Impact of PAH on biological health parameters of soils of an Indian refinery and adjoining agricultural area—a case study

The present study is aimed at analysing and comparing different soil enzymes in soil samples of native contaminated sites of a Mathura refinery and adjoining agricultural land. Enzyme activities are considered as indicators of soil quality and changes in biogeochemical function due to management or perturbations. Soil samples were collected from the premises and nearby area of Mathura refinery, India. Biological health parameters (dehydrogenase, aryl esterase, aryl sulphatase, \upbeta \upbeta -glucosidase, alkaline phosphatase, acid phosphatase, lipase, laccase and catalase activity) were estimated in the soil samples. Among all the samples, sewage sludge soil showed maximum activity of enzymes, microbial biomass carbon and most probable number of polycyclic aromatic hydrocarbon (PAH) degraders in soils spiked with three- to four-ring PAHs at 50 ppm. Available phosphorus, potassium and nitrogen was also exceptionally high in this sample, indicating maximum microbial bioconversion due to presence of nutrients stimulating potent PAH-degrading microorganisms.     

Clastogenicity of landfarming soil treated with sugar cane vinasse

The addition of nutrients and/or soil bulking agents is used in bioremediation to increase microbial activity in contaminated soils. For this purpose, some studies have assessed the effectiveness of vinasse in the bioremediation of soils contaminated with petroleum waste. The present study was aimed at investigating the clastogenic/aneugenic potential of landfarming soil from a petroleum refinery before and after addition of sugar cane vinasse using the Allium cepa bioassay. Our results show that the addition of sugar cane vinasse to landfarming soil potentiates the clastogenic effects of the latter probably due the release of metals that were previously adsorbed into the organic matter. These metals may have interacted synergistically with petroleum hydrocarbons present in the landfarming soil treated with sugar cane vinasse. We recommend further tests to monitor the effects of sugar cane vinasse on soils contaminated with organic wastes.     

Effects of lead and zinc mining contamination on bacterial community diversity and enzyme activities of vicinal cropland

In the process of mining activity, many kinds of heavy metals enter into soils with dust, causing serious contamination to the environment. In this study, six soils were sampled from cropland at different distances from a lead/zinc mine in Heilongjiang Province, China. The total contents of lead and zinc in the vicinal cropland exceeded the third level of environmental quality standard for soil in China, which indicated that soils in this area were moderately contaminated. Bacterial community diversity and population were greatly decreased when the concentrations of lead and zinc were beyond 1,500 and 995 mg kg(-1), respectively, as analyzed by plate counting and polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). The bands of DGGE patterns varied with the degree of contamination. The activities of soil urease, phosphatase, and dehydrogenase were negatively correlated with the concentrations of lead and zinc. The highest inhibitory effect of heavy metals on soil enzyme activities was observed in urease. It was noted that PCR-DGGE patterns combined with soil enzyme activity analysis can be indices for the soil quality assessment by heavy metal contamination.     

Response of ammonia-oxidizing archaea and bacteria to long-term industrial effluent-polluted soils,Gujarat, Western India

Soil nitrifiers have been showing an important role in assessing environmental pollution as sensitive biomarkers. In this study, the abundance and diversity of ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB) were investigated in long-term industrial waste effluent (IWE) polluted soils. Three different IWE polluted soils characterized as uncontaminated (R1), moderately contaminated (R2), and highly contaminated (R3) were collected in triplicate along Mahi River basin, Gujarat, Western India. Quantitative numbers of ammonia monooxygenase α-subunit (amoA) genes as well as 16S rRNA genes indicated apparent deleterious effect of IWE on abundance of soil AOA, AOB, bacteria, and archaeal populations. Relatively, AOB was more abundant than AOA in the highly contaminated soil R3, while predominance of AOA was noticed in uncontaminated (R1) and moderately contaminated (R2) soils. Soil potential nitrification rate (PNR) significantly (P?<?0.05) decreased in polluted soils R2 and R3. Reduced diversity accompanied by apparent community shifts of both AOB and AOA populations was detected in R2 and R3 soils. AOB were dominated with Nitrosospira-like sequences, whereas AOA were dominated by Thaumarchaeal “group 1.1b (Nitrososphaera clusters).” We suggest that the significant reduction in abundance and diversity AOA and AOB could serve as relevant bioindicators for soil quality monitoring of polluted sites. These results could be further useful for better understanding of AOB and AOA communities in polluted soils.     

Impact of Zn and Cu on the development of phenanthrene catabolism in soil

Mixtures of polycyclic aromatic hydrocarbons (PAHs) and heavy metals are of major concern in contaminated soil. Biodegradation of PAHs in metal-contaminated soils is complicated because metals are toxic and cannot be degraded by biological processes. This investigation considered the effects of Zn and Cu (50, 100, 500 and 1,000 mg/kg) on ¹⁴C-phenanthrene biodegradation in soil over 60-day contact time. The presence of Zn at all concentrations and low concentrations of Cu (50 and 100 mg/kg) had no significant effect (p?>?0.05) on the development of phenanthrene catabolism; however, at higher Cu concentrations, the development of phenanthrene catabolism and bacterial cell numbers were significantly reduced (p?<?0.05). This suggests that Cu is more toxic than Zn to soil microbial PAH catabolic activity. Metal/PAH-contaminated soils represent one of the most difficult remedial challenges and insights into PAH biodegradation in the presence of metals is necessary in order to assess the potential for bioremediation.     

铅锌矿区土壤微生物区系及酶活性调查

通过对四川省汉源县富泉乡万顺铅锌矿区土壤微生物区系及微生物活性的调查，结果表明，铅锌矿区土壤几种重金属含量明显高于临近非矿区土壤。该矿区土壤微生物区系组成和微生物活性显著不同于临近非矿区土壤，随着重金属含量的增加，土壤微生物数量、微生物多样性指数、微生物生物量碳以及纤维素分解强度均显著降低，但土壤基础呼吸却明显升高；土壤脲酶、碱性磷酸酶、多酚氧化酶对铅锌矿较为敏感，而蔗糖酶和过氧化氢酶受到的抑制作用不明显。     

Ecotoxicological and microbiological characterization of soils from heavy-metal- and hydrocarbon-contaminated sites

The aims of this study were to characterize soils from industrial sites by combining physicochemical, microbiological, and ecotoxicological parameters and to assess the suitability of these assays for evaluation of contaminated sites and ecological risk assessment. The soil samples were taken from long-term contaminated sites containing high amounts of heavy metals (sites 1 and 2) or petroleum hydrocarbons (site 3) located in the upper Silesia Industrial Region in southern Poland. Due to soil heterogeneity, large differences between all investigated parameters were measured. Microbiological properties revealed the presence of high numbers of viable hetrotrophic microorganisms. Soil enzyme activities were considerably reduced or could not be detected in contaminated soils. Activities involved in N turnover (N mineralization and nitrification) were significantly (P?<?0.05) higher in samples from the metal-contaminated sites than in samples from the hydrocarbon-contaminated site, whereas the opposite was observed for phosphatase activity. The Microtox test system appeared to be the most appropriate to detect toxicity and significant differences in toxicity between the three sites. The Ostracodtoxkit test was the most appropriate test system to detect toxicity in the hydrocarbon-contaminated soil samples. Correlation analysis between principal components (obtained from factor analysis) determined for physicochemical, microbiological, and ecotoxicological soil properties demonstrated the impact of total and water-extractable contents of heavy metals on toxicity.     

In situ phytoremediation of a soil historically contaminated by metals, hydrocarbons and polychlorobiphenyls

In the past several years, industrial and agricultural activities have led to serious environmental pollution, resulting in a large number of contaminated sites. As a result, much recent research activity has focused on the application of bioremediation technologies as an environmentally friendly and economically feasible means for decontamination of polluted soil. In this study horse manure and Populus nigra (var. italica) (HM + P treatment) have been used, at real scale level, as an approach for bioremediation of a soil historically contaminated by metals (Pb, Cr, Cd, Zn, Cu and Ni) and organic contaminants, such as polychlorobiphenyls and petroleum hydrocarbon. After one year, the HM + P phytotreatment was effective in the reclamation of the polluted soil from both organic and inorganic contaminants. A reduction of about 80% in total petroleum hydrocarbon (TPH), and 60% in polychlorobiphenyls (PCBs) and total metals was observed in the HM + P treatment. In contrast, in the horse manure (HM) treatment, used as control, a reduction of only about 30% of TPH was obtained. In order to assess both effectiveness and evolution of the remediation system to a biologically active soil ecosystem, together with the pollution parameters, the parameters describing the evolution of the soil functionality (enzymatic activities and protein SDS-PAGE pattern) were investigated. A stimulation of the metabolic soil processes (increase in dehydrogenase activity) was observed in the HM + P compared to the HM treatment. Finally, preliminary protein SDS-PAGE results have permitted the identification of proteins that have been recovered in the HM + P soil with respect to the HM; this may become a basic tool for improving the biogeochemical status of soil during the decontamination through the identification of microbial populations that are active in soil decontamination.     

Prediction of PAH biodegradation in field contaminated soils using a cyclodextrin extraction technique

Biodegradation has been identified as a major loss process for organic contaminants in soils and, as a result, microbial strategies have been developed for the remediation of contaminated land. Prediction of the biodegradable fraction would be important for determining bioremediation end-points in the clean-up of contaminated land. The aim of this study was to investigate the ability of a cyclodextrin extraction to predict the extent to which polycyclic aromatic hydrocarbons (PAHs) would be degraded microbiologically in field contaminated soils; further testing the robustness and reproducibility of this extraction in chemically complex systems. Dichloromethane and hydroxypropyl-beta-cyclodextrin (HPCD) extractable fractions were measured together with the PAH biodegradable fraction in each of the six field contaminated soils. The amounts of PAHs degraded by the catabolic activity of the indigenous microflora in each of the soils were correlated with HPCD-extractable PAH concentrations. The regressions showed that the amounts of lower molecular weight PAHs extracted by the HPCD were not significantly (P > 0.05) different to the amounts that were degraded. However, higher molecular weight PAHs that were extracted by HPCD did differ significantly (P < 0.05) from the amounts degraded. Although the HPCD extraction did overestimate the microbially degradable fraction of the higher molecular weight PAHs, overall the correlations between the HPCD extractable fraction and the microbially degradable fraction were very close, with mean values of the slope of line for the six soils equalling 1. This study further describes the robust and reproducible nature of the aqueous-based soil extraction technique reliably measuring the extent to which PAHs will be microbially degraded in soil.     

Impacts of epigeic, anecic and endogeic earthworms on metal and metalloid mobility and availability

The introduction of earthworms into soils contaminated with metals and metalloids has been suggested to aid restoration practices. Eisenia veneta (epigeic), Lumbricus terrestris (anecic) and Allolobophora chlorotica (endogeic) earthworms were cultivated in columns containing 900 g soil with 1130, 345, 113 and 131 mg kg(-1) of As, Cu, Pb and Zn, respectively, for up to 112 days, in parallel with earthworm-free columns. Leachate was produced by pouring water on the soil surface to saturate the soil and generate downflow. Ryegrass was grown on the top of columns to assess metal uptake into biota. Different ecological groups affected metals in the same way by increasing concentrations and free ion activities in leachate, but anecic L. terrestris had the greatest effect by increasing leachate concentrations of As by 267%, Cu by 393%, Pb by 190%, and Zn by 429% compared to earthworm-free columns. Ryegrass grown in earthworm-bearing soil accumulated more metal and the soil microbial community exhibited greater stress. Results are consistent with earthworm enhanced degradation of organic matter leading to release of organically bound elements. The degradation of organic matter also releases organic acids which decrease the soil pH. The earthworms do not appear to carry out a unique process, but increase the rate of a process that is already occurring. The impact of earthworms on metal mobility and availability should therefore be considered when inoculating earthworms into contaminated soils as new pathways to receptors may be created or the flow of metals and metalloids to receptors may be elevated.     

Potential Applications for Pyrolysis-GC/MS in Bioremediation

Preliminary data are presented from a set of experiments designed to promote the use of pyrolysis-GC/MS in bioremediation. Studies were designed to aid researchers in developing a pyrolysis-GC/MS method and identifying how the method could help characterize bioremediation, particularly in organic soils. Since sample size affects the results of pyrolytic analyses, the first experiment demonstrated how an appropriate sample size might be selected for pyrolysis-GC/MS testing. In order to show how quantitative results can be obtained from pyrolysis-GC/MS, a second experiment determined the 'goodness of fit' for a standard curve relating the chromatographic area under a pyrogram to actual mass units. The third experiment investigated ways in which pyrolysis-GC/MS analyses could improve our understanding of bioremediation in organic soils contaminated with crude oil. Experimental results confirmed that differences in analyte mass affect the extent of pyrolytic cracking. In pyrograms of the test soil, the ratio of toluene to total product showed that for a sample mass between 1.8 and 2.0 mg, variability in the cracking pattern was minimized. Unlike deviations outside this range, small deviations within the range did not appreciably effect the toluene ratio. A standard curve was prepared for pyrolytic analyses by plotting the total chromatographic area of all pyrolysis products versus the mass of organic material pyrolyzed. These data were fit with a straight line having an 'R²' of 0.73. Based on a bench scale bioremediation experiment, preliminary pyrolysis-GC/MS results were used to predict that compounds derived from lignin and carbohydrates would be degraded faster in uncontaminated than contaminated soils. Appreciable degradation of both compounds, however, occurred in contaminated soils. In addition, results suggested that using pyrolysis-GC/MS to quantify the sum of all n-alkanes and the ratio of odd to even chain n-alkanes could help researchers distinguish between the degradation of petroleum and non-petroleum hydrocarbons in contaminated and uncontaminated soils.     

Use of zeolite to neutralise nickel in a soil environment

Nickel is a heavy metal which is a stable soil pollutant which is difficult to remediate. An attempt to reduce its impact on the environment can be made by changing its solubility. The right level of hydrogen ions and the content of mineral and organic colloids are crucial in this regard. Therefore, methods to neutralise heavy metals in soil are sought. There are no reports in the literature on the possibility of using minerals in the detoxication of a soil environment contaminated with metals. It is important to fill the gap in research on the effect of zeolites on the microbiological, biochemical and physicochemical properties of soils under pressure from heavy metals. Therefore, a pot experiment was conducted on two soils which examined the effect of various levels of contamination of soil with nickel on the activity of soil enzymes, physical and chemical properties and growth and development of plants. An alleviating effect of zeolite Bio.Zeo.S.01 on the negative impact of nickel on the soil and a plant (oats) was examined. The enzyme activity and the oat yield were found to be significantly and negatively affected by an excess of nickel in the soil, regardless of the soil type. The metal was accumulated more in the oat roots than in the above-ground parts. An addition of zeolite decreased the level of accumulation of nickel in oats grown only on sandy-silty loam. Zeolite Bio.Zeo.S.01 used in the study only slightly alleviated the negative effect of nickel on the biochemical properties of soil. Therefore, its usability in the remediation of soil contaminated with nickel is small.     

Metal Contamination in Urban Soils of Coastal Tuscany (Italy)

Urban soils are often contaminated by metals deriving from human activities. Urban polluted soils can affect human health through direct physical contact or through the food chain. The behaviour of metals is affected by chemical and physical soil properties. Heavy metal concentration was determined in some soils of three medium sized towns of coastal Tuscany, Central Italy. Soil samples were collected in roadsides, urban agricultural soils (allotments), playgrounds and public parks. The analysis included total metal content (Pb, Cu, Zn, Ni, Cd), and sequential extraction. Lead reached the highest levels in the soils and was higher near roads. In urban agricultural soils and in allotments Cu was present in noticeable quantities (300 mg kg⁻¹). The presence of Cu in urban soils seems to be typical of soils used for a long period as agricultural land, especially vineyards in the area covered by this study. Sequential extraction was performed to evaluate the mobility of the metals and to better understand the impact of the anthropogenic activity on urban sites. In the present study principal components analysis (PCA) is employed to describe the characteristics of urban soils.     

Petroleum contaminated soil in Oman: Evaluation of bioremediation treatment and potential for reuse in hot asphalt mix concrete

This paper presents a study that aims at evaluating the leaching characteristics of petroleum contaminated soils as well as their application in hot mix asphalt concrete. Soil samples are environmentally characterized in terms of their total heavy metals and hydrocarbon compounds and leachability. The total petroleum hydrocarbon (TPH) present in the PCS before and after treatment was determined to be 6.8% and 5.3% by dry weight, indicating a reduction of 1% in the TPH of PCS due to the current treatment employed. Results of the total heavy metal analysis on soils indicate that the concentrations of heavy metals are lower when extraction of the soil samples is carried out using hexane in comparison to TCE. The results show that the clean soils present in the vicinity of contaminated sites contain heavy metals in the following decreasing order: nickel (Ni), followed by chromium (Cr), zinc (Zn), copper (Cu), lead (Pb), and vanadium (V). The current treatment practice employed for remediation of the contaminated soil reduces the concentrations of nickel and chromium, but increases the concentrations of all remaining heavy metals.     

Restoration of taxonomic and functional genes after bioaugmentation of petroleum contaminated soil

Soil microbial ecosystems are responsive to environmental changes that underpin the biological functions of the soil. The present study was conducted to profile variations in the microbial ecological system of remediated soil (R) and petroleum contaminated soil (P) based on comparisons with soil that had not been contaminated (N), using a cloning library of taxonomic genes (16S rRNA gene for bacteria and 18S rRNA gene for eukaryotes) and functional genes (nifH, amoA and narG). The results showed that N and R had a similar distribution in both the taxonomic genes and functional genes for bacteria and eukaryotes, which were dominated by Proteobacteria and Arthropoda, respectively. Phylogenetic analysis based on the nifH gene showed that the sequences from the three soils were clustered into six taxonomic groups, Actinobacteridae, and Alpha-, Beta-, Gamma- and Delta-proteobacteria, as well as an unclassified group. Evaluation of the amoA gene revealed that all sequences derived from the three samples belonged to Betaproteobacteria. The R and N soil had similar Shannon-Wiener diversity index (H') values, both of which were significantly higher than that of the P soil. The most abundant bacterial phylotype identified in the N and R soils were the same and were related to an uncultured bacterial clone (GAN-SB17, FN423475). None of the narG genes were found in the P soil. Similar results in terms of distribution, composition and the related index were obtained for nifH and amoA. These parameters may comprise a biological ecology index that may be applied to aid the design, implementation and evaluation of soil bioremediation.     

Effect of biosludge and biofertilizer amendment on growth of Jatropha curcas in heavy metal contaminated soils

The pot experiments were conducted to evaluate the effect of different concentrations of arsenic, chromium and zinc contaminated soils, amended with biosludge and biofertilizer on the growth of Jatropha curcas which is a biodiesel crop. The results further showed that biosludge alone and in combination with biofertilizer significantly improved the survival rates and enhanced the growth of the plant. With the amendments, the plant was able to grow and survive upto 500, 250 and 4,000 mg kg(-1) of As, Cr and Zn contaminated soils, respectively. The results also showed that zinc enhanced the growth of J. curcas more as compared to other metals contaminated soils. The heavy metal accumulation in plant increased with increasing concentrations of heavy metals in soil, where as a significant reduction in the metal uptake in plant was observed, when amended with biosludge and biofertilizer and biosludge alone. It seems that the organic matter present in the biosludge acted as metal chelator thereby reducing the toxicity of metals to the plant. Findings suggest that plantation of J. curcas may be promoted in metal contaminated soils, degraded soils or wasteland suitably after amending with organic waste.     

Effect of heavy metal contaminated shooting range soils on mycorrhizal colonization of roots and metal uptake by leek

We grew leek (Allium porrum) in soils of two shooting ranges heavily contaminated with heavy metals in the towns of Zuchwil and Oberuzwil in Switzerland as a bioassay to test theactivity of arbuscular mycorrhizal (AM) fungi in these soils.Soil samples were taken from (1) front of the shooting house(HOUSE), (2) the area between house and target (FIELD) and (3) the berm (BACKSTOP). Samples of Ribwort plantain (Plantagolanceolata) growing naturally within the shooting ranges werealso collected and the colonization of its roots by mycorrhizalfungi was measured. The number of AM spores in the soils wassignificantly reduced concomitant with the increase in thedegree of soil contamination with metals. In Zuchwil,mycorrhizal fungi equally colonized roots of Ribwort plantainsampled from BACKSTOP and HOUSE. In Oberuzwil, however, plantsfrom BACKSTOP had lower colonization when compared with thosesampled from HOUSE. Colonization of leek was strongly reducedin the BACKSTOP soil of Zuchwil and slightly reduced in theBACKSTOP soil of Oberuzwil when compared with plants grown inrespective HOUSE soil. Concentrations of Cd, Cr, Cu, Ni, Pb andZn in the leaves of leek grown in the BACKSTOP soil was withinthe range considered toxic for human consumption. This pointsto the high degree of bio-availability of these metal in thesesoils. Significant decrease in the number of mycorrhizal sporesin the BACKSTOP soils in Zuchwil and the low colonization ofleek roots grown in these soils point to possible changes inthe species diversity of mycorrhizal fungi in these soils.     

Characterization and validation of a Portuguese natural reference soil to be used as substrate for ecotoxicological purposes

This study describes the first attempt to validate a Portuguese natural soil (PTRS1) to be used as reference soil for ecotoxicological purposes, aimed to both: (i) obtain ecotoxicological data for the derivation of Soil Screening Values (SSVs) with regional relevance, acting as a substrate to be spiked with ranges of concentrations of the chemicals under evaluation and (ii) act as control and as substrate for the dilution of contaminated soils in ecotoxicological assays performed to evaluate the ecotoxicity of contaminated soils, in tier 2 of risk assessment frameworks, applied to contaminated lands. The PTRS1 is a cambisol from a granitic area integrated in the Central Iberian Zone. After chemical characterization of the soil in terms of pseudo-total metals, PAHs, PCBs and pesticide contents, it was possible to perceive that some metals (Ba, Be, Co, Cr and V) surpass the Dutch Target Values (Dtvs) corrected for the percentage of organic matter and clay of the PTRS1. Nevertheless, these metals displayed total concentrations below the background total concentrations described for Portuguese soils in general. The same was observed for aldrin, endosulfan I, endosulfan II, heptachlor epoxide, and heptachlor; however the Dtvs corrected become negligible. The performance of invertebrate and plant species, commonly used in standard ecotoxicological assays, was not compromised by both soil properties and soil metal contents. The results obtained suggest that the PTRS1 can be used as a natural reference soil in ecotoxicological assays carried out under the scope of ecological risk assessment.     

Fingerprinting of petroleum hydrocarbons (PHC) and other biogenic organic compounds (BOC) in oil-contaminated and background soil samples

Total petroleum hydrocarbons (TPH) or petroleum hydrocarbons (PHC) are one of the most widespread soil contaminants in Canada, the United States and many other countries worldwide. Clean-up of PHC-contaminated soils costs the Canadian economy hundreds of millions of dollars annually. In Canada, most PHC-contaminated site evaluations are based on the methods developed by the Canadian Council of the Ministers of the Environment (CCME). However, the CCME method does not differentiate PHC from BOC (the naturally occurring biogenic organic compounds), which are co-extracted with petroleum hydrocarbons in soil samples. Consequently, this could lead to overestimation of PHC levels in soil samples. In some cases, biogenic interferences can even exceed regulatory levels (300 μg g(-1) for coarse soils and 1300 μg g(-1) for fine soils for Fraction 3, C(16)-C(34) range, in the CCME Soil Quality Level). Resulting false exceedances can trigger unnecessary and costly cleanup or remediation measures. Therefore, it is critically important to develop new protocols to characterize and quantitatively differentiate PHC and BOC in contaminated soils. The ultimate objective of this PERD (Program of Energy Research and Development) project is to correct the misconception that all detectable hydrocarbons should be regulated as toxic petroleum hydrocarbons. During 2009-2010, soil and plant samples were collected from over forty oil-contaminated and paired background sites in various provinces. The silica gel column cleanup procedure was applied to effectively remove all target BOC from the oil-contaminated sample extracts. Furthermore, a reliable GC-MS method in combination with the derivatization technique, developed in this laboratory, was used for identification and characterization of various biogenic sterols and other major biogenic compounds in these oil-contaminated samples. Both PHC and BOC in these samples were quantitatively determined. This paper reports the characterization results of this set of 21 samples. In general, the presence of petroleum-characteristic alkylated PAH homologues and biomarkers can be used as unambiguous indicators of the contamination of oil and petroleum product hydrocarbons; while the absence of petroleum-characteristic alkylated PAH homologues and biomarkers and the presence of abundant BOC can be used as unambiguous indicators of the predominance of natural organic compounds in soil samples.     

Toxicity assessment for petroleum-contaminated soil using terrestrial invertebrates and plant bioassays

The assessment of soil quality after a chemical or oil spill and/or remediation effort may be measured by evaluating the toxicity of soil organisms. To enhance our understanding of the soil quality resulting from laboratory and oil field spill remediation, we assessed toxicity levels by using earthworms and springtails testing and plant growth experiments. Total petroleum hydrocarbons (TPH)-contaminated soil samples were collected from an oilfield in Sfax, Tunisia. Two types of bioassays were performed. The first assessed the toxicity of spiked crude oil (API gravity 32) in Organization for Economic Co-operation and Development artificial soil. The second evaluated the habitat function through the avoidance responses of earthworms and springtails and the ability of Avena sativa to grow in TPH-contaminated soils diluted with farmland soil. The EC₅₀ of petroleum-contaminated soil for earthworms was 644 mg of TPH/kg of soil at 14 days, with 67 % of the earthworms dying after 14 days when the TPH content reached 1,000 mg/kg. The average germination rate, calculated 8 days after sowing, varied between 64 and 74 % in low contaminated soils and less than 50 % in highly contaminated soils.