethanesulfonic sodium on methanogenesis: Methanogen metabolism and community structure.

Production of medium-chain carboxylic acids (MCCAs) by chain elongation (CE) presents a competitive alternative to conventional products of methane in anaerobic digestion treating organic waste streams, considering energy recovery, economic, and environmental profits. However, the system stability and performance largely rely on the selective suppression of methanogens while stimulation of CE bacteria. Commercial inhibitors such as 2-bromoethanesulfonic sodium (BES) was shown to be effective, but controversial conclusions exist on its inhibition characteristics and the inhibition mechanism remains unclear. Therefore, this study systematically investigated the responses of methanogenesis in granular sludge to various BES levels, focusing on methane production, methanogenic pathway, dynamic populations, electron transport and energy metabolism. Results showed that compared with the control, 3.0 g/L BES was sufficient to induce a 72.9% reduced level on accumulative methane production by the end of 4 cycles (28 days), which was likely to be attributed to the significantly suppressed metabolic pathways and intracellular regulations. Specifically, BES suppressed the electron transport via unproper electron carriers and reduced electron amount as indicated by the decreased level of enzymes and genes involved such as coenzyme F420, CO dehydrogenase and NADH:ubiquinone reductase (H+-translocating). Moreover, BES regulated the intracellular energy metabolism, leading to the impeded ATP synthesis but enhanced ATP consumption as evidenced by the variations on the activity or abundance of acetate kinase, A1Ao-ATP synthase, nitrogenase and ATP citrate synthase. Additionally, BES enriched hydrogenotrophic methanogenesis over acetoclastic one as supported by variations on the archaeal community structures and regulations of differentially expressed genes involved. Moreover, BES also reduced the contents of both protein and carbohydrate in extracellular polymeric substances (EPS). This study is expected to enhance understanding of BES contribution to methanogenesis inhibition but MCCAs production in CE bioreactors.

Effect of free ammonia shock on Chlorella sp. in wastewater: Concentration-dependent activity response and enhanced settleability.

The unstable microbial activity and unsatisfactory settling performance impede the development and implementation of microalgal wastewater treatment, especially in high-ammonium wastewater in the presence of free ammonia (FA). The shock of FA due to the nutrient fluctuation in wastewater was demonstrated as the primary stress factor suppressing microalgal activities. Recent study has clearly revealed the inhibition mechanism of FA at a specific high level (110.97 mg/L) by inhibiting the genetic information processing, photosynthesis, and nutrient metabolism. However, the effects of various FA shock concentrations on microalgal activities and settling performance remain unknown, limiting the wastewater bioremediation efficiencies improvement and the process development. Herein, a concentration-dependent shock FA (that was employed on microalgae during their exponential growth stages) effect on microalgal growth and photosynthesis was observed. Results showed that the studied five FA shock concentrations ranging from 25 to 125 mg/L significantly inhibited biomass production by 14.7-57.0%, but sharp reductions in photosynthesis with the 36.0-49.0% decreased Fv/Fm values were only observed when FA concentration was above 75.0 mg/L. On the other hand, FA shock enhanced microalgal settling efficiency by 12.8-fold, which was believed to be due to the stimulated intra- and extracellular protein contents and thereby the enhanced extracellular polymer substances (EPS) secretion. Specifically, FA shock induced 40.2 ± 2.3% higher cellular protein content at the cost of the decreased carbohydrates (22.6 ± 1.3%) and fatty acid (39.0 ± 0.8%) contents, further improving the protein secretion by 1.21-fold and the EPS production by 40.2 ± 2.3%. These FA shock-induced variations in intra- and extracellular biomolecules were supported by the up-regulated protein processing and export at the assistance of excessive energy generated from fatty acid degradation and carbohydrates consumption. In addition, FA shock significantly decreased the biomass nutritional value as indicated by the 1.86-fold lower essential amino acid score and nearly 50% reduced essential to non-essential amino acids ratio, while slightly decreased the biodiesel quality. This study is expected to enrich the knowledge of microalgal activities and settling performance in response to fluctuant ammonium concentrations in wastewater and to promote the development of microalgal wastewater treatment.

Understanding the influence of free nitrous acid on microalgal-bacterial consortium in wastewater treatment: A critical review.

Microalgal-bacterial consortium (MBC) constitutes a sustainable and efficient alternative to the conventional activated sludge process for wastewater treatment (WWT). Recently, integrating the MBC process with nitritation (i.e., shortcut MBC) has been proposed to achieve added benefits of reduced carbon and aeration requirements. In the shortcut MBC system, nitrite or free nitrous acid (FNA) accumulation exerts antimicrobial influences that disrupt the stable process performance. In this review, the formation and interactions that influence the performance of the MBC were firstly summarized. Then the influence of FNA on microalgal and bacterial monocultures and related mechanisms together with the knowledge gaps of FNA influence on the shortcut MBC were highlighted. Other challenges and future perspectives that impact the scale-up of the shortcut MBC for WWT were illustrated. A potential roadmap is proposed on how to maximize the stable operation of the shortcut MBC system for sustainable WWT and high-value biomass production.

Instant Inhibition and Subsequent Self-Adaptation of Chlorella sp. Toward Free Ammonia Shock in Wastewater: Physiological and Genetic Responses.

Free ammonia (FA) has been recently demonstrated as the primary stress factor suppressing microalgal activities in high-ammonium wastewater. However, its inhibition mechanism and microalgal self-adaptive regulations remain unknown. This study revealed an initial inhibition and subsequent self-adaptation of a wastewater-indigenous Chlorella sp. exposed to FA shock. Mutual physiological and transcriptome analysis indicated that genetic information processing, photosynthesis, and nutrient metabolism were the most influenced metabolic processes. Specifically, for the inhibition behavior, DNA damage was indicated by the significantly up-regulated related genes, leading to the activation of cell cycle checkpoints, programmed apoptosis, and suppressed microalgal growth; FA shock inhibited the photosynthetic activities including both light and dark reactions and photoprotection through non-photochemical quenching; ammonium uptake was also suppressed with the inhibited glutamine synthetase/glutamine α-oxoglutarate aminotransferase cycle and the inactivated glutamate dehydrogenase pathway. With respect to microalgal self-adaptation, DNA damage possibly enhanced overall cell viability through reprogramming the cell fate; recovered nutrient uptake provided substances for self-adaptation activities including amino acid biosynthesis, energy production and storage, and genetic information processing; elevated light reactions further promoted self-adaptation through photodamage repair, photoprotection, and antioxidant systems. These findings enrich our knowledge of microalgal molecular responses to FA shock, facilitating the development of engineering optimization strategies for the microalgal wastewater bioremediation system.

Granular indigenous microalgal-bacterial consortium for wastewater treatment: Establishment strategy, functional microorganism, nutrient removal, and influencing factor.

Granular indigenous microalgal-bacterial consortium (G-IMBC) system integrates the advantages of the MBC and granular activated sludge technologies, also with superior microalgal wastewater adaptation capacity. In this review, the concept of IMBC was firstly described, followed by its establishment and acclimation strategies. Characteristics and advantages of G-IMBC system compared to other IMBC systems (i.e., attached and floc IMBC systems) were then introduced. Moreover, the involved functional microorganisms and their interactions, as well as nutrient removal mechanisms were systematically and critically reviewed. Finally, the influencing factors including wastewater characteristics and operation factors were discussed. This study aims to provide a comprehensive up-to-date summary of the G-IMBC system for sustainable wastewater treatment.

Effect of short-term light irradiation with varying energy densities on the activities of nitrifiers in wastewater.

Microalgal-bacterial consortium (MBC) process has been proposed as an alternative to conventional activated sludge process for nitrogen removal from wastewater. As one of the most influencing parameters, light irradiation effects on microalgae have been extensively investigated. However, light influence on the performance of nitrifiers in activated sludge and its mechanism remains unclear. In this study, the effects of three factors (light irradiation power, irradiation time and sludge concentration) on activities and physiological characteristics of ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB) were systematically studied through both the Design of Experiments driven response surface methodology (RSM) approach and light-nitrification kinetic modeling. Results indicated that light irradiation with the specific light energy density (Es) at 0.0203-0.1571 kJ·mg-1 VSS (80-160 W/400-1000 µmol·m-2·s-1, 2.0-5.0 h and 2750-4250 mg·L-1) stimulated the relative AOB activities (rAOB) by 120.0%. This was supported by the increased electron transport system activity, key enzyme activity (AMO) , gene expression (amoA) and energy generation (ATP consumption) in the light treatment. Moreover, further Es increasing up to 0.18 kJ·mg-1 VSS inhibited both AOB and NOB activities. The inhibition was ascribed to the joint light responses of metabolic disorders and lipid peroxidation. The findings enhance our understanding of nitrifiers' physiological responses to short-term light irradiation, and promote the development of MBC as a sustainable approach for wastewater treatment.

Response of substrate kinetics and biological mechanisms to various pH constrains for cultured Nitrobacter and Nitrospira in nitrifying bioreactor.

Nitrite (NO2-) oxidation is an essential step of biological nitrogen cycling in natural ecosystems, and is performed by chemolithoautotrophic nitrite-oxidizing bacteria (NOB). Although Nitrobacter and Nitrospira are regarded as representative NOB in nitrification systems, little attention has focused on kinetic characterisation of the coexistence of Nitrobacter and Nitrospira at various pH values. Here, we evaluate the substrate kinetics, biological mechanism and microbial community dynamics of an enrichment culture including Nitrobacter (17.5 ± 0.9%) and Nitrospira (7.2 ± 0.6%) in response to various pH constrains. Evaluation of the Monod equation at pH 6.0, 6.5, 7.0, 7.5, 8.0 and 8.5 showed that the enrichment had maximum rate (rmax) and maximum substrate affinity (KS) for NO2- oxidation at pH 7.0, which was also supported by the largest absolute abundance of Nitrobacter nxrA (5.26 × 107 copies per g wet sludge) and Nitrospira nxrB (1.975 × 109 copies per g wet sludge) genes. Moreover, the predominant species for the Nitrobacter-like nxrA were N. vulgaris and N. winogradskyi, while for the Nitrospira-like nxrB, the predominant species were N. japonica, N. calida and Ca. N. bockiana. Furthermore, the rmax was strongly and positively correlated with the abundance of the Nitrobacter nxrA or Nitrospira nxrB genes, or N. winogradsk, whereas KS was positively correlated with the abundance of Nitrobacter nxrA or Nitrospira nxrB genes or Ca. N. bockiana. Overall, this study could improve basis kinetic parameters and biological mechanism of NO2- oxidation in WWTPs.

Unravelling multiple removal pathways of oseltamivir in wastewater by microalgae through experimentation and computation.

Increased worldwide consumption of antiviral drugs (AVDs) amid COVID-19 has induced enormous burdens to the existing wastewater treatment systems. Microalgae-based bioremediation is a competitive alternative technology due to its simultaneous nutrient recovery and sustainable biomass production. However, knowledge about the fate, distribution, and interaction of AVDs with microalgae is yet to be determined. In this study, a concentration-determined influence of AVD oseltamivir (OT) was observed on the biochemical pathway of Chlorella sorkiniana (C.S-N1) in synthetic municipal wastewater. The results showed that high OT concentration inhibited biomass growth through increased oxidative stress and restrained photosynthesis. Nevertheless, complete OT removal was achieved at its optimized concentration of 10 mg/L by various biotic (82%) and abiotic processes (18.0%). The chemical alterations in three subtypes of extracellular polymeric substances (EPS) were primarily investigated by electrostatic (OT +8.22 mV vs. C.S-N1 -18.31 mV) and hydrophobic interactions between EPS-OT complexes supported by secondary structure protein analysis. Besides, six biodegradation-catalyzed transformation products were identified by quadrupole-time-of-flight mass spectrometer and by density functional theory. Moreover, all the TPs exhibited log Kow ≤ 5 and bioconcentration factor values of < 5000 L/kg, meeting the practical demands of environmental sustainability. This study broadens our understanding of microalgal bioadsorption and biodegradation, promoting microalgae bioremediation for nutrient recovery and AVDs removal.

Denitrifying anaerobic methane-oxidizing bacteria in river networks of the Taihu Basin: Community dynamics and assembly process.

Denitrifying anaerobic methane-oxidizing bacteria (DAMO bacteria) plays an important role in reducing methane emissions from river ecosystems. However, the assembly process of their communities underlying different hydrologic seasons remains unclarified. In this study, the dynamics of DAMO bacterial communities in river networks of the Taihu Basin were investigated by amplicon sequencing across wet, normal, and dry seasons followed by multiple statistical analyses. Phylogenetic analysis showed that Group B was the major subgroup of DAMO bacteria and significant dynamics for their communities were observed across different seasons (constrained principal coordinate analysis, p = 0.001). Furthermore, the neutral community model and normalized stochasticity ratio model were applied to reveal the underlying assembly process. Stochastic process and deterministic process dominated the assembly process in wet season and normal season, respectively and similar contributions of deterministic and stochastic processes were observed in dry season. Meanwhile, abundant (relative abundance >0.1%) and rare (relative abundance <0.01%) DAMO bacterial communities were found to be shaped via distinct assembly processes. Deterministic and stochastic processes played a considerable role in shaping abundant DAMO bacterial communities, while deterministic process mainly shaped rare DAMO bacterial communities. Results of this study revealed the dynamics of DAMO bacterial communities in river networks and provided a theoretical basis for further understanding of the assembly process.

Insights into the multi-targeted effects of free nitrous acid on the microalgae Chlorella sorokiniana in wastewater.

Microalgal-bacterial consortium process (MBCP) proposed as an alternative to the activated sludge process contains free nitrous acid (FNA). FNA antimicrobial influences on nitrifiers have been demonstrated. However, its influence on microalgae is largely unknown, limiting the system stability of MBCP. This study revealed the multi-targeted responses of a model wastewater microalgae, Chlorella sorokiniana, to FNA exposure through physiological and transcriptomic analyses. Results showed a concentration-dependent FNA-influence as both microalgal growth and photosynthesis (Fv/Fm, rETR, Y(II), NPQ) inversely correlated with FNA doses. Increased ROS, MDA content (5.0-fold), SOD (2.7-fold), and LDH (12.0-fold) activities in the treatments revealed FNA-induced oxidative pressure. Moreover, RNA-sequencing results revealed significantly downregulated genes related to photosynthesis, respiration, nitrogen metabolism, and tricarboxylic acid cycle. Comparatively, peroxisome, chlorophyll, and carotenoid genes were upregulated. These findings elucidate the inhibitory mechanisms of FNA on microalgae and contribute towards the prospective practical application of the MBCP system for sustainable wastewater treatment.

Microalgal Activity and Nutrient Uptake from Wastewater Enhanced by Nanoscale Zerovalent Iron: Performance and Molecular Mechanism.

Microalgae-based bioremediation presents an alternative to traditional biological wastewater treatment. However, its efficiency is still challenging due to low microalgal activities and growth rate in wastewater. Iron plays an important role in microbial metabolism and is effective to stimulate microbial growth. In this study, a novel approach was proposed to simultaneously promote microalgal activity and nutrient uptake from wastewater using nanoscale zerovalent iron (nZVI), and the underlying molecular mechanism was explored. Compared to the control, 0.05 mg/L of nZVI significantly enhanced biomass production by 113.3% as well as NH4+-N and PO43--P uptake rates by 32.2% and 75.0%, respectively. These observations were attributed to the enhanced metabolic pathways and intracellular regulations. Specifically, nZVI alleviated the cellular oxidative stress via decreased peroxisome biogenesis as indicated by reduced reactive oxygen species, enzymes, and genes involved. nZVI promoted ammonium assimilation, phosphate metabolism, carbon fixation, and energy generation. Moreover, nZVI regulated the biosynthesis and conversions of intracellular biocomposition, leading to increased carotenoid, carbohydrate, and lipid productions and decreased protein and fatty acid yields. The above metabolisms were supported by the regulations of differentially expressed genes involved. This study provided an nZVI-based approach and molecular mechanism for enhancing microalgal activities and nutrient uptake from wastewater.