Tingting Ma

Oxidative Stress, Cytotoxicity and Genotoxicity in Earthworm Eisenia fetida at Different Di-n-Butyl Phthalate Exposure Levels

Recognized as ubiquitous contaminants in soil, the environmental risk of phthalic acid esters (PAEs) is of great concern recently. Effects of di-n-butyl phthalate (DnBP), an extensively used PAE compound to Eisenia fetida have been investigated in spiked natural brown yellow soil (Alfisol) for soil contact test. The toxicity of DnBP to E. fetida on the activity of superoxide dismutase (SOD) activity, peroxidase (POD), reactive oxygen species (ROS) content, and the apoptosis of coelomocytes and DNA damage at the 7th, 14th, 21st and 28th day of the incubation have been paid close attention to. In general, SOD activity and ROS content were significantly induced, opposite to total protein content and POD activity, during the toxicity test of 28 days especially under concentrations higher than 2.5 mg kg-1. The reduction in neutral red retention (NRR) time along with the increase of dead coelomocytes as the increasing of DnBP concentrations, indicating severe damage to cell viability under varying pollutant stress during cultivation, which could also be proved by comet assay results for exerting evident DNA damage in coelomocytes. DnBP in spiked natural soil could indeed cause damage to tissues, coelomocytes and the nucleus of E. fetida. The key point of the apparent change in different indices presented around 2.5 mg DnBP kg-1 soil, which could be recommended as the threshold of DnBP soil contamination, so that further investigation on threshold values to other soil animals or microorganisms could be discussed.

A new procedure combining GC-MS with accelerated solvent extraction for the analysis of phthalic acid esters in contaminated soils

An optimized procedure based on gas chromatography-mass spectrometry (GC-MS) combined with accelerated solvent extraction (ASE) is developed for the analysis of six phthalic acid esters (PAEs), which are priority soil pollutants nominated by United States Environmental Protection Agency (USEPA). Quantification of PAEs in soil employs ultrasonic extraction (UE) (USEPA 3550) and ASE (USEPA 3545), followed by clean up procedures involving three different chromatography columns and two combined elution methods. GC-MS conditions under selected ion monitoring (SIM) mode are described and quality assurance and quality control (QA/QC) criteria with high accuracy and sensitivity for target analytes were achieved. Method reliability is assured with the use of an isotopically labeled PAE, di-n-butyl phthalate-d4 (DnBP-D4), as a surrogate, and benzyl benzoate (BB) as an internal standard, and with the analysis of certified reference materials (CRM). QA/QC for the developed procedure was tested in four PAE-spiked soils and one PAE-contaminated soil. The four spiked soils were originated from typical Chinese agricultural fields and the contaminated soil was obtained from an electronic waste dismantling area. Instrument detection limits (IDLs) for the six PAEs ranged 0.10–0.31 μg·L−1 and method detection limits (MDLs) of the four spiked soils varied from a range of 20–70 μg·kg−1 to a range of 90–290 μg·kg−1. Linearity of response between 20 μg·L−1 and 2 mg·L−1 was also established and the correlation coefficients (R) were all>0.998. Spiked soil matrix showed relative recovery rates between 75 and 120% for the six target compounds and about 93% for the surrogate substance. The developed procedure is anticipated to be highly applicable for field surveys of soil PAE pollution in China.

Oxytetracycline Toxicity and Its Effect on Phytoremediation by Sedum plumbizincicola and Medicago sativa in Metal-Contaminated Soil

Excessive use of antibiotics potentially threatens human health, agricultural production, and soil phytoremediation. This arouses concern over the potential adverse effects of a commonly used antibiotic, oxytetracycline (OTC), on plants used for soil remediation and possible stimulation of antibiotic resistance genes in soils. A greenhouse experiment was conducted to investigate different rates (0, 1, 5, and 25 mg kg-1) and frequencies (one single high and daily low application) of OTC addition to soil on phytoremediation of a heavy metal contaminated soil by Sedum plumbizincicola and/or Medicago sativa (alfalfa). After 90 days both Cd and Zn were substantially removed by phytoextraction into S. plumbizincicola shoots especially at the high OTC (25 mg kg-1) treatment which also led to inhibition of antioxidative enzyme activities in both plant species. Soil microbial activity decreased significantly with the addition of OTC, and this was ameliorated by planting alfalfa and S. plumbizincicola together. OTC at <5 mg kg-1 increased the biomass of both plant species, but the frequency of OTC addition had no effect on the rate of metal removal. Alfalfa exhibited greater detoxification ability and effectiveness in soil microbial activity promotion than S. plumbizincicola with intercropping. Phytoremediation by alfalfa and S. plumbizincicola in association can both promote the removal of heavy metals and also alleviate the toxic effects of pollutants on plants and soil microbes even at relatively high soil OTC concentrations.

Efficiency of Repeated Phytoextraction of Cadmium and Zinc from an Agricultural Soil Contaminated with Sewage Sludge

Long-term application of sewage sludge resulted in soil cadmium (Cd) and zinc (Zn) contamination in a pot experiment conducted to phytoextract Cd/Zn repeatedly using Sedum plumbizincicola and Apium graceolens in monoculture or intercropping mode eight times. Shoot yields and soil physicochemical properties changed markedly with increasing number of remediation crops when the two plant species were intercropped compared with the unplanted control soil and the two monoculture treatments. Changes in soil microbial indices such as average well colour development, soil enzyme activity and soil microbial counts were also significantly affected by the growth of the remediation plants, especially intercropping with S. plumbizincicola and A. graveolens. The higher yields and amounts of Cd taken up indicated that intercropping of the hyperaccumulator and the vegetable species may be suitable for simultaneous agricultural production and soil remediation, with larger crop yields and higher phytoremediation efficiencies than under monoculture conditions.

Toxicity effects of di-(2-ethylhexyl) phthalate to Eisenia fetida at enzyme, cellular and genetic levels

Di-(2-ethylhexyl) phthalate (DEHP) is a dominant phthalic acid ester (PAE) that has aroused public concern due to its resistance to degradation and its toxicity as an endocrine-disrupting compound. Effects of different concentrations of DEHP on Eisenia fetida in spiked natural soil have been studied in the body of the earthworm by means of soil cultivation tests 7, 14, 21 and 28 days after exposure. The results indicated that, in general, superoxide dismutase (SOD) activity, malondialdehyde (MDA) content, metallothionein (MT) content, the expression of heat shock protein 70 (HSP 70) and all the tested geno-toxicity parameters are promoted as time elapses and with increasing concentration of DEHP. However, peroxidase (POD) activity, neutral red retention time (NRRT) and mitochondrial membrane potential difference values were found to decrease even at a low concentration of DEHP of 1 mg kg-1 soil (p<0.05). Clear toxic effects of DEHP on E. fetida have been generally recognized by means of the disturbance of antioxidant enzyme activity/content and critical proteins, cell membrane and organelle disorder and DNA damage estimated by length of tail, tail DNA ratio, and tail moment parameters. A concentration of DEHP of 3 mg kg-1 may be recommended as a precaution against the potential risk of PAEs in soils and for indicating suitable threshold values for other soil animals and soil micro-organisms.

Phytotoxicity in seven higher plant species exposed to di-n-butyl phthalate or bis （2-ethylhexyl） phthalate

We investigated phytotoxicity in seven plant species exposed to a range of concentrations (0–500 mg·kg−1 soil) of di-n-butyl phthalate (DnBP) or bis (2-ethylhexyl) phthalate (DEHP), two representative phthalate esters (PAEs) nominated by USEPA as priority pollutants and known environmental estrogens. We studied seed germination, root elongation, seedling growth, biomass (fresh weight, FW) and malondialdehyde (MDA) content of shoots and roots of wheat (Triticum aestivum L.), alfalfa (Medicago sativa L.), perennial ryegrass (Lolium perenne), radish (Raphanus sativus L.), cucumber (Cucumis sativus L.), oat (Avena sativa) and onion (Allium cepa L.), together with monitoring of plant pigment content (chlorophyll a, b and carotinoids) in alfalfa, radish and onion shoots. Root elongation, seedling growth and biomass of the test species were generally inhibited by DnBP but not by DEHP, indicating a lower level of phytotoxicity of DEHP than of DnBP. MDA contents of four species were promoted by PAE exposure, but not in alfalfa, ryegrass or onion shoots, indicating lower sensitivity of these three species to PAE pollutants. Plant pigment contents were clearly affected under the stress of both pollutants, implying the potential damage to the photosynthetic system of test plants, mainly by decreasing the content of chlorophyll a and b. Results of DnBP and DEHP phytotoxicity to the primary growth of test plants has provided information for the assessment of their environmental risk in the soil and also forms a basis for the further analysis of their toxic effects over the whole growth period of different plant species.

Uptake of silver by brown rice and wheat in soils repeatedly amended with biosolids

There have been numerous studies simulating the behaviour and fate of silver (Ag) applied to soils in biosolids in recent decades but the results remain to be verified under actual farming conditions. Here, we report a study of the transfer of Ag along the biosolids-soil-crop pathway with repeated biosolid applications over a four-year period to three contrasting soil types under rice-wheat rotation cultivation. A systematic investigation of Ag concentrations in biosolids throughout China shows Ag concentrations ranging from 0.64 to 7.47mgkg-1 with a mean value of 3.58mgkg-1 and a median of 3.02mgkg-1 on a dry weight basis. Silver concentrations were significantly higher in industrial biosolids than in domestic or mixed flow biosolids. Biosolids application enhanced Ag accumulation in whole wheat. Silver concentrations in whole wheat increased to 20.8, 20.5 and 4.87μgkg-1 after four years of high-metal industrial biosolids application to an acid Typic Ali-Perudic Aragsol, a neutral Typic Hapli-Stagnic Anthrosol, and a calcareous Typic Carbonati-Perudic Ferrosol, respectively. Moreover, the Ag translocation factor also increased in wheat following biosolids application with values of 5.6, 3.1 and 1.4, respectively. However, Ag accumulation in rice was found only in the acid soil with no discernible increase (p>0.05) in the translocation factor. The seasonal redox cycle may contribute to this phenomenon. A seedling incubation experiment confirms the influence of soil water regime on Ag bioavailability with a higher Ag translocation factor during the wheat growing season than the rice growth period. Incorporating a fallow period during the wheat (winter crop) season might be a suitable strategy for repeated biosolids application.

Modulation of the efficiency of trace metal phytoremediation by Sedum plumbizincicola by microbial community structure and function

AimsRecent research on the rhizosphere microbiome indicates that plants may benefit more from microbial communities than from individual members in a community. The link between soil microbiome and plant performance remains poorly understood during trace metal phytoremediation by hyperaccumulator plants. The effect of different microbiomes on the cadmium (Cd)/zinc (Zn) phytoextraction efficiency of Sedum plumbizincicola was therefore explored.MethodsDifferent microbiomes were set up using γ-irradiation and vancomycin addition and a pot experiment was conducted. The microbial communities were characterized using high-throughput sequencing and their functions were predicted.ResultsPlant Cd and Zn contents in the vancomycin-amended treatment were about 38–46% and 35%–53% respectively lower than those in the other treatments. Bacterial composition and diversity were substantially changed by γ-irradiation and vancomycin addition but plant trace metal accumulation did not change subsequently. The bacterial functional profiles were very different following treatment with vancomycin compared with the other treatments at KEGG orthology hierarchy level 3. Genes related to up-regulated biofilm formation and down-regulated siderophore synthesis may partly contribute together to the low plant trace metal accumulation following vancomycin treatment.ConclusionsSpecific bacterial community function rather than bacterial community diversity and composition may affect trace metal uptake by hyperaccumulator plants.