Chiral Inversion of 2-Arylpropionic Acid Enantiomers under Anaerobic Conditions.

This work examined the chiral inversion of 2-arylpropionic acids (2-APAs) under anaerobic conditions and the associated microbial community. The anaerobic condition was simulated by two identical anaerobic digesters. Each digester was fed with the substrate containing 11 either pure (R)- or pure (S)-2-APA enantiomers. Chiral inversion was evidenced by the concentration increase of the other enantiomer in the digestate and the changes in the enantiomeric fraction between the two enantiomers. Both digesters showed similar and poor removal of 2-APAs (≤30%, except for naproxen) and diverse chiral inversion behaviors under anaerobic conditions. Four compounds exhibited (S → R) unidirectional inversion [flurbiprofen, ketoprofen, naproxen, and 2-(4-tert-butylphenyl)propionic acid], and the remaining seven compounds showed bidirectional inversion. Several aerobic and facultative anaerobic bacterial genera (Candidatus Microthrix, Rhodococcus, Mycobacterium, Gordonia, and Sphingobium) were identified in both digesters and predicted to harbor the 2-arylpropionyl-CoA epimerase (enzyme involved in chiral inversion) encoding gene. These genera presented at low abundances, <0.5% in the digester dosed with (R)-2-APAs and <0.2% in the digester dosed with (S)-2-APAs. The low abundances of these genera explain the limited extent of chiral inversion observed in this study.

Ecological impact of the antibiotic ciprofloxacin on microbial community of aerobic activated sludge.

This study investigated the effects and fate of the antibiotic ciprofloxacin (CIP) at environmentally relevant levels (50-500 µg/L) in activated sludge (AS) microbial communities under aerobic conditions. Exposure to 500 µg/L of CIP decreased species diversity by about 20% and significantly altered the phylogenetic structure of AS communities compared to those of control communities (no CIP exposure), while there were no significant changes upon exposure to 50 µg/L of CIP. Analysis of community composition revealed that exposure to 500 µg/L of CIP significantly reduced the relative abundance of Rhodobacteraceae and Nakamurellaceae by more than tenfold. These species frequently occur in AS communities across many full-scale wastewater treatment plants and are involved in key ecosystem functions (i.e., organic matter and nitrogen removal). Our analyses showed that 50-500 µg/L CIP was poorly removed in AS (about 20% removal), implying that the majority of CIP from AS processes may be released with either their effluents or waste sludge. We therefore strongly recommend further research on CIP residuals and/or post-treatment processes (e.g., anaerobic digestion) for waste streams that may cause ecological risks in receiving water bodies.

Antiseptic chlorhexidine in activated sludge: Biosorption, antimicrobial susceptibility, and alteration of community structure.

Chlorhexidine (CHX) is a broad-spectrum antimicrobial, which may pose environmental health risks. This study examined the removal potential and the mechanisms regulating the fate of CHX in activated sludge (AS). Bioreactors inoculated with AS removed 74 ±â€¯8% and 81 ±â€¯6% of CHX at steady state while receiving 0.5 and 1 mg/L CHX, respectively. Analysis of the removal pathways showed that biosorption, rather than biological breakdown or other abiotic losses, largely (>70%) regulated the removal of CHX. 16S rRNA gene-based analysis revealed that CHX selected for Luteolibacter (4.3-10.1-fold change) and Runella (6.2-14.1-fold change) with potential multi-drug resistance mechanisms (e.g., efflux pumps). In contrast, it significantly reduced core members (Comamonadaceae and Flavobacteriaceae) of AS, playing a key role in contaminant removal and floc formation directly associated with the performance of WWTPs (e.g., wastewater effluent quality). Antimicrobial susceptibility testing showed that 0.4-1.3 mg/L of CHX can be sublethal to AS. Our work provided new insights into the fate of CHX in urban waste streams and the potential toxicity and effects on the structure and function of AS, which has practical implications for the management of biological WWTPs treating CHX.

Application of a novel molecular technique to characterise the effect of settling on microbial community composition of activated sludge.

Activated sludge (AS) and return activated sludge (RAS) microbial communities from three full-scale municipal wastewater treatment plants (denoted plant A, B and C) were compared to assess the impact of sludge settling (i.e. gravity thickening in the clarifier) and profile microorganisms responsible for nutrient removal and reactor foaming. The results show that all three plants were dominated with microbes in the phyla of Proteobacteria, Bacteroidetes, Verrucomicrobia, Actinobacteria, Chloroflexi, Firmicutes, Nitrospirae, Spirochaetae, Acidobacteria and Saccharibacteria. AS and RAS shared above 80% similarity in the microbial community composition, indicating that sludge thickening does not significantly alter the microbial composition. Autotrophic and heterotrophic nitrifiers were present in the AS. However, the abundance of autotrophic nitrifiers was significantly lower than that of the heterotrophic nitrifiers. Thus, ammonium removal at these plants was achieved mostly by heterotrophic nitrification. Microbes that can cause foaming were at 3.2% abundance, and this result is well corroborated with occasional aerobic biological reactor foaming. By contrast, these microbes were not abundant (<2.1%) at plant A and C, where aerobic biological reactor foaming has not been reported.

Continuous transformation of chiral pharmaceuticals in enzymatic membrane bioreactors for advanced wastewater treatment.

This study demonstrates continuous enantiomeric inversion and further biotransformation of chiral profens including ibuprofen, naproxen and ketoprofen by an enzymatic membrane bioreactor (EMBR) dosed with laccase. The EMBR showed non-enantioselective transformations, with high and consistent transformation of both (R)- and (S)-ibuprofen (93 ± 6%, n = 10), but lower removals of both enantiomers of naproxen (46 ± 16%, n = 10) and ketoprofen (48 ± 17%, n = 10). Enantiomeric analysis revealed a bidirectional but uneven inversion of the profens, for example 14% inversion of (R)- to (S)- compared to 4% from (S)- to (R)-naproxen. With redox-mediator addition, the enzymatic chiral inversion of both (R)- and (S)-profens remained unchanged, although the overall conversion became enantioselective; except for (S)-naproxen, the addition of redox mediator promoted the degradation of (R)-profens only.

Coupling granular activated carbon adsorption with membrane bioreactor treatment for trace organic contaminant removal: breakthrough behaviour of persistent and hydrophilic compounds.

This study investigated the removal of trace organic contaminants by a combined membrane bioreactor - granular activated carbon (MBR-GAC) system over a period of 196 days. Of the 22 compounds investigated here, all six hydrophilic compounds with electron-withdrawing functional groups (i.e., metronidazole, carbamazepine, ketoprofen, naproxen, fenoprop and diclofenac) exhibited very low removal efficiency by MBR-only treatment. GAC post-treatment initially complemented MBR treatment very well; however, a compound-specific gradual deterioration of the removal of the above-mentioned problematic compounds was noted. While a 20% breakthrough of all four negatively charged compounds namely ketoprofen, naproxen, fenoprop and diclofenac occurred within 1000-3000 bed volumes (BV), the same level of breakthrough of the two neutral compounds metronidazole and carbamazepine did not occur until 11,000 BV. Single-solute isotherm parameters did not demonstrate any discernible correlation individually with any of the parameters that may govern adsorption onto GAC, such as log D, number of hydrogen-bond donor/acceptor groups, dipole moment or aromaticity ratio of the compounds. The isotherm data, however, could differentiate the breakthrough behaviour between negatively charged and neutral trace organic contaminants.

Microalgae-bacteria consortium for wastewater treatment and biomass production.

The diversity of microalgae and bacteria allows them to form a complementary consortium for efficient wastewater treatment and nutrient recovery. This review highlights the potential of wastewater-derived microalgal biomass as a renewable feedstock for producing animal feed, biofertilisers, biofuel, and many valuable biochemicals. Data corroborated from this review shows that microalgae and bacteria can thrive in many environments. Microalgae are especially effective at utilising nutrients from the water as they grow. This review also consolidates the current understanding of microalgae characteristics and their interactions with bacteria in a consortium system. Recent studies on the performance of only microalgae and microalgae-bacteria wastewater treatment are compared and discussed to establish a research roadmap for practical implementation of the consortium systems for various wastewaters (domestic, industrial, agro-industrial, and landfill leachate wastewater). In comparison to the pure microalgae system, the consortium system has a higher removal efficiency of up to 15% and shorter treatment time. Additionally, this review addresses a variety of possibilities for biomass application after wastewater treatment.

Bioremoval of estrogens by laccase immobilized onto polyacrylonitrile/polyethersulfone material: Effect of inhibitors and mediators, process characterization and catalytic pathways determination.

The presence of micropollutants in water, wastewater and soil are a global problem due to their persistent effect on ecosystems and human health. Although there are many methods of removal of environmental pollutants, they are often ineffective for degradation of pharmaceuticals, including estrogens. In presented work we proposed fabrication of electrospun material from polyacrylonitrile/polyethersulfone (PAN/PES) as a support for laccase immobilization by covalent binding. Oxidoreductase was attached to the electrospun fibers using polydopamine as a linker and produced system was used for degradation of two estrogens: 17ß-estradiol (E2) and 17α-ethynylestradiol (EE2). It was shown that 92% of E2 and 100% of EE2 were degraded after 24 h of the process. Moreover, the effect of surfactants, metal ions and mediators on conversion efficiencies of estrogens was investigated and it was confirmed that immobilized enzyme possessed higher resistance to inhibitory agents as well as thermal and storage stability, compared to its native form. Finally, estrogenic activities of E2 and EE2 solutions decreased around 99% and 87%, respectively, after enzymatic conversion, that corresponds to significant reduction of the total organic carbon and formation of low-toxic final products of estrogens degradation.

Selection of microalgae strains for sustainable production of aviation biofuel.

This study develops and applies the PROMETHEE-GAIA method as a new tool to select microalgae strains for aviation fuel production. Assessment involves 19 criteria with equal weighting in three aspects, namely biomass production, lipid quality, and fatty acid methylester properties. Here, the method is demonstrated for evaluating 17 candidate microalgae strains. Chlorella sp. NT8a is assessed as the most suitable strain for aviation fuel production. The results also show that unmodified biofuel from the most suitable strain could not meet all jet fuel standards. In particular, microalgae-based fuel could not satisfy the required density, heating value and freezing points of the international jet fuel standards. These results highlight the need for a broad action plan including improvement in the processing or modification of biofuel produced from microalgae and revision of the current jet fuel standards to facilitate the introduction of microalgae-based biofuel for the aviation industry.

Effects of harvesting on morphological and biochemical characteristics of microalgal biomass harvested by polyacrylamide addition, pH-induced flocculation, and centrifugation.

The effects of microalgae harvesting methods on microalgal biomass quality were evaluated using three species namely the freshwater green alga Chlorella vulgaris, marine red alga Porphyridium purpureum and marine diatom Phaeodactylum tricornutum. Harvesting efficiencies of polyacrylamide addition, alkaline addition, and centrifugation ranged from 85 to 95, 59-92 and 100%, respectively, across these species. Morphology of the harvested cells (i.e. compromised cell walls) was significantly impacted by alkaline pH-induced flocculation for all three species. Over 50% of C. vulgaris cells were compromised with alkaline pH compared to < 10% with polyacrylamide and centrifugation. The metabolic profiles varied depending on harvesting methods. Species-specific decrease of certain metabolites was observed. These results suggest that the method of harvest can alter the metabolic profile of the biomass amongst the three harvesting methods, polyacrylamide addition showed higher harvesting efficiency with less compromised cells and higher retention of industry important biochemicals.

Hydrogen sulphide management in anaerobic digestion: A critical review on input control, process regulation, and post-treatment.

Hydrogen sulphide (H2S) in biogas is a problematic impurity that can inhibit methanogenesis and cause equipment corrosion. This review discusses technologies to remove H2S during anaerobic digestion (AD) via: input control, process regulation, and post-treatment. Post-treatment technologies (e.g. biotrickling filters and scrubbers) are mature with >95% removal efficiency but they do not mitigate H2S toxicity to methanogens within the AD. Input control (i.e. substrate pretreatment via chemical addition) reduces sulphur input into AD via sulphur precipitation. However, available results showed <75% of H2S removal efficiency. Microaeration to regulate AD condition is a promising alternative for controlling H2S formation. Microaeration, or the use of oxygen to regulate the redox potential at around -250 mV, has been demonstrated at pilot and full scale with >95% H2S reduction, stable methane production, and low operational cost. Further adaptation of microaeration relies on a comprehensive design framework and exchange operational experience for eliminating the risk of over-aeration.

Free and immobilized biocatalysts for removing micropollutants from water and wastewater: Recent progress and challenges.

Enzymatic conversion of micropollutants into less-toxic derivatives is an important bioremediation strategy. This paper aims to critically review the progress in water and wastewater treatment by both free and immobilized enzymes presenting this approach as highly efficient and performed under environmentally benign and friendly conditions. The review also summarises the effects of inorganic and organic wastewater matrix constituents on enzymatic activity and degradation efficiency of micropollutants. Finally, application of enzymatic reactors facilitate continuous treatment of wastewater and obtaining of pure final effluents. Of a particular note, enzymatic treatment of micropollutants from wastewater has been mostly reported by laboratory scale studies. Thus, this review also highlights key research gaps of the existing techniques and provides future perspectives to facilitate the transfer of the lab-scale solutions to a larger scale and to improve operationability of biodegradation processes.

Simultaneous nutrient recovery and algal biomass production from anaerobically digested sludge centrate using a membrane photobioreactor.

This study aims to evaluate the performance of C. vulgaris microalgae to simultaneously recover nutrients from sludge centrate and produce biomass in a membrane photobioreactor (MPR). Microalgae growth and nutrient removal were evaluated at two different nutrient loading rates (sludge centrate). The results show that C. vulgaris microalgae could thrive in sludge centrate. Nutrient loading has an indiscernible impact on biomass growth and a notable impact on nutrient removal efficiency. Nutrient removal increased as the nutrient loading rate decreased and hydraulic retention time increased. There was no membrane fouling observed in the MPR and the membrane water flux was fully restored by backwashing using only water. However, the membrane permeability varies with the hydraulic retention time (HRT) and biomass concentration in the reactor. Longer HRT offers higher permeability. Therefore, it is recommended to operate the MPR system in lower HRT to improve the membrane resistance and energy consumption.

Monitoring antibiotic resistance genes in wastewater treatment: Current strategies and future challenges.

Antimicrobial resistance (AMR) is a growing threat to human and animal health. Progress in molecular biology has revealed new and significant challenges for AMR mitigation given the immense diversity of antibiotic resistance genes (ARGs), the complexity of ARG transfer, and the broad range of omnipresent factors contributing to AMR. Municipal, hospital and abattoir wastewater are collected and treated in wastewater treatment plants (WWTPs), where the presence of diverse selection pressures together with a highly concentrated consortium of pathogenic/commensal microbes create favourable conditions for the transfer of ARGs and proliferation of antibiotic resistant bacteria (ARB). The rapid emergence of antibiotic resistant pathogens of clinical and veterinary significance over the past 80 years has re-defined the role of WWTPs as a focal point in the fight against AMR. By reviewing the occurrence of ARGs in wastewater and sludge and the current technologies used to quantify ARGs and identify ARB, this paper provides a research roadmap to address existing challenges in AMR control via wastewater treatment. Wastewater treatment is a double-edged sword that can act as either a pathway for AMR spread or as a barrier to reduce the environmental release of anthropogenic AMR. State of the art ARB identification technologies, such as metagenomic sequencing and fluorescence-activated cell sorting, have enriched ARG/ARB databases, unveiled keystone species in AMR networks, and improved the resolution of AMR dissemination models. Data and information provided in this review highlight significant knowledge gaps. These include inconsistencies in ARG reporting units, lack of ARG/ARB monitoring surrogates, lack of a standardised protocol for determining ARG removal via wastewater treatments, and the inability to support appropriate risk assessment. This is due to a lack of standard monitoring targets and agreed threshold values, and paucity of information on the ARG-pathogen host relationship and risk management. These research gaps need to be addressed and research findings need to be transformed into practical guidance for WWTP operators to enable effective progress towards mitigating the evolution and spread of AMR.

A comprehensive analysis of an effective flocculation method for high quality microalgal biomass harvesting.

Flocculation is a low-cost harvesting technique for microalgae biomass production, but flocculation efficiency is species dependent. In this study, we investigated the efficacy of two synthetic (polyacrylamide) and one natural (chitosan) flocculants against three algal species: the cyanobacterium Synechocystis sp., the freshwater Chlorella vulgaris, and the marine Phaeodactylum tricornutum at laboratory and pilot scales to evaluate harvesting efficiency, biomass integrity and media recycling. Growth phase affected the harvesting efficiency of the eukaryotic microalgae. The flocculation was optimal at stationary phase with high flocculation efficiency achieved using polyacrylamides at 24-36 mg/g dry weight. The effect of the flocculants on the harvested biomass was investigated. The flocculated Synechocystis sp. showed a higher proportion of compromised cells compared to C. vulgaris and P. tricornutum likely due to differences in cell walls composition. Compromised cells could lead to the release of valuable products into the surrounding growth media during flocculation. The residual culture media was recycled once with no impact on cell growth for all treatments and algal species. The flocculation technique was demonstrated at pilot-scale using 350 L microalgal suspension, showing an efficiency of 82-90% at a polyacrylamide dosage of 6.5-10 mg/L. This efficiency and the biomass quality are comparable to the laboratory-scale results. Overall, results indicate that polyacrylamide flocculants work on a wide range of species without the need for pre-treatment. The information generated in this study can contribute to making the microalgae industry more competitive.

Factors governing microalgae harvesting efficiency by flocculation using cationic polymers.

This study aims to elucidate the mechanisms governing the harvesting efficiency of Chlorella vulgaris by flocculation using a cationic polymer. Flocculation efficiency increased as microalgae culture matured (i.e. 35-45, 75, and > 97% efficiency at early, late exponential, and stationary phase, respectively. Unlike the negative impact of phosphate on flocculation in traditional wastewater treatment; here, phosphorous residue did not influence the flocculation efficiency of C. vulgaris. The observed dependency of flocculation efficiency on growth phase was driven by changes in microalgal cell properties. Microalgal extracellular polymeric substances (EPS) in both bound and free forms at stationary phase were two and three times higher than those at late and early exponential phase, respectively. Microalgae cells also became more negatively charged as they matured. Negatively charged and high EPS content together with the addition of high molecular weight and positively charged polymer could facilitate effective flocculation via charge neutralisation and bridging.

Promotion of direct interspecies electron transfer and potential impact of conductive materials in anaerobic digestion and its downstream processing - a critical review.

Addition of conductive materials (CMs) has been reported to facilitate direct interspecies electron transfer (DIET) and improved anaerobic digestion (AD) performance. This review summarises the benefits and outlines remaining research challenges of the addition of CMs with a focus on the downstream processing of AD. CM addition may alter biogas quality, digestate dewaterability, biosolids volume, and centrate quality. Better biogas quality has been observed due to the adsorption of H2S to metallic CMs. The addition of CMs results in an increase in solid content of the digestate and thus an additional requirement for sludge dewatering and handling and the final biosolids volume for disposal. This review highlights the need for more research at pilot scale to validate the benefits of CM addition and to evaluate CM selection, doses, material costs, and the impact on downstream processes. The lack of research on the impact of CMs on the downstream process of AD is highlighted.

Significance of the presence of antibiotics on the microbial consortium in wastewater - The case of nitrofurantoin and furazolidone.

Antibiotics in wastewater leads to migration of pollutants and disrupts natural processes of mineralization of organic matter. In order to understand the mechanism of this, research was undertaken on the influence of nitrofurantoin (NFT) and furazolidone (FZD), on the behaviour of a consortium of microorganisms present in a model wastewater in a bioreactor. Our study confirmed biodegradation of the antibiotics by the microbial consortium, with the degradation efficiency within 10 days of 65% for FZD, but only 20% for NFT. The kinetic study proved that the presence of analysed antibiotics had no adverse effect on the microbes, but the consortium behaviour differ significantly with the NFT reducing the consumption of organic carbon in wastewater and increasing the production of extracellular biopolymeric and volatile organic compounds, and the FZD reducing assimilation of other carbon sources to a less extent, at the expense of cellular focus on biodegradation of this antibiotic.

Chiral inversion of 2-arylpropionoic acid (2-APA) enantiomers during simulated biological wastewater treatment.

This study examined the removal and enantio­specific fate of a suite of eleven chiral 2-arylpropionic acids (2-APAs) during biological wastewater treatment simulated in a laboratory-scale membrane bioreactor (MBR). Using pure (R)- and (S)- enantiomers in the MBR influent, chiral inversion was determined through the increase in the concentration of the non-dominant enantiomer and changes in the enantiomeric fraction (EF) between the two enantiomers during the treatment process. Effective (>90%) and similar removal rates between (R)- and (S)- enantiomers were confirmed for eight 2-APAs. In this study, 2-APAs exhibited diverse and distinctive chiral inversion behaviours: two 2-APAs showed (R→S) unidirectional inversion, three 2-APAs showed (S→R) unidirectional inversion, and six 2-APAs showed bidirectional inversion. This is the first study to report chiral inversion behaviours of a comprehensive suite of 2-APAs with a variety of functional groups substituted onto the aryl ring. A decrease in effluent EF over time was observed for two 2-APAs. This study shows that chiral inversion of 2-APAs varies significantly from compound to compound, despite the high similarity in their chemical structures.

Harvesting Porphyridium purpureum using polyacrylamide polymers and alkaline bases and their impact on biomass quality.

This study aims to examine the flocculation efficiency of Porphyridium purpureum (i.e. a red marine microalga with high content of pigments and fatty acids) grown in seawater medium using polyacrylamide polymers and alkaline flocculation. Polymers Flopam™ and FO3801 achieved the highest flocculation efficiency of over 99% at the optimal dose of 21 mg per g of dry biomass through charge neutralisation and bridging mechanism. The addition of sodium hydroxide, potassium hydroxide, and sodium carbonate also achieved flocculation efficiency of 98 and 91%, respectively, but high doses were required (i.e. > 500 mg per g of dry biomass). Calcium hydroxide was not as effective and could only achieve 75% flocculation efficiency. Precipitation of magnesium hydroxide was identified as the major cause of hydroxide-induced flocculation. On the other hand, sodium carbonate addition induced flocculation via both magnesium and calcium carbonate co-precipitation. The large mass of precipitates caused a sweeping effect and enmeshed the microalgal cells to trigger sedimentation. Cell membrane integrity analysis of flocculated P. purpureum indicated that polyacrylamide polymers led to significant compromised cells (i.e. 96%), compared to the alkaline bases (70-96% compromised cells). These results appear to be the first to demonstrate the high efficiency of polyacrylamide polymer and alkaline flocculation of P. purpureum but at the expense of the biomass quality.