The unexpected effect of the compound microbial agent NP-M2 on microbial community dynamics in a nonylphenol-contaminated soil: the self-stability of soil ecosystem.

Background: Nonylphenol (NP) is widely recognized as a crucial environmental endocrine-disrupting chemical and persistent toxic substance. The remediation of NP-contaminated sites primarily relies on biological degradation. Compound microbial products, as opposed to pure strains, possess a greater variety of metabolic pathways and can thrive in a wider range of environmental conditions. This characteristic is believed to facilitate the synergistic degradation of pollutants. Limited research has been conducted to thoroughly examine the potential compatibility of compound microbial agents with indigenous microflora, their ability to function effectively in practical environments, their capacity to enhance the dissipation of NP, and their potential to improve soil physicochemical and biological characteristics. Methods: In order to efficiently eliminate NP in contaminated soil in an eco-friendly manner, a simulation study was conducted to investigate the impact of bioaugmentation using the functional compound microbial agent NP-M2 at varying concentrations (50 and 200 mg/L) on the dynamics of the soil microbial community. The treatments were set as follows: sterilized soil with 50 mg/kg NP (CK50) or 200 mg/kg NP (CK200); non-sterilized soil with 50 mg/kg NP (TU50) or 200 mg/kg NP (TU200); non-sterilized soil with the compound microbial agent NP-M2 at 50 mg/kg NP (J50) or 200 mg/kg NP (J200). Full-length 16S rRNA analysis was performed using the PacBio Sequel II platform. Results: Both the indigenous microbes (TU50 and TU200 treatments) and the application of NP-M2 (J50 and J200 treatments) exhibited rapid NP removal, with removal rates ranging from 93% to 99%. The application of NP-M2 further accelerated the degradation rate of NP for a subtle lag period. Although the different treatments had minimal impacts on the soil bacterial α-diversity, they significantly altered the ß-diversity and composition of the bacterial community. The dominant phyla were Proteobacteria (35.54%-44.14%), Acidobacteria (13.55%-17.07%), Planctomycetes (10.78%-11.42%), Bacteroidetes (5.60%-10.74%), and Actinobacteria (6.44%-8.68%). The core species were Luteitalea_pratensis, Pyrinomonas_methylaliphatogenes, Fimbriiglobus_ruber, Longimicrobium_terrae, and Massilia_sp003590855. The bacterial community structure and taxon distribution in polluted soils were significantly influenced by the activities of soil catalase, sucrase, and polyphenol oxidase, which were identified as the major environmental factors. Notably, the concentration of NP and, to a lesser extent, the compound microbial agent NP-M2 were found to cause major shifts in the bacterial community. This study highlights the importance of conducting bioremediation experiments in conjunction with microbiome assessment to better understand the impact of bioaugmentation/biostimulation on the potential functions of complex microbial communities present in contaminated soils, which is essential for bioremediation success.

[Effect of Manure Application on the Adsorption of Antibiotics to Soil].

Sulfonamide antibiotics and florfenicol(FFC) are commonly used antibiotics in Zhejiang Province. They have weak adsorption on soil and are easy to migrate, with high environmental risks. In recent years, most of the studies on the potential risk of fecal-derived antibiotics to farmland soil were conducted by adding manure under laboratory conditions; therefore, it is impossible to assess the risk of antibiotic pollution under natural fertilization. Therefore, batch balance experiments were conducted to explore the effects of different soil types and manure types on the adsorption of antibiotics in the soil, in which five types of dryland farmland soils[Lin'an(LA), Jiashan(JS), Longyou(LY), Kaihua(KH), and Jinhua(JH)]in Zhejiang Province that have been used with different fertilizers(chicken manure, pig manure, and chemical fertilizer) for a long time were chosen, and four types of commonly used antibiotics[sulfadiazine(SD), sulfamethazine(SMT), sulfamethoxazole(SMZ), and FFC]were selected. The results showed that the adsorption of the four antibiotics in the experimental soil was weak, and the adsorption capacity decreased in the order of:SMT(1.44-13.23 mg1-(1/n)·L1/n·kg-1)>SMZ(0.73-6.05 mg1-(1/n)·L1/n·kg-1)>SD(0.16-5.57 mg1-(1/n)·L1/n·kg-1)>FFC(0.27-3.81 mg1-(1/n)·L1/n·kg-1). The Freundlich model was superior to the linear model in fitting the isotherm adsorption of SD, SMT, and FFC, in which SD and FFC belonged to "S" type adsorption, and SMT belonged to "L" type adsorption. For SMZ, the fitting effect of the linear model was better than that of the Freundlich model. The contents of total organic carbon(TOC) and dissolved organic carbon(DOC) could better predict the adsorption capacity of the four antibiotics(r=0.548-0.808), and the values of cation exchange capacity(CEC) and electrical conductivity(EC) could better predict the adsorption capacity of SMT and FFC(r=0.758-0.841). Compared with the application of chemical fertilizer, manure application increased the values of TOC, DOC, CEC, and EC in acidic and neutral soils, which was conducive to the adsorption of antibiotics on the soil. Meanwhile, manure application also increased pH in acidic and neutral soils, which was not conducive to the adsorption of antibiotics on the soil. In addition, manure application reduced the values of TOC, DOC, CEC, EC, and pH in alkaline soils. The lower pH was conducive to antibiotic adsorption on the soil, whereas the lower content of the other four was not conducive to antibiotic adsorption on the soil. For the acidic soil with low fertility, the application of manure increased soil fertility and thus increased the adsorption of antibiotics on the soil, such as the LA soil with chicken manure, the LY(1) soil with pig manure, and the JH soil with chicken manure and pig manure. However, for the acidic and neutral soils with high fertility, the application of manure had significantly increased soil pH and thus reduced the adsorption of antibiotics on the soil, such as the JS soil with chicken manure and pig manure and the LY(2) soil with chicken manure. For calcareous soil with high fertility and pH(such as KH soil), the adsorption profiles of the four types of antibiotics on the soil showed diversity after the application of manure:the adsorption capacity of SD increased significantly after the application of chicken manure and pig manure, whereas the adsorption capacity of SMT and SMZ decreased significantly, and the adsorption capacity of FFC declined significantly after the application of chicken manure. Therefore, manure application according to soil fertility could effectively control the environmental risk of fecal antibiotics.

Aging behavior of microplastics accelerated by mechanical fragmentation: alteration of intrinsic and extrinsic properties.

Microplastics (MPs) inevitably undergo multiple aging processes during their life cycle in the environment. However, the information regarding the mechanical fragmentation behavior of MPs remained unclear, including the changes in the intrinsic properties of aged MPs, the measurement of aging degree, the underlying mechanism, and the interaction with heavy metals. Here, MPs (PS, PP, PET) were aged by crushing (-CR) and ball-milling (-BM) to simulate mild and severe mechanical fragmentation, respectively. Our results indicated that mechanical fragmentation significantly affected the morphology of MPs. The aging degree of MP-BM was deeper compared to MP-CR owing to smaller particle size, larger specific surface area, poorer heat resistance, better hydrophilicity, and richer oxygen-containing functional groups. The carbonyl index (CI) and O/C ratio were used to measure the aging degree of the two mechanical aging treatments. Besides, the mechanism was proposed and the discrepancy between the two treatments was elaborated from three aspects including the excitation energy source, reaction interface, and reaction dynamics. Furthermore, the extrinsic properties of MPs altered with the increase of aging degree; specifically, the adsorption capacities of heavy metals were enhanced. Meanwhile, it was unveiled that the CI value and O/C ratio played a vital role in estimating the adsorption ability of heavy metals. The findings not only reveal the mechanical fragmentation behavior of MPs but also provide new insights into the assessment of the potential risks of the aged MPs via chemical indexes.

Fate of 14C-acetyl sulfamethoxazole during the activated sludge process.

Compared to antibiotic parent molecule, human metabolites are generally more polar and sometimes not less toxic in wastewater. However, most researches focus on the fate of parent molecule. Therefore, behaviors of human metabolites are little known. Moreover, though much has been done on the fate of antibiotics during activated sludge process, there are still some limitations and gaps. In the present study, [Ring-14C] acetyl sulfamethoxazole (14C-Ac-SMX) was used to investigate the fate of human metabolite of SMX during activated sludge process at environmentally relevant concentration. At the end of 216 h, 3.1% of the spiked activity in the initial aqueous phase was mineralized, 50% was adsorbed onto the solid phase, and 36.5% still remained in the aqueous phase, indicating that adsorption, not biodegradation, was the main dissipation pathway. In the existence of microbial activities, accumulation into the solid phase was much higher, which was less bioavailable by chemical sequential extraction. The multimedia kinetic model simultaneously depicted the fate of Ac-SMX in the gas, aqueous, and solid phases, and demonstrated that microbially accelerated accumulation onto the solid phase was attributed to lower desorption rate from the solid phase to the aqueous phase, where adsorption rate was not the key factor. Therefore, Ac-SMX cannot be efficiently mineralized and remain in the aqueous or the solid phases. The accumulation in the solid phase is less bioavailable and is hard to be desorbed in the existence of microbial activities, and should not be easily degraded, and may lead to the development of antibiotic-resistant bacteria and genes after discharge into the environment.

Impact of sludge treatments on the extractability and fate of acetyl sulfamethoxazole residues in amended soils.

Sludge recycled in agriculture may bring antibiotics into cropped soils. The nature, total amount, and availability of the antibiotics in soil partly depend on the sludge treatments. Our paper compares the fate of N-acetyl sulfamethoxazole (AC-SMX) residues between soils incubated with the same sludge but submitted to different processes before being added in soil. The fate of 14C-AC-SMX residues was studied in mixtures of soil and sludges at different treatment levels: 1) activated and 2) centrifuged sludges, both enriched with 14C-AC-SMX, and 3) limed and 4) heat-dried sludges obtained by treating the previously contaminated centrifuged sludge. The evolution of the extractability of 14C residues (CaCl2, methanol) and their mineralization were followed during 119 days. More than 80% of the initial 14C-activity was no longer extractable after 14 days, except in soil with limed sludge. Liming and drying the centrifuged sludge decreased the mineralized 14C fraction from 5.7-6.4% to 1.2-1.8% and consequently, the corresponding soils contained more 14C residues after 119 days. Although 14C residues were more CaCl2-extractable in soil with limed sludge, they seemed to be poorly bioavailable for biodegradation. For all solid sludges, the mineralization rate of 14C-AC-SMX residues was strongly correlated to that of sludge organic carbon, with a coefficient three times lower for the limed and dried sludges than for the centrifuged sludge after 14 days.

Impact of liming and drying municipal sewage sludge on the amount and availability of (14)C-acetyl sulfamethoxazole and (14)C-acetaminophen residues.

Acetyl Sulfamethoxazole (AC-SMX) and acetaminophen (ACM) can be found in municipal sewage sludge, and their content and availability may be influenced by sludge treatments, such as drying and liming. A sludge similarly centrifuged with/without a flocculant was spiked with (14)C-labelled AC-SMX or ACM. Then, it was either limed (20% CaO) or/and dried under different laboratory conditions (1 week at ambient temperature; and 48 h at 40 or 80 °C). The total amount and distribution of the (14)C-compounds among several chemical fractions, based on the sludge floc definition, were assessed at the end of the treatments. All the (14)C-activity brought initially was recovered in the limed and/or dried sludges for AC-SMX but only between 44.4 and 84.9% for ACM, with the highest rate obtained for the limed sludge. Drying at 80 °C or liming increased the percentage of the sludge total organic carbon recovered in the extracts containing soluble extracellular polymeric substances (S-EPS) and the percentage of the total (14)C-activity extracted simultaneously. The non-extractable residues represented only 3.9-11.6% of the total (14)C-activity measured in the treated sludges for AC-SMX and 16.9-21.8% for ACM. The presence of AC-SMX and ACM residues in the treated sludges, after liming and drying under different conditions, was shown using some (14)C-labelled molecules. At this time scale and according to the extraction method selected, most of the (14)C-residues remained soluble and easily extractable for both compounds. This result implies that certain precautions should be taken when storing sludges before being spread on the field. Sludge piles, particularly the limed sludge, should be protected from rain to limit the production of lixiviates, which may contain residues of AC-SMX and ACM.

Modeling the release of organic contaminants during compost decomposition in soil.

Composts, incorporated in soils as amendments, may release organic contaminants during their decomposition. COP-Soil is presented here as a new model to simulate the interaction between organic contaminants and compost, using one module for organic matter and one for organic pollutants, with these modules being linked by several assumptions. Published results of laboratory soil incubations using labeled carbon pollutants from compost were used to test the model for one polycyclic aromatic hydrocarbon (PAH), two surfactants and one herbicide. Several simulation scenarios were tested using (i) the organic pollutant module either alone or coupled to the organic matter module, (ii) various methods to estimate the adsorption coefficients (Kd) of contaminants on organic matter and (iii) different degrading biomasses. The simulations were improved if the organic pollutant module was coupled with the organic matter module. Multiple linear regression model for Kd as a function of organic matter quality yielded the most accurate simulation results. The inclusion of specific biomass in the model made it possible to successfully predict the PAH mineralization.

Characterization of a novel melamine-degrading bacterium isolated from a melamine-manufacturing factory in China.

Melamine (2,4,6-triamino-1,3,5-triazine, C3H6N6), belonging to the s-triazine family, is an anthropogenic and versatile raw material for a large number of consumer products and its extensive use has resulted in the contamination of melamine in the environment. A novel melamine-degrading bacterium strain CY1 was isolated from a melamine-manufacturing factory in China. The strain is phylogenetically different from the known melamine-degrading bacteria. Approximately, 94 % melamine (initial melamine concentration 4.0 mM, initial cell OD 0.05) was degraded in 10 days without the addition of additional carbon source. High-performance liquid chromatography showed the production of degradation intermediates including ammeline, ammelide, cyanuric acid, biuret, and urea. Kinetic simulation analysis indicated that transformation of urea into ammonia was the rate-limiting step for the degradation process. The melamine-cyanurate complex was formed due to self-assembly of melamine and cyanuric acid during the degradation. The tracking experiment using CY1 cells and (13)C3-melamine showed that the CY1 could mineralize s-triazine ring carbon to CO2. The strain CY1 could also catalyze partial transformation of cyromazine, a cyclopropyl derivative of melamine, to 6-(cyclopropylamino)-[1,3,5]triazine-2,4-diol.

[Application of solidification technology in ecological protection of rural riverbank].

A self-developed binder was used for the solidification of construction refuse piles and whole soil matrix, and a technology of this solidification combining with grass-planting was adopted to ecologically protect the rural riverbanks at Tianshan Village of Shanghai. This technology and other ecological engineering techniques were also employed to reconstruct the ecological environment of a sewage pond at the Village. The results showed that the solidified piles had an anti-compression strength of up to 7.3 MPa, with good hydraulic permeability, fast hardening rate, and low drying shrinkage, which met the requirements for ecological safety. The solidified stakes could be used at a low temperature of above -18 degrees C with addition of certain anti-freezing agents. The riverbank underpinned with the solidified stakes had higher anti-compressive strength, higher ability of anti-soil erosion, and better hydraulic permeability; and its soil had the similar moisture content to bare riverbank soil, with no detrimental effects on the root growth of planted grass. After soil solidification, the shearing strength of the riverbank increased by 50 times, and its soil loss was only 5% of the bare riverbank. In the first 10 days after adopting this technology, parts of Cynodon dactylon roots on the surface of solidified soil matrix began to extend into soil; after one month, 60% of the roots penetrated into deeper soil layer; and 11 months later, the grass roots completely grew in-depth in the soil. The combination of our solidification technique with vegetation reconstruction satisfied the requirements of both stabilizing riverbank and improving riparian habitat.

Phosphate (Pi) and arsenate uptake by two wheat (Triticum aestivum) cultivars and their doubled haploid lines.

BACKGROUND AND AIMS: Arsenic accumulation in cereal crops represents an important pathway for human exposure to arsenic from the environment. The objectives of the present work were to find whether the relationship between arsenate and phosphate (Pi) uptake rate differs among genotypes and to select genotypes with a low arsenate uptake rate with the aim of improving food safety and human health. METHODS: A hydroponic experiment was conducted using two wheat (Triticum aestivum) cultivars (Hanxuan 10 and Lumai 14) and ten doubled haploid (DH) lines derived from them to investigate Pi and arsenate uptake over 48 h. Ten plants were transferred to bottles containing 50 mL of pre-treatment solution containing 0.5 mM CaCl2 and 5 mM MES set at pH 6.0 with 330 microM Pi as KH2PO4 and 7.33 microM arsenate. The solutions were aerated continuously. At 8, 24 and 48 h after uptake, 1 mL of test solution was sampled for determination of Pi and arsenate concentrations. KEY RESULTS AND CONCLUSIONS: For each wheat line, Pi and arsenate concentrations in the test solution decreased with uptake time. Exponential (for Pi) or polynomial (for arsenate) regression plots fitted the data closely. For all genotypes, net Pi uptake rates decreased with time (from 0 to 48 h). However, net arsenate uptake rates decreased with time for D5, changed little with time for the male parent, D4 and D6, and increased with time for the others. An inflexion of about 25 microm Pi was observed for the relationship between arsenate and Pi concentrations in the test solution, indicating that 25 microm could be the point where the high-affinity uptake system 'switches on', or dominates over low-affinity uptake. In addition, the male parent, D1, D6 and D10 were considered ideal genotypes because they possess Pi transporters that discriminate strongly against arsenate and are expected to accumulate less arsenate in the field.

Arsenate (As) uptake by and distribution in two cultivars of winter wheat (Triticum aestivum L.).

Two cultivars of winter wheat (Triticum aestivum L.) (Jing 411 and Lovrin 10) were used to investigate arsenate (As) uptake and distribution in plants grown in hydroponic culture and in the soil. Results showed that without As addition, Lovrin 10 had higher biomass than Jing 411 in the soil pot experiment; in the hydroponic experiment Lovrin 10 had similar root biomass to and lower shoot biomass than Jing 411. Increasing P supply from 32 to 161 microM resulted in lower tissue As concentrations, and increasing As supply from 0 to 2,000 microM resulted in lower tissue P concentrations. Increasing P supply tended to increase shoot-to-root ratios of As concentrations, and increasing As supply tended to decrease shoot-to-root ratios of As concentrations. Both cultivars invested more in root production under P deficient conditions than under P sufficient conditions. Lovrin 10 invested more biomass production to roots than Jing 411, which might be partly responsible for higher shoot P and As concentrations and higher shoot-to-root ratios of As concentrations. Moreover, Lovrin 10 allocated less As to roots than Jing 411 and the difference disappeared with decreasing P supply.

Arsenate and phosphate interaction in Saccharomyces cerevisiae.

In the present study, arsenate (As(V)) and phosphate (P(V)) interactions were investigated in growth, uptake and RNA content in yeast (Saccharomyces cerevisiae). Yeast grew slowly with As(V) concentrations increasing in the medium. However, the maximal population density was almost the same among different As(V) treatments. It was in the late log phase that yeast growth was augmented by low As(V), which was maybe due to the fact that methionine metabolism was stressed by vitamin B6 deprivation, so As(V) treatments did not affect maximal population density. However, with P (V) concentrations increasing, the maximal population density increased. Therefore, the maximal population density was determined by P (V) concentrations in the medium but not by As (V) concentrations in the medium. Ycfl p(a tonoplast transpor) transports As(GS)3 into the vacuole, but arsenic(As) remaining in the thalli was 1.27% with As(V) exposure for 60 h, from which it can be speculated that the percentage of As transported into vacuole should be lower than 1.27%. However, the percentage of As pumped out of cell was 71.49% with As (V) exposure for 68 h. Although two pathways (extrusion and sequestration) were involved in As detoxification in yeast, the extrusion pathway played a major role in As detoxification. RNA content was the highest in the early-log phase and was reduced by As(V).

[Effects of different arbuscular mycorrhizal fungi on Tagetes erecta growth and diesel degradation].

The effects of G. mosseae, G. geospora, G. constrictum and bacteria on diesel tolerance of Tagetes erecta were investigated under greenhouse conditions. The results showed that AM fungi could still develop mycorrhizal assosiations with mum when the diesel concentration was 5,000 mg.kg-1. White mum was better than yellow mum in diesel tolerance, with 63.1% total biomass increased. The colonization rate of inoculating AM fungi treatment was 3.5%-29.9% higher than the control. G. mosseae and G. geospora were better strains, their biomass increasing 9.0% and 42.7% than the control, respectively, while the effect of inoculating mixed AM fungi was not obvious. Bacteria inhibited the colonization of arbuscular mycorrhizal fungi on white mum, but promoted the vegetative and reproductive growth of mycorrhizal mum. Among 5 inoculation treatments, treatments of inoculating G. geospora and inoculating mixed AM fungi and bacteria were better, with 16.51% and 14.05% more diesel degradation rate than that of the control, respectively.