Bioremediation and phytoremediation of total petroleum hydrocarbons (TPH) under various conditions

Remediation of contaminated soils is often studied using fine-textured soils rather than low-fertility sandy soils, and few studies focus on recontamination events. This study compared aerobic and anaerobic treatments for remediation of freshly introduced used motor oil on a sandy soil previously phytoremediated and bioacclimated (microorganisms already adapted in the soil environment) with some residual total petroleum hydrocarbon (TPH) contamination. Vegetated and unvegetated conditions to remediate anthropogenic fill containing residual TPH that was spiked with nonaqueous phase liquids (NAPLs) were evaluated in a 90-day greenhouse pot study. Vegetated treatments used switchgrass (Panicum virgatum). The concentration of aerobic bacteria were orders of magnitude higher in vegetated treatments compared to unvegetated. Nevertheless, final TPH concentrations were low in all saturated soil treatments, and high in the presence of switchgrass. Concentrations were also low in unvegetated pots with fertilizer. Acclimated indigenous microbial communities were shown to be more effective in breaking down hydrocarbons than introducing microbes from the addition of plant treatments in sandy soils. Remediation of fresh introduced NAPLs on pre-phytoremediated and bioacclimated soil was most efficient in saturated, anaerobic environments, probably due to the already pre-established microbial associations, easily bioavailable contaminants, and optimized soil conditions for microbial establishment and survival.     

Clonal variation in survival and growth of hybrid poplar and willow in an in situ trial on soils heavily contaminated with petroleum hydrocarbons

Species and hybrids between species belonging to the genera Populus (poplar) and Salix (willow) have been used successfully for phytoremediation of contaminated soils. Our objectives were to: 1) evaluate the potential for establishing genotypes of poplar and willow on soils heavily contaminated with petroleum hydrocarbons and 2) identify promising genotypes for potential use in future systems. We evaluated height, diameter, and volume after first year budset by testing 20 poplar clones and two willow clones. Unrooted cuttings, 20 cm long, were planted in randomized complete blocks at 0.91- x 0.91-m spacing at Gary, IN, USA (41.5 degrees N, 87.3 degrees W). Four commercial poplar clones (NM6, DN5, DN34, and DN182) were planted as 20- and 60-cm cuttings. Sixty-cm cuttings exhibited greater height and diameter than 20-cm cuttings; however, we recommend continued use and testing of different combinations of genotype and cutting length. We identified promising genotypes for potential use in future systems and we recommend allocating the majority of resources into commercial poplar clones, given their generalist growth performance. However, further utilization and selection of experimental clones is needed. Specific clones rather than genomic groups should be selected based on the geographic location and soil conditions of the site.     

Availability and toxicity of cadmium to forage grasses grown in contaminated soil

It is important to know the mechanisms for forage development, especially those related to the tolerance of potentially toxic elements, when considering their use in phytoremediation in heavy metal contaminated areas. In this study, we evaluated plant growth, concentration, and the availability of cadmium (Cd) for forage grasses (Panicum maximum Jacq. cv. Aruana and cv. Tanzânia; Brachiaria decumbens cv. Basilisk; Brachiaria brizantha cv. Xaraés and cv. Marandu) cultivated in Cd contaminated soils. The experiments were performed under greenhouse conditions over a 90-day evaluation period, and the Cd rates were 2, 4, and 12 mg/kg of soil. The relative growth rate of the forage grasses decreased as Cd rates increased, and the following descending order of susceptibility was observed: Marandu > Xaraés > Aruana > Tanzânia > Basilisk, with regard to phytotoxicity in these plants. The forage Cd concentration increased in line with increases in the Cd rates. Cd contents extracted by Mehlich-1 and by diethylenetriaminepentaacetic acid presented high positive correlation with forage relative growth. The forage plants did not block Cd entry into the food chain because they were not capable of limiting Cd absorption.     

Microbial expression profiles in the rhizosphere of willows depend on soil contamination

The goal of phytoremediation is to use plants to immobilize, extract or degrade organic and inorganic pollutants. In the case of organic contaminants, plants essentially act indirectly through the stimulation of rhizosphere microorganisms. A detailed understanding of the effect plants have on the activities of rhizosphere microorganisms could help optimize phytoremediation systems and enhance their use. In this study, willows were planted in contaminated and non-contaminated soils in a greenhouse, and the active microbial communities and the expression of functional genes in the rhizosphere and bulk soil were compared. Ion Torrent sequencing of 16S rRNA and Illumina sequencing of mRNA were performed. Genes related to carbon and amino-acid uptake and utilization were upregulated in the willow rhizosphere, providing indirect evidence of the compositional content of the root exudates. Related to this increased nutrient input, several microbial taxa showed a significant increase in activity in the rhizosphere. The extent of the rhizosphere stimulation varied markedly with soil contamination levels. The combined selective pressure of contaminants and rhizosphere resulted in higher expression of genes related to competition (antibiotic resistance and biofilm formation) in the contaminated rhizosphere. Genes related to hydrocarbon degradation were generally more expressed in contaminated soils, but the exact complement of genes induced was different for bulk and rhizosphere soils. Together, these results provide an unprecedented view of microbial gene expression in the plant rhizosphere during phytoremediation.     

蚯蚓在植物修复芘污染土壤中的作用

采用盆栽试验法,研究了蚯蚓(Pheretima hupeiensis)在植物修复芘污染土壤中的作用。结果显示,试验浓度(20.24-321.42 mg/kg) 范围内,蚯蚓活动促进了芘污染土壤中修复植物黑麦草(Lolium multiforum)黑麦草的生长,其根冠比明显增大。添加蚯蚓72 d后,种植黑麦草的土壤中芘的去除率高达60.01%-86.26%,其平均去除率(74.66%)比无蚯蚓活动的土壤-植物系统(64.55%)提高10.11%,比无植物对照组(18.24%)提高56.42%。各种生物、非生物修复因子中,植物-微生物交互作用对芘去除的平均贡献率(51.75%)最为突出,比无蚯蚓活动时(44.94%)提高6.81%。说明蚯蚓活动可强化土壤-植物系统对土壤芘污染的修复作用。     

Variation in the uptake of Arsenic, Cadmium, Lead, and Zinc by different species of willows Salix spp. grown in contaminated soils

The experiment assessed the variability of in seven clones of willow plants of high biomass production (Salix smithiana S-218, Salix smithiana S-150, Salix viminalis S-519, Salix alba S-464, Salix ’Pyramidalis’ S-141, Salix dasyclados S-406, Salix rubens S-391). They were planted in a pots for three vegetation periods in three soils differing in the total content of risk elements. Comparing the calculated relative decrease of total metal contents in soils, the phytoextraction potential of willows was obtained for cadmium (Cd) and zinc (Zn), moderately contaminated Cambisol and uncontaminated Chernozem, where aboveground biomass removed about 30% Cd and 5% Zn of the total element content, respectively. The clones showed variability in removing Cd and Zn, depending on soil type and contamination level: S. smithiana (S-150) and S. rubens (S-391) demonstrated the highest phytoextraction effect for Cd and Zn. For lead (Pb) and arsenic (As), the ability to accumulate the aboveground biomass of willows was found to be negligible in both soils. The results confirmed that willow plants show promising results for several elements, mainly for mobile ones like cadmium and zinc in moderate levels of contamination. The differences in accumulation among the clones seemed to be affected more by the properties of clones, not by the soil element concentrations or soil properties. However, confirmation and verification of the results in field conditions as well as more detailed investigation of the mechanisms of cadmium uptake in rhizosphere of willow plants will be determined by further research.     

Remediation of contaminated soils by biotechnology with nanomaterials: bio-behavior,applications, and perspectives

Soil contamination caused by heavy metals and organic pollutants has drawn world-wide concern. Biotechnology has been applied for many years to the decontamination of soils polluted with organic and inorganic contaminants, and novel nanomaterials (NMs) has attracted much concern due to their high capacity for the removal/stabilization/degradation of pollutants. Recently, developing advanced biotechnology with NMs for the remediation of contaminated soils has become a hot research topic. Some researchers found that bioremediation efficiency of contaminated soils was enhanced by the addition of NMs, while others demonstrated that the toxicity of NMs to the organism negatively influenced the repair capacity of polluted soils. This paper reviews the application of biotechnology and NMs in soil remediation, and further provides a critical view of the effects of NMs on the phytoremediation and micro-remediation of contaminated soils. This review also discusses the future research needs for the combined application of biotechnology and NMs in soil remediation.     

Phytoremediation of arsenic and lead in contaminated soil using Chinese brake ferns (Pteris vittata) and Indian mustard (Brassica juncea)

Field and greenhouse experiments were performed to assess the performance of phytoremediation of arsenic and lead from contaminated soil at an EPA Superfund site (Barber Orchard). Chinese Brake ferns (Pteris vittata) were used to extract arsenic. On average, fern shoot arsenic concentrations were as high as 20 times the soil arsenic concentrations under field conditions. It was estimated that 8 years would be required to reduce the acid-extractable portion of soil arsenic to safe levels (40 mg/kg). The effect of soil pH on arsenic extraction was also investigated. Results indicate that increasing soil pH may improve arsenic removal. Indian mustard plants (Brassica juncea) were used under greenhouse conditions to phytoextract soil lead. EDTA was applied to soil and was found to improve lead extraction. When the EDTA concentration was 10 mmol EDTA/kg soil in soil containing 338 mg Pb/kg soil, mustard plants extracted approximately 32 mg of lead. In conclusion, phytoremediation would be a suitable alternative to conventional remediation techniques, especially for soils that do not require immediate remediation.     

Uptake and Bioaccumulation of Pentachlorophenol by Emergent Wetland Plant Phragmites australis (Common Reed) in Cadmium Co-contaminated Soil

Despite many studies on phytoremediation of soils contaminated with either heavy metals or organics, little information is available on the effectiveness of phytoremediation of co-occurring metal and organic pollutants especially by using wetland species. Phragmites australis is a common wetland plant and its potential for phytoremediation of cadmium pentachlorophenol (Cd-PCP) co-contaminated soil was investigated. A greenhouse study was executed to elucidate the effects of Cd (0, 10, and 20 mg kg^?1) without or with PCP (0, 50, and 250 mg kg^?1) on the growth of the wetland plant P. australis and its uptake, accumulation and removal of pollutant from soils. After 75 days, plant biomass was significantly influenced by interaction of Cd and PCP and the effect of Cd on plant growth being stronger than that of PCP. Coexistence of PCP at low level lessened Cd toxicity to plants, resulting in improved plant growth and increased Cd accumulation in plant tissues. The dissipation of PCP in soils was significantly influenced by interactions of Cd, PCP and plant presence or absence. As an evaluation of soil biological activities after remediation soil enzyme was measured.     

Potential for phytoremediation of polychlorinated biphenyl-(PCB-)contaminated soil

Weathered soils contaminated with commercial-grade Aroclor 1260 from three sites in Canada were used to investigate the polychlorinated biphenyl (PCB) phytoextraction potential of nine plant species (Festuca arundinacea, Glycine max, Medicago sativa, Phalaris arundinacea, Lolium multiflorum, Carex normalis, and three varieties of Cucurbita pepo ssp. pepo) under controlled greenhouse conditions. The soils used varied in PCB concentration (90-4200 microg/g) and total organic content (0.06-2.02%). Greenhouse experiments controlled for PCB volatilization through the use of a vented enclosure and by isolating the contaminated soils with parafilm. After 8 wks, PCB concentrations of 47-6700 microg/g were observed in root tissues. Although PCB concentrations in shoot tissues were lower (< 1-470 microg/g), the absolute amounts of PCBs observed in shoot tissue were significant (1.7-290 microg) once shoot biomass was accounted for. Congener signatures indicated that tetra- to hexa-chlorobiphenyls contributed the largest proportions to shoot tissues, but hepta-to nona-chorobiphenyls were also present in measurable amounts. Overall, the results indicate that varieties of C. pepo were more effective at extracting PCBs from soil than other plants screened The evidence suggests that this was mainly due to root uptake of PCBs and tranlocation to the shoots, rather than volatilization of PCBs from soil. All plants screened showed signs of stress in the most highly contaminated soil (4200 microg/g), but not in the two lower PCB contaminated soils (250 and 90 microg/g, respectively). No detectable decreases in soil PCB concentrations were observed in these short-term greenhouse experiments, but the results suggest that this may be achievable through multiple plantings.     

Methodology for using contaminated soil leachability testing to determine soil cleanup levels at contaminated petroleum underground storage tank (UST) sites

The costs of environmental remediation at leaking petroleum underground storage tank (UST) sites are influenced significantly by soil cleanup levels. The use of conservative generic soil cleanup levels may be inappropriate at some sites contaminated by leaking petroleum USTs. At many contaminated sites, a primary objective of site remediation is long‐term protection of water resources (e.g., groundwater) from pollution. Leaching of pollutants from residual soil contamination to groundwater is a primary consideration in establishing site‐specific soil cleanup levels at fuel‐contaminated sites. The use of laboratory soil leachability testing methods may be useful in objectively evaluating the leaching potential of contaminants from residual soil contamination and estimating potential groundwater impacts. Developing soil cleanup levels that are protective of water resources must include a technically sound integration of site‐specific soil leachability data and contaminant attenuation factors. Evaluation of the leaching potentials of soil contaminants may also provide essential supplementary information for other site characterization methods that may be used to evaluate risks to human health. Contaminant leachability testing of soils may provide a cost‐effective and technically based method for determining soil cleanup levels that are protective of groundwater resources at contaminated petroleum UST sites.     

有机污染土壤和地下水生物修复研究热点和趋势——基于Web of Science数据库的文献计量学分析

生物修复作为经济有效、绿色可持续的修复技术,在有机污染土壤和地下水修复上具有广阔的应用前景。基于WebofScience核心数据库,通过文献计量可视化应用软件VOSviewer和CiteSpace,分析了1990–2020年有机污染土壤和地下水生物修复领域的研究热点及趋势。结果表明,有机污染土壤和地下水生物修复领域的论文发表数量呈增长趋势,发文总量最多的国家是美国和中国,但是2012年后中国年发文量快速增加,并位居第一。该领域的相关研究主要发表在Chemosphere、Environmental ScienceTechnology、Science of the Total Environment等top期刊上。全球研究机构中中国科学院发文量最多,但是来自美国加州大学的总被引频次和h-index最高。发文量最多的是来自英国兰卡斯特大学的学者Semple教授,我国发文量最多的是来自中国科学院南京土壤研究所的骆永明研究员。下一步研究重点和热点:针对复合污染土壤和地下水,研发新型耦合强化生物修复技术,采用先进的分子生物学方法探索功能微生物及其功能基因,阐明生物降解机理,明确原位污染土壤和地下水的靶向性调控机制。     

2020污染土壤的生物修复专刊序言

生物修复技术,作为可持续发展的重要方向,因其环境友好、高效且无二次污染并能从根本上解决土壤污染问题而受到关注,已经在土壤污染治理中得到了广泛的应用。为了梳理和凝练生物修复技术的发展状况,本专刊收录了该研究领域的16篇论文,分别从植物修复、微生物修复、联合修复、重金属吸收积累的相关分子机制、资源化再利用等方面,详细阐述生物修复技术的发展动态,展望未来的发展趋势,为促进生物修复技术的发展提供参考。     

Biodegradation of petroleum hydrocarbons by filamentous fungi (Aspergillus ustus and Purpureocillium lilacinum) isolated from used engine oil contaminated soil

The use of microorganisms for remediation and restoration of hydrocarbons contaminated soils is an effective and economic solution. The current study aims to find out efficient telluric filamentous fungi to degrade petroleum hydrocarbons pollutants. Six fungal strains were isolated from used engine (UE) oil contaminated soil. Fungi were screened for their ability to degrade crude oil, diesel and UE oil using 2.6-dichlorophenol indophenol (DCPIP). Two isolates were selected, identified and registered at NCBI as Aspergillus ustus HM3.aaa and Purpureocillium lilacinum HM4.aaa. Fungi were tested for their tolerance to different concentration of petroleum oils using radial growth diameter assay. Hydrocarbons removal percentage was evaluated gravimetrically. The degradation kinetic of crude oil was studied at a time interval of 10 days. A.ustus was the most tolerant fungi to high concentration of petroleum oils in solid medium. Quantitative analysis showed that crude oil was the most degraded oil by both isolate; P. lilacinium and A. ustus removed 44.55% and 30.43% of crude oil, respectively. The two fungi were able to degrade, respectively, 27.66 and 21.27% of diesel and 14.39 and 16.00% of UE oil. As compared to the controls, these fungi accumulated high biomass in liquid medium with all petroleum oils. Likewise, crude oil removal rate constant (K) and half-lives (t_1/2) were 0.02 day⁻¹, 34.66 day and 0.015 day⁻¹, 46.21 day for P. lilacinium and A. ustus, respectively. The selected fungi appear interesting for petroleum oils biodegradation and their application for soil bioremediation require scale-up studies.     

Phytoremediation of soil contaminated with PCBs using different plants and their associated microbial communities

In this work, we evaluate the abilities of the plants Brassica juncea, Avena sativa, Brachiaria decumbens, and Medicago sativa to uptake polychlorinated biphenyls (PCBs) and induce degradation of soil microorganisms from contaminated soil. Removal of PCBs 44, 66, 118, 153, 170, and 180 was evaluated in both rhizospheric and nonrhizospheric soils. Microbial and bphA1 gene quantifications were performed by real-time PCR. The PCB concentrations in plant tissues and soil were determined, and a fluorescein diacetate (FDA) hydrolysis assay was used to measure microbial activity in soil. The removal percentages for all PCB congeners in planted soil versus unplanted control soil were statistically significant and varied between 45% and 63%. PCBs 118, 153, 138, and 170 were detected in Brachiaria decumbens roots at different concentrations. In planted soil, an increase in the concentration of bacteria was observed compared to the initial concentration and the concentration in unplanted control soil; however, no significant differences were identified between plants. The number of copies of the bphA1 gene was higher in rhizospheric versus non- rhizospheric soil for all plants at the end of the experiment. However, alfalfa and oat rhizospheric soil showed significant differences in the copy number of the bphA1 gene. In general, the concentration of fluorescein in the rhizospheric soil was greater than that in the nonrhizospheric soil. Although the plants had a positive effect on PCB removal, this effect varied depending on the type of PCB, the plant, and the soil.     

Phytoextraction and Accumulation of Mercury in Three Plant Species: Indian Mustard (Brassica Juncea), Beard Grass (Polypogon monospeliensis), and Chinese Brake Fern (Pteris vittata)

The objective of this research was to screen and search for suitable plant species to phytoextract mercury-contaminated soil. Our effort focused on using some of the known metal-accumulating wild-type plants since no natural plant species with mercury-hyperaccumulat ing properties has yet been identified. Three plant species were evaluated for their uptake efficiency for mercury: Indian mustard (Brassica juncea), beard grass (Polypogon monospeliensis), and Chinese brake fern (Pteris vittata). Four sets of experiments were conducted to evaluate the phytoremediation potential of these three plant species: a pot study with potting mix where mercury was provided daily as HgCl₂ solution; experiments with freshly mercury-spiked soil; and a study with aged soils contaminated with different mercury sources (HgCl₂, Hg(NO₃)₂, and HgS). Homemade sunlit chambers were also used to study foliar uptake of Hg from ambient air. Among the three plant species, Chinese brake fern showed the least stress symptoms resulting from mercury exposure and had the highest mercury accumulation. Our results indicate that Chinese brake fern may be a potential candidate for mercury phytoextraction. We found that mercury contamination is biologically available for plant uptake and accumulation, even if the original and predominating mercury form is HgS, and also after multiple phytoremediation cycles.     

Responses and roles of roots,microbes, and degrading genes in rhizosphere during phytoremediation of petroleum hydrocarbons contaminated soil

Abstract

Rhizodegradation performed by plant roots and the associated bacteria is one of the major mechanisms that contribute to removal of petroleum hydrocarbons (PHCs) during phytoremediation. In this study, the pot-culture experiment using wild ornamental Hylotelephium spectabile (Boreau) H. Ohba was designed to explore responses and roles of roots, microbes, and degrading genes in the rhizodegradation process. Results showed that PHCs degradation rate by phytoremediation was up to 37.6–53.3% while phytoaccumulation accounted for a low proportion, just at 0.3–13.3%. A total of 37 phyla were classified through the high throughput sequencing, among which Proteobacteria, Actinobacteria, and Acidobacteria were the three most dominant phyla, accounting for >60% of the phylum frequency. The selective enrichment of PHC degraders with high salt-tolerance, including Alcanivorax and Bacteroidetes, was induced. Generally, relative abundance of the PHC degrading genes increased significantly with an increase in PHCs concentrations, and the gene copy number in the phytoremediation group was 1.46–14.44 times as much as that in the unplanted controls. Overall, the presence of PHCs and plant roots showed a stimulating effect on the development of specific degraders containing PHC degrading genes, and correspondingly, a biodegradation-beneficial community structure had been constructed to contribute to PHCs degradation in the rhizosphere.     

有机物及重金属植物修复研究进展

植物修复技术是近年来发展起来的一种非常有前途的生物治理技术 ,也是当前学术界研究的热点领域 .本文对植物修复的类型 ,植物修复土壤、水体有机污染物、重金属和某些放射性核素的过程、机理及可能影响因子作了概括和详尽讨论 ,并就国内外近年来植物修复技术在污染环境中的应用和研究成果进行了综述     

植物-固定化菌剂联合修复多环芳烃污染土壤

以火凤凰根际土壤中发现的3种优势菌[分枝杆菌(Ⅰ)、产黄纤维单胞菌(Ⅱ)、少动鞘氨醇单胞菌(Ⅲ)]构建的多菌剂体系为供试菌剂,针对大港油田原油污染土壤,将固定化供试菌剂接种于修复植物火凤凰根际,探讨供试菌剂强化火凤凰修复多环芳烃(PAHs)污染土壤的效果.结果 表明:处理ⅠⅢ(有效活茵数为109 cfu·mL-1)和Ⅰ...     

Potential of wheat (Triticum aestivum L.) and pea (Pisum sativum) for remediation of soils contaminated with bromides and PAHs

The aim of the research was to study a removal of polycyclic aromatic hydrocarbons (PAHs) and phytoextraction of bromine (Br) from contaminated soils. The experiments using pea and wheat seedlings as potential candidates for soil remediation were performed. The soil for the experiments was collected from a site slightly contaminated by some PAHs. Before planting, the soil was exposed to 20 mg of Br/kg of soil. In the soil taken from rhizosphere of pea and wheat, the concentrations of many PAHs decreased up to 7 times compared to the concentrations of the compounds in the initial soil. Pea was capable of more effectively influencing the soil PAHs than wheat. The growth of pea and wheat in the soil spiked with Br resulted in a significant increase of Br concentration in a plant. Concentration of Br in roots of pea and wheat increased 21 and 3 times, respectively. Bromine content in leaves of wheat and pea increased 10 and 4.5 times. This accumulation of Br in the plants led to a decrease of its concentration in the rhizosphere soil. The experimental results demonstrated a good ability of the plants to cleanup the soils contaminated with organic and inorganic compounds.