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.

Heterogeneous Antibiotic Resistance Gene Removal Impedes Evaluation of Constructed Wetlands for Effective Greywater Treatment.

Microorganisms carrying antimicrobial resistance genes are often found in greywater. As the reuse of greywater becomes increasingly needed, it is imperative to determine how greywater treatment impacts antimicrobial resistance genes (ARGs). Using qPCR and SmartChip™ qPCR, we characterized ARG patterns in greywater microbial communities before, during, and after treatment by a recirculating vertical flow constructed wetland. In parallel, we examined the impact of greywater-treated irrigation on soil, including the occurrence of emerging micropollutants and the taxonomic and ARG compositions of microbial communities. Most ARGs in raw greywater are removed efficiently during the winter season, while some ARGs in the effluents increase in summer. SmartChip™ qPCR revealed the presence of ARGs, such as tetracycline and beta-lactam resistance genes, in both raw and treated greywater, but most abundantly in the filter bed. It also showed that aminoglycoside and vancomycin gene abundances significantly increased after treatment. In the irrigated soil, the type of water (potable or treated greywater) had no specific impact on the total bacterial abundance (16S rRNA gene). No overlapping ARGs were found between treated greywater and greywater-irrigated soil. This study indicates ARG abundance and richness increased after treatment, possibly due to the concentration effects of the filter beds.

Mitigation of antimicrobial resistance genes in greywater treated at household level.

Greywater often contains microorganisms carrying antimicrobial resistance genes (ARGs). Reuse of greywater thus potentially facilitates the enrichment and spread of multidrug resistance, posing a possible hazard for communities that use it. As water reuse becomes increasingly necessary, it is imperative to determine how greywater treatment impacts ARGs. In this study, we characterize ARG patterns in greywater microbial communities before and after treatment by a recirculating vertical flow constructed wetland (RVFCW). This greywater recycling method has been adopted by some small communities and households for greywater treatment; however, its ability to remove ARGs is unknown. We examined the taxonomic and ARG compositions of microbial communities in raw and treated greywater from five households using shotgun metagenomic sequencing. Total ARGs decreased in abundance and diversity in greywater treated by the RVFCW. In parallel, the microbial communities decreased in similarity in treated greywater. Potentially pathogenic bacteria associated with antimicrobial resistance and mobile genetic elements were detected in both raw and treated water, with a decreasing trend after treatment. This study indicates that RVFCW systems have the potential to mitigate antimicrobial resistance-related hazards when reusing treated greywater, but further measures need to be taken regarding persistent mobile ARGs and potential pathogens.

Anthropogenic antimicrobial micropollutants and their implications for agriculture.

Antibiotics and disinfectants have saved millions of human lives and cured uncountable animal diseases, but their activity is not limited to the site of application. Downstream, these chemicals become micropollutants, contaminating water at trace levels, resulting in adverse impacts on soil microbial communities and threatening crop health and productivity in agricultural settings and perpetuating the spread of antimicrobial resistance. Especially as resource scarcity drives increased reuse of water and other waste streams, considerable attention is needed to characterize the fate of antibiotics and disinfectants and to prevent or mitigate environmental and public health impacts. In this review, we hope to provide an overview of why increasing concentrations of micropollutants such as antibiotics are concerning in the environment, how they can pose health risks for humans, and how they can be countered using bioremediation strategies.

Clinically relevant pathogens on surfaces display differences in survival and transcriptomic response in relation to probiotic and traditional cleaning strategies.

Indoor surfaces are paradoxically presumed to be both colonized by pathogens, necessitating disinfection, and "microbial wastelands." In these resource-poor, dry environments, competition and decay are thought to be important drivers of microbial community composition. However, the relative contributions of these two processes have not been specifically evaluated. To bridge this knowledge gap, we used microcosms to evaluate whether interspecies interactions occur on surfaces. We combined transcriptomics and traditional microbiology techniques to investigate whether competition occurred between two clinically important pathogens, Acinetobacter baumannii and Klebsiella pneumoniae, and a probiotic cleaner containing a consortium of Bacillus species. Probiotic cleaning seeks to take advantage of ecological principles such as competitive exclusion, thus using benign microorganisms to inhibit viable pathogens, but there is limited evidence that competitive exclusion in fact occurs in environments of interest (i.e., indoor surfaces). Our results indicate that competition in this setting has a negligible impact on community composition but may influence the functions expressed by active organisms. Although Bacillus spp. remained viable on surfaces for an extended period of time after application, viable colony forming units (CFUs) of A. baumannii recovered following exposure to a chemical-based detergent with and without Bacillus spp. showed no statistical difference. Similarly, for K. pneumoniae, there were small statistical differences in CFUs between cleaning scenarios with or without Bacillus spp. in the chemical-based detergent. The transcriptome of A. baumannii with and without Bacillus spp. exposure shared a high degree of similarity in overall gene expression, but the transcriptome of K. pneumoniae differed in overall gene expression, including reduced response in genes related to antimicrobial resistance. Together, these results highlight the need to fully understand the underlying biological and ecological mechanisms for community assembly and function on indoor surfaces, as well as having practical implications for cleaning and disinfection strategies for infection prevention.

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 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.

A review on the important aspects of lipase immobilization on nanomaterials.

Lipase is one of the most widely used enzymes and plays an important role in biotechnological and industrial processes including food, paper, and oleochemical industries, as well as in pharmaceutical applications. However, its aqueous solubility and instability make its application relatively difficult and expensive. The immobilization technique is often used to improve lipase performance, and the strategy has turned out to be a promising method. Immobilized lipase on nanomaterials (NMs) has shown superiority to the free lipase, such as improved thermal and pH stability, longer stable time, and the capacity of being reused. However, immobilization of lipase on NMs also sometimes causes activity loss and protein loading is relatively lowered under some conditions. The overall performance of immobilized lipase on NMs is influenced by mechanisms of immobilization, type of NMs being used, and physicochemical features of the used NMs (such as particle size, aggregation behavior, NM dimension, and type of coupling/modifying agents being used). Based on the specific features of lipase and NMs, this review discusses the recent developments, some mechanisms, and influence of NMs on lipase immobilization and their activity. Multiple application potential of the immobilized lipases has also been considered.

Extraction of rare earth elements from a contaminated cropland soil using nitric acid, citric acid, and EDTA.

Rare earth elements (REEs) contamination to the surrounding soil has increased the concerns of health risk to the local residents. Soil washing was first attempted in our study to remediate REEs-contaminated cropland soil using nitric acid, citric acid, and ethylene diamine tetraacetic acid (EDTA) for soil decontamination and possible recovery of REEs. The extraction time, washing agent concentration, and pH value of the washing solution were optimized. The sequential extraction analysis proposed by Tessier was adopted to study the speciation changes of the REEs before and after soil washing. The extract containing citric acid was dried to obtain solid for the X-ray fluorescence (XRF) analysis. The results revealed that the optimal extraction time was 72 h, and the REEs extraction efficiency increased as the agent concentration increased from 0.01 to 0.1 mol/L. EDTA was efficient to extract REEs over a wide range of pH values, while citric acid was around pH 6.0. Under optimized conditions, the average extraction efficiencies of the major REEs in the contaminated soil were 70.96%, 64.38%, and 62.12% by EDTA, nitric acid, and citric acid, respectively. The sequential extraction analyses revealed that most soil-bounded REEs were mobilized or extracted except for those in the residual fraction. Under a comprehensive consideration of the extraction efficiency and the environmental impact, citric acid was recommended as the most suitable agent for extraction of the REEs from the contaminated cropland soils. The XRF analysis revealed that Mn, Al, Si, Pb, Fe, and REEs were the major elements in the extract indicating a possibile recovery of the REEs.