Bacterial degradation of perfluoroalkyl acids.

Advances in biological degradation of per- and polyfluoroalkyl substances (PFAS) have shown that bioremediation is a promising method of PFAS mineralization; however, most of these studies focus on remediation of more reactive polyfluorinated compounds. This review focuses on the defluorination of the more recalcitrant perfluorinated alkyl acids (PFAAs) by bacteria. We highlight key studies that report PFAA degradation products, specific bacteria, and relevant genes. Among these studies, we discuss trends in anaerobic versus aerobic conditions with specific bacterial species or consortia. This holistic review seeks to elucidate the state of PFAA biodegradation research and discuss the need for future research for environmental application.

Defluorination of PFAS by Acidimicrobium sp. strain A6 and potential applications for remediation.

Acidimicrobium sp. strain A6 is a recently discovered autotrophic bacterium that is capable of oxidizing ammonium while reducing ferric iron and is relatively common in acidic iron-rich soils. The genome of Acidimicrobium sp. strain A6 contains sequences for several reductive dehalogenases, including a gene for a previously unreported reductive dehalogenase, rdhA. Incubations of Acidimicrobium sp. strain A6 in the presence of perfluorinated substances, such as PFOA (perfluorooctanoic acid, C8HF15O2) or PFOS (perfluorooctane sulfonic acid, C8HF17O3S), have shown that fluoride, as well as shorter carbon chain PFAAs (perfluoroalkyl acids), are being produced, and the rdhA gene is expressed during these incubations. Results from initial gene knockout experiments indicate that the enzyme associated with the rdhA gene plays a key role in the PFAS defluorination by Acidimicrobium sp. strain A6. Experiments focusing on the defluorination kinetics by Acidimicrobium sp. strain A6 show that the defluorination kinetics are proportional to the amount of ammonium oxidized. To explore potential applications for PFAS bioremediation, PFAS-contaminated biosolids were augmented with Fe(III) and Acidimicrobium sp. strain A6, resulting in PFAS degradation. Since the high demand of Fe(III) makes growing Acidimicrobium sp. strain A6 in conventional rectors challenging, and since Acidimicrobium sp. strain A6 was shown to be electrogenic, it was grown in the absence of Fe(III) in microbial electrolysis cells, where it did oxidize ammonium and degraded PFAS.

Enhanced Feammox activity and perfluorooctanoic acid (PFOA) degradation by Acidimicrobium sp. Strain A6 using PAA-coated ferrihydrite as an electron acceptor.

Acidimicrobium sp. Strain A6 (A6) can degrade perfluoroalkyl acids (PFAAs) by oxidizing NH4+ while reducing Fe(Ⅲ). However, supplying and distributing Fe(III) phases in sediments is challenging since surface charges of Fe(III)-phases are typically positive while those of sediments are negative. Therefore, ferrihydrite particles were coated with polyacrylic acid (PAA) with four different molecular weights, resulting in a negative zeta potential on their surface. Zeta potential was determined as a function of pH and PAA loading, with the lowest value observed when the PAA/ferrihydrite ratio was > 1/5 (w/w) at a pH of 5.5. Several 50-day incubations with an A6-enrichment culture were conducted to determine the effect of PAA-coated ferrihydrite as the electron acceptor of A6 on the Feammox activity and PFOA degradation. NH4+ oxidation, PFOA degradation, production of shorter-chain PFAS, and F- were observed in all PAA-coated samples. The 6 K and 450 K treatments exhibited significant reductions in PFOA concentration and substantial F- production compared to incubations with bare ferrihydrite. Electrochemical impedance spectroscopy showed lowered charge transfer resistance in the presence of PAA-coated ferrihydrite, indicating that PAAs facilitated electron transfer to ferrihydrite. This study highlights the potential of PAA-coated ferrihydrite in accelerating PFAS defluorination, providing novel insights for A6-based bioremediation strategies.

Modeling the kinetics of perfluorooctanoic and perfluorooctane sulfonic acid biodegradation by Acidimicrobium sp. Strain A6 during the feammox process.

Per- and polyfluoroalkyl substances (PFAS) are emerging contaminants of concern due to their health effects and persistence in the environment. Although perfluoroalkyl acids (PFAAs), such as perfluorooctanoic acid (PFOA) and perfluorooctanesulfonic acid (PFOS) are very difficult to biodegrade because they are completely saturated with fluorine, it has recently been shown that Acidimicrobium sp. A6 (A6), which oxidizes ammonium under iron reducing conditions (Feammox process), can defluorinate PFAAs. A kinetic model was developed and tested in this study using results from previously published laboratory experiments, augmented with results from additional incubations, to couple the Feammox process to PFAS defluorination. The experimental results show higher Feammox activity and PFAS degradation in the A6 enrichment cultures than in the highly enriched A6 cultures. The coupled experimental and modeling results show that the PFAS defluorination rate is proportional to the rate of ammonium oxidation. The ammonium oxidation rate and the defluorination rate increase monotonically, but not linearly, with increasing A6 biomass. Given that different experiments had different level of Feammox activity, the parameters required to simulate the Feammox varied between A6 cultures. Nonetheless, the kinetic model was able to simulate an anaerobic incubation system and show that PFAS defluorination is proportional to the Feammox activity.

Biodegradation of PFOA in microbial electrolysis cells by Acidimicrobiaceae sp. strain A6.

Acidimicrobiaceae sp. strain A6 (A6), is an anaerobic autotrophic bacterium capable of oxidizing ammonium (NH4+) while reducing ferric iron and is also able to defluorinate PFAS under these growth conditions. A6 is exoelectrogenic and can grow in microbial electrolysis cells (MECs) by using the anode as the electron acceptor in lieu of ferric iron. Therefore, cultures of A6 amended with perfluorooctanoic acid (PFOA) were incubated in MECs to investigate its ability to defluorinate PFAS in such reactors. Results show a significant decrease in PFOA concentration after 18 days of operation, while producing current and removing NH4+. The buildup of fluoride and shorter chain perfluorinated products was detected only in MECs with applied potential, active A6, and amended with PFOA, confirming the biodegradation of PFOA in these systems. This work sets the stage for further studies on the application of A6-based per- and polyfluorinated alkyl substances (PFAS) bioremediation in microbial electrochemical systems for water treatment.

Anaerobic degradation of perfluorooctanoic acid (PFOA) in biosolids by Acidimicrobium sp. strain A6.

Anaerobic incubations were performed with biosolids obtained from an industrial wastewater treatment plant (WWTP) that contained perfluorooctanoic acid (PFOA), and with per- and polyfluoroalkyl substances- (PFAS) free, laboratory-generated, biosolids that were spiked with PFOA. Biosolid slurries were incubated for 150 days as is, after augmenting with either Acidimicrobium sp. Strain A6 or ferrihydrite, or with both, Acidimicrobium sp. Strain A6 and ferrihydrite. Autoclaved controls were run in parallel. Only the biosolids augmented with both, Acidimicrobium sp. Strain A6 and ferrihydrite showed a decrease in the PFOA concentration, in excess of 50% (total, dissolved, and solid associated). Higher concentrations of PFOA in the biosolids spiked with PFOA and no previous PFAS exposure allowed to track the production of fluoride to verify PFOA defluorination. The buildup of fluoride over the incubation time was observed in these biosolid incubations spiked with PFOA. A significant increase in the concentration of perfluoroheptanoic acid (PFHpA) over the incubations of the filter cake samples from the industrial WWTP was observed, indicating the presence of a non-identified precursor in these biosolids. Results show that anaerobic incubation of PFAS contaminated biosolids, after augmentation with Fe(III) and Acidimicrobium sp. Strain A6 can result in PFAS defluorination.

Tropical cyclone effects on water and sediment chemistry and the microbial community in estuarine ecosystems.

Frequent and intense storm disturbances can have widespread and strong effects on the nitrogen and iron cycles and their associated microbial communities in estuary systems. A three-year investigation was conducted in the Pearl River and Zhanjiang estuaries in Guangdong Province, China through repeated sampling at three timepoints, defined as pre-storm (<1 month before storm), post-storm (<1 month after storm), and non-storm (6-8 months after storm). Increased nutrient concentrations (total organic carbon, nitrate, nitrite, ammonium, and sulfate) in both the sediment and water column were observed immediately after storm. The microbial community experienced extensive and immediate changes determined by an observed composition shift in the nitrogen and iron-cycling microbiomes. Analysis of sediment samples displayed a shift from nitrogen-to sulfur-cycling microorganisms and an increase in microbial interactions that were not observed in the water column. The chemical profile and microbial community components both returned to baseline conditions 6-8 months following storm disturbance. Finally, significant correlations were found between chemical and microbial data, suggesting that niche-sharing microbes may respond similarly to stimuli that impact their ecosystem. Increases in nutrient availability can favor the abundance of specific taxa, as demonstrated by an increase in Acidimicrobium that affect both nitrogen and iron cycling.

A critical review of modeling Poly- and Perfluoroalkyl Substances (PFAS) in the soil-water environment.

Due to their health effects and the recalcitrant nature of their CF bonds, Poly- and Perfluoroalkyl Substances (PFAS) are widely investigated for their distribution, remediation, and toxicology in ecosystems. However, very few studies have focused on modeling PFAS in the soil-water environment. In this review, we summarized the recent development in PFAS modeling for various chemical, physical, and biological processes, including sorption, volatilization, degradation, bioaccumulation, and transport. PFAS sorption is kinetic in nature with sorption equilibrium commonly quantified by either a linear, the Freundlich, or the Langmuir isotherms. Volatilization of PFAS depends on carbon chain length and ionization status and has been simulated by a two-layer diffusion process across the air water interface. First-order kinetics is commonly used for physical, chemical, and biological degradation processes. Uptake by plants and other biota can be passive and/or active. As surfactants, PFAS have a tendency to be sorbed or concentrated on air-water or non-aqueous phase liquid (NAPL)-water interfaces, where the same three isotherms for soil sorption are adopted. PFAS transport in the soil-water environment is simulated by solving the convection-dispersion equation (CDE) that is coupled to PFAS sorption, phase transfer, as well as physical, chemical, and biological transformations. As the physicochemical properties and concentration vary greatly among the potentially thousands of PFAS species in the environment, systematic efforts are needed to identify models and model parameters to simulate their fate, transport, and response to remediation techniques. Since many process formulations are empirical in nature, mechanistic approaches are needed to further the understanding of PFAS-soil-water-plant interactions so that the model parameters are less site dependent and more predictive in simulating PFAS remediation efficiency.

Performance of sulfur-based autotrophic denitrification and denitrifiers for wastewater treatment under acidic conditions.

Autotrophic denitrification under acidic conditions using sulfide (S2-), elemental sulfur (S0), and thiosulfate (S2O32-) as electron donors are evaluated. Results from batch and column experiments show that when different S species were supplied, different pH conditions and denitrifier communities were required for denitrification to occur. Nitrate and nitrite were removed via autotrophic denitrification at pH ranging from 4 to 8, when S2- or S2O32- was the electron donor, while with S0 denitrification was only observed at pH > 6. When S2- was used as electron donor, it was converted to S0, and S0 was not used while S2- was available. When addition of S2- was discontinued, or S2- depleted, S0 that had accumulated was used as electron donor for denitrification. These findings demonstrate that sulfur-based autotrophic denitrification can proceed under acidic conditions, but that the addition of appropriate S species and the presence of an effective denitrifier community are required.

Defluorination of Perfluorooctanoic Acid (PFOA) and Perfluorooctane Sulfonate (PFOS) by Acidimicrobium sp. Strain A6.

Incubations with pure and enrichment cultures of Acidimicrobium sp. strain A6 (A6), an autotroph that oxidizes ammonium to nitrite while reducing ferric iron, were conducted in the presence of PFOA or PFOS at 0.1 mg/L and 100 mg/L. Buildup of fluoride, shorter-chain perfluorinated products, and acetate was observed, as well as a decrease in Fe(III) reduced per ammonium oxidized. Incubations with hydrogen as a sole electron donor also resulted in the defluorination of these PFAS. Removal of up to 60% of PFOA and PFOS was observed during 100 day incubations, while total fluorine (organic plus fluoride) remained constant throughout the incubations. To determine if PFOA/PFOS or some of their degradation products were metabolized, and since no organic carbon source except these PFAS was added, dissolved organic carbon (DOC) was tracked. At concentrations of 100 mg/L, PFOA/PFOS were the main contributors to DOC, which remained constant during the pure A6 culture incubations. Whereas in the A6 enrichment culture, DOC decreased slightly with time, indicating that as defluorination of PFOS/PFOA occurred, some of the products were being metabolized by heterotrophs present in this culture. Results show that A6 can defluorinate PFOA/PFOS while reducing iron, using ammonium or hydrogen as the electron donor.

Coupled fenton-denitrification process for the removal of organic matter and total nitrogen from coke plant wastewater.

This work assesses the feasibility of applying a Coupled Fenton-Denitrification (CFD) process for the treatment of wastewater from a coking plant. This highly toxic effluent is characterized by comparable carbon and nitrogen contents and it is usually released into the treatment system at well above room temperature. Recalcitrant organic matter can be easily removed in a first step using Fenton treatment. Working at 50 °C, pH0: 3, and a wastewater obtained from a coking plant, the stoichiometric amount of H2O2 relative to COD and a H2O2/Fe2+ weight ratio of 50, around 60% of carbon load was mineralized whereas H2O2 was completely depleted. However, no changes were observed in the total nitrogen content. A subsequent denitrification stage led to an additional 80% TOC (overall above 90%) and 75% Total Nitrogen removal. This was done in a batch bioreactor at room temperature over 72 h, using a 40-day pre-acclimated denitrifying biomass. These results point to the possibility of designing a combined chemical oxidation and biological treatment to deal with complex effluents containing refractory organic matter including high concentrations of nitrogen species.

Degradation of tetra- and trichloroethylene under iron reducing conditions by Acidimicrobiaceae sp. A6.

The degradation of trichloroethylene (TCE) and tetrachloroethylene (PCE), in incubations where ammonium was oxidized while iron was being reduced indicates that these compounds can be degraded during the Feammox process by Acidimicrobiaceae sp. A6 (ATCC, PTA-122488). None of these compounds were degraded in incubations to which no ammonium was added, indicating that they were degraded during the oxidation of ammonium. Degradation of TCE and PCE (ranging between 32% and 55%) was observed in incubations with a pure Acidimicrobiaceae sp. A6 culture as well as an Acidimicrobiaceae sp. A6 enrichment culture over a 2-week period. In addition to these batch studies, a column study, with a 5-h hydraulic residence time, was conducted contrasting the degradation of TCE in iron-rich soil columns that were either seeded with a pure or an enrichment culture of Acidimicrobiaceae sp. A6 to achieve ammonium oxidation under iron reduction, and a control column that was initially not seeded and later seeded with Geobacter metallireducens. While there was ∼22% TCE removal in the columns seeded with Acidimicrobiaceae sp. A6, there was no removal in the unseeded column or the column seeded with G. metallireducens which was being operated under iron reducing conditions. Feammox is an anoxic process that requires acidic conditions. Hence, these results indicate that this process might be harnessed where other bioremediation strategies are difficult, since many require neutral or alkaline conditions, and supplying ammonium to an anoxic aquifer is relatively easy, since there are not many processes that will oxidize ammonium in the absence of dissolved oxygen.

Anaerobic ammonium oxidation coupled to iron reduction in constructed wetland mesocosms.

Acidimicrobiaceae sp. A6 (referred to as A6) was recently identified as playing a key role in the Feammox process (ammonium oxidation coupled to iron reduction). Two constructed wetlands (CW) were built and bioaugmented with A6 to determine if, under the right conditions, Feammox can be enhanced in CWs by having strata with higher iron content. Hence, the solid stratum in the CWs was sand, and one CW was augmented with ferrihydrite. Vertical ammonium (NH4+) concentration profiles in the CW mesocosms were monitored regularly. After four months of operation, when reducing conditions were established in the CWs, they were inoculated with an enrichment culture containing A6 and monitored for an additional four months, after which they were dismantled and analyzed. During the four-month period after the A6 enrichment culture injection, 25.0 ±â€¯7.3% of NH4+ was removed from the CW with the high iron substrate whereas 11.0 ±â€¯9.7% was removed from the CW with the low iron substrate on average. Since the CW with high NH4+ removal had the same plant density, same bacterial biomass, same fraction of ammonium oxidizing bacteria (AOB), a higher biomass of A6, and a higher pH (NH4+ oxidation by Feammox raises pH, whereas NH4+ oxidation by aerobic AOB decreases pH), this difference in NH4+ removal is attributed to the Feammox process, indicating that wetlands can be constructed to take advantage of the Feammox process for increased NH4+ removal.

Electrode Colonization by the Feammox Bacterium Acidimicrobiaceae sp. Strain A6.

Acidimicrobiaceae sp. strain A6 (A6), from the Actinobacteria phylum, was recently identified as a microorganism that can carry out anaerobic ammonium (NH4+) oxidation coupled to iron reduction, a process also known as Feammox. Being an iron-reducing bacterium, A6 was studied as a potential electrode-reducing bacterium that may transfer electrons extracellularly onto electrodes while gaining energy from NH4+ oxidation. Actinobacteria species have been overlooked as electrogenic bacteria, and the importance of lithoautotrophic iron reducers as electrode-reducing bacteria at anodes has not been addressed. By installing electrodes in the soil of a forested riparian wetland where A6 thrives, in soil columns in the laboratory, and in A6-bioaugmented constructed wetland (CW) mesocosms and by operating microbial electrolysis cells (MECs) with pure A6 culture, the characteristics and performances of this organism as an electrode-reducing bacterium candidate were investigated. In this study, we show that Acidimicrobiaceae sp. strain A6, a lithoautotrophic bacterium, is capable of colonizing electrodes under controlled conditions. In addition, A6 appears to be an electrode-reducing bacterium, since current production was boosted shortly after the CWs were seeded with enrichment A6 culture and current production was detected in MECs operated with pure A6, with the anode as the sole electron acceptor and NH4+ as the sole electron donor.IMPORTANCE Most studies on electrogenic microorganisms have focused on the most abundant heterotrophs, while other microorganisms also commonly present in electrode microbial communities, such as Actinobacteria strains, have been overlooked. The novel Acidimicrobiaceae sp. strain A6 (Actinobacteria) is an iron-reducing bacterium that can colonize the surface of anodes in sediments and is linked to electrical current production, making it an electrode-reducing bacterium. Furthermore, A6 can carry out anaerobic ammonium oxidation coupled to iron reduction. Therefore, findings from this study open the possibility of using electrodes instead of iron as electron acceptors, as a means to promote A6 to treat NH4+-containing wastewater more efficiently. Altogether, this study expands our knowledge of electrogenic bacteria and opens the possibility of developing Feammox-based technologies coupled to bioelectric systems for the treatment of NH4+ and other contaminants in anoxic systems.

Seasonal distribution of nitrifiers and denitrifiers in urban river sediments affected by agricultural activities.

Nitrifiers and denitrifiers play a critical role in nitrogen removal in urban river sediments that are also affected by agricultural activities. However, the seasonal variations and vertical profile of these organisms in these river sediments are not well understood. In this study, the seasonal and depth (0 to 30 cm) distributions of the abundance and activity of nitrifiers and denitrifiers in sediments of the Pearl River in Guangzhou city were quantifying via qPCR and RT-qPCR according to various nitrifying and denitrifying functional genes, and their diversities were analyzed via high-throughput sequencing on an Illumina MiSeq platform. Results show that the distribution of nitrifiers and denitrifiers in these urban sediments were more abundant and active during the summer than winter; had distinct vertical distributions in the bacterial numbers and activity, with higher activity of the nirS gene (yearly averaged RNA:DNA 2.5% at 18 to 22 cm, vs. a yearly-depth average of 0.65%) but with lower overall numbers (yearly averaged 2.1 × 106 copies g-1 at 18 to 22 cm, vs. a yearly-depth average of 12.5 × 106 copies g-1); and their amoA and nosZ gene diversities in the sediments exhibited a correlation with the communities in nearby agricultural soils.

Isolation and characterization of an ammonium-oxidizing iron reducer: Acidimicrobiaceae sp. A6.

Acidimicrobiaceae sp. A6 (ATCC, PTA-122488), a strain that has been previously reported to play a key role in the oxidation of ammonium (NH4+) under iron reducing conditions, has now been isolated from riparian wetland soils in New Jersey, USA. Incubations of this strain in a medium containing ferrihydrite as the ferric iron [Fe(III)] source, CO2 as the carbon source, under room temperature, and a pH of 4.5, resulted in 52% of NH4+ removal over a 20-day incubation period, while reducing Fe(III) in the expected stoichiometric ratio when NH4+ was oxidized to nitrite with Fe(III) as the electron acceptor. This study demonstrates that this new isolated strain is capable of oxidizing NH4+ while reducing iron under anaerobic conditions.

Simultaneous measurements of dissolved CH4 and H2 in wetland soils.

Biogeochemical processes in wetland soils are complex and are driven by a microbiological community that competes for resources and affects the soil chemistry. Depending on the availability of various electron acceptors, the high carbon input to wetland soils can make them important sources of methane production and emissions. There are two significant pathways for methanogenesis: acetoclastic and hydrogenotrophic methanogenesis. The hydrogenotrophic pathway is dependent on the availability of dissolved hydrogen gas (H2), and there is significant competition for available H2. This study presents simultaneous measurements of dissolved methane and H2 over a 2-year period at three tidal marshes in the New Jersey Meadowlands. Methane reservoirs show a significant correlation with dissolved organic carbon, temperature, and methane emissions, whereas the H2 concentrations measured with dialysis samplers do not show significant relationships with these field variables. Data presented in this study show that increased dissolved H2 reservoirs in wetland soils correlate with decreased methane reservoirs, which is consistent with studies that have shown that elevated levels of H2 inhibit methane production by inhibiting propionate fermentation, resulting in less acetate production and hence decreasing the contribution of acetoclastic methanogenesis to the overall production of methane.

Phosphate enhanced abiotic and biotic arsenic mobilization in the wetland rhizosphere.

Although abiotic process of competitive sorption between phosphate (P) and arsenate (As(V)), especially onto iron oxides, are well understood, P-mediated biotic processes of Fe and As redox transformation contributing to As mobilization and speciation in wetlands remain poorly defined. To gain new insights into the effects of P on As mobility, speciation, and bioavailability in wetlands, well-controlled greenhouse experiments were conducted. As expected, increased P levels contributed to more As desorption, but more interestingly the interactions between P and wetland plants played a synergistic role in the microbially-mediated As mobilization and enhanced As uptake by plants. High levels of P promoted plant growth and the exudation of labile organic carbon from roots, enhancing the growth of heterotrophic bacteria, including As and Fe reducers. This in turn resulted in both, more As desorption into solution due to reductive iron dissolution, and a higher fraction of the dissolved As in the form of As(III) due to the higher number of As(V) reducers. Consistent with the dissolved As results, arsenic-XANES spectra from solid medium samples demonstrated that more As was sequestered in the rhizosphere as As(III) in the presence of high P levels than for low P levels. Hence, increased P loading to wetlands stimulates both abiotic and biotic processes in the wetland rhizosphere, resulting in more As mobilization, more As reduction, as well as more As uptake by plants. These interactions are important to be taken into account in As fate and transport models in wetlands and management of wetlands containing As.

Effect of dissimilatory iron and sulfate reduction on arsenic dynamics in the wetland rhizosphere and its bioaccumulation in wetland plants (Scirpus actus).

Microbial redox transformations of arsenic (As) are coupled to dissimilatory iron and sulfate reduction in the wetlands, however, the processes involved are complex and poorly defined. In this study, we investigated the effect of dissimilatory iron and sulfate reduction on As dynamics in the wetland rhizosphere and its bioaccumulation in plants using greenhouse mesocosms. Results show that high Fe (50µM ferrihydrite/g solid medium) and SO42- (5mM) treatments are most favorable for As sequestration in the presence of wetland plants (Scirpus actus), probably because root exudates facilitate the microbial reduction of Fe(III), SO42-, and As(V) to sequester As(III) by incorporation into iron sulfides and/or plant uptake. As retention in the solid medium and accumulation in plants were mainly controlled by SO42- rather than Fe levels. Compared to the low SO42- (0.1mM) treatment, high SO42- resulted in 2 times more As sequestered in the solid medium, 30 times more As in roots, and 49% less As in leaves. An As speciation analysis in pore water indicated that 19% more dissolved As was reduced under high SO42- than low SO42- levels, which is consistent with the fact that more dissimilatory arsenate-respiring bacteria were found under high SO42- levels.