Schisandrin B ameliorates adjuvant-induced arthritis in rats via modulation of inflammatory mediators, oxidative stress, and HIF-1α/VEGF pathway.

OBJECTIVES: Schisandrin B (Sch B) has been shown to possess anti-inflammatory and antioxidant properties, however, its antirheumatoid arthritis properties and potential mechanism remain unexplored. This study evaluated the potential of Sch B in adjuvant-induced arthritic (AIA) rats. METHODS: AIA was induced by injecting 0.1 ml of CFA into the paw of rats and the animals were administered with Sch B (50 mg/kg) for 28 days. The effects of Sch B were evaluated using arthritis severity, serum levels of oxido-inflammatory, and metabolic index parameters. KEY FINDINGS: Sch B eased arthritic symptoms by significantly reducing paw swelling and arthritic score and increased body weight gain. Moreover, Sch B alleviated the levels of oxido-inflammatory markers including interleukin-1 beta, interleukin-6, tumor necrosis factor alpha, nuclear factor kappa B, transforming growth factor ß1, inducible nitric oxide synthase and malonaldehyde, as well as increased the levels of superoxide dismutase, glutathione, and Nrf2. Sch B also remarkably restored the altered levels of triglyceride, aspartate aminotransferase, lactic acid, pyruvate, phosphoenolpyruvate carboxylase, glucose, hypoxia inducible factor-1 alpha, and vascular endothelial growth factor. In addition, Sch B markedly alleviated p65 expression in the treated AIA rats. CONCLUSION: This study suggests that Sch B alleviated AIA by reducing oxidative stress, inflammation, and angiogenesis.

Rethinking Object Saliency Ranking: A Novel Whole-Flow Processing Paradigm.

Existing salient object detection methods are capable of predicting binary maps that highlight visually salient regions. However, these methods are limited in their ability to differentiate the relative importance of multiple objects and the relationships among them, which can lead to errors and reduced accuracy in downstream tasks that depend on the relative importance of multiple objects. To conquer, this paper proposes a new paradigm for saliency ranking, which aims to completely focus on ranking salient objects by their "importance order". While previous works have shown promising performance, they still face ill-posed problems. First, the saliency ranking ground truth (GT) orders generation methods are unreasonable since determining the correct ranking order is not well-defined, resulting in false alarms. Second, training a ranking model remains challenging because most saliency ranking methods follow the multi-task paradigm, leading to conflicts and trade-offs among different tasks. Third, existing regression-based saliency ranking methods are complex for saliency ranking models due to their reliance on instance mask-based saliency ranking orders. These methods require a significant amount of data to perform accurately and can be challenging to implement effectively. To solve these problems, this paper conducts an in-depth analysis of the causes and proposes a whole-flow processing paradigm of saliency ranking task from the perspective of "GT data generation", "network structure design" and "training protocol". The proposed approach outperforms existing state-of-the-art methods on the widely-used SALICON set, as demonstrated by extensive experiments with fair and reasonable comparisons. The saliency ranking task is still in its infancy, and our proposed unified framework can serve as a fundamental strategy to guide future work. The code and data will be available at https://github.com/MengkeSong/Saliency-Ranking-Paradigm.

Resveratrol-induced SIRT1 activation inhibits glycolysis-fueled angiogenesis under rheumatoid arthritis conditions independent of HIF-1α.

OBJECTIVE: This study investigated the impacts of SIRT1 activation on rheumatoid arthritis (RA)-related angiogenesis. METHODS: HUVECs were cultured by different human serum. Intracellular metabolites were quantified by UPLC-MS. Next, HUVECs and rat vascular epithelial cells under different inflammatory conditions were treated by a SIRT1 agonist resveratrol (RSV). Cytokines and biochemical indicators were detected by corresponding kits. Protein and mRNA expression levels were assessed by immunoblotting and PCR methods, respectively. Angiogenesis capabilities were evaluated by migration, wound-healing and tube-formation experiments. To down-regulate certain signals, gene-specific siRNA were applied. RESULTS: Metabolomics study revealed the accelerated glycolysis in RA serum-treated HUVECs. It led to ATP accumulation, but did not affect GTP levels. RSV inhibited pro-angiogenesis cytokines production and glycolysis in both the cells, and impaired the angiogenesis potentials. These effects were mimicked by an energy metabolism interrupter bikini in lipopolysaccharide (LPS)-primed HUVECs, largely independent of HIF-1α. Both RSV and bikinin can inhibit the activation of the GTP-dependent pathway Rho/ROCK and reduce VEGF production. Abrogation of RhoA signaling reinforced HIF-1α silencing-brought changes in LPS-stimulated HUVECs, and overshadowed the anti-angiogenesis potentials of RSV. CONCLUSION: Glycolysis provides additional energy to sustain Rho/ROCK activation in RA subjects, which promotes VEGF-driven angiogenesis and can be inhibited by SIRT1 activation.

Assessing the bioavailability of antibiotics in soil with the diffusive gradients in thin films (DGT).

The diffusive gradients in thin films (DGT) technique is an excellent method for investigating the dynamic processes of antibiotics in soils. However, whether it is applicable in antibiotic bioavailability assessment is yet to be disclosed. This study employed DGT to determine the antibiotic bioavailability in soil, and compared the results with plant uptake, soil solutions, and solvent extraction methods. DGT exhibited predictive capability for plant taking in antibiotics proved by the significant linear relationship between the DGT based concentration (CDGT) and antibiotic concentration in roots and shoots. Although the performance of soil solution was acceptable based on linear relationship analysis, its stability was weaker than DGT. The results based on plant uptake and DGT indicated the bioavailable antibiotic contents in different soils were inconsistent because of the distinct mobility and resupply of sulphonamides and trimethoprim in different soils, as represented by Kd and Rds, which were affected by soil properties. Plant species played an important role in antibiotic uptake and translocation. Antibiotic uptake by plants depends on antibiotic, plant and soil. These results confirmed the capability of DGT in determining antibiotic bioavailability for the first time. This work provided a simple and powerful tool for environmental risk evaluation of antibiotics in soils.

The synergistic toxicity effect of di(2-ethylhexyl)phthalate and plant growth disturbs the structure and function of soil microbes in the rhizosphere.

Di-(2-ethylhexyl) phthalate (DEHP) is a dominant phthalic acid ester in the environment and commonly occurs at high concentration in agricultural soils. Its influence on the soil microbial community has been widely reported, while research related to its effects on microbial structure, function, and interactions in the rhizosphere, a microbial hotspot region in the terrestrial ecosystem, is still limited. This study investigated the response of microbes in the rhizosphere to DEHP contamination. DEHP reduced microbial quantity, shifted the microbial community structure, and enriched the soil bacteria with potential DEHP degraders. Although the rhizosphere can alleviate DEHP toxicity, DEHP still played an important role in microbial community construction in the rhizosphere. Interestingly, some microbes were influenced by the synergistic toxicity effect of DEHP addition and plant growth, and there were significant differences in their relative abundance and alpha diversity in soil treated with both DEHP and planting compared to soils with just DEHP spiking or planting. The genes related to cell motility, metabolism of terpenoids and polyketides, protein families, genetic information processing, and replication and repair pathways changed only in soil treated with both DEHP and planting further proved the existence of synergistic toxicity. Anyway, the impact of DEHP on microbial function in the rhizosphere was important with 52.42‰ of the genes being changed. The change in cell motility, biofilm formation, and genes related to the quorum sensing pathway might affect the relationship between microbes, which play a crucial role in ecosystem function. This was further proven by changes in the microbial co-occurrence pattern. Our results can benefit risk evaluation of DEHP to microbial community in the rhizosphere, which is important for the effective function of terrestrial ecosystems and soil health.

Improving RGB-D Salient Object Detection via Modality-Aware Decoder.

Most existing RGB-D salient object detection (SOD) methods are primarily focusing on cross-modal and cross-level saliency fusion, which has been proved to be efficient and effective. However, these methods still have a critical limitation, i.e., their fusion patterns - typically the combination of selective characteristics and its variations, are too highly dependent on the network's non-linear adaptability. In such methods, the balances between RGB and D (Depth) are formulated individually considering the intermediate feature slices, but the relation at the modality level may not be learned properly. The optimal RGB-D combinations differ depending on the RGB-D scenarios, and the exact complementary status is frequently determined by multiple modality-level factors, such as D quality, the complexity of the RGB scene, and degree of harmony between them. Therefore, given the existing approaches, it may be difficult for them to achieve further performance breakthroughs, as their methodologies belong to some methods that are somewhat less modality sensitive. To conquer this problem, this paper presents the Modality-aware Decoder (MaD). The critical technical innovations include a series of feature embedding, modality reasoning, and feature back-projecting and collecting strategies, all of which upgrade the widely-used multi-scale and multi-level decoding process to be modality-aware. Our MaD achieves competitive performance over other state-of-the-art (SOTA) models without using any fancy tricks in the decoder's design. Codes and results will be publicly available at https://github.com/MengkeSong/MaD.

Elsholtzia splendens promotes phenanthrene and polychlorinated biphenyl degradation under Cu stress through enrichment of microbial degraders.

Co-contamination of heavy metals and organic pollutants is widespread in the environment. Metal-tolerant/hyperaccumulating plants have the advantage of enhancing co-operation between plants and rhizospheric microbes under heavy metal stress, but the underlying mechanism remains unclear. In the present study, the effects of Elsholtzia splendens and Lolium perenne on the rhizospheric microbial community and degraders of phenanthrene (PHE) and polychlorinated biphenyls (PCBs) were investigated. The results showed E. splendens could tolerate high Cu concentrations, while L. perenne was sensitive to Cu toxicity. Although Cu played the most important role in microbial community construction, both E. splendens and L. perenne caused shifts in the rhizospheric microbial community. For PHE and PCB degradation, L. perenne was more efficient under low Cu concentrations, whereas E. splendens performed better under high Cu concentrations. This difference can be attributed to shifts in the degrader community and key degradation genes identified by stable isotope probing. Moreover, higher abundances of various genes for organic pollutant degradation were observed in the rhizosphere of E. splendens than L. perenne based on gene prediction under high Cu stress. Our study reveals underlying mechanism of the advantages of heavy metal-tolerant plants for organic pollutant removal in soils co-contaminated with heavy metals.

New insight into the mechanism underlying the effect of biochar on phenanthrene degradation in contaminated soil revealed through DNA-SIP.

Biochar has been widely used for the remediation of polycyclic aromatic hydrocarbon (PAH)-contaminated soil, but its mechanism of influencing PAH biodegradation remains unclear. Here, DNA-stable isotope probing coupled with high-throughput sequencing was employed to assess its influence on phenanthrene (PHE) degradation, the active PHE-degrading microbial community and PAH-degradation genes (PAH-RHDα). Our results show that both Low-BC and High-BC (soils amended with 1 % and 4 % w/w biochar, respectively) treatments significantly decreased PHE biodegradation and bioavailable concentrations with a dose-dependent effect compared to Non-BC treatment (soils without biochar). This result could be attributed to the immobilisation of PHE and alteration of the composition and abundance of the PHE-degrading microbial consortium by biochar. Active PHE degraders were identified, and those in the Non-BC, Low-BC and High-BC microcosms differed taxonomically. Sphaerobacter, unclassified Diplorickettsiaceae, Pseudonocardia, and Planctomyces were firstly linked with PHE biodegradation. Most importantly, the abundances of PHE degraders and PAH-RHDα genes in the 13C-enriched DNA fractions of biochar-amended soils were greatly attenuated, and were significantly positively correlated with PHE biodegradation. Our findings provide a novel perspective on PAH biodegradation mechanisms in biochar-treated soils, and expand the understanding of the biodiversity of microbes involved in PAH biodegradation in the natural environment.

Per- and polyfluoroalkyl substances (PFASs) in the soil-plant system: Sorption, root uptake, and translocation.

Per- and polyfluoroalkyl substances (PFASs) are ubiquitous in the environment but pose potential risks to ecosystems and human health. The soil-plant system plays an important role in the bioaccumulation of PFASs. Because most PFASs in the natural environment are anionic and amphiphilic (both lipophilic and hydrophilic), their sorption and accumulation behaviors differ from those of neutral organic and common ionic compounds. In this review, we discuss processes affecting the availability of PFASs in soil after analyzing the potential mechanisms underlying the sorption and uptake of PFASs in the soil-plant system. We also summarize the current knowledge on root uptake and translocation of PFASs in plants. We found that the root concentration factor of PFASs for plants grown in soil was not significantly correlated with hydrophobicity, whereas the translocation factor was significantly and negatively correlated with PFAS hydrophobicity regardless of whether plants were grown hydroponically or in soil. Further research on the cationic, neutral, and zwitterionic forms of diverse PFASs is urgently needed to comprehensively understand the environmental fates of PFASs in the soil-plant system. Additional research directions are suggested, including the development of more accurate models and techniques to evaluate the bioavailability of PFASs, the effects of root exudates and rhizosphere microbiota on the bioavailability and plant uptake of PFASs, and the roles of different plant organelles, lipids, and proteins in the accumulation of PFASs by plants.

Uptake, Acropetal Translocation, and Enantioselectivity of Perfluorooctane Sulfonate in Maize Coexisting with Copper.

Plant uptake and translocation of perfluorooctane sulfonate (PFOS) are critical for food safety and raise major concerns. However, those processes are associated with many undisclosed mechanisms, especially when PFOS coexist with heavy metals. In this study, we investigated the effect of copper (Cu) on PFOS distribution in maize tissues by assessing the PFOS concentration and enantioselectivity. The presence of <100 µmol/L Cu exerted a limited effect on PFOS bioaccumulation, while >100 µmol/L Cu damaged the root cell membrane and increased root permeability, resulting in a higher PFOS concentration in roots. The suppression of acropetal translocation might be attributed to Cu inhibition of carrier proteins. The enantiomer fraction (EF) of 1m-PFOS at <100 µmol/L Cu was higher than that in a commercial product (0.5). Racemic PFOS was detected at >100 µmol/L Cu in roots and the EF variation changed from positive to negative in shoots. These EF results evidenced the existence of a protein-mediated uptake pathway. Besides, this study indicated the challenge of chiral signature application in PFOS source identification, given the effects of heavy metals and plants on PFOS enantioselectivity. The findings provide insight into PFOS bioaccumulation in plants cocontaminated with Cu and will facilitate environmental risk assessment.

Shifts in a Phenanthrene-Degrading Microbial Community are Driven by Carbohydrate Metabolism Selection in a Ryegrass Rhizosphere.

Plants usually promote pollutant bioremediation by several mechanisms including modifying the diversity of functional microbial species. However, conflicting results are reported that root exudates have no effects or negative effects on organic pollutant degradation. In this study, we investigated the roles of ryegrass in phenanthrene degradation in soils using DNA stable isotope probing (SIP) and metagenomics to reveal a potential explanation for conflicting results among phytoremediation studies. Phenanthrene biodegradation efficiency was improved by 8% after 14 days of cultivation. Twelve and ten operational taxonomic units (OTUs) were identified as active phenanthrene degraders in non-rhizosphere and rhizosphere soils, respectively. The active phenanthrene degraders exhibited higher average phylogenetic distances in rhizosphere soils (0.33) than non-rhizosphere soils (0.26). The Ka/Ks values (the ratio of nonsynonymous to synonymous substitutions) were about 10.37% higher in the rhizosphere treatment among >90% of all key carbohydrate metabolism-related genes, implying that ryegrass may be an important driver of microbial community variation in the rhizosphere by relieving the carbohydrate metabolism pressure and improving the survival ability of r-strategy microbes. Most Ka/Ks values of root-exudate-related metabolism genes exhibited little change, except for fumarate hydratase that increased 13-fold in the rhizosphere compared to that in the non-rhizosphere treatment. The Ka/Ks values of less than 50% phenanthrene-degradation-related genes were affected, 30% of which increased and 70% behaved oppositely. Genes with altered Ka/Ks values had a low percentage and followed an inconsistent changing tendency, indicating that phenanthrene and its metabolites are not major factors influencing the active degraders. These results suggested the importance of carbohydrate metabolism, especially fumaric acid, in rhizosphere community shift, and hinted at a new hypothesis that the rhizosphere effect on phenanthrene degradation efficiency depends on the existence of active degraders that have competitive advantages in carbohydrate and fumaric acid metabolism.

Large-scale biogeographical patterns of antibiotic resistome in the forest soils across China.

Soil is a reservoir of environmental resistomes. Information about their distribution, profiles, and driving forces in undisturbed environments is essential for understanding and managing modern antibiotic resistance genes (ARGs) in human disturbed environments. However, knowledge about the resistomes in pristine soils is limited, particularly at national scale. Here, we conducted a national-scale investigation of soil resistomes in pristine forests across China. Although the antibiotics content was low and ranged from below limit of detection (LOD) to 0.290 µg/kg, numerous detected ARGs conferring resistance to major classes of modern antibiotics were identified and indicated forest soils as a potential source of resistance traits. ARGs ranged from 6.20 × 10-7 to 2.52 × 10-3 copies/16S-rRNA and were predominated by those resisting aminoglycoside and encoding deactivation mechanisms. Low abundance of mobile genetic elements (MGEs) and its scarcely positive connections with ARGs suggest the low potential of horizontal gene transfer. The geographic patterns of ARGs and ARG-hosts in pristine forest soils were mainly driven by soil physiochemical variables and followed a distance-decay relationship. This work focusing on pristine soils can provide valuably new information for our understanding of the ARGs in human disturbed environments.

Interactions between microplastics and organic compounds in aquatic environments: A mini review.

Microplastics (MPs) are widely distributed in aquatic environments. They may release toxic substances or act as carriers for other organic compounds and pathogens, with potential to cause harm to the ecological environment and human health. A key concern is how MPs interact with organic compounds. We reviewed related works conducted under both laboratory conditions and in field aquatic environments to investigate the mechanisms of interactions between MPs and organic compounds from three perspectives: MPs, organic compounds, and environmental factors. The crystallinity and specific surface area of the MPs, and the functional groups, ionic form and strength of both MPs and organic compounds are key factors affecting their interactions. Environmentally realistic concentration settings for both MPs and organic compounds are critical for interpretation of the results of sorption experiments. The effect of salinity on interactions is mainly due to changes in pH. These results contribute to a better understanding of the environmental behavior, and potential ecological and human health risks of microplastics.

Land-use changes alter soil bacterial composition and diversity in tropical forest soil in China.

Tropical forests, under pressure from human activities, are important reservoirs of biodiversity and regulators of global biogeochemical cycles. Land-use and management are influential drivers of environmental change and ecosystem sustainability. However, only limited studies have analysed the impacts of planting age and vegetation type under land-use change on soil microbial community in tropical forests simultaneously. Here, we assessed soil bacterial community composition and diversity under different land-use in Hainan Province, China, using high-throughput sequencing combined with PICRUSt analysis. Land-use included natural forest, 5-year-old cropland, young (5-year-old) rubber tree plantation, and old (30-year-old) rubber tree plantation. Land-use changes altered the soil bacterial community composition but had a non-significant influence on alpha diversity (P > .05). We found that bacterial beta-diversity significantly decreased in young rubber tree plantation soils and cropland soils compared to natural forest as a control. In contrast, soil bacterial beta-diversity increased in old rubber tree plantation soils, indicating the effects of time since planting. There was no difference in microbial beta-diversity between soils from cropland and young rubber tree plantation. Soil bulk density and moisture, not pH, were the main environmental factors explaining the variability in microbial diversity. PICRUSt analysis of soil bacterial predicted gene abundances within metabolic pathways and indicated that land-use change altered soil functional traits, e.g., amino acid-related enzymes, ribosomes, DNA repair/recombination proteins and oxidative phosphorylation. Also, vegetation type, not planting age, had significant impacts on soil functional traits. Overall, planting age had the greatest influence on soil bacterial beta-diversity, while vegetation type was more crucial for soil functional traits (P < .05).

Alleviated Antibiotic-Resistant Genes in the Rhizosphere of Agricultural Soils with Low Antibiotic Concentration.

The influence of the rhizosphere on the abundance and diversity of antibiotic resistance genes (ARGs) has been recognized but there is a lack of consensus because of broad ranges of plant species and antibiotic concentrations across different habitats and the elusive underlying mechanisms. Here, we profiled antibiotic concentrations and resistomes in the rhizosphere and bulk soils by cultivating 10 types of crops in manure-amended agricultural soils. Rhizosphere effects altered the antibiotic resistome structure, significantly increased the absolute abundance of the antibiotic resistome, and decreased their relative abundance, contrasting previous studies. Such plantation-driven variation in ARGs resulted from the boost of bacterial lineages with negative relationships with ARGs and the constraint of the potential ARG-hosts in the rhizosphere of plants cultivated in soils with low antibiotic concentrations as the selective pressure. This mechanism is not reported previously and deepens our understanding about the rhizosphere effects on ARGs.

The complex interactions between novel DEHP-metabolising bacteria and the microbes in agricultural soils.

The indigenous microorganisms with the ability of metabolising di-(2-ethylhexyl) phthalate (DEHP) in agricultural soils and their interactions with non-degrading microbes were revealed by DNA-based stable isotope probing coupled with molecular ecological network. Aside from the previously reported DEHP degraders (family Planococcaceae and genus Sphingobacterium), five OTUs representing bacteria affiliated with genus Brevundimona, class Spartobacteria, genus Singulisphaera, genus Dyella and class Ktedonobacteria were linked with DEHP biodegradation. The analysis of the constructed ecological network based on soil microbial communities demonstrated the negative relationships between DEHP degraders and the dominant family Oxalobacteraceae in soils. Additionally, two cultivable bacteria isolated from the same soils, Rhizobium-1 and Ensifer-1, had strong capabilities in degrading DEHP but their involvement in in situ DEHP degradation was questioned, as their DNA was not labelled with 13C from DEHP. These findings provide deeper understanding on the indigenous DEHP-degrading communities and will benefit the remediation of phthalate esters contaminated soils.

The presence of in situ sulphamethoxazole degraders and their interactions with other microbes in activated sludge as revealed by DNA stable isotope probing and molecular ecological network analysis.

Wastewater treatment plants (WWTPs) are the main hotspots for the release of antibiotics, including the widely used sulphonamides. Microbes play important roles in eliminating sulphonamides in WWTPs, and knowledge about these degraders and their interactions within the microbial community is crucial for operating and optimising WWTPs. In the present study, stable isotope probing (SIP) coupled with high-throughput sequencing as culture-independent approach revealed four operational taxonomic units (OTUs) involved in sulphamethoxazole (SMX) degradation in activated sludge. Except for the OTU affiliated with Gammaproteobacteria, the others have not been previously reported to possess the ability to metabolise SMX. The isolated SMX degrader by culture-dependent method did not participate in SMX biodegradation in situ according to the SIP analysis, and showed weak correlations with other members in the activated sludge. The complex interactions between in situ active SMX degraders and non-degrading microbes might explain our failure to isolate these degraders. In addition, sul1 genes associated with SMX resistance were also labelled with 13C, suggesting that they might benefit from SMX degradation and/or originate from the active SMX degraders. These findings broaden our understanding of the diversity of SMX-degrading microbes and their associated characteristics in WWTPs.

Biphenyl-Metabolizing Microbial Community and a Functional Operon Revealed in E-Waste-Contaminated Soil.

Primitive electronic waste (e-waste) recycling activities release massive amounts of persistent organic pollutants (POPs) and heavy metals into surrounding soils, posing a major threat to the ecosystem and human health. Microbes capable of metabolizing POPs play important roles in POPs remediation in soils, but their phylotypes and functions remain unclear. Polychlorinated biphenyls (PCBs), one of the main pollutants in e-waste contaminated soils, have drawn increasing attention due to their high persistence, toxicity, and bioaccumulation. In the present study, we employed the culture-independent method of DNA stable-isotope probing to identify active biphenyl and PCB degraders in e-waste-contaminated soil. A total of 19 rare operational taxonomic units and three dominant bacterial genera ( Ralstonia, Cupriavidus, and uncultured bacterium DA101) were enriched in the 13C heavy DNA fraction, confirming their functions in PCBs metabolism. Additionally, a 13.8 kb bph operon was amplified, containing a bphA gene labeled by 13C that was concentrated in the heavy DNA fraction. The tetranucleotide signature characteristics of the bph operon suggest that it originated from Ralstonia. The bph operon may be shared by horizontal gene transfer because it contains a transposon gene and is found in various bacterial species. This study gives us a deeper understanding of PCB-degrading mechanisms and provides a potential resource for the bioremediation of PCBs-contaminated soils.

Identification of biphenyl-metabolising microbes in activated biosludge using cultivation-independent and -dependent approaches.

Microbes have important roles in removing organic pollutants in wastewater treatment plants (WWTPs), especially in mineralising recalcitrant persistent organic pollutants (POPs) such as polychlorinated biphenyls (PCBs). However, the majority of the microorganisms that metabolise these pollutants in situ remain elusive owing to barriers of traditional techniques in unravel yet-to-be-cultivated microbes. In this study, DNA stable-isotope probing (SIP) coupled with high-throughput sequencing was applied to identify the microbes responsible for PCB degradation in the activated biosludge of a WWTP using 13C-labelled biphenyl (BP). Results of time-course SIP revealed different bacteria and archaea involved in BP metabolism, which dominated the BP-degrading community at different time points. BP degradation by the genera Spartobacteria, Alicyclobacillus, Flavobacterium and the order Cenarchaeales has not been reported previously. The abundance of biphenyl dioxygenase (bphA) genes increased over time and a novel bphA gene was identified from the 13C-heavy DNA fraction. In addition, three cultivable BP degraders were isolated, but did not participate in BP degradation in situ or contain the identified bphA genes. Taken together, these data reveal the huge potential and important roles of yet-to-be-cultivated microbes responsible for PCB degradation in activated biosludge, providing fundamental knowledge on WWTP management to remove POPs.

Autochthonous Bioaugmentation-Modified Bacterial Diversity of Phenanthrene Degraders in PAH-Contaminated Wastewater as Revealed by DNA-Stable Isotope Probing.

To reveal the mechanisms of autochthonous bioaugmentation (ABA) in wastewater contaminated with polycyclic aromatic hydrocarbons (PAHs), DNA-stable-isotope-probing (SIP) was used in the present study with the addition of an autochthonous microorganism Acinetobacter tandoii LJ-5. We found LJ-5 inoculum produced a significant increase in phenanthrene (PHE) mineralization, but LJ-5 surprisingly did not participate in indigenous PHE degradation from the SIP results. The improvement of PHE biodegradation was not explained by the engagement of LJ-5 but attributed to the remarkably altered diversity of PHE degraders. Of the major PHE degraders present in ambient wastewater ( Rhodoplanes sp., Mycobacterium sp., Xanthomonadaceae sp. and Enterobacteriaceae sp.), only Mycobacterium sp. and Enterobacteriaceae sp. remained functional in the presence of strain LJ-5, but five new taxa Bacillus, Paenibacillus, Ammoniphilus, Sporosarcina, and Hyphomicrobium were favored. Rhodoplanes, Ammoniphilus, Sporosarcina, and Hyphomicrobium were directly linked to, for the first time, indigenous PHE biodegradation. Sequences of functional PAH-RHDα genes from heavy fractions further proved the change in PHE degraders by identifying distinct PAH-ring hydroxylating dioxygenases between ambient degradation and ABA. Our findings indicate a new mechanism of ABA, provide new insights into the diversity of PHE-degrading communities, and suggest ABA as a promising in situ bioremediation strategy for PAH-contaminated wastewater.