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Polybrominated diphenyl ethers (PBDEs) are persistent, highly toxic, and widely distributed environmental pollutants. The microbial populations and functional reductive dehalogenases (RDases) responsible for PBDE debromination in anoxic systems remain poorly understood, which confounds bioremediation of PBDE-contaminated sites. Here, we report a PBDE-debrominating enrichment culture dominated by a previously undescribed Dehalococcoides mccartyi population. A D. mccartyi strain, designated TZ50, whose genome contains 25 putative RDase-encoding genes, was isolated from the debrominating enrichment culture. Strain TZ50 dehalogenated a mixture of pentabrominated diphenyl ether (penta-BDE) and tetra-BDE congeners (total BDEs, 1.48 μM) to diphenyl ether within 2 weeks (0.58 μM Br-/day) via ortho- and meta-bromine elimination; strain TZ50 also dechlorinated tetrachloroethene (PCE) to vinyl chloride and ethene (260.2 μM Cl-/day). Results of native PAGE, proteomic profiling, and in vitro enzymatic activity assays implicated the involvement of three RDases in PBDE and PCE dehalogenation. TZ50_0172 (PteATZ50) and TZ50_1083 (TceATZ50) were responsible for the debromination of penta- and tetra-BDEs to di-BDE. TZ50_0172 and TZ50_1083 were also implicated in the dechlorination of PCE to trichloroethene (TCE) and of TCE to vinyl chloride/ethene, respectively. The other expressed RDase, TZ50_0090 (designated BdeA), was associated with the debromination of di-BDE to diphenyl ether, but its role in PCE dechlorination was unclear. Comparatively few RDases are known to be involved in PBDE debromination, and the identification of PteATZ50, TceATZ50, and BdeA provides additional information for evaluating debromination potential at contaminated sites. Moreover, the ability of PteATZ50 and TceATZ50 to dehalogenate both PBDEs and PCE makes strain TZ50 a suitable candidate for the remediation of cocontaminated sites. IMPORTANCE The ubiquity, toxicity, and persistence of polybrominated diphenyl ethers (PBDEs) in the environment have drawn significant public and scientific interest to the need for the remediation of PBDE-contaminated ecosystems. However, the low bioavailability of PBDEs in environmental compartments typically limits bioremediation of PBDEs and has long impeded the study of anaerobic microbial PBDE removal. In the current study, a novel Dehalococcoides mccartyi strain, dubbed strain TZ50, that expresses RDases that mediate organohalide respiration of both PBDEs and chloroethenes was isolated and characterized. Strain TZ50 could potentially be used to remediate multiple cooccurring organohalides in contaminated systems.

Fluoroquinolone antibiotics like ofloxacin (OFL) have been frequently detected in the aquatic environment. Recently manganese-oxidizing bacteria (MOB) have attracted research efforts on the degradation of recalcitrant pollutants with the aid of their biogenic manganese oxides (BioMnOx). Herein, the degradation of OFL with a strain of MOB (Pseudomonas sp. F2) was investigated for the first time. It was found that the bacteria can degrade up to 100% of 5 μg/L OFL. BioMnOx and Mn(III) intermediates significantly contributed to the degradation. Moreover, the degradation was clearly declined when the microbial activity was inactivated by heat or ethanol, indicating the importance of bioactivity. Possible transformation products of OFL were identified by HPLC-MS and the degradation pathway was proposed. In addition, the toxicity of OFL was reduced by 66% after the degradation.

Water is essential for life on Earth, and an important medium for microbial energy and metabolism. Dormancy is a state of low metabolic activity upon unfavorable conditions. Many microorganisms can switch to a metabolically inactive state after water shortage, and recover once the environmental conditions become favorable again. Here, we resuscitated dormant anammox bacteria from dry terrestrial ecosystems after a resting period of >10 ka by addition of water without any other substrates. Isotopic-tracer analysis showed that water induced nitrate reduction yielding sufficient nitrite as substrate and energy for activating anammox bacteria. Subsequently, dissimilatory nitrate reduction to ammonium (DNRA) provided the substrate ammonium for anammox bacteria. The ammonium and nitrite formed were used to produce dinitrogen gas. High throughput sequencing and network analysis identified Brocadia as the dominant anammox species and a Jettenia species seemed to connect the other community members. Under global climate change, increasing precipitation and soil moisture may revive dormant anammox bacteria in arid soils and thereby impact global nitrogen and carbon cycles.

An integrated investigation to document high anammox abundance, activity and diversity in upflow anaerobic sludge blanket (UASB) reactor treating low-strength ammonium loading sewage was performed and showed that the optimal anammox granular sludge sizes could mitigate undesirable N 2 O emission. The enhanced anammox bacterial abundance, activity and specific anammox rate were achieved with optimal granules sludge sizes of 0.5–0.9 mm with multiple “ Jettenia ”, “ Brocadia ”, and “ Anammoxoglobus ” species. The tightly-bound extracellular polymeric substance (TB-EPS) was the main EPS layer found in anammox granular sludge, in which polysaccharides play an important structural role. Over this granular sludge sizes, the anammox bacterial abundance and activity did not significantly decrease, but N 2 O emission significantly increased. High throughput sequencing and ecological networks demonstrated the patterns of anammox and their co-occurring bacteria, with availability N 2 O-producer and N 2 O-reducer functional genes. Incomplete denitrification and insufficient carbon source mainly contributed to N 2 O production in granular sludge, as supported by results of stratification analysis.

The distribution and importance of anaerobic ammonium oxidation (anammox) and nitrite-dependent anaerobic methane oxidation (n-damo) have been identified in aquatic ecosystems; their role in agricultural upland soils however has not yet been well investigated. In this study, we examined spatio-temporal distributions of anammox and n-damo bacteria in soil profiles (300 cm depth) from an agricultural upland. Monitoring nitrogen (N) conversion activity using isotope-tracing techniques over the course of one year showed denitrification (99.0% N-loss in the winter and 85.0% N-loss in the summer) predominated over anammox (1.0% N-loss in the winter and 14.4% N-loss in the summer) and n-damo (0.6% N-loss in the winter) in surface soils (0-20 cm). While below 20 cm depth, N-loss was dominated by anammox (79.4 ± 14.3% in the winter and 65.4 ± 12.5% in the summer) and n-damo was not detected. Phylogenetic analysis showed that Candidatus Brocadia anammoxidans dominated the anammox community in the surface soil and Candidatus Brocadia fulgida dominated below 20 cm depth. Dissimilatory nitrate reduction to ammonium (DNRA), another nitrite reduction process, was found to play a limited role (4.9 ± 3.5%) in the surface soil compared with denitrification; below 80 cm DNRA rates were much higher than rates of anammox and denitrification. Ammonium oxidation was the main source of NO2- above 80 cm (70.9 ± 23.3%), the key influencing factor on anammox rates, and nitrate reduction (100%) was the main NO2- source below 80 cm. Considering the anammox, n-damo and denitrification rates as a whole in the sampled soil profile, denitrification is still the main N-loss process in upland soils.

TetraBromoBisphenol-A (TBBPA) is a widely used brominated flame retardant and an emerging contaminant that has amassed significant environmental impacts. Though there are a few studies that report the bioremediation of TBBPA, there is no direct evidence to suggest a metabolic use of TBBPA as the sole electron acceptor, which offers an advantage in the complete and energy-efficient process of debromination under anaerobic conditions. In this study, Dehalococcoides mccartyi strain CG1 was identified to be capable of utilizing TBBPA as the sole electron acceptor at its maximum soluble concentrations (7.3 μM) coupled with cell growth. A previously characterized reductive dehalogenase (RDase), PcbA1, and six other RDases of strain CG1 were detected during TBBPA debromination via transcriptional and proteomic analyses. Furthermore, as a commonly co-contaminated brominated flame retardant of TBBPA, penta-BDEs were debrominated synchronously with TBBPA by strain CG1. This study provides deeper insights into the versatile dehalogenation capabilities of D. mccartyi strain CG1 and its role in in situ remediations of persistent organic pollutants in the environment.

Bioremediation of polychlorinated biphenyls (PCBs) is impeded by difficulties in massively cultivating bioinoculant. Meanwhile, sewage sludge is rich in pollutant-degrading microorganisms and nutrients, drawing our attention to investigate their potential to be used as a supplement for bioremediation of PCBs. Here we reported extensive microbial reductive dechlorination of PCBs by waste activated sludge (WAS) and digestion sludge (DS), which were identified to harbor multiple putative organohalide-respiring bacteria (i.e., Dehalococcoides, Dehalogenimonas, Dehalobacter, and uncultivated Dehalococcoidia) and PCB reductive dehalogenase genes (i.e., pcbA4 and pcbA5). Consequently, amendment of 1-20% (w/w) fresh WAS/DS enhanced the attenuation of PCBs by 126-544% in a soil microcosm compared with the control soil, with the fastest dechlorination of PCBs being achieved when spiked with 20% fresh WAS. Notably, dechlorination pathways of PCBs were also changed by sludge amendment. Microbial and physicochemical analyses revealed that the enhanced dechlorination of PCBs by sludge amendment was largely attributed to the synergistic effects of sludge-derived nutrients, PCB-dechlorinating bacteria, and stimulated growth of beneficial microorganisms (e.g., fermenters). Finally, risk assessment of heavy metals suggests low potential ecological risks of sludge amendment in soil. Collectively, our study demonstrates that sewage sludge amendment could be an efficient, cost-effective and environment-friendly approach for in situ bioremediation of PCBs.

Trihalomethanes such as chloroform and bromoform, although well-known as a prominent class of disinfection by-products, are ubiquitously distributed in the environment due to widespread industrial usage in the past decades. Chloroform and bromoform are particularly concerning, of high concentrations detected and with long half-lives up to several hundred days in soils and groundwater. In this study, we report a Dehalobacter- and Desulfovibrio-containing co-culture that exhibits dehalogenation of chloroform (~0.61 mM) to dichloromethane and bromoform (~0.67 mM) to dibromomethane within 10-15 days. This co-culture was further found to dechlorinate 1,1,1-trichloroethane (1,1,1-TCA) (~0.65 mM) to 1,1-dichloroethane within 12 days. The Dehalobacter species present in this co-culture, designated Dehalobacter sp. THM1, was found to couple growth with dehalogenation of chloroform, bromoform, and 1,1,1-TCA. Strain THM1 harbors a newly identified reductive dehalogenase (RDase), ThmA, which catalyzes chloroform, bromoform, and 1,1,1-TCA dehalogenation. Additionally, based on the sequences of thmA and other identified chloroform RDase genes, ctrA, cfrA, and tmrA, a pair of chloroform RDase gene-specific primers were designed and successfully applied to investigate the chloroform dechlorinating potential of microbial communities. The comparative analysis of chloroform RDases with tetrachloroethene RDases suggests a possible approach in predicting the substrate specificity of uncharacterized RDases in the future.

Anthropogenic organohalide contaminants present in wastewater treatment plants (WWTPs) often remain untreated and can be discharged into the environment. Although organohalide respiring bacteria (OHRB) contribute to the elimination of anthropogenic organohalides in natural anaerobic environments, reductive dehalogenation by OHRB in mainstream WWTPs remains poorly understood. In this study, we quantified OHRB during a long-term operation of a municipal WWTP with short hydraulic and sludge retention times (3 h and 1.5–5 days, respectively). The obligate OHRB were detected at high levels (averaging 2.56 ± 1.73 × 107 and 3.11 ± 1.16 × 107 16S rRNA gene copies/ml MLSS sludge in anoxic and aerobic zones, respectively) over the entire sampling period and throughout the wastewater treatment train. Microcosms derived from mainstream activated sludge contained an unidentified member of the Dehalococcoides genus that metabolically dechlorinated triclosan, used as a representative emerging organohalide antimicrobial, to diclosan, suggesting the potential of anaerobic degradation of emerging contaminants in WWTPs. To further understand the mechanisms for such antimicrobials’ removal, an investigation of dechlorination of triclosan by Dehalococcoides strains was conducted. Dechlorination of environmentally relevant concentrations of triclosan to diclosan was observed in Dehalococcoides mccartyi strain CG1, yielding 4.59 ± 0.34 × 108 cells/μmole Cl− removed at a rate of 0.062 μM/day and a minimal inhibitory concentration of 0.5 mg/L. Notably, both the tolerance of strain CG1 to triclosan and the rate of triclosan dechlorination increased when CG1 was cultured in the presence of both triclosan and tetrachloroethene. Taken together, our results suggest that anaerobic degradation of organohalide antimicrobials might be more prevalent in mainstream WWTPs than previously speculated, though the low growth yields that are supported by triclosan dechlorination seem to indicate that other organohalide substrates could be necessary to sustain OHRB populations in these systems.

NH4+ removal at low temperature (

Chloroethenes are common groundwater and soil contaminants due to extensive historic utilization and inappropriate discharge. The tendency for chloroethenes to become sequestered as dense non-aqueous phase liquids (DNAPL)-a point source to groundwater contamination and causing high concentrations of chloroethenes in proximal aquifers poses a great challenge for remediation of chloroethene contaminated sites. In this study, we report isolation and characterization of a Dehalococcoides mccartyi strain 11G which couples growth with reductive dechlorination of trichloroethenes (TCE), dichloroethene (DCE) isomers and vinyl chloride (VC) to ethene at a growth yield ranging from 2.47 ± 0.23 × 108 to 5.64 ± 0.43 × 108 cells/µmoles Cl- released and co-metabolically dechlorinates tetrachloroethene (PCE) in the presence of TCE. Compared with previous D. mccartyi strains showing dechlorination of TCE at up to 2.0 mM, strain 11G is distinguished by its capacity to dechlorinate chloroethenes at initial concentrations of DCE isomers as high as 4 mM and TCE as high as 3.5 mM to ethene. Bioaugmentation of a contaminated microcosm with strain 11G resulted in complete detoxification of a mixture of 5 mM chloroethenes (2.5 mM of each TCE and cis-DCE) after 40 days. Strain 11G is a promising candidate for in situ bioremediation of high-concentration-chloroethene contaminated sites.

Anaerobic ammonium oxidation (anammox) widely occurs in marine ecosystems, and it plays an important role in the global nitrogen cycle. But in freshwater ecosystems its occurrence, distribution and contribution, especially in extreme environments, are still not well known. In this study, anammox process was investigated in some extreme environments of freshwater ecosystems, such as those with high (above 75°C) and low (below -35°C) temperature, high (pH > 8) and low (pH 300 mg kg(-1) ). The polymerase chain reaction (PCR) screening results showed that anammox bacteria were widespread in the examined sediments from freshwater extreme environments. Quantitative PCR showed that the abundance of anammox bacteria ranged from 6.94 × 10(4) to 8.05 × 10(6) hydrazine synthase (hzsB) gene copies g(-1) dry soil. (15) N-labelled incubation experiments indicated the occurrence of anammox in all examined sediments and the potential anammox rates ranged from 0.02 to 6.24 nmol N g(-1) h(-1) , with a contribution of 3.45-58.74% of the total N2 production. In summary, these results demonstrate the occurrence of anammox in these extreme environments, inferring that anammox may harbour a wide ecological niche in the freshwater ecosystems.

Fastidious anaerobic bacteria play critical roles in environmental bioremediation of halogenated compounds. However, their characterization and application have been largely impeded by difficulties in growing them in pure culture. Thus far, no pure culture has been reported to respire on the notorious polychlorinated biphenyls (PCBs), and functional genes responsible for PCB detoxification remain unknown due to the extremely slow growth of PCB-respiring bacteria. Here we report the successful isolation and characterization of three Dehalococcoides mccartyi strains that respire on commercial PCBs. Using high-throughput metagenomic analysis, combined with traditional culture techniques, tetrachloroethene (PCE) was identified as a feasible alternative to PCBs to isolate PCB-respiring Dehalococcoides from PCB-enriched cultures. With PCE as an alternative electron acceptor, the PCB-respiring Dehalococcoides were boosted to a higher cell density (1.2 × 10(8) to 1.3 × 10(8) cells per mL on PCE vs. 5.9 × 10(6) to 10.4 × 10(6) cells per mL on PCBs) with a shorter culturing time (30 d on PCE vs. 150 d on PCBs). The transcriptomic profiles illustrated that the distinct PCB dechlorination profile of each strain was predominantly mediated by a single, novel reductive dehalogenase (RDase) catalyzing chlorine removal from both PCBs and PCE. The transcription levels of PCB-RDase genes are 5-60 times higher than the genome-wide average. The cultivation of PCB-respiring Dehalococcoides in pure culture and the identification of PCB-RDase genes deepen our understanding of organohalide respiration of PCBs and shed light on in situ PCB bioremediation.

Summary 1,1,1-Trichloroethane (TCA) and chloroform are two notorious groundwater pollutants. Here we report the isolation and characterization of Desulfitobacterium sp. strain PR that rapidly dechlorinates both compounds. In pyruvate-amended medium, strain PR reductively dechlorinates ∼ 1.0 mM TCA completely to monochloroethane within 15 days. Under the same conditions, strain PR dechlorinates ∼ 1.2 mM chloroform to predominantly dichloromethane (∼ 1.14 mM) and trace amount of monochloromethane (∼ 0.06 mM) within 10 days. Strain PR shares 96.7% 16S rRNA gene sequence similarity with its closest relative – Desulfitobacterium metallireducens strain 853-15; however, it distinguishes itself from known Desulfitobacterium strains by its inability of utilizing several of their commonly shared substrates such as lactate, thiosulfate and sulfite. A reductive dehalogenase gene (ctrA) in strain PR was identified to be responsible for dechlorination of both TCA and chloroform, showing a maximum expression level of 5.95 ∼ 6.25 copies of transcripts cell-1. CtrA shares 94% amino acid sequence identity with CfrA in Dehalobacter sp. strain CF50 and DcrA in Dehalobacter sp. strain DCA. Interestingly, strain PR could tolerate high aqueous concentrations (up to 0.45 mM) of trichloroethene, another groundwater pollutant that often coexists with TCA/chloroform. As the first chloroform-respiring and the second TCA-respiring isolate that has been identified, Desulfitobacterium sp. strain PR may prove useful in remediation of halogenated alkanes with trihalomethyl (-CX3) groups.

Soil nitrogen (N) deficiency is the major factor contributing to low primary productivity on the Qinghai-Tibet Plateau. However, most of our understanding of N cycling is still based on human disturbed environments, and the microbial mechanisms governing N loss in low primary productivity environment remain unclear. This study explores three microbial N loss pathways in eight wetland and dryland soil profiles from the Qinghai-Tibet Plateau, at an elevation of above 5000 m with little human activity, using 15N isotopic tracing slurry technology, quantitative PCR, and high-throughput sequencing. No denitrifying anaerobic methane oxidation was detected. Anammox occurred in two of the wetland (n = 4) and dryland (n = 4) soil profiles, while denitrification widely occurred and was the dominant N loss pathway in all samples. Where denitrification and anammox co-occurred, both abundance and activity were higher in wetland than in dryland soils and higher in surface than in subsurface soils. In comparison with non-anammox sites, nitrate levels initiate anammox-related N cycling. High-throughput sequencing and network analysis of nirK, nirS, nosZ, and hzsB gene communities showed that Bradyrhizobiaceae (a family of rhizobia) may play a dominant role in N loss pathways in this region. Given the geological evolution and relatively undisturbed habitat, these findings strongly suggest that denitrification is the dominant N loss pathway in terrestrial habitats of the Qing-Tibet Plateau with minimal anthropogenic activity.

Denitrification has long been regarded as the only pathway for terrestrial nitrogen (N) loss to the atmosphere. Here we demonstrate that large-scale anaerobic ammonium oxidation (anammox), an overlooked N loss process alternative to denitrification which bypasses nitrous oxide (N2O), is ubiquitous in inland waters of China and contributes significantly to N loss. Anammox rates in aquatic systems show different levels (1.0–975.9 μmol N m−2 h−1, n = 256) with hotspots occurring at oxic-anoxic interfaces and harboring distinct biogeochemical and biogeographical features. Extrapolation of these results to the China-national level shows that anammox could contribute about 2.0 Tg N yr−1, which equals averagely 11.4% of the total N loss from China’s inland waters. Our results indicate that a significant amount of the nitrogen lost from inland waters bypasses denitrification, which is important for constructing more accurate climate models and may significantly reduce potential N2O emission risk at a large scale.

This study investigated the diversity, community composition, and abundance of anaerobic ammonium oxidation (anammox) bacteria along the altitudinal gradient in Qinghai–Tibet Plateau. Two types of soil samples (wetland and dryland soils, n = 123) were collected from 641 m to 5,033 m altitudes. Polymerase chain reaction (PCR) screening showed that anammox were not widespread, and were only detected in 9 sampling sites of the 50 sites tested by amplifying the 16S rRNA genes. Then, only samples collected from Linzhi (2,715 m), Rikaze (4,030 m), and Naqu (5,011 m), which were positive for the presence of anammox, were further processed to explore the biogeography of anammox bacteria in Qinghai–Tibet Plateau. Results of high‐throughput sequencing targeting the hydrazine synthesis β‐subunit (hzsB) gene revealed the presence of three known anammox genera (Candidatus Brocadia, Candidatus Jettenia, and Candidatus Kuenenia) in both soil types. Their diversity, community composition, and abundance did not show significant variation with altitude at large scale. However, it was the small‐scale environmental heterogeneities between wetland and dryland soils that determined their biogeographical distribution. Specifically, the dryland soils had higher diversity of anammox bacteria than the wetland soils, but their abundance patterns varied. The community composition of anammox bacteria were found to be influenced by soil nitrate content.

Dehalococcoides mccartyi exhibits versatile capabilities to respire halogenated compounds under anaerobic conditions. In this study, we report the assembly and annotation of the complete genome of a chloroethene dechlorinating D. mccartyi strain 11a5. Bearing a 1,461,973 base-pair chromosome, strain 11a5 distinguishes itself from other D. mccartyi strains by possessing a 5,940 base-pair circular extrachromosomal genetic element which contains a reductive dehalogenase homolog. The whole genome of strain 11a5 harbors 31 putative reductive dehalogenase genes. Through transcriptional and proteomic analyses, we identified a new tetrachloroethene (PCE) reductive dehalogenase, PteA (11a5_1355), which catalyzes reductive dechlorination from PCE to trichloroethene (TCE) and shares only 38% similarity in amino acid sequence with its closest relative PceA in D. mccartyi strain 195. The acquisition of the genome of strain 11a5 enlarged the database of D. mccartyi and enriched our understanding of this unique species, among which, the identification of a new PCE reductive dehalogenase can assist in understanding PCE dechlorination process. In addition, the discovery of the circular extrachromosomal genetic element in strain 11a5 may provide insights to investigate how reductive dehalogenase homologous genes are transferred and carried by organohalide respiring bacteria.