Aerobic Biotransformation Research Articles

Promoting nitrogen conversion in aerobic biotransformation of swine slurry with the co-application of manganese sulfate and biochar

This study aimed to explore the co-application of MnSO4 (Mn) and biochar (BC) in nitrogen conversion during the composting process. A 70-day aerobic composting was conducted using swine slurry, supplemented with different levels of Mn (0, 0.25%, and 0.5%) and 5% BC. The results demonstrated that the treatment with 0.5MnBC had the highest levels of NH4+-N (3.07 g kg−1), TKN (29.90 g kg−1), and NO3−-N (1.94 g kg−1) among all treatments. Additionally, the 0.5MnBC treatment demonstrated higher urease, protease, nitrate reductase, and nitrite reductase activities than the other treatments, with the peak values of 18.12, 6.96, 3.57, and 15.14 mg g−1 d−1, respectively. The addition of Mn2+ increased the total organic nitrogen content by 29.59%–47.82%, the acid hydrolyzed ammonia nitrogen (AN) content by 13.84%–57.86% and the amino acid nitrogen (AAN) content by 55.38%–77.83%. The richness of Chloroflexi and Ascomycota was also enhanced by the simultaneous application of BC and Mn. Structural equation modeling analysis showed that Mn2+ can promote the conversion of Hydrolyzed Unknown Nitrogen (HUN) into AAN, and there is a positive association between urease and NH4+-N according to redundancy analysis. Firmicutes, Basidiomycota, and Mortierellomycota showed significant positive correlations with ASN, AN, and NH4+-N, indicating their crucial roles in nitrogen conversion. This study sheds light on promoting nitrogen conversion in swine slurry composting through the co-application of biochar and manganese sulfate.

Aerobic biotransformation of 6:2 fluorotelomer sulfonate in soils from two aqueous film-forming foam (AFFF)-impacted sites

Although 6:2 fluorotelomer sulfonate (6:2 FTS) is a common ingredient in aqueous film-forming foam (AFFF) formulations, its environmental fate at AFFF-impacted sites remains poorly understood. This study investigated the biotransformation of 6:2 FTS in microcosms prepared with soils collected from two AFFF-impacted sites; the former Loring Air Force Base (AFB) and Robins AFB. The half-life of 6:2 FTS in Loring soil was 43.3 days; while >60 mol% of initially spiked 6:2 FTS remained in Robins soil microcosms after a 224-day incubation. Differences in initial sulfate concentrations and the depletion of sulfate over the incubation likely contributed to the different 6:2 FTS biotransformation rates between the two soils. At day 224, stable transformation products, i.e., C4C7 perfluoroalkyl carboxylates, were formed with combined molar yields of 13.8 mol% and 1.2 mol% in Loring and Robins soils, respectively. Based on all detected transformation products, the biotransformation pathways of 6:2 FTS in the two soils were proposed. Microbial community analysis suggests that Desulfobacterota microorganisms may promote 6:2 FTS biotransformation via more efficient desulfonation. In addition, species from the genus Sphingomonas, which exhibited higher tolerance to elevated concentrations of 6:2 FTS and its biotransformation products, are likely to have contributed to 6:2 FTS biotransformation. This study demonstrates the potential role of biotransformation processes on the fate of 6:2 FTS at AFFF-impacted sites and highlights the need to characterize site biogeochemical properties for improved assessment of 6:2 FTS biotransformation behavior.

Aerobic biotransformation of Sargassum fluitans in combination with sheep manure: optimization of control variables

ABSTRACT Sargassum fluitans was composted alongside sheep manure, in a transformative process that significantly enhanced the algal material’s properties. Post the screening/washing/screening pretreatment, the content of total volatile solids escalated to 73.20%, while ash content reduced to 16.45%. Concurrently, lignin values surged to 30.12% as the biodegradability factor declined to 21%. The pretreatment decreased electrical conductivity from 11.60 to 1.32 DS/m. Employing a central composite design and response surface analysis pinpointed the optimal substrate combinations for carbon/nitrogen ratios of 35:1 and 25:1. The chosen combinations presented a high coefficient of determination (R2 = 0.9589, carbon/nitrogen ratio; R2 = 0.6584, pH), indicative of a robust statistical fit. Over a 45-day period, composting was conducted using bioreactors or biopiles, maintaining near-neutral pH values and temperatures slightly above ambient levels. The composting process reduced up to 94% of fecal coliforms in the 1:1 combination. Physicochemical analyses confirmed that the final product is a valuable compost-soil improver, with great potential for usage in organic agriculture, reforestation, and urban green spaces. Hence, this research underscores composting as an efficient technique in managing organic waste, including the emergent and seasonal Sargassum fluitans, thus addressing a pressing environmental concern with an innovative, effective solution.

Aerobic Biotransformation and Defluorination of Fluoroalkylether Substances (ether PFAS): Substrate Specificity, Pathways, and Applications.

Fluoroalkylether substances (ether PFAS) constitute a large group of emerging PFAS with uncertain environmental fate. Among them, GenX is the well-known alternative to perfluorooctanoic acid and one of the six proposed PFAS to be regulated by the U.S. Environmental Protection Agency. This study investigated the structure-biodegradability relationship for 12 different ether PFAS with a carboxylic acid headgroup in activated sludge communities. Only polyfluorinated ethers with at least one -CH2- moiety adjacent to or a C=C bond in the proximity of the ether bond underwent active biotransformation via oxidative and hydrolytic O-dealkylation. The bioreactions at ether bonds led to the formation of unstable fluoroalcohol intermediates subject to spontaneous defluorination. We further demonstrated that this aerobic biotransformation/defluorination could complement the advanced reduction process in a treatment train system to achieve more cost-effective treatment for GenX and other recalcitrant perfluorinated ether PFAS. These findings provide essential insights into the environmental fate of ether PFAS, the design of biodegradable alternative PFAS, and the development of cost-effective ether PFAS treatment strategies.

Roles of various enzymes in the biotransformation of 6:2 fluorotelomer alcohol (6:2 FTOH) by a white-rot fungus

Six-carbon-chained polyfluoroalkyl substances, such as 6:2 fluorotelomer alcohol (6:2 FTOH), are being used to replace longer chained compounds in the manufacture of various commercial products. This study examined the effects of growth substrates and nutrients on specific intracellular and extracellular enzymes mediating 6:2 FTOH aerobic biotransformation by the white-rot fungus, Phanerochaete chrysosporium. Cellulolytic conditions with limited glucose were a suitable composition, resulting in high 5:3 FTCA yield (37 mol%), which is a key intermediate in 6:2 FTOH degradation without forming significant amounts of terminal perfluorocarboxylic acids (PFCAs). Sulfate and ethylenediaminetetraacetic acid (EDTA) were also essential for 5:3 FTCA production, but, at lower levels, resulted in the buildup of 5:2 sFTOH (52 mol%) and 6:2 FTUCA (20 mol%), respectively. In non-ligninolytic nutrient-rich medium, 45 mol% 6:2 FTOH was transformed but produced only 12.7 mol% 5:3 FTCA. Enzyme activity studies imply that cellulolytic conditions induce the intracellular cytochrome P450 system. In contrast, extracellular peroxidase synthesis is independent of 6:2 FTOH exposure. Gene expression studies further verified that peroxidases were relevant in catalyzing the downstream transformations from 5:3 FTCA. Collectively, the identification of nutrients and enzymatic systems will help elucidate underlying mechanisms and biogeochemical conditions favorable for fungal transformation of PFCA precursors in the environment.

Methods involved in the treatment of four representative pharmaceuticals in hospital wastewater using sonochemical and biological processes

A primary pollution source by pharmaceuticals is hospital wastewater (HWW). Herein, the methods involved in the action of a biological system (BS, aerobic activated sludge) or a sonochemical treatment (US, 375 kHz and 30.8 W), for degrading four relevant pharmaceuticals (azithromycin, ciprofloxacin, paracetamol, and valsartan) in HWW, are shown. Before treatment of HWW, the correct performance of BS was assessed using glucose as a reference substance, monitoring oxygen consumption, and organic carbon removal. Meanwhile, for US, a preliminary test using ciprofloxacin in distilled water was carried out. The determination of risk quotients (RQ) and theoretical analyses about reactive moieties on these target substances are also presented. For both, the degradation of the pharmaceuticals and the calculation of RQ, analyses were performed by LC-MS/MS. The BS action decreased the concentration of paracetamol and valsartan by ∼96 and 86%, respectively. However, a poor action on azithromycin (2% removal) was found, whereas ciprofloxacin concentration increased ∼20%; leading to an RQ value of 1.61 (high risk) for the pharmaceuticals mixture. The analyses using a biodegradation pathway predictor (EAWAG-BDD methodology) revealed that the amide group on paracetamol and alkyl moieties on valsartan could experience aerobic biotransformations. In turn, US action decreased the concentration of the four pharmaceuticals (removals > 60% for azithromycin, ciprofloxacin, and paracetamol), diminishing the environmental risk (RQ: 0.51 for the target pharmaceuticals mixture). Atomic charge analyses (based on the electronegativity equalization method) were performed, showing that the amino-sugar on azithromycin; piperazyl ring, and double bond close to the two carbonyls on ciprofloxacin, acetamide group on paracetamol, and the alkyl moieties bonded to the amide group of valsartan are the most susceptible moieties to attacks by sonogenerated radicals. The LC-MS/MS analytical methodology, RQ calculations, and theoretical analyses allowed for determining the degrading performance of BS and US toward the target pollutants in HWW.•Biological and sonochemical treatments as useful methods for degrading 4 representative pharmaceuticals are presented.•Sonochemical treatment had higher degrading action than the biological one on the target pharmaceuticals.•Methodologies for risk environmental calculation and identification of moieties on the pharmaceuticals susceptible to radical attacks are shown.

Assessing aerobic biotransformation of 8:2 fluorotelomer alcohol in aqueous film-forming foam (AFFF)-impacted soils: Pathways and microbial community dynamics

Production of 8:2 fluorotelomer alcohol (8:2 FTOH) for industrial and consumer products, including aqueous film-forming foams (AFFFs) used for firefighting, has resulted in its widespread occurrence in the environment. However, the fate of 8:2 FTOH at AFFF-impacted sites remains largely unknown. Using AFFF-impacted soils from two United States Air Force Bases, microcosm experiments evaluated the aerobic biotransformation of 8:2 FTOH (extent and byproduct formation) and the dose-response on microbial communities due to 8:2 FTOH exposure. Despite different microbial communities, rapid transformation of 8:2 FTOH was observed during a 90-day incubation in the two soils, and 7:2 secondary fluorotelomer alcohol (7:2 sFTOH) and perfluorooctanoic acid (PFOA) were detected as major transformation products. Novel transformation products, including perfluoroalkane-like compounds (1H-perfluoroheptane, 1H-perfluorohexane, and perfluoroheptanal) were identified by liquid chromatography-high resolution mass spectrometry (LC-HRMS) and used to develop aerobic 8:2 FTOH biotransformation pathways. Microbial community analysis suggests that species from genus Sphingomonas are potential 8:2 FTOH degraders based on increased abundance in both soils after exposure, and the genus Afipia may be more tolerant to and/or involved in the transformation of 8:2 FTOH at elevated concentrations. These findings demonstrate the potential role of biological processes on PFAS fate at AFFF-impacted sites through fluorotelomer biotransformation.

Transformation mechanism of methylphosphonate to methane by Burkholderia sp: Insight from multi-labeled water isotope probing and transcriptomic

Methylphosphonate (MPn), has been identified as a likely source of methane in aerobic ocean and may be responsible for the “ocean methane paradox”, that is oversaturation of dissolved methane in oxic sea waters. However, the mechanism underlying the cleavage of C–P bonds during microbial degradation is not well understood. Using multi-labeled water isotope probing (MLWIP) and transcriptome analysis, we investigated the phosphate oxygen isotope systematics and mechanisms of microbial-mediated degradation of MPn in this study. In the aerobic culture containing MPn as the only phosphorus source, there was a significant release of inorganic phosphate (149.4 μmol/L) and free methane (268.3 mg/L). The oxygen isotopic composition of inorganic phosphorus (δ18OP) of accumulated released phosphate was 4.50‰, 23.96‰, and 40.88‰, respectively, in the corresponding 18O-labeled waters of −10.3‰, 9.9‰, and 30.6‰, and the slope obtained in plots of δ18OP versus the oxygen isotopic composition of water (δ18OW) was 0.89. Consequently, 89% of the oxygen atoms (Os) in phosphate (PO4) were exchanged with 18O-labeled waters in the medium, while the rest were exchanged with intracellular metabolic water. It has been confirmed that the C–P bond cleavage of MPn occurs in the cell with both ambient and metabolic water participation. Moreover, phn gene clusters play significant roles to cleave the C–P bond of MPn for Burkholderia sp. HQL1813, in which phnJ, phnM and phnI genes are significantly up-regulated during MPn decomposition to methane. In conclusion, the aerobic biotransformation of MPn to free methane by Burkholderia sp. HQL1813 has been elucidated, providing new insights into the mechanism that bio-cleaves C–P bonds to produce methane aerobically in aqueous environments for representative phosphonates.

Biological and Chemical Transformation of the Six-Carbon Polyfluoroalkyl Substance N-Dimethyl Ammonio Propyl Perfluorohexane Sulfonamide (AmPr-FHxSA).

Sites impacted by aqueous film-forming foam (AFFF) contain co-contaminants that can stimulate biotransformation of polyfluoroalkyl substances. Here, we compare how microbial enrichments from AFFF-impacted soil amended with diethyl glycol monobutyl ether (found in AFFF), aromatic hydrocarbons (present in co-released fuels), acetate, and methane (substrates used or formed during bioremediation) impact the aerobic biotransformation of an AFFF-derived six-carbon electrochemical fluorination (ECF) precursor N-dimethyl ammonio propyl perfluorohexane sulfonamide (AmPr-FHxSA). We found that methane- and acetate-oxidizing cultures resulted in the highest yields of identifiable products (38 and 30%, respectively), including perfluorohexane sulfonamide (FHxSA) and perfluorohexane sulfonic acid (PFHxS). Using these data, we propose and detail a transformation pathway. Additionally, we examined chemical oxidation products of AmPr-FHxSA and FHxSA to provide insights on remediation strategies for AmPr-FHxSA. We demonstrate mineralization of these compounds using the sulfate radical and test their transformation during the total oxidizable precursor (TOP) assay. While perfluorohexanoic acid accounted for over 95% of the products formed, we demonstrate here for the first time two ECF-based precursors, AmPr-FHxSA and FHxSA, that produce PFHxS during the TOP assay. These findings have implications for monitoring poly- and perfluoroalkyl substances during site remediation and application of the TOP assay at sites impacted by ECF-based precursors.

Potential of biological sulphur recovery from thiosulphate by haloalkaliphilic Thioalkalivibrio denitrificans

ABSTRACT The aim of this study was to investigate the potential for elemental sulphur recovery from sulphurous solutions under aerobic and anoxic conditions by haloalkalophilic Thioalkalivibrio denitrificans at 0.8–19.6 g S2O3 2−-S L−1 and 0.2–0.58 g NO2 L−1, respectively. The experiments were conducted as batch assays with haloalkaline (pH 10 and ≥ 14 g Na+ L−1) thiosulphate solution. Aerobically, the highest biotransformation rate of thiosulphate obtained was 0.03 h−1 at 8.5 g L S2O3 2−-S. Based on Monod model, the maximum substrate utilisation rate (qm) was 0.024 h−1 with half saturation constant (Ks) 0.42 g S2O3 2−-S L−1 at initial [S2O3 2--S] of 14 g L−1. S0 accumulated at [S2O3 2−-S] ≥ 1.5 g L−1 (10% yield at initial 9.5 g S2O3 2−-S L−1) and the highest S0 yield estimated with the model was 61% with initial [S2O3 2--S] of 16.5 g L−1. Anoxically, the maximum nitrite removal rate based on Monod modelling was 0.011 h−1 with Ks = 0.84 g NO2 − L−1. Aerobically and anoxically the maximum specific growth rates (µm) were 0.046 and 0.022 h−1, respectively. In summary, high-rate aerobic biotransformation kinetics of thiosulphate were demonstrated, whereas the rates were slower and no S0 accumulated under anoxic conditions. Thus, future developments of biotechnical applications for the recovery of S0 from haloalkaline streams from the process industry should focus on aerobic treatment. Highlights Haloalkaline S2O3 2− biotransformations kinetics by Thioalkalivibrio denitrificans Aerobic thiosulphate-S bioconversion up to 0.024 h−1 with Ks = 0.42 g S2O3 2−-S L−1 10% S0 yield with initial 9.5 g S2O3 2--S L−1 in aerobic condition Anoxic NO2 removal up to 0.01 h−1 with Ks = 0.84 g NO2 − L−1

Neutral Per- and Polyfluoroalkyl Substances, Butyl Carbitol, and Organic Corrosion Inhibitors in Aqueous Film-Forming Foams: Implications for Vapor Intrusion and the Environment.

Per- and polyfluoroalkyl substances (PFAS), butyl carbitol, and corrosion inhibitors are components of aqueous film-forming foams (AFFFs). Volatile (neutral) fluorotelomerization (FT)- and electrochemical fluorination (ECF)-based PFAS, butyl carbitol, and organic corrosion inhibitors were quantified in 39 military specification (MilSpec), non-MilSpec, and alcohol resistant-AFFF concentrates (undiluted) from 1974 to 2010. Fluorotelomer alcohols were found only in FT-based AFFFs and N-methyl- and N-ethyl-perfluoroalkyl sulfonamides, and sulfonamido ethanols were found only in ECF-based AFFFs. Neutral PFAS and benzotriazole, 4-methylbenzotriazole, and 5-methybenzotriazole occurred at mg/L levels in the AFFFs, while butyl carbitol occurred at g/L levels. Neutral PFAS concentrations in indoor air due to vapor intrusion of a nearby undiluted AFFF release are estimated to be anywhere from 2 to >10 orders of magnitude higher than documented background indoor air concentrations. Estimated butyl carbitol and organic corrosion inhibitor concentrations were lower than and comparable to indoor concentrations recently measured, respectively. The wide range of neutral PFAS concentrations and Henry's law constants indicate that field, soil-gas measurements are needed to validate the estimations. Co-discharged butyl carbitol likely contributes to oxygen depletion in AFFF-impacted aquifers and may hinder the natural PFAS aerobic biotransformation. Organic corrosion inhibitors in AFFFs indicate that these are another source of corrosion inhibitors in the environment.

Efficient biodegradation of phenanthrene using Pseudomonas stutzeri LSH-PAH1 with the addition of sophorolipids: Alleviation of biotoxicity and cometabolism studies

Phenanthrene (PHE) is widely distributed, and it can cause genotoxicity in humans by interacting with enzymes in the body. A current challenge for PHE bioremediation is the inhibitory effect of biotoxic intermediates on bacterial growth. Notably, the aerobic biotransformation processes for PHE in the presence of sophorolipids have been poorly studied. Here, a PHE-degrading strain was isolated from sediments and identified as Pseudomonas stutzeri and named LSH-PAH1. It was observed that 1-naphthol (a biotoxic substance that can inhibit strain growth) was produced during the PHE metabolism process of LSH-PAH1. The biodegradation ratio increased from 21.4% to 91.7% within 48 h after the addition of sophorolipids. Unexpectedly, this addition accelerated the metabolic process for 1-naphthol rather than causing its accumulation. The cometabolism of 1-naphthol and sophorolipids alleviated the biotoxic effects for the strain, which was verified by gene expression analysis. We identified a new PHE-degrading strain and provided a mechanism for PHE biodegradation using LSH-PAH1 with the addition of sophorolipids, which provides a reference for practical applications of the bioremediation of PHE and study of the cometabolism of biotoxic intermediates.

Aerobic biotransformation of a novel highly functionalized polyfluoroether-based surfactant using activated sludge from a wastewater treatment plant.

A replacement fluorosurfactant has been recently introduced to the European market as an alternative to other per- and polyfluoroalkyl substances (PFAS) that have been phased-out or banned. Here, we incubated this novel fluorosurfactant (diFESOS, [F7C3OCHFCF2SCH2CH2OC(O)]2C2H3SO3-) which contains ether and thioether insertions, and its known polyfluoroalkyl degradation products, an alcohol (FESOH) and carboxylic acid (FESCA), with activated sludge from a wastewater treatment plant under sulfur-limited conditions. Dosed chemicals and their transformation products were monitored using ultra-high performance liquid chromatography-tandem mass spectrometry, and gas chromatography-mass spectrometry. In addition to FESOH and FESCA, two smaller metabolites were identified: C3F7OCHFCOO- (2H-3:2 PFECA) and perfluoropropanoic acid (PFPrA). 2H-3:2 PFECA presumably was a result of S-dealkylation of FESCA, which then resulted in the abiotic cleavage of two C-F bonds; no S-oxygenation was observed. Overall, the terminal products of this biotransformation likely have lower bioaccumulation potential than the parent fluorosurfactant based on comparison to other similar PFAS.

Correlating bacterial and archaeal community with efficiency of a coking wastewater treatment plant employing anaerobic-anoxic-oxic process in coal industry

Coking wastewater (CWW) contains various complex pollutants, and biological treatment processes are frequently applied in the coking wastewater treatment plants (CWWTPs). The present work is to evaluate the contaminants removal of a full-scale CWWTP with an anaerobic-anoxic-oxic process (A/A/O), to reveal function of bacterial and archaeal community involved in different bioreactors, and to clarify the relationship between the performance and microbial community. Illumina Miseq sequencing of bacteria showed that β-proteobacteria dominated in three bioreactors with relative abundance of 60.2%~81.7%. 75.2% of sequences were assigned to Petrobacter in the bioreactor A1, while Thiobacillus dominated in A2 and O with relative abundance of 31.8% and 38.7%, respectively. Illumina Miseq sequencing of archaea revealed a high diversity of methanogens existed in A1 and A2 activated sludge. Moreover, Halostagnicola was the dominant archaea in A1 and A2 activated sludge with relative abundance of 41.8% and 66.5%, respectively. Function predicted analysis explored that function of bacteria was similar to that of archaea but the relative abundance differed from each other. A putative biodegradation model of CWW treatment in A/A/O process indicated that A1 and A2 activated sludge mainly reduced carbohydrate, protein, TN, phenol and cyanide, as well as methane production. Bacteria in the bioreactor O were responsible for aerobic biotransformation of residual carbohydrates, refractory organics and nitrification. The redundancy analysis (RDA) further revealed that removal of COD, TN, and NO3−-N, phenol and cyanides were highly correlated with some anaerobic bacteria and archaea, whereas the transformation of NH4+-N was positively correlated with some aerobic bacteria.

Impact of elevated phosphogypsum on soil fertility and its aerobic biotransformation through indigenous microorganisms (IMO's) based technology

Phosphogypsum (PG) is a waste by-product of phosphate fertilizer industry, produced in huge amount during the manufacture of phosphoric acid by economic wet process. Assessment of PG toxicity on soil has been poorly emphasized, therefore an efficient methods needs to be adopted to assess its toxic effect on soil fertility. We also need an effective eco-technological strategies for better waste PG management in order to improve the environmental health. The present study aimed to investigate the impact of PG toxicity on fertile soil and utilization of indigenous microorganisms for aerobic detoxification of PG contaminated soil to evaluate the scope for biostimulation based in situ bioremediation. In this study it is evident that application of PG to fertile soil in certain concentration results highly acidic, sulfate rich, aerobic environment, thus severely weakens the metabolic activity of the indigenous microorganisms. This investigation via microcosm based study further evaluated the intrinsic biotransformation ability of these microorganisms and found that was enhanced significantly (>95% reduction in sulfate concentration in 180 days) with carbon, nitrogen and phosphate amendments. Community level physiological profiling analyses indicated distinct shift in metabolic abilities following carbon amendments. Our study for the first time may help to formulate a strategy in aerobic biotransformation of PG contaminated soil environment, yet appreciable rate by supplying adequate nutrients.

Evidence for cometabolic transformation of weathered toxaphene under aerobic conditions using camphor as a co-substrate.

Toxaphene is a persistent organic pollutant, composed of approximately 1000 highly chlorinated bicyclic terpenes. The purpose of this study was to evaluate if camphor, a structural analogue of toxaphene, could stimulate aerobic biotransformation of weathered toxaphene. Two enrichment cultures that degrade camphor as the sole carbon source were established from contaminated soil and biosolids. These cultures were used to evaluate aerobic transformation of weathered toxaphene. Only the biosolids culture could transform compounds of technical toxaphene (CTTs) in the presence of camphor, while no transformation was observed in the presence of glucose or with toxaphene as a sole carbon source. The transformed toxaphene had lower concentration of CTTs with longer retention times, and higher concentration of compounds with lower retention times. Gas chromatography with electron capture negative ion mass spectrometry (GC/ECNI-MS) showed that aerobic biotransformation mainly occurred with Cl8 - and Cl9 -CTTs compounds. The patterns of Cl6 - and Cl7 -CTTs were also simplified albeit to a much lesser extent. Seven camphor-degrading bacteria were isolated from the enrichment culture but none of them could degrade toxaphene. Camphor degrading culture can aerobically transform CCTs via reductive pathway probably by co-metabolism using camphor as a co-substrate. Since camphor is naturally produced by different plants, this study suggests that stimulation of aerobic transformation of toxaphene may occur in nature. Moreover plants, which produce camphor or similar compounds, might be used in bioremediation of contaminated soils.

Aerobic degradation of nonhalogenated organophosphate flame esters (OPEs) by enriched cultures from sludge: Kinetics, pathways, bacterial community evolution, and toxicity evaluation

The degradation by bacteria has been considered the main process for eliminating nonhalogenated organophosphate esters (OPEs) from wastewater treatment plants (WWTPs), but limited research has reported the biodegradation processes and clarified the microbial-mediated mechanisms for nonhalogenated OPE degradation in WWTPs. The aim of this study was to monitor the biodegradation of the most common nonhalogenated OPEs, namely, tris(2-butoxyethyl) phosphate (TBOEP), tris (n-butyl) phosphate (TNBP) and trisphenyl phosphate (TPHP), under aerobic conditions by sludge cultures from a conventional sewage plant. The microbial cultures were enriched separately with each OPE from activated sludge cultures, and the presence of glucose significantly enhanced degradation of the OPEs during the enrichment. The removal ratios for the three OPEs reached 29.3–89.9% after 5 cycles (25 days) of cultivation, and the first-order degradation kinetics followed the order of TPHP > TBOEP > TNBP, with their half-lives ranging between 12.8 and 99.0 h. Pathways of hydrolysis, hydroxylation, methoxylation, and substitution were confirmed for the aerobic biodegradation of these nonhalogenated OPEs, but only di-alkyl phosphates (DAPs) largely accumulated in culture medium as the most predominant transformation products. Phylotypes in Klebsiella were significantly more abundant during OPE biodegradation than in the initial sludge, which indicated that these microorganisms are associated with the biodegradation of nonhalogenated OPEs in sludge culture. Biodegradation of all investigated nonhalogenated OPEs was associated with a significant reduction in the residual toxicity to Vibrio fischeri, indicating a rather positive ecotoxicological outcome of the aerobic biotransformation processes achieved by the enriched sludge culture.

Effect of substrate concentrations on aerobic biotransformation of 6:2 fluorotelomer sulfonate (6:2 FTS) in landfill leachate

Biotransformation of 6:2 fluorotelomer sulfonate (FTS) results in the formation of short-chain (C4 – C6) perfluorocarboxylic acids (PFCAs) in landfill leachate. Although leachate substrate concentrations (i.e., organic carbon, ammonia) vary widely, their effects on 6:2 FTS biotransformation and PFCAs formation are unknown. This study investigated the effect of organic carbon and ammonia concentration in 6:2 FTS aerobic biotransformation and PFCA formation in leachate. Biotransformation experiments were conducted with sediment collected from a landfill leachate ditch, to which deionized (DI) water and various amounts of leachate were added. Microbial community analysis using 16S rRNA indicated that while phylum Proteobacteria dominated the bacterial composition throughout the 60 days, Actinobacteria increased with time. Many genera from Proteobacteria and Actinobacteria can synthesize a wide array of enzymes, indicating that these phyla are likely to play an important role in 6:2 FTS biotransformation. Higher biotransformation of 6:2 FTS was observed in leachate-added microcosms (∼21%), compared to DI water microcosm (∼14%), likely reflecting the substrate dependency of 6:2 FTS biotransformation. Substrate limiting conditions in DI water microcosm resulted in slightly greater formation of ∑(C4 – C6) PFCAs (∼14 mol%), compared with leachate added microcosms (10–13 mol%). The findings suggest that dilution of landfill leachate, (e.g., during wet seasons), likely results in reduced 6:2 FTS biotransformation and increased PFCAs formation compared to dry conditions. Observed formation of C7 – C8 PFCAs in the live microcosms suggested that landfills act as secondary sources of legacy PFCAs (e.g., perfluorooctanoic acid) in the environment.

Aerobic biotransformation of fluorotelomer compounds in landfill leachate-sediment

Consumer products containing fluorotelomer polymers are a source of fluorotelomer compounds to the environment following their disposal at landfills. The fate and transformation of fluorotelomer compounds are unknown in landfill leachates. This study investigates the aerobic biotransformation of 8:2 fluorotelomer alcohol (FTOH) and 6:2 fluorotelomer sulfonate (FTS) in landfill leachate-sediment microcosms using batch tests. Spiked 8:2 FTOH, 6:2 FTS and their known biotransformation products were quantified in sediment-leachate and headspace over 90 days under aerobic conditions. 8:2 FTOH and 6:2 FTS biotransformation was slow (half-life >>30 d) in landfill leachate-sediment microcosm, suggesting persistence of fluorotelomer compounds under the conditions investigated. Significant volatilization (>20%) of 8:2 FTOH was observed in the microcosm headspace after 90 days. C6 – C8 and C4 – C6 perfluorocarboxylic acids (PFCAs) were the most abundant products for 8:2 FTOH and 6:2 FTS, respectively. PFCAs accounted for 4–9 mol% of the initially spiked parent compounds at 90 days. Perfluorooctanoic acid (PFOA) was the single most abundant product of 8:2 FTOH (>2.8 mol% at 90 days). The unaccounted mass (20 to 35 mol%) of the initially spiked parent compounds indicated formation of fluorotelomer intermediates and sediment-bound residue. Overall the findings suggest that aerobic biotransformation of fluorotelomer compounds acts as a secondary source of long- and short-chain (≤C7) PFCAs in the environment. Partitioning of semi-volatile fluorotelomer compounds (e.g., 8:2 FTOH) to the gas-phase indicates possible long-range transport and subsequent release of PFCAs in pristine environments. Short-chain fluorotelomer replacements (e.g., 6:2 FTS) result in a higher abundance of short-chain PFCAs in landfill leachate. Future research is needed to understand the long-term exposure effects of short-chain PFCAs to humans, aquatic life and biota.

Aerobic biotransformation of 6:2 fluorotelomer sulfonic acid in soil

Per- and polyfluoroalkyl substances (PFASs) have been widely used in numerous industrial and household products due to its hydrophobic and lipophobic nature. As the main active ingredient, PFASs have been widely used in aqueous film forming foam (AFFF) used to fight liquid-fueled fires. As the electrochemical fluorination (ECF)-based PFASs were gradually phased out by major PFAS manufacturers worldwide, fluorotelomer-based PFASs, such as 6:2 fluorotelomer sulfonic acid (6:2 FTSA, [F(CF2)6CH2CH2SO3H]) and its precursors, have been widely detected in soil and groundwater effected by firefighting training activities. A few precursors (e.g., 6:2 fluorotelomer alcohol, 6:2 FTOH, [F(CF2)6CH2CH2OH]) have been well studied, but there are still leaving major gaps in understanding of 6:2 FTSA fate and aerobic biotransformation in soil. Soil collected from forest area was dosed with 6:2 FTSA to investigate its biotransformation rate and to identify major transformation products in a flow-through soil incubation system over 90 d. The overall mass balance at day 90 in sterile and live treatments averaged 97.4% and 76.8%, respectively, suggesting that there may be soil-bound residues and conjugates between fluorinated transformation products and dissolved soil components only in live test system not in the sterile soil. The 6:2 FTSA biotransformation was relatively low with a half-life more than 56 d, and 33% of initial 6:2 FTSA remained at day 90. According to the transformation products and the results in 6:2 FTSA biodegradation, the pathways of 6:2 FTSA aerobic biodegradation in the soil were proposed. The volatile 5:2 secondary alcohol (5:2 sFTOH, [F(CF2)5CH(OH)CH3]) and 5:2 fluorotelomer ketone (5:2 FT ketone, [F(CF2)5COCH3]) were the most abundant intermediate transformation products at the beginning, and reached highest at day 7 and 14, respectively. Perfluoropentanoic acid (PFPeA, [F(CF2)4COOH]) and perfluorohexanoic acid (PFHxA, [F(CF2)5COOH]) were produced as end products by 6:2 FTSA through desulfonation, oxidation and other reactions. In another minor pathway, 5:3 polyfluorinated acid (5:3 acid, [F(CF2)5CH2CH2COOH]) and perfluorobutanoic acid (PFBA, [F(CF2)3COOH]) were also formed. The concentrations of these end products, including PFBA, PFPeA, PFHxA, and 5:3 acid, increased in all experiment period. At day 90, the generations of PFPeA, PFHxA, 5:3 acid, and PFBA at a yield of 20.4 mol%, 6.5 mol%, 9.5 mol%, and 1.0 mol% were observed. Perfluoroheptanoic acid (PFHpA, [F(CF2)6COOH]) was not detected in the soil, suggesting that there was no α-oxidation in the process of 6:2 FTSA biotransformation in live soil. NaOH-acetonitrile extraction plus ENVI-Carb™ clean up could breakdown the binding between 5:3 acid and organic matter of the soil, and an enhancement factor up to 18 could be achieved for 5:3 acid by this cleanup method. The sorption of 6:2 FTSA to soil could low the biotransformation. The enzymatic desulfonation step may limit 6:2 FTSA biotransformation in aerobic soil. This study indicates that 6:2 FTSA may be persistent in aerobic soil, and a potential source of short chain perfluorocarboxylic acids (PFCAs) in soil. More studies need to be done to understanding the fate of AFFF-related polyfluoroalkyl precursors in soil environment in the future work.