H/D Exchange Coupled with 2H-labeled Stable Isotope-Assisted Metabolomics Discover Transformation Products of Contaminants of Emerging Concern.

Stable isotope-assisted metabolomics (SIAM) is a powerful tool for discovering transformation products (TPs) of contaminants. Nevertheless, the high cost or lack of isotope-labeled analytes limits its application. In-house H/D (hydrogen/deuterium) exchange reactions enable direct 2H labeling to target analytes with favorable reaction conditions, providing intuitive and easy-to-handle approaches for environmentally relevant laboratories to obtain cost-effective 2H-labeled contaminants of emerging concern (CECs). We first combined the use of in-house H/D exchange and 2H-SIAM to discover potential TPs of 6PPD (N-1,3-dimethylbutyl-N'-phenyl-p-phenylenediamine), providing a new strategy for finding TPs of CECs. 6PPD-d9 was obtained by in-house H/D exchange with favorable reaction conditions, and the impurities were carefully studied. Incomplete deuteride, for instance, 6PPD-d8 in this study, constitutes a major part of the impurities. Nevertheless, it has few adverse effects on the 2H-SIAM pipeline in discovering TPs of 6PPD. The 2H-SIAM pipeline annotated 9 TPs of 6PPD, and commercial standards further confirmed the annotated 6PPDQ (2-anilino-5-(4-methylpentan-2-ylamino)cyclohexa-2,5-diene-1,4-dione) and PPPD (N-phenyl-p-phenylenediamine). Additionally, a possible new formation mechanism for 6PPDQ was proposed, highlighting the performance of the strategy. In summary, this study highlighted a new strategy for discovering the TPs of CECs and broadening the application of SIAM in environmental studies.

SMRT and Illumina RNA sequencing reveal novel insights into the heat stress response and crosstalk with leaf senescence in tall fescue.

BACKGROUND: As a cool-season grass species, tall fescue (Festuca arundinacea) is challenged by increasing temperatures. Heat acclimation or activation of leaf senescence, are two main strategies when tall fescue is exposed to heat stress (HS). However, lacking a genome sequence, the complexity of hexaploidy nature, and the short read of second-generation sequencing hinder a comprehensive understanding of the mechanism. This study aims to characterize the molecular mechanism of heat adaptation and heat-induced senescence at transcriptional and post-transcriptional levels. RESULTS: Transcriptome of heat-treated (1 h and 72 h) and senescent leaves of tall fescue were generated by combining single-molecular real-time and Illumina sequencing. In total, 4076; 6917, and 11,918 differentially expressed genes (DEGs) were induced by short- and long-term heat stress (HS), and senescence, respectively. Venn and bioinformatics analyses of DEGs showed that short-term HS strongly activated heat shock proteins (Hsps) and heat shock factors (Hsfs), as well as specifically activated FK506-binding proteins (FKBPs), calcium signaling genes, glutathione S-transferase genes, photosynthesis-related genes, and phytohormone signaling genes. By contrast, long-term HS shared most of DEGs with senescence, including the up-regulated chlorophyll catabolic genes, phytohormone synthesis/degradation genes, stress-related genes, and NACs, and the down-regulated photosynthesis-related genes, FKBPs, and catalases. Subsequently, transient overexpression in tobacco showed that FaHsfA2a (up-regulated specifically by short-term HS) reduced cell membrane damages caused by HS, but FaNAC029 and FaNAM-B1 (up-regulated by long-term HS and senescence) increased the damages. Besides, alternative splicing was widely observed in HS and senescence responsive genes, including Hsps, Hsfs, and phytohormone signaling/synthesis genes. CONCLUSIONS: The short-term HS can stimulate gene responses and improve thermotolerance, but long-term HS is a damage and may accelerate leaf senescence. These results contribute to our understanding of the molecular mechanism underlying heat adaptation and heat-induced senescence.

Neural network establishes co-occurrence links between transformation products of the contaminant and the soil microbiome.

It remains challenging to establish reliable links between transformation products (TPs) of contaminants and corresponding microbes. This challenge arises due to the sophisticated experimental regime required for TP discovery and the compositional nature of 16S rRNA gene amplicon sequencing and mass spectrometry datasets, which can potentially confound statistical inference. In this study, we present a new strategy by combining the use of 2H-labeled Stable Isotope-Assisted Metabolomics (2H-SIAM) with a neural network-based algorithm (i.e., MMvec) to explore links between TPs of pyrene and the soil microbiome. The links established by this novel strategy were further validated using different approaches. Briefly, a metagenomic study provided indirect evidence for the established links, while the identification of pyrene degraders from soils, and a DNA-based stable isotope probing (DNA-SIP) study offered direct evidence. The comparison among different approaches, including Pearson's and Spearman's correlations, further confirmed the superior performance of our strategy. In conclusion, we summarize the unique features of the combined use of 2H-SIAM and MMvec. This study not only addresses the challenges in linking TPs to microbes but also introduces an innovative and effective approach for such investigations. Environmental Implication: Taxonomically diverse bacteria performing successive metabolic steps of the contaminant were firstly depicted in the environmental matrix.

Microcalorimetric investigation of the effect of non-ionic surfactant on biodegradation of pyrene by PAH-degrading bacteria Burkholderia cepacia.

Polycyclic aromatic hydrocarbons (PAHs) are widespread in various ecosystems and are pollutants of great concern due to their potential toxicity, mutagenecity and carcinogenicity. Surfactant has become a hot topic for its wide application in the bioremediation of PAHs. The aim of this work is to explore a microcalorimetric method to determine the toxic effect of pyrene on Bacillus subtilis (B. subtilis) and the PAH-degrading bacteria Burkholderia cepacia (B. cepacia) and to evaluate the effect of Tween 80 on biodegradation of pyrene. Power-time curves were studied and calorimetric parameters including the growth rate constant (k), half inhibitory concentration (IC50), and total thermal effect (Q(T)) were determined. B. subtilis, B. cepacia and B. cepacia with Tween 80 were completely inhibited when the concentration of pyrene were 200, 800 and 1600 µg mL⁻¹, respectively. B. cepacia shows better tolerance to pyrene than B. subtilis. Tween 80 significantly improves the biodegradation of pyrene by increasing the bioavailability of pyrene. In addition, the expression of catechol 2,3-dioxygenase (C23O) in B. cepacia is responsible for the degradation of pyrene and plays an important role in improving the biodegradation of pyrene. Moreover, the activity of C23O increases with the application of Tween 80. The enhanced bioavailability and biodegradation of pyrene by Tween 80 shows the potential use of Tween 80 in the PAHs bioremediation.

A comparative study on the impact of phthalate esters on soil microbial activity.

In this study, an isothermal microcalorimetric technique was used to demonstrate the impact of dimethyl phthalate (DMP), di-n-octyl phthalate (DOP) and diethyl phthalate (DEP) on soil microbial activity. The effect of these phthalate esters (≤100 µg g⁻¹ soil) follows the order: DEP > DMP > DOP but changed to DMP > DEP > DOP when the concentrations of the phthalate esters are above 100 µg g⁻¹ soil. DMP, DEP and DOP significantly decreased (p < 0.05) the soil urease activity. The effect of phthalate esters on soil microbial activity is not the same with those on enzymatic activity. In addition, DEP, which has a lower bioavailability, is less toxic to soil microbes than that of DMP when its concentration is above 100 µg g⁻¹ soil.

Bioaccumulation, internal distribution and toxicity of bisphenol S in the earthworm Eisenia fetida.

Due to the strict rules and restrictions on the utilization of bisphenol A (BPA) around the world, an emerging endocrine disrupting chemical, bisphenol S (BPS) has been widely utilized as a substitute and frequently detected in the environment, even in the human body. Although it has been widely studied in the aquatic systems, the fate and toxicological effect of BPS in soil invertebrates are poorly known. This study presented a comprehensive exploration into the attenuation, bioaccumulation, and physiological distribution of BPS in an ecologically significant soil invertebrate, as well as its subsequent ecotoxicological effect to earthworm for the first time. The E. fetida could promote the BPS attenuation in soil, with degradation rates of 92.8 ± 1.6 % and 98.6 ± 1.1 % at dosage of 1.0 mg/kg dry weight soil (DWS) and 0.1 mg/kg DWS, respectively. The bioaccumulation of BPS in the earthworm was up to 111.6 ± 6.0 mg/kg lipid and 12.9 ± 2.9 mg/kg lipid with the initial dosage of 1.0 mg/kg DWS and 0.1 mg/kg DWS, respectively. Furthermore, BPS could induce oxidative stress and the process of antioxidant defense in earthworm cells at relatively high dose (1.0 mg/kg DWS and 10.0 mg/kg DWS), suggesting potential risks of BPS to the soil environment. This study could contribute to a more in-depth understanding of the fate of BPS in soil-earthworm system, and indicate a necessity for better understanding the environmental fate and ecological risks of BPA substitutes in the future.

Bisphenol S degradation in soil and the dynamics of microbial community associated with degradation.

Bisphenol S (BPS) has been widely applied as a replacement for BPA in industrial application, leading to the frequent detection in the environment. However, its impact on soil microbial communities has not been well reported. Here, effects of BPS exposure on soil microbial communities in the presence of polystyrene (PS) microplastics were revealed. Rapid degradation of BPS occurred with a degradation rate of up to 98.9 ± 0.001 % at 32 d. The presence of BPS reduced the diversity of soil microbial communities, and changed community structures. After BPS treatment, Proteobacteria, and its members Methylobacillus, Rhodobacteraceae and Mesorhizobium became dominant, and were considered as potential biomarkers indicating BPS contamination. Co-occurrence network analysis revealed the increased relationships of certain groups of microbes after BPS treatment. The resultant low stability and resilience towards environment disturbance of microbial community networks implied the biotoxicity of BPS towards soil ecosystems. The degradation and biotoxicity of BPS (p > 0.05) in soil was not affected by the presence of PS. Our findings showed that exposure to BPS could reshape soil microbial communities and impair the robustness of microbial co-occurrence networks.

Phytotoxicity and accumulation of BPS to Pistia stratiotes under the influence of microplastics.

Bisphenol S (BPS) is a contaminant of emerging concern, its exposure and phytotoxicity towards plants, however, is scarce. This study aimed at revealing the BPS translocation in plants and phytotoxicity in the presence of Polystyrene (PS) microplastics. Results found that BPS and PS showed no effect on plant growth, indicating the tolerance of plants towards BPS and PS co-contamination. In addition, plants enriched BPS from soil, and a major part of absorbed BPS was accumulated in roots, as supported by the higher BCF value in roots compared with leaves. Besides, the low TF (<1) suggested the capacity of plants to accumulate BPS in roots, and less translocation to leaves. PS negatively affected the translocation of BPS in plants. PS with large size (5 µm) also increased the distribution of BPS in organelles. Exposure risk assessment suggested low concern of BPS carried in plants to human health. This study underlines the bioaccumulation of BPS in plants, and the effects of PS in the translocation process.

Occurrence and removal of bisphenol analogues in wastewater treatment plants and activated sludge bioreactor.

The occurrence and removal of ten bisphenol analogues (BPs) in municipal wastewater treatment plants (WWTPs) and laboratory scale activated sludge bioreactor (ASBR) were studied. All targeted BPs except for tetrachlorobisphenol A (TCBPA) were detected in the four WWTPs (W1, W2, W3 and W4) in the ng/L and ng/g dry weight range in wastewater and activated sludge, respectively, indicating that in addition to BPA, the BPA substitutes were widely used in our daily life and industrial production. Discrepant results regarding the removal efficiencies of BPs by different wastewater treatment processes were obtained. The removal rates were 55.6%, 24.4%, -10.1%, 71.4%, 38.9%, 58.0%, 39.1% and 6.4% in W1, 65.4%, 32.8%, 44.7, -13.5%, 20.1%, -29.6%, -25.1% and 99.4% in W2, 11.6%, 48.8%, 38.9%, 22.0%, 99.0%, -29.2%, -56.5% and 32.6% in W3, 33.9%, 30.5%, 17.4%, -47.6%, 62.9%, 83.0%, 4.4% and -4.3% in W4, for BPA, BPB, BPE, BPF, BPS, BPZ, BPAF and BPAP, respectively. The removal of ten targeted BPs in lab-scale continuous flow conventional ASBR and the key factors were investigated. The simulated laboratory-scale ASBR were highly effective in removing BPA, BPB, BPE, BPF, BPM and BPS with removal efficiencies of >94.3%, while BPZ, BPAP, BPAF and TCBPA were recalcitrant to elimination in the stimulated bioreactor with removal efficiencies of 71.3 ± 13.7%, 55.1 ± 21.2%, 47.4 ± 9.5% and 45.3 ± 16.6%, respectively. Protonation, hydrophobicity and molecular features of BPs were critical for their elimination in wastewater.

Lead-induced oxidative stress triggers root cell wall remodeling and increases lead absorption through esterification of cell wall polysaccharide.

Tall fescue (Festuca arundinacea Schreb) shows remarkable tolerance to lead (Pb), but the mechanisms involved in metal tolerance are not yet well understood. Here, tall fescue were firstly cultivated hydroponically with Pb2+ (0, 50, 200 and 1000 mg/L) for 14 days. The results showed that remodeling of root architecture plays important roles in tolerance of tall fescue to Pb2+ stress. Increased cell wall (CW) components contribute to restrict high amount of Pb2+ in roots. Additionally, the uronic acid contents of pectin, hemicellulose 1 (HC1) and hemicellulose 2 (HC2) increased under Pb2+ stress. We further observed that tall fescue cultivated with H2O2 showed similar remodeling of root architecture as Pb2+ treatment. Furthermore, pectin, HC1 and HC2 fractions were sequentially extracted from 0 and 10 mM H2O2 treated roots, and Pb2+ adsorption capacity and contents of carboxyl groups of pectin and HC2 fractions were steadily increased under H2O2 treatment in vitro. Our results suggest that degrees of esterification of pectin and HC2 are regulated by H2O2. High amount of low-esterified pectin and HC2 offer more carboxyl groups, provide more Pb2+ binding sites, and restrict more Pb2+ in the CW, which may enhance tolerance of tall fescue to Pb2+ stress.

[Advances in biodegradation of pharmaceuticals and personal care products].

Pharmaceuticals and personal care products (PPCPs) are a group of emerging environmental micropollutants, including prescription drugs and over-the-counter drugs (e.g., antibiotics, synthetic musk, painkiller, depressor, contraceptive drugs, soporific and weight-loss drug), and personal care products (e.g., cosmetics, synthetic perfume, sunscreen, hair spray, tint and fungicide). Extensive attention has been paid to PPCPs because of their potential negative effects on the environments and human health. Abundant researches have focused on the biodegradation of PPCPs. This review summarizes and discusses the biodegradation method, the diversity of PPCPs-degrading microorganisms, the degradation ability, metabolites and proposed pathways as well as the mechanisms of PPCPs' biodegradation. In addition, prospects for further research on biodegradation of PPCPs are also discussed.

Assessment of hexachlorcyclohexane biodegradation in contaminated soil by compound-specific stable isotope analysis.

Compound-specific isotope analysis (CSIA) was firstly applied to explore the biodegradation of hexachlorcyclohexane (HCH) isomers in contaminated soil. Concentrations and compound-specific carbon isotope ratio profiles of HCH in different specific ex-situ pilot-scale contaminated soil mesocosms were determined. The addition of nutrients and Sphingobium spp. significantly enhanced the degradation of HCH in contaminated soils within 90 days. Isomer specific biodegradation of HCHs was observed with α- and γ-HCH being more degradable than ß and δ-HCH. Stable carbon isotope fractionation of HCH was observed and the δ13C values shifted from -28.8 ±â€¯0.3‰ to -24.8 ±â€¯0.7‰ upon 87.3% removal, -27.9 ±â€¯0.2‰ to -25.9 ±â€¯0.5‰ upon 72.8% removal, -29.4 ±â€¯0.3‰ to -19.9 ±â€¯0.6‰ upon 95.8% removal, and -27.8 ±â€¯0.5‰ to -23.6 ±â€¯0.7‰ after 96.9% removal for α, ß, γ, and δ-HCH, respectively. Furthermore, the enrichment factor ε for α, ß, γ, and δ-HCH biodegradation in soil was obtained for the first time as -2.0‰, -1.5‰, -3.2‰, and -1.4‰, which could play a critical role in assessing in situ biodegradation of HCH isomers in field site soil. Results from ex-situ pilot-scale experiments clearly demonstrated that CSIA could be a promising tool to qualitatively and quantitatively evaluate in situ biodegradation of HCH in contaminated field site.

An innovative approach for sequential extraction of phosphorus in sediments: Ferrous iron P as an independent P fraction.

Accurate identification of phosphorus (P) forms is crucially important for understanding the geochemical cycle of P; however, until now the role of ferrous iron P (Fe(II)-P) buried in sediments has been completely ignored in nearly all sequential extraction procedures developed. Using sediment cores sampled from Donghu Lake in Wuhan, China, this study explored a modified version of widely used sequential P extraction method (SEDEX; Ruttenberg, 1992) in which Fe(II)-P was identified as an independent fraction. Based on the high selectivity of the extractant (0.2% 2,2'-bipyridine+0.1 M KCl) and the dissolution equilibrium of P, procedures for extracting Fe(II)-P were optimized using a 1:100 solid:liquid ratio and extraction at 50 ± 1 °C for 24 h. The sedimentary P extracted was divided into five fractions: loosely-bound P, Fe(II)-P, CDB-P, Ca-P and O-P. Fe(II)-P was the predominant fraction in fresh sediments in Donghu Lake, accounting for 15.7-49.9% of TP, with a mean of 31.6%. The mean values of Ca-P, O-P, CDB-P and loosely-bound P were 28.4%, 22.7%, 17.1% and 4.3%, respectively. Combined with component analysis of extracts and recovery experiments of standard reference minerals (vivianite, Fe3(PO4)2·8H2O) in natural sediments, extraction of Fe(II)-P with 0.2% 2,2-bipridine and 0.1 M KCl was robust, with a good recovery rate (88.7-100.6%) and little of the Ca-P dissolved. It is possible to use this innovative SEDEX not only to distinguish the contribution of different P matrices in fresh sediments, but also to investigate the transformation of sedimentary P under different redox conditions. Therefore, greater focus on Fe(II)-P is necessary, because it is a major sink for the geochemical process of sedimentary P.

Characterization of green synthesized nano-formulation (ZnO-A. vera) and their antibacterial activity against pathogens.

The application of nanotechnology in medicine has recently been a breakthrough in therapeutic drugs formulation. This paper presents the structural and optical characterization of a new green nano-formulation (ZnO-Aloe vera) with considerable antibacterial activity against pathogenic bacteria. Its particle structure, size and morphology were characterized by XRD, TEM and SEM. And optical absorption spectra and photoluminescence were measured synchronously. Their antibacterial activity against Escherichia coli and Staphylococcus aureus was also investigated using thermokinetic profiling and agar well diffusion method. The nano-formulation is spherical shape and hexagonal with a particle size ranging from 25 to 65 nm as well as an increased crystallite size of 49 nm. For antibacterial activity, the maximum inhibition zones of ZnO and ZnO+A. vera are 18.33 and 26.45 mm for E. coli, 22.11 and 28.12 mm for S. aureus (p<0.05). Considering Pmax, Qt and k, ZnO+A. vera nano-formulation has a significant (p < 0.05) antibacterial effect against S. aureus almost at all concentration and against E. coli at 15 and 25mg/L. ZnO+A. vera nano-formulation is much more toxic against S. aureus than E. coli, with an IC50 of 13.12 mg/L and 21.31 mg/L, respectively. The overall results reveal that the ZnO-A. vera nano-formulation has good surface energy, crystallinity, transmission, and enriched antibacterial activities. Their antibacterial properties are possibly relevant to particle size, microstructural ionization, the crystal formation and the Gram property of pathogens. This ZnO-A. vera nano-formulation could be utilized effectively as a spectral and significant antibacterial agent for pathogens in future medical and environmental concerns.

Short-term effect of aniline on soil microbial activity: a combined study by isothermal microcalorimetry, glucose analysis, and enzyme assay techniques.

The accidents of aniline spill and explosion happened almost every year in China, whereas the toxic effect of aniline on soil microbial activity remained largely unexplored. In this study, isothermal microcalorimetric technique, glucose analysis, and soil enzyme assay techniques were employed to investigate the toxic effect of aniline on microbial activity in Chinese soil for the first time. Soil samples were treated with aniline from 0 to 2.5 mg/g soil to tie in with the fact of aniline spill. Results from microcalorimetric analysis showed that the introduction of aniline had a significant adverse effect on soil microbial activity at the exposure concentrations ≥0.4 mg/g soil (p < 0.05) and ≥0.8 mg/g soil (p < 0.01), and the activity was totally inhibited when the concentration increased to 2.5 mg/g soil. The glucose analysis indicated that aniline significantly decreased the soil microbial respiratory activity at the concentrations ≥0.8 mg/g soil (p < 0.05) and ≥1.5 mg/g soil (p < 0.01). Soil enzyme activities for ß-glucosidase, urease, acid-phosphatase, and dehydrogenase revealed that aniline had a significant effect (p < 0.05) on the nutrient cycling of C, N, and P as well as the oxidative capacity of soil microorganisms, respectively. All of these results showed an intensively toxic effect of aniline on soil microbial activity. The proposed methods can provide toxicological information of aniline to soil microbes from the metabolic and biochemical point of views which are consistent with and correlated to each other.

Effects of petroleum contamination on soil microbial numbers, metabolic activity and urease activity.

The influence of petroleum contamination on soil microbial activities was investigated in 13 soil samples from sites around an injection water well (Iw-1, 2, 3, 4) (total petroleum hydrocarbons (TPH): 7.5-78 mg kg(-1)), an oil production well (Op-1, 2, 3, 4, 5) (TPH: 149-1110 mg kg(-1)), and an oil spill accident well (Os-1, 2, 3, 4) (TPH: 4500-34600 mg kg(-1)). The growth rate constant (µ) of glucose stimulated organisms, determined by microcalorimetry, was higher in Iw soil samples than in Op and Os samples. Total cultivable bacteria and fungi and urease activity also decreased with increasing concentration of TPH. Total heat produced demonstrated that TPH at concentrations less than about 1 g kg(-1) soil stimulated anaerobic respiration. A positive correlation between TPH and soil organic matter (OM) and stimulation of fungi-bacteria-urease at low TPH doses suggested that TPH is bound to soil OM and slowly metabolized in Iw soils during OM consumption. These methods can be used to evaluate the potential of polluted soils to carry out self-bioremediation by metabolizing TPH.

Short-time effect of heavy metals upon microbial community activity.

Microcalorimetry was applied to assess and compare the toxic effect of heavy metals, such as As, Cu, Cd, Cr, Co, Pb and Zn, on the soil microbial activities and community. About 1.0 g soil spiked 5.0mg glucose and 5.0mg ammonium sulfate, the microbial activities were recorded as power-time curves, and their indices, microbial growth rate constant k, total heat evolution Q(T), metabolic enthalpy Delta H(met) and mass specific heat rate J(Q/S), were calculated. Comparing these thermodynamic parameters associated with growth yield, a general order of toxicity to the soil was found to be Cr>Pb>As>Co>Zn>Cd>Cu. When soil was exposed to heavy metals, the amount of bacteria and fungi decreased with the incubation time, and the bacterial number diminished sharply. It illustrates that fungi are more tolerant, and bacteria-fungi ratio would be altered under metal stress. To determine the status of the glucose consumed, a glucose biosensor with eggshell membrane was used to measure the remaining glucose in soil sample. Results showed that the time at which glucose was consumed completely was agreed with the microcalorimetric time to a large extent, and depended on the toxicity of heavy metals as well.