Combined effects of oxytetracycline and microplastic on wheat seedling growth and associated rhizosphere bacterial communities and soil metabolite profiles.

The widespread application of antibiotics and plastic films in agriculture leads to new characteristics of soil pollution with the coexistence of antibiotics and microplastics. However, their combined effects on wheat seedling growth and associated rhizosphere bacterial communities and soil metabolite profiles remain unclear. Here, in the potted experiment, wheat was treated with individual oxytetracycline (0, 5.0, 50.0, and 150.0 mg kg-1) and the combination of oxytetracycline and polyethylene microplastic (0.2%). Results showed that 150 mg kg-1 oxytetracycline combined with microplastic significantly reduced the biomass and height of the plant. Compared with CK, all the treatments exposed to the combination of oxytetracycline and polyethylene microplastic significantly promoted carotenoid content and peroxidase activity in wheat leaves. Soil dehydrogenase and urease activities were more sensitive to current pollutant exposure than sucrase activity. Oxytetracycline (150 mg kg-1) alone and in combination with polyethylene significantly decreased the abundances of certain genera belonging to plant growth-promoting rhizobacteria (PGPR) in soil, such as Arthrobacter, Gemmatimonas, Massilia, and Sphingomonas. Combined exposure of 150 mg kg-1 oxytetracycline and polyethylene microplastic significantly altered multiple metabolites including organic acids and sugars. Network analysis indicated that co-exposure of 150 mg kg-1 oxytetracycline and microplastic may affect the colonization and succession of PGPR by regulating soil metabolites, thereby indirectly inhibiting wheat seedling growth. The results help to elucidate the potential mechanisms of phytotoxicity of the combination of oxytetracycline and polyethylene microplastic.

Stress response to oxytetracycline and microplastic-polyethylene in wheat (Triticum aestivum L.) during seed germination and seedling growth stages.

Much efforts have been devoted to clarify the phytotoxicity of individual contaminants in plants, such as individual antibiotic and microplastic; however, little is known about the phytotoxicity of their combined exposure. Here, we investigated the effects of individual and combined exposure of wheat (Triticum aestivum L.) (Xiaoyan 22) to oxytetracycline (OTC) and polyethylene (PE) microplastics using physiological and metabolic profilings. During the seed germination stage, OTC induced phytotoxicity, as observed through the changes of root elongation, sprout length, fresh weight and the vitality index, with significant effect at the 50 and 150 mg·L-1 levels; the effect of PE microplastics depended on the OTC level in the combined exposure groups. During seedling cultivation, catalase (CAT) and ascorbate peroxidase (APX), as antioxidant enzyme indices, were sensitive to OTC exposure stress, although OTC was not determined in leaves. Untargeted metabolomics of wheat leaves revealed OTC concentration-, metabolite class- and PE-dependent metabolic responses. Dominant metabolites included carboxylic acids, alcohols, and amines in the control group and all treatment groups. Compared to only OTC treatment, PE reprogrammed carboxylic acid and alcohol profiles in combined exposure groups with obvious separation in PLS-DA. Combined exposure induced fewer metabolites than OTC exposure alone at the 5 and 50 mg·L-1 levels. The shared metabolite numbers were higher in the OTC groups than in the PE-OTC groups. Pathway enrichment analysis showed a drift in metabolic pathways between individual and combined exposure to OTC and PE, which included glyoxylate and dicarboxylate metabolism, amino acid metabolism and isoquinoline alkaloid biosynthesis. Among metabolites, aromatic acids and amino acids were more sensitive to combined exposure than individual exposure. These results contribute to clarifying the underlying mechanisms of phytotoxicity of individual and combined exposure to OTC and PE.

Linkage of antibiotic resistance genes, associated bacteria communities and metabolites in the wheat rhizosphere from chlorpyrifos-contaminated soil.

Rhizosphere is a crucial site for the proliferation of antibiotic resistance genes (ARGs) in agricultural soil. Pesticide contamination is ubiquitous in soil, such as chlorpyrifos as one of the most commonly used pesticides. However, limited knowledge is reported about ARGs profiles changes and the driving mechanism of ARGs prevalence in rhizosphere soil after adding pesticide. In this study, irrespective of chlorpyrifos presence, the abundances of ARGs (tetM, tetO, tetQ, tetW, tetX, sul1 and sul2) and intI1 in rhizosphere soil of wheat were obviously higher than those in bulk soil. 20.0 mg·kg-1 chlorpyrifos significantly increased the abundance of total ARGs and intI1 in bulk soil, respectively, at day 50 and 100, but not in rhizosphere soil. Rhizosphere influence on ARGs was far greater than chlorpyrifos. ARGs and intI1 abundances were higher at day 50 than ones at day 100. C/N ratio and NO3--N content, which were affected by rhizosphere and cultivation time, significantly explained the increased ARGs. Compared to bulk soil, rhizosphere shifted host bacteria of tetracycline resistance genes (TRGs), intI1 at genus level, and host bacteria of sul1, sul2 at phylum level. Rhizosphere simplified the linkage of ARGs, host bacteria and metabolites. Bacterial communities played important roles in the variation of ARGs and intI1, and the difference in the distribution of potential hosts between bulk and rhizosphere soil was related to metabolites abundance and composition. These results provide valuable information for understanding the linkage of ARGs, associated bacteria communities and metabolites in the wheat rhizosphere soil.

Environmentally Relevant-Level CeO2 NP with Ferrous Amendment Alters Soil Bacterial Community Compositions and Metabolite Profiles in Rice-Planted Soils.

The environmental risks and benefits associated with the introduction of CeO2 nanoparticle (NP) in agricultural soil must be carefully assessed. The ferrous ion is rich in rhizosphere soil of rice due to the reduction states underground. The aim of this study was to investigate the effects of environmentally relevant-level CeO2 NP (25 mg·kg-1) in the absence or presence of ferrous (30 mg·kg-1) amendment on soil bacterial communities and soil metabolomics in rice-planted soil over 150 days. Results showed that CeO2 NP exposure changed soil bacterial community compositions and soil metabolomics, and the above changes were further shifted with the ferrous amendment. Several functionally significant bacterial phyla containing Proteobacteria and Bacteroidetes abundances, which were associated with carbon and nitrogen cycling, were promoted after CeO2 NP exposure with ferrous amendment. However, CeO2 NP inhibited plant-growth-promoting rhizobacteria containing genera Bacillus and Arthrobacter irrespective of the presence or absence of ferrous. Among rhizosphere soil enzyme activities, cellulose activity was the most sensitive for CeO2 NP exposure. NP decreased Firmicutes and increased Chloroflexi, Rokubacteria, and Thaumarchaeota abundances at the phylum level, which contributed to reduce soil cellulose activity. Additionally, CeO2 NP positively or negatively affected soil pH, Ce accumulation in root, and rice physiological properties (root-POD, stem-POD). As a result, the above factors were related to the changes of Chloroflexi, Gemmatimonadetes, Rokubacteria, Thaumarchaeota, and Nitrospirae at the phylum level. After adding CeO2 NP with ferrous or not, the main metabolic changes were concentrated on fluctuations in starch and sucrose metabolism, nitrogen metabolism, sulfur metabolism, propanoate metabolism, fatty acid metabolism, and urea cycle. The eight changed metabolites containing glycerol monstearate, boric acid, monopalmitin, palmitic acid, alkane, ethanol, dicarboximide, and stearic acid accounted for the separation of different treatments with CeO2 NP exposure. Activities of soil enzymes (urease, invertase, and cellulose), pH, and soil organic matter affected dominant metabolites containing fatty acids, inorganic acid, and sugar. Network analysis showed that the influence of soil bacterial community on metabolites varied with metabolites and bacteria species. The presence of CeO2 NP mainly promoted fatty acids (hexanoic acid, nonanoic acid) and amino acid (oxoproline) and amine (diethanolamine) concentrations, which could be from the increased Proteobacteria abundance after CeO2 NP exposure. Phylum Proteobacteria had the most genus species containing 13 genera affecting soil metabolite profiles. These results provide valuable information for understanding the impact of environmentally relevant-level CeO2 NP exposure on soil microbial communities and metabolites with or without ferrous, which is needed to understand the ecological risk posed by long-term CeO2 NP exposure in rice-planted soil with rich ferrous.

Citric acid enhances Ce uptake and accumulation in rice seedlings exposed to CeO2 nanoparticles and iron plaque attenuates the enhancement.

To assess the role of citric acid, as a typical low-molecular-weight organic acid from root exudates, on cerium (Ce) uptake, accumulation and translocation in rice seedlings (Oryza sativa L.) exposed to two CeO2 nanoparticles (NPs) (14 nm and 25 nm). A hydroponic experiment was performed under two citric acid levels (0.01 and 0.04 mmol L-1) combined with iron plaque presence. Citric acid significantly enhanced surface-Ce, root-Ce and shoot-Ce accumulation, irrespective of NPs size and iron plaque presence. The increased surface-Ce was associated with the promoted interactive attraction between NPs and root surface, and the enhanced NPs dissolution. Surface-Ce (containing crystalline and amorphous fractions of iron plaque) accumulation increased with the increase of citric acid concentrations. However, the enhancement influence of 0.01 mmol L-1 citric acid on root-Ce, shoot-Ce accumulations, rice-Ce distribution and TFroot-shoot was more remarkable than citric acid (0.04 mmol L-1), which suggested higher food security risk for human health with environment-level citric acid. Iron plaque presence attenuated the enhancement effect of citric acid on rice-Ce accumulation and distribution (containing surface-Ce, root-Ce and shoot-Ce) due to the reduced attractive interaction between NPs and root surface from the effect of Fe2+ being dissolved by iron plaque. Above effect of citric acid and iron plaque was more remarkable in 25 nm NP than 14 nm NP.

Citric acid and glycine reduce the uptake and accumulation of Fe2O3 nanoparticles and oxytetracycline in rice seedlings upon individual and combined exposure.

Uptake of nanoparticles and antibiotics by plants is root exudates-dependent, however, the underlying influence processes and mechanisms from different root exudates are rarely investigated. A hydroponic experiment was conducted to investigate the accumulation of Fe2O3 nanoparticle (NP) and oxytetracycline (OTC) in rice seedlings, in the absence or presence of citric acid or glycine, acting as components of root exudates. Irrespective of individual or combined exposure of Fe2O3 NP and OTC, citric acid and glycine both reduced surface-Fe, surface-OTC, root-OTC, shoot-OTC accumulations with dose-effect relationship. Two exudates increased |ζ| values of NP, which weakened the interactive attraction between NP and root surface and then decreased surface-Fe accumulation. Citric acid and glycine binding with OTC in solution decreased surface-OTC accumulation, and further decreased root-OTC and shoot-OTC accumulations. Combined exposure of two pollutants alleviated the reduction effect of citric acid and glycine on surface-Fe/surface-OTC/root-OTC accumulations due to their high accumulations in combined exposure compared to individual exposure. Although citric acid and glycine promoted TFroot-shoot and TFsurface-root of two pollutants, respectively, they always decreased total rice-Fe and rice-OTC accumulations. Therefore, the presence of root exudates decreased the bioaccumulation of Fe2O3 NP and OTC in rice upon their individual and combined exposure through changing their environmental behaviors in rhizosphere.

Iron plaque reduces cerium uptake and translocation in rice seedlings (Oryza sativa L.) exposed to CeO2 nanoparticles with different sizes.

This study aims to assess the role of iron plaque (IP) on cerium (Ce) uptake and translocation by rice after CeO2 nanoparticles (NPs) exposure over a 4 days period. A hydroponic experiment was performed under two IP levels (low and high) combined with two CeO2 NPs size (14 nm and 25 nm). It was found that CeO2 NPs as the main form was absorbed by rice due to limited NPs dissolution in hydroponic solution. IP significantly reduced surface-Ce, root-Ce and shoot-Ce accumulation, irrespective of CeO2 NPs sizes. The reduced uptake of Ce was more obvious in NP25 than NP14. Ce accumulations decreased with increasing IP amounts. In IP treatments, the interactive attraction between NPs and root surface was weakened through the enhancement of hydrodynamic diameters and the reduction of ζ-potential of CeO2 NPs in solution, as well as the reduction of |ζ| values of rice root, which reduced the Ce bioaccumulation in rice. PCA indicated the negative correlation between surface-Ce (IP-C-Ce and IP-A-Ce) and NPs size, and between shoot-Ce/root-Ce and IP-Fe/tissue-Fe. IP also decreased Ce translocation from root to shoot. A full life study indicated the reduction effect of IP on surface-Ce, root-Ce, shoot-Ce and grain-Ce accumulations. These findings are significant as they imply that the IP formation is a promising approach for preventing Ce accumulation in rice, which would regulate Ce uptake by rice in the following growth stages and decrease the health risk of CeO2 NPs exposure in agricultural environment.

Influence of the root plaque formation with different species on oxytetracycline accumulation in rice (Oryza sativa L.) and its elimination in culture solution.

Hydroponic experiments were conducted to investigate the role of different root plaque formation on oxytetracycline (OTC) uptake/translocation by rice seedlings (Oryza sativa L.) and solution-OTC elimination at two initial OTC concentrations (10 and 30 mg L-1). The results indicated OTC accumulation in rice was always in the order root surface > shoot > inside root whether plaques were formed or not. It demonstrated that Fe-Mn-Mt (montmorillonite) treatment was easier to promote significantly (p < 0.05) OTC accumulation in the underground part (root surface and inside root) and decrease significantly (p < 0.05) OTC translocation from the root to the shoot in rice compared to no plaque treatments (CK), especially for OTC 30 mg L-1 level with the lowest shoot-OTC accumulation in Fe-Mn-Mt treatment. Plaque treatments increased half-life of solution-OTC elimination in the order Fe-Mn-Mt > Fe-Mn > Fe > CK, which was caused mainly by OTC degradation from Fe2+-binding influence in solution, not by the enhancement of OTC accumulation on the root surface and inside root. And solution-OTC elimination increased with decreasing initial OTC concentrations, the drop of Fe2+ and the increment of Fe3+ and pH during the experiment. These findings are useful for reducing OTC accumulation and translocation in rice aboveground parts and eliminating OTC contamination in agricultural environment simultaneously through complicated plaque formation under higher OTC concentration exposure (30 mg L-1) in the future design.

[Fraction distribution and risk assessment of heavy metals in stream sediments from a typical nonferrous metals mining city].

A modified Tessier's sequential extraction procedure was used to investigate the fraction of seven types of heavy metals (Cd, Cr, Cu, Zn, Ni, Pb, As) in the surface sediments from Huixi Stream in Tongling City, a typical nonferrous metals mining city, China. Based on speciation distribution analysis of these metals, contamination degree and ecological risk assessment of heavy metals were conducted by means of risk assessment code (RAC) and mean sediment quality guideline quotient (SQG-Q). The results show that: (1) Cr and As are major composed with residual fractions, Zn, Ni and Pb are mainly constituted of residual and bound to iron and manganese oxides fractions, and Cu is dominated by bounding to organic matter, while Cd exists in approximate mass fractions of exchangeable, bound to carbonates, bound to iron and manganese oxides, and residue. (2) Carbonate and exchangeable mass fractions of Cd, Cr, Cu, Zn, Ni, Pb and As reach 46.48%, 4.62%, 4.05%, 4.12%, 9.17%, 0.97% and 0.03%, respectively. According to the RAC, Cd is of high risk to the environment, Cr, Cu, Zn and Ni are of low risk to the environment, while Pb and As pose extreme low risk to the environment. (3) The SQG index, calculated with SQG-Q, is 10.42, which is far higher than the threshold value 1.0, indicating that the sediment in Huixi Stream has a very high potential for biological toxicity effect. The PEL-Q indexes corresponding to Cd, Cr, Cu, Zn, Ni, Pb and As approach 4.23, 1.14, 20.75, 6.04, 2.33, 4.58 and 41.71, respectively, suggesting that all these metals have great potentials for biological toxicity and the adverse effects will frequently occur.

[Contamination and health risk for heavy metals via consumption of vegetables grown in fragmentary vegetable plots from a typical nonferrous metals mine city].

A systematic survey of As, Ni, Cu, Pb, Cd and Zn concentrations in eight kinds of vegetables (involving 226 samples) and their corresponding soils at 35 sampling sites in the fragmentary vegetable plots of a typical nonferrous metals mine city, Tongling, was carried out for assessing heavy metal pollution, bio-accumulation ability and potential health risk to local inhabitants due to exposure via consumption of vegetables. The results showed that: (1) The soils of the studied vegetable plots were seriously contaminated by heavy metals and the mean concentrations of As, Ni, Cu, Pb, Cd and Zn reached 96.96, 56.64, 1 247.82, 313.59, 6.743 and 600.96 mg x kg(-1), respectively, all significantly exceeding the soil background value of Tongling city; (2) The mean values of integrated pollution index corresponding to eight varieties of vegetables were all higher than the threshold value (i. e. 3.0) of heavy pollution; (3) In general, the largest bioaccumulation factor of heavy metals in vegetables was As, followed by Ni and Cu, and the order of pollution degree of heavy metals in vegetables was Ni > Zn > Cu > Pb > As > Cd; (4) The target hazard quotients (THQs) of As, Ni, Cu, Pb, Cd and Zn were 17.92, 1.01, 10.14, 0.73, 0.21 and 1.93, respectively. Arsenic and copper were the major risk contributors for inhabitants since the THQs of them respectively mounted to 56.10% and 31.75% of the total THQ value according to the average vegetable consumption; (5) The estimated daily intake (DI) of As, Ni, Cu, Pb, Cd and Zn from vegetables was 324.38, 1 211.25, 24 326.25, 176.25, 12.75 and 34 800 microg x d(-1) for adult residents, respectively; and (6) The target cancer risk (TR) of vegetables polluted by As to individual human health was 8.06 x 10(-3), significantly higher than the management standard (i. e. 10(-6) - 10(-4)) of United States Environmental Protection Agency (US EPA) and the standard (i. e. 5.0 x 10(-5)) of International Commission on Radiological Protection (ICRP), indicating that it was quite unsafe for the general population to consume vegetables from the studied fragmentary plots.