Microalgae: A potential bioagent for treatment of emerging contaminants from domestic wastewater.

Water crisis around the world leads to a growing interest in emerging contaminants (ECs) that can affect human health and the environment. Research showed that thousands of compounds from domestic consumers, such as endocrine disrupting chemicals (EDCs), personal care products (PCPs), and pharmaceuticals active compounds (PhAcs), could be found in wastewater in concentration mostly from ng L-1 to µg L-1. However, generally, wastewater treatment plants (WWTPs) are not designed to remove these ECs from wastewater to their discharge levels. Scientists are looking for economically feasible biotreatment options enabling the complete removal of ECs before discharge. Microalgae cultivation in domestic wastewater is likely a feasible approach for removing emerging contaminants and simultaneously removing any residual organic nutrients. Microalgal growth rate and contaminants removal efficiency could be affected by various factors, including light intensity, CO2 addition, presence of different nutrients, etc., and these parameters could greatly help make microalgae treatment more efficient. Furthermore, the algal biomass harvests could be repurposed to produce various bulk chemicals such as sustainable aviation fuel, biofuel, bioplastic, and biochar; this could significantly enhance the economic viability. Therefore, this review summarizes the microalgae-based bioprocess and their mechanisms for removing different ECs from different wastewaters and highlights the different strategies to improve the ECs removal efficiency. Furthermore, this review shows the role of different ECs in biomass profile and the relevance of using ECs-treated microalgae biomass to produce green products, as well as highlights the challenges and future research recommendations.

Development of economical and sustainable cultivation system for biomass production and simultaneous treatment of municipal wastewater using Tetraselmis indica BDUG001.

Microalgal-based bioprocess offers several advantages including wastewater reclamations, therefore present study assessed the usability of the combination of untreated municipal sewage wastewater (UTMSWW) and secondary treated municipal sewage wastewater (STSWW) for nutrient removal and recovery by Tetraselmis indica (T. indica) BDUG001. The present study optimized the additional nutrient supplementations (e.g. ASN-III) percentage and day-night cycle, pH and pH with aeration for monitoring high-rate biomass production and nutrient recovery. The study results showed that the combination of 75% UTMSWW + 25% ASN-III supported maximum biomass production (2.65 ± 0.07 g/L). In the optimized day-night cycle (12:12 h), T. indica BDUG001 showed improved biomass production (2.75 ± 0.07 g/L), biomass productivity (165.63 ± 4.42 mg/L/d), and photosynthetic pigments production. Under optimized pH∼ 7.0 with aeration, maximum total nitrate (TN) removal efficiency (87.67 ± 3.08-91.55 ± 1.92%) was observed, while COD and TP removal was maximum at pH ∼ 9.0. The maximum biomass production (2.35 ± 0.07-2.77 ± 0.04 g/L) with biomass productivity (93.75 ± 167.19 ± 2.21 mg/L/d) and lipid content (42.98 ± 1.86-47.85 ± 0.21% DCW) were also at pH 7.0. with aeration. The present study verified the utilization of UTMSWW with the combination of conventional medium, optimized day-night cycle, pH with aeration along with designing low-cost PBR. It was the ideal system for the cultivation of T. indica BDUG001 for the recovery of nutrients from wastewater, production of biofuels and value-added feedstock.

Agriculture Waste Biomass Repurposed into Natural Fibers: A Circular Bioeconomy Perspective.

Fibers come from natural and fossil resources and are an essential commodity widely used by textile industries. Considering current supply and future demands, the repurposing of agricultural residues into fibers is an eco-friendly, attractive option that might mitigate environmental pollution. In this review, we have summarized multiple alternate secondary sources for fiber production, with a case study using banana plant residual biomass, a common agricultural waste in many developing countries. Specifically, in this review we have compared the different processing methods, e.g., chemical, mechanical, or biological methods, for repurposing agricultural residual biomass (including banana waste) into fibers. The development and analysis of an integrated biorefinery approach is needed to promote the fiber production from various agro-residual biomasses within the framework of circular bioeconomic concepts.

Influence of Carbon Sources on Biomass and Biomolecule Accumulation in Picochlorum sp. Cultured under the Mixotrophic Condition.

The major downfalls of the microalgal biorefinery are low volume of high value product accumulation, low biomass productivity and high cultivation costs. Here, we aimed to improve the biomass productivity of the industrially relevant Picochlorum sp. BDUG 100241 strain. The growth of Picochlorum sp. BDUG 100241 was investigated under different cultivations conditions, including photoautotrophic (with light), mixotrophic (1% glucose, with light) and heterotrophic (1% glucose, without light). Among them, Picochlorum sp. BDUG100241 showed the highest growth in the mixotrophic condition. Under different (1%) carbon sources' supplementation, including glucose, sodium acetate, glycerol, citric acid and methanol, Picochlorum sp. BDUG100241 growth was tested. Among them, sodium acetate was found to be most suitable carbon source for Picochlorum sp. BDUG 100241 growth, biomass (1.67 ± 0.18 g/L) and biomolecule productivity. From the different concentrations of sodium acetate (0, 2.5, 5.0, 7.5 and 10 g/L) tested, the maximum biomass production of 2.40 ± 0.20 g/L with the biomass productivity of 95 ± 5.00 mg/L/d was measured from 7.5 g/L in sodium acetate. The highest total lipid (53.50 ± 1.70%) and total carotenoids (0.75 ± 0.01 µg/mL) contents were observed at the concentration of 7.5 g/L and 5.0 g/L of sodium acetate as a carbon source, respectively. In conclusion, the mixotrophic growth condition containing 7.5 g/L of sodium acetate showed the maximum biomass yield and biomolecule accumulation compared to other organic carbon sources.

Two-phase anaerobic digestion of food waste: Effect of semi-continuous feeding on acidogenesis and methane production.

In present investigation, effect of diverting acidogenic off-gas from leached bed reactor (LBR) to up-flow anaerobic sludge blanket (UASB) reactor during semi-continuous food waste (FW) anaerobic digestion was evaluated. In test LBR headspace pressure (3.3 psi) was maintained with intermittent headspace gas transfer into UASB. In control, same headspace pressure was maintained without gas transfer. The semi-continuous FW addition affected the characteristics and production of leachate in control and test LBR. The cumulative COD, total soluble products and methane yields were 1.26, 1.37 and 3 times higher in the test LBR than the control. The acetate and methane yields from test LBR were 697.8 g·kgVSadded-1 and 167.55 mL·gCOD-1feeding. Acidogenic gas transfer maintained low partial pressure of hydrogen and the hydrogen to carbon-di-oxide ratio in the headspace of LBR, which were thermodynamically favorable for microbial metabolism and concomitant high-rate production of acetate-rich volatile fatty acid and methane-rich biogas from FW.

Biomass pre-treatments of the N2-fixing cyanobacterium Tolypothrix for co-production of methane.

Tolypothrix, a self-flocculating, fast growing, CO2 and nitrogen-fixing cyanobacterium, can be cultivated in nutrient-poor ash dam waters of coal-fired power stations, converting CO2 emissions into organic biomass. Therefore, the biomass of Tolypothrix sp. is a promising source for bio-fertiliser production, providing micro- and macronutrients. Energy requirements for production could potentially be offset via anaerobic digestion (AD) of the produced biomass, which may further improve the efficiency of the resulting biofertilizer. The aim of this study was to evaluate the effectiveness of pre-treatment conditions and subsequent methane (CH4) production of Tolypothrix under out-door cultivation conditions. Pre-treatments on biogas and methane production for Tolypothrix sp. biomass investigated were: (1) thermal at 95 °C for 10 h, (2) hydrothermal by autoclave at 121 °C at 1013.25 hPa for 20 min, using a standard moisture-heat procedure, (3) microwave at an output power of 900 W and an exposure time of 3 min, (4) sonication at an output power of 10 W for 3.5 h at 10 min intervals with 20 s breaks and (5) freeze-thaw cycles at -80 °C for 24 h followed by thawing at room temperature. Thermal, hydrothermal and sonication pre-treatments supported high solubilization of organic compounds up to 24.40 g L-1. However, higher specific CH4 production of 0.012 and 0.01 L CH4 g-1 volatile solidsadded. was achieved for thermal and sonic pre-treatments, respectively. High N- and low C-content of the Tolypothrix biomass affected CH4 recovery, while pre-treatment accelerated production of volatile acids (15.90 g L-1) and ammonia-N-accumulation (1.41 g L-1), leading to poor CH4 yields. Calculated theoretical CH4 yields based on the elemental composition of the biomass were ~55% higher than actual yields. This highlights the complexity of interactions during AD which are not adequately represented by elemental composition.

Metal(loid) speciation and transformation by aerobic methanotrophs.

Manufacturing and resource industries are the key drivers for economic growth with a huge environmental cost (e.g. discharge of industrial effluents and post-mining substrates). Pollutants from waste streams, either organic or inorganic (e.g. heavy metals), are prone to interact with their physical environment that not only affects the ecosystem health but also the livelihood of local communities. Unlike organic pollutants, heavy metals or trace metals (e.g. chromium, mercury) are non-biodegradable, bioaccumulate through food-web interactions and are likely to have a long-term impact on ecosystem health. Microorganisms provide varied ecosystem services including climate regulation, purification of groundwater, rehabilitation of contaminated sites by detoxifying pollutants. Recent studies have highlighted the potential of methanotrophs, a group of bacteria that can use methane as a sole carbon and energy source, to transform toxic metal (loids) such as chromium, mercury and selenium. In this review, we synthesise recent advances in the role of essential metals (e.g. copper) for methanotroph activity, uptake mechanisms alongside their potential to transform toxic heavy metal (loids). Case studies are presented on chromium, selenium and mercury pollution from the tanneries, coal burning and artisanal gold mining, respectively, which are particular problems in the developing economy that we propose may be suitable for remediation by methanotrophs. Video Abstract.

Bacterial community analysis of biofilm on API 5LX carbon steel in an oil reservoir environment.

This study aimed to characterize the biofilm microbial community that causes corrosion of API 5LX carbon steel. API 5LX carbon steel coupons were incubated with raw produced water collected from two oil reservoir stations or filter-sterilized produced water. Biofilm 16S rRNA amplicon sequencing revealed that the bacterial community present in the biofilm was dominated by Proteobacteria, including Marinobacter hydrocarbonoclaustics and Marinobacter alkaliphilus. Electrochemical analysis such as impedance and polarization results indicated that Proteobacteria biofilm accelerated corrosion by ~ twofold (2.1 ± 0.61 mm/years) or ~ fourfold (~ 3.7 ± 0.42 mm/years) when compared to the control treatment (0.95 ± 0.1 mm/years). Scanning electron and atomic force microscopy revealed the presence of a thick biofilm and pitting corrosion. X-ray diffraction revealed higher amounts of the corrosion products Fe2O3, γ-FeOOH, and α-FeOOH, and confirmed that the microbial biofilm strongly oxidized the iron and contributed to the acceleration of corrosion of carbon metal API 5LX.

Food waste treatment by anaerobic co-digestion with saline sludge and its implications for energy recovery in Hong Kong.

Potential of methane production by co-digestion of food waste with saline sludge produced from sewage receiving seawater toilet flushing was investigated to determine its suitability for food waste management in Hong Kong by making use of excess design capacity of sludge digesters. High salinity of saline sludge (12.8 mS/cm) affected degradation of organic compounds resulting in an increase in sCOD by 135% as compared to an increase by 283% in treatments with non-saline sludge (4.2 mS/cm) co-digestion. This inhibitory effect was also evident by lower VS removal efficiency of 32.65% for saline versus 54.23% for non-saline sludge based co-digestion. Furthermore, non-saline sludge gave a 3.4-fold higher methane yield than saline sludge co-digestion. It is concluded that co-digestion of food waste with both sludges could be adopted as a potential strategy to make use of excess digestion capacity of existing wastewater treatment facilities but is more viable for non-saline sludge.

Lipid accumulation potential of oleaginous yeasts: A comparative evaluation using food waste leachate as a substrate.

In present study, the efficiency of three oleaginous yeasts i.e., Yarrowia lipolytica, Rhodotorula glutinis and Cryptococcus curvatus were compared for their lipid assimilation capacities using three different FW-leachates as a medium. The FW-leachates were collected from dry anaerobic digesters and diluted to achieve carbohydrate content of 25gL-1 prior to yeast inoculations. Around 5% of yeast cultures were individually mixed in three different FW-leachate mediums and incubated under 30°C and 150rpm agitation for 6days. The Y. lipolytica produced high biomass with lipid contents of 49.0±2% on dry weight basis. Whereas, the acetic acid concentration of >6gL-1 inhibited the growth of R. glutinis. The study observed that the selection of appropriate FW-leachate composition is highly important for biolipid accumulation by oleaginous yeasts.

Enzyme-mediated biodegradation of long-chain n-alkanes (C32 and C40) by thermophilic bacteria.

Removal of long-chain hydrocarbons and n-alkanes from oil-contaminated environments are mere important to reduce the ecological damages, while bio-augmentation is a very promising technology that requires highly efficient microbes. In present study, the efficiency of pure isolates, i.e., Geobacillus thermoparaffinivorans IR2, Geobacillus stearothermophillus IR4 and Bacillus licheniformis MN6 and mixed consortium on degradation of long-chain n-alkanes C32 and C40 was investigated by batch cultivation test. Biodegradation efficiencies were found high for C32 by mixed consortium (90%) than pure strains, while the pure strains were better in degradation of C40 than mixed consortium (87%). In contrast, the maximum alkane hydroxylase activities (161 µmol mg-1 protein) were recorded in mixed consortium system that had supplied with C40 as sole carbon source. Also, the alcohol dehydrogenase (71 µmol mg-1 protein) and lipase activity (57 µmol mg-1 protein) were found high. Along with the enzyme activities, the hydrophobicity natures of the bacterial strains were found to determine the degradation efficiency of the hydrocarbons. Thus, the study suggested that the hydrophobicity of the bacteria is a critical parameter to understand the biodegradation of n-alkanes.

Responses of mixed methanotrophic consortia to variable Cu2+/Fe2+ ratios.

Methane mitigation in landfill top cover soils is mediated by methanotrophs whose optimal methane (CH4) oxidation capacity is governed by environmental and complex microbial community interactions. Optimization of CH4 remediating bio-filters need to take microbial responses into account. Divalent copper (Cu2+) and iron (Fe2+) are present in landfills at variable ratios and play a vital role in methane oxidation capacity and growth of methanotrophs. This study, as a first of its kind, therefore quantified effects of variable Cu2+ and Fe2+ (5:5, 5:25 and 5:50 µM) ratios on mixed methanotrophic communities enriched from landfill top cover (LB) and compost soils (CB). CH4 oxidation capacity, CH4 removal efficiencies, fatty acids content/profiles and polyhydroxybutyrate (PHB; a biopolymer) contents were also analysed to quantify performance and potential co-product development. Mixed methanotroph cultures were raised in 10 L continuous stirred tank reactors (CSTRs, Bioflo® & Celligen® 310 Fermentor/Bioreactor; John Morris Scientific, Chatswood, NSW, Australia). Community structure was determined by amplifying the V3-V4 region of 16s rRNA gene. Community structure and, consequently, fatty acid-profiles changed significantly with increasing Cu2+/Fe2+ ratios, and responses were different for LB and CB. Effects on methane oxidation capacities and PHB content were similar in the LB- and CB-CSTR, decreasing with increasing Cu2+/Fe2+ ratios, while biomass growth was unaffected. In general, high Fe2+ concentration favored growth of the type -II methanotroph Methylosinus in the CB-CSTR, but methanotroph abundances decreased in the LB-CSTR. Increase in Cu2+/Fe2+ ratio increased the growth of Sphingopyxis in both systems, while Azospirllum was co-dominant in the LB- but absent in the CB-CSTR. After 13 days, methane oxidation capacities and PHB content decreased by ∼50% and more in response to increasing Fe2+ concentrations. Although methanotroph abundance was ∼2% in the LB- (compared to >50% in CB-CSTR), methane oxidation capacities were comparable in the two systems, suggesting that methane oxidation capacity was maintained by the dominant Azospirllum and Sphingopyxis in the LB-CSTR. Despite similar methanotroph inoculum community composition and controlled environmental variables, increasing Cu2+/Fe2+ ratios resulted in significantly different microbial community structures in the LB- and CB-CSTR, indicative of complex microbial interactions. In summary, our results suggest that a detailed understanding of allelopathic interactions in mixed methanotrophic consortia is vital for constructing robust bio-filters for CH4 emission abatement.

Application of recombinant Pediococcus acidilactici BD16 (fcs +/ech +) for bioconversion of agrowaste to vanillin.

Biotechnological production of vanillin is gaining momentum as the natural synthesis of vanillin that is very expensive. Ferulic acid (FA), a costly compound, is used as the substrate to produce vanillin biotechnologically and the making process is still expensive. Therefore, this study investigated the practical use of an agrobiomass waste, rice bran, and provides the first evidence of a cost-effective production of vanillin within 24 h of incubation using recombinant Pediococcus acidilactici BD16 (fcs +/ech +). Introduction of two genes encoding feruloyl CoA synthetase and enoyl CoA hydratase into the native strain increased vanillin yield to 4.01 g L-1. Bioconversion was monitored through the transformation of phenolic compounds. A hypothetical metabolic pathway of rice bran during the vanillin bioconversion was proposed with the inserted pathway from ferulic acid to vanillin and compared with that of other metabolic engineered strains. These results could be a gateway of using recombinant lactic acid bacteria for industrial production of vanillin from agricultural waste.

Culture scale-up and immobilisation of a mixed methanotrophic consortium for methane remediation in pilot-scale bio-filters.

Robust methanotrophic consortia for methane (CH4) remediation and by-product development are presently not readily available for industrial use. In this study, a mixed methanotrophic consortium (MMC), sequentially enriched from a marine sediment, was assessed for CH4 removal efficiency and potential biomass-generated by-product development. Suitable packing material for bio-filters to support MMC biofilm establishment and growth was also evaluated. The enriched MMC removed ∼7-13% CH4 under a very high gas flow rate (2.5 L min-1; 20-25% CH4) in continuous-stirred tank reactors (∼10 L working volume) and the biomass contained long-chain fatty acids (i.e. C16 and C18). Cultivation of the MMC on plastic bio-balls abated ∼95-97% CH4 in pilot-scale non-sterile outdoor-operated bio-filters (0.1 L min-1; 1% CH4). Contamination by cyanobacteria had beneficial effects on treating low-level CH4, by providing additional oxygen for methane oxidation by MMC, suggesting that the co-cultivation of MMC with cyanobacterial mats does not interfere with and may actually be beneficial for remediation of CH4 and CO2 at industrial scale.

Influence of nutrients on oxidation of low level methane by mixed methanotrophic consortia.

Low-level methane emissions from coal mine ventilation air (CMV-CH4; i.e., 1 % CH4) can significantly contribute to global climate change, and therefore, treatment is important to reduce impacts. To investigate CMV-CH4 abatement potential, five different mixed methanotrohic consortia (MMCs) were established from soil/sediment sources, i.e., landfill top cover soil, bio-solid compost, vegetated humus soil, estuarine and marine sediments. Enrichment conditions for MMCs were as follows: nitrate mineral salt (NMS) medium, pH ~ 6.8; 25 °C; 20-25 % CH4; agitation 200 rpm; and culture period 20 days, in mini-bench-top bioreactors. The enriched cultures were supplemented with extra carbon (methanol 0.5-1.5 %, formate 5-15 mM, and acetate 5-15 mM), nitrogen (nitrate 0.5-1.5 g L(-1), ammonium 0.1-0.5 g L(-1), or urea: 0.1-0.5 g L(-1)), and trace elements (copper 1-5 µM, iron 1-5 µM, and zinc 1-5 µM) in different batch experiments to improve low-level CH4 abatement. Average CH4 oxidation capacities (MOCs) of MMCs varied between 1.712 ± 0.032 and 1.963 ± 0.057 mg g(-1)DWbiomass h(-1). Addition of formate improved the MOCs of MMCs, but the dose-response varied for different MMCs. Acetate, nitrate and copper had no significant effect on MOCs, while addition of methanol, ammonium, urea, iron and zinc impacted negatively. Overall, MMCs enriched from marine sediments and landfill top cover soil showed high MOCs which were largely resilient to nutrient supplementation, suggesting a strong potential for biofilter development for industrial low-level CH4 abatement, such as those present in CMV.

Hydrolysis-acidogenesis of food waste in solid-liquid-separating continuous stirred tank reactor (SLS-CSTR) for volatile organic acid production.

The use of conventional continuous stirred tank reactor (CSTR) can affect the methane (CH4) recovery in a two-stage anaerobic digestion of food waste (FW) due to carbon short circuiting in the hydrolysis-acidogenesis (Hy-Aci) stage. In this research, we have designed and tested a solid-liquid-separating CSTR (SLS-CSTR) for effective Hy-Aci of FW. The working conditions were pH 6 and 9 (SLS-CSTR-1 and -2, respectively); temperature-37°C; agitation-300rpm; and organic loading rate (OLR)-2gVSL(-1)day(-1). The volatile fatty acids (VFA), enzyme activities and bacterial population (by qPCR) were determined as test parameters. Results showed that the Hy-Aci of FW at pH 9 produced ∼35% excess VFA as compared to that at pH 6, with acetic and butyric acids as major precursors, which correlated with the high enzyme activities and low lactic acid bacteria. The design provided efficient solid-liquid separation there by improved the organic acid yields from FW.