Biodegradation Of Total Petroleum Hydrocarbons Research Articles

Bioremediation of petroleum hydrocarbons polluted soil by spent mushroom substrates: Microbiological structure and functionality

Spent mushroom substrate (SMS) holds valuable microbiota that can be useful in remediating polluted soils with hydrocarbons. However, the microorganisms behind the bioremediation process remain uncertain. In this work, a bioremediation assay of total petroleum hydrocarbons (TPHs) polluted soil by SMS application was performed to elucidate the microorganisms and consortia involved in biodegradation by a metabarcoding analysis. Untreated polluted soil was compared to seven bioremediation treatments by adding SMS of Agaricus bisporus, Pleurotus eryngii, Pleurotus ostreatus, and combinations. Soil microbial activity, TPH biodegradation, taxonomic classification, and predictive functional analysis were evaluated in the microbiopiles at 60 days. Different metagenomics approaches were performed to understand the impact of each SMS on native soil microbiota and TPHs biodegradation. All SMSs enhanced the degradation of aliphatic and aromatic hydrocarbons, being A. bisporus the most effective, promoting an efficient consortium constituted by the bacterial families Alcanivoraceae, Alcaligenaceae, and Dietziaceae along with the fungal genera Scedosporium and Aspergillus. The predictive 16 S rRNA gene study partially explained the decontamination efficacy by observing changes in the taxonomic structure of bacteria and fungi, and changes in the potential profiles of estimated degradative genes across the different treatments. This work provides new insights into TPHs bioremediation.

Unveiling the capacity of bioaugmentation application, in comparison with biochar and rhamnolipid for TPHs degradation in aged hydrocarbons polluted soil

Persistent, aged hydrocarbons in soil hinder remediation, posing a significant environmental threat. While bioremediation offers an environmentally friendly and cost-effective approach, its efficacy for complex contaminants relies on enhancing pollutant bioavailability. This study explores the potential of immobilized bacterial consortia combined with biochar and rhamnolipids to accelerate bioremediation of aged total petroleum hydrocarbon (TPH)-contaminated soil. Previous research indicates that biochar and biosurfactants can increase bioremediation rates, while mixed consortia offer sequential degradation and higher hydrocarbon mineralization. The present investigation aimed to assess whether combining these strategies could further enhance degradation in aged, complex soil matrices. The bioaugmentation (BA) with bacterial consortium increased the TPHs degradation in aged soil (over 20% compared to natural attenuation - NA). However, co-application of BA with biochar and rhamnolipid higher did not show a statistically prominent synergistic effect. While biochar application facilitated the maintenance of hydrocarbon degrading bacterial consortium in soil, the present study did not identify a direct influence in TPHs degradation. The biochar application in contaminated soil contributed to TPHs adsorption. Rhamnolipid alone slightly increased the TPHs biodegradation with NA, while the combined bioaugmentation treatment with rhamnolipid and biochar increased the degradation between 27.5 and 29.8%. These findings encourage further exploration of combining bioaugmentation with amendment, like biochar and rhamnolipid, for remediating diverse environmental matrices contaminated with complex and aged hydrocarbons.

Bioaugmentation and vermicompost facilitated the hydrocarbon bioremediation: scaling up from lab to field for petroleum-contaminated soils.

The biodegradation of total petroleum hydrocarbon (TPH) in soil is very challenging due to the complex recalcitrant nature of hydrocarbon, hydrophobicity, indigenous microbial adaptation and competition, and harsh environmental conditions. This work further confirmed that limited natural attenuation of petroleum hydrocarbons (TPHs) (15% removal) necessitates efficient bioremediation strategies. Hence, a scaling-up experiment for testing and optimizing the use of biopiles for bioremediation of TPH polluted soils was conducted with three 500-kg pilots of polluted soil, and respective treatments were implemented: including control soil (CT), bioaugmentation and vermicompost treatment (BAVC), and a combined application of BAVC along with bioelectrochemical snorkels (BESBAVC), all maintained at 40% field capacity. This study identified that at pilot scale level, a successful application of BAVC treatment can achieve 90.3% TPH removal after 90 days. BAVC's effectiveness stemmed from synergistic mechanisms. Introduced microbial consortia were capable of TPH degradation, while vermicompost provided essential nutrients, enhanced aeration, and, potentially, acted as a biosorbent. Hence, it can be concluded that the combined application of BAVC significantly enhances TPH removal compared to natural attenuation. While the combined application of a bioelectrochemical snorkel (BES) with BAVC also showed a significant TPH removal, it did not differ statistically from the individual application of BAVC, under applied conditions. Further research is needed to optimize BES integration with BAVC for broader applicability. This study demonstrates BAVC as a scalable and mechanistically sound approach for TPH bioremediation in soil.

Performance evaluation of electrokinetic bioremediation for weathered petroleum hydrocarbon-contaminated soil

This study investigates the impact of Electrokinetic Remediation (EKR) time on total petroleum hydrocarbons (TPH) removal from soil, focusing on electroosmotic phenomena guided by Helmholtz-Smoluchowski theory. Soil samples were exposed to a constant 2 V/cm voltage gradient for 8, 16, and 24 hours, utilizing a 0.05 M Na2SO4 solution as an electrolyte to enhance soil conductivity. Biostimulation was combined with EKR, and TPH levels and microbial colonies (TPC) were monitored over a 4-week period. Maintaining optimal conditions (25-40°C, pH 6-8, 30-60% water content) was crucial for effective biodegradation. Results showed that a 24-hour EKR duration was most effective, achieving an 87.9% removal rate. The 16-hour duration closely followed at 85.7%, while the 8-hour duration exhibited lower effectiveness at 62.5%. TPC counts increased in the fourth week for the 16-hour and 24-hour durations but decreased for the 8-hour duration. Post-EKR, a gradual decline in TPH levels indicated electroosmotic flow’s (EOF) positive impact on TPH desorption and biodegradation. Electroosmosis influence was evident in varying TPH concentrations among segments after EKR, with higher levels near the cathode in the 16-hour and 24-hour durations. Lower TPH near the cathode in the 8-hour variation suggested electroosmosis-induced desorption, while reduced concentrations in regions A (near anode) and B (middle) resulted from biodegradation and microbial mobility due to electrophoresis. Confirmation of electroosmosis across all durations was supported by observed water content and EOF volume. The 24-hour duration exhibited the highest EOF proportion in the cathode chamber at 22.2%, followed by 16 hours at 16.1%, and 8 hours at 8.6%.

Potential natural attenuation of petroleum hydrocarbons in fuel contaminated soils: Focusing on anaerobic fuel biodegradation involving microbial Fe(III) reduction

Liquid fossil fuels, collectively known as total petroleum hydrocarbons (TPHs), are highly toxic and frequently leak into subsurface environments due to anthropogenic activities. As an in-situ biological remedial option for TPH contamination, aerobic TPH biodegradation is limited due to oxygen's low solubility in water, and because it is consumed quickly by aerobic bacteria. Thus, we investigated the potential of anaerobic TPH degradation by indigenous fermenting bacteria and Fe(III)-reducing bacteria. Twenty 6–10 m soil cores were collected from a closed military base subject to ongoing TPH contamination since the 1980s. Physicochemical and microbial properties were determined at 0.5-m intervals in each core. To assess the relationship between TPH degradation and microbial Fe(III) reduction, soil samples were grouped into high-TPH (>500 mg kg−1) and high-Fe(II) (>450 mg kg−1), high-TPH and low-Fe(II), low-TPH and high-Fe(II), and low-TPH and low-Fe(II) groups. Alpha diversity was significantly lower in high-TPH groups than in low-TPH groups, suggesting that high TPH concentrations exerted a strong selective pressure on bacterial communities. In the high-TPH and low-Fe(II) group, fermenting bacteria, including Microgenomatia and Chlamydiae, were more abundant, suggesting that TPH biodegradation occurred via fermentation. In the high-TPH and high-Fe(II) group, Fe(III)-reducing bacteria, including Geobacter and Zoogloea, were more abundant, suggesting that microbial Fe(III) reduction enhances TPH biodegradation. In contrast, the fermenting and/or Fe(III)-reducing bacteria were not statistically abundant in the low-TPH groups.

Ribosomal RNA gene operon copy number, a functional trait indicating the hydrocarbon degradation level of bacterial communities

The lack of universal indicators for predicting microbial biodegradation potential and assessing remediation effects limits the generalization of bioremediation. The community-level ribosomal RNA gene operon (rrn) copy number, an important functional trait, has the potential to serve as a key indicator of the bioremediation of organic pollutants. A meta-analysis based on 1275 samples from 26 hydrocarbon-related studies revealed a positive relationship between the microbial hydrocarbon biodegradation level and the community-level rrn copy number in soil, seawater and culture. Subsequently, a microcosm experiment was performed to decipher the community-level rrn copy number response mechanism during total petroleum hydrocarbon (TPH) biodegradation. The treatment combining straw with resuscitation-promoting factor (Rpf) exhibited the highest community-level rrn copy number and the most effective biodegradation compared with other treatments, and the initial TPH content (20,000 mg kg-1) was reduced by 67.67% after 77 days of incubation. TPH biodegradation rate was positively correlated with the average community-level rrn copy number (p = 0.001, R2 = 0.5781). Both meta and community analyses showed that rrn copy number may reflect the potential of hydrocarbon degradation and microbial dormancy. Our findings provide insight into the applicability of the community-level rrn copy number to assess bacterial biodegradation for pollution remediation.

Efficacy of bioadmendments in reducing the influence of salinity on the bioremediation of oil-contaminated soil

This study aimed to investigate the potential of three bioamendments (rice husk biochar, wheat straw biochar, and spent mushroom compost) to enhance microbial degradation of crude oil in saline soil. A soil microcosm experiment was conducted, comparing the response of soil microorganisms to crude oil under saline (1 % NaCl) and non-saline conditions. The soils were amended with different bioamendments at varying concentrations (2.5 % or 5 %), and degradation rates were monitored over a 120-day period at 20 °C. The results showed that the bioamendments significantly influenced the degradation of total petroleum hydrocarbons (TPH) in both non-saline and saline soils by 67 % and 18 % respectively. Non-saline soils exhibited approximately four times higher TPH biodegradation compared to saline soils. Among the bioamendments, rice husk biochar and spent mushroom compost had the greatest impact on biodegradation in saline soil, while wheat straw and rice husk biochar combined with spent mushroom compost showed the most significant effects in non-saline soil. The study also revealed that the bioamendments facilitated changes in the microbial community structure, particularly in the treatments with rice husk biochar and wheat straw biochar. Actinomycetes and fungi were found to be more tolerant to soil salinity, especially in the treatments with rice husk biochar and wheat straw biochar. Additionally, the production of CO2, indicating microbial activity, was highest (56 % and 60 %) in the treatments combining rice husk biochar or wheat straw biochar with spent mushroom compost in non-saline soil, while in saline soil rice husk biochar treatment (50 %) was the highest. Overall, this research demonstrates that the application of bioamendments, particularly rice husk biochar and wheat straw biochar combined with spent mushroom compost, can effectively enhance the biodegradation of crude oil in saline soil. These findings highlight the potential of such bioamendments as green and sustainable solutions for soil pollution, especially in the context of climate change-induced impacts on high-salinity soils, including coastal soils.

New insight into the effect of nitrogen on hydrocarbon degradation in petroleum-contaminated soil revealed through 15N tracing and flow cytometry

Nitrogen (N) has been widely used to dissipate total petroleum hydrocarbons (TPH) in the oil-contaminated soil, but the relationships of hydrocarbon transformation, N cycling and utilization, and microbial characteristics during TPH biodegradation still remain unclear. In this study, 15N tracers (K15NO3 and 15NH4Cl) were used as stimulants for TPH degradation to compare the bioremediation potential of TPH in the historically (5 a) and freshly (7 d) petroleum-contaminated soils. During bioremediation process, TPH removal and carbon balance, N transformation and utilization, as well as microbial morphologies were investigated using 15N tracing and flow cytometry. Results showed that TPH removal rates were higher in the freshly polluted soils (61.59 % for K15NO3 amendment and 48.55 % for 15NH4Cl amendment) than in the historically polluted soils (35.84 % for K15NO3 amendment and 32.30 % for 15NH4Cl amendment), and TPH removal rate through K15NO3 amendment was higher than that of 15NH4Cl in the freshly polluted soils. This result was attributed to the higher N gross transformation rates in the freshly contaminated soils (0.0034–0.432 mmol N kg−1 d−1) when compared with that in the historically contaminated soils (0.009–0.04 mmol N kg−1 d−1), which led to more TPH transformation to residual carbon (51.84 %–53.74 %) in the freshly polluted soils than that in the historically polluted soils (24.67 %–33.47 %). Based on the fluorescence intensity displayed by the combination of stains and cellular components to indicate microbial morphology and activity, flow cytometry analysis showed that nitrogen addition was beneficial for the membrane integrity of TPH-degrading bacteria, and nitrogen also enhanced DNA synthesis and activity of TPH-degrading fungi in freshly polluted soil. Correlation and structural equation modeling analysis identified that K15NO3 was beneficial to synthesize DNA of the TPH-degrading fungi but not the bacteria, which contributed to enhance TPH bio-mineralization in the soils with K15NO3 amendment.

MODELLING OF THE TOTAL PETROLEUM HYDROCARBON (TPH) DEGRADATION DURING THE BIOREMEDIATION OF CRUDE OIL POLLUTED SOIL USING COW DUNG AND WIRE CROTONS

The contamination of soil due to activities involved in crude oil exploration and production, and oil spillage emanating from pipeline vandalization, sabotage, theft, corrosion of pipeline and the enterprises of artisanal refineries pose health and ecological problems. This is especially as crude oil contains chemical compounds which are grouped into the Total Petroleum Hydrocarbons (TPH), and known to be toxic, mutagenic, and carcinogenic. Bioremediation is currently being explored as a cost effective and environmentally friendly methods of remediating the soil from this contaminant. Thus, the current work is aimed at developing a mathematical model for the bioremediation process. This was carried out while assuming that the process of degradation is a first order reaction, and the principle of the law of conservation of mass was applied. A linear model ( ) was developed relating the instantaneous concentration of TPH, initial concentration of TPH and time duration of the bioremediation. The model fitted well to the experimental data as shown by the result of the statistical analysis where the Analysis of Variance (ANOVA) showed a p-value of 0.004 suggesting that the model is statistically significant. The p-values of the model variables such as time duration of the bioremediation was 0.004, this suggested that the variable is significant. The model can be used as a supporting tool in the management of the biodegradation of TPH to predict the degradation concentration of TPH during the bioremediation process at ambient condition of temperature and pressure using cow dung, wire croton and their combination as a carbon source. KEYWORDS: Bioremediation; cow dung; wire croton; degradation; Mathematical modelling

Field scale biodegradation of total petroleum hydrocarbons and soil restoration by Ecopiles: microbiological analysis of the process.

Ecopiling is a method for biodegradation of hydrocarbons in soils. It derives from Biopiles, but phytoremediation is added to biostimulation with nitrogen fertilization and bioaugmentation with local bacteria. We have constructed seven Ecopiles with soil heavily polluted with hydrocarbons in Carlow (Ireland). The aim of the study was to analyze changes in the microbial community during ecopiling. In the course of 18 months of remediation, total petroleum hydrocarbons values decreased in 99 and 88% on average for aliphatics and aromatics, respectively, indicating a successful biodegradation. Community analysis showed that bacterial alfa diversity (Shannon Index), increased with the degradation of hydrocarbons, starting at an average value of 7.59 and ending at an average value of 9.38. Beta-diversity analysis, was performed using Bray-Curtis distances and PCoA ordination, where the two first principal components (PCs) explain the 17 and 14% of the observed variance, respectively. The results show that samples tend to cluster by sampling time instead of by Ecopile. This pattern is supported by the hierarchical clustering analysis, where most samples from the same timepoint clustered together. We used DSeq2 to determine the differential abundance of bacterial populations in Ecopiles at the beginning and the end of the treatment. While TPHs degraders are more abundant at the start of the experiment, these populations are substituted by bacterial populations typical of clean soils by the end of the biodegradation process. Similar results are found for the fungal community, indicating that the microbial community follows a succession along the process. This succession starts with a TPH degraders or tolerant enriched community, and finish with a microbial community typical of clean soils.

Remediation of petroleum contaminated soil by persulfate oxidation coupled with microbial degradation

This study was conducted on soil polluted with high concentrations of crude oil (12,835 ± 572.76 mg/kg) with different dosages of persulfate (PS) coupled with hydrocarbon-degrading mixed bacteria (including Enterobacteriaceae, Stenotrophomonas, Pseudomonas, Acinetobacter, and Achromobacter) obtained from long-term oil-contaminated soil. The interaction among total petroleum hydrocarbons (TPH), soil parameters, and microbial communities during a 187-d combined remediation process was evaluated. The result showed that the TPH removal of 1% PS oxidation combined with bioremediation was 80.05%; this was 4.88% and 20.94% higher than that of bioremediation and 1% PS oxidation combined with natural attenuation. This is a result of 1%PS stimulated enzyme secretion and dissolved organic carbon content, especially fulvic acid, improving TPH mobility and bioavailability in soil. Furthermore, introducing mixed bacteria after oxidation further increased enzyme activities and TPH-degrading ability, thus promoting carbon substrate conversion and biodegradation rate. Conversely, the excessive oxidation of 3% and 5% PS had a negative impact on the soil environment and microbial diversity resulting in lower TPH degradation (36%-57%). High-throughput sequencing revealed that the abundance and diversity of soil bacteria decreased in all remediation groups compared to those in the pristine soil. Redundancy analysis showed that the main factors affecting 1% PS oxidation combined with bioremediation during subsequent microbial remediation were pH, cation exchange capacity, and lipase activity, which may indirectly influence TPH biodegradation by increasing the abundance of genus Immundisolibacteraceae, Comamonadaceae, and Acinetobacter. This study provides data on in situ chemical oxidation integrated with the bioremediation of petroleum hydrocarbon-contaminated soils.

Biostimulation of Petroleum-Contaminated Soil Using Organic and Inorganic Amendments.

The most common approaches for the in-situ bioremediation of contaminated sites worldwide are bioaugmentation and biostimulation. Biostimulation has often proved more effective for chronically contaminated sites. This study examined the effectiveness of optimized water hyacinth compost in comparison with other organic and inorganic amendments for the remediation of crude oil-polluted soils. Water hyacinth was found to be rich in nutrients necessary to stimulate microbial growth and activity. An organic geochemical analysis revealed that all amendments in this study increased total petroleum hydrocarbon (TPH) biodegradation by ≥75% within 56 days, with the greatest biodegradation (93%) occurring in sterilized soil inoculated with optimized water hyacinth compost. This was followed by polluted soil amended with a combination of spent mushroom and water hyacinth composts (SMC + WH), which recorded a TPH biodegradation of 89%. Soil amendment using the inorganic fertilizer NPK (20:10:10) resulted in 86% TPH biodegradation. On the other hand, control samples (natural attenuation) recorded only 4% degradation. A molecular analysis of residual polycyclic aromatic hydrocarbons (PAHs) showed that the 16 PAHs designated by the US EPA as priority pollutants were either completely or highly degraded in the combined treatment (SMC + WH), indicating the potential of this amendment for the environmental remediation of soils contaminated with recalcitrant organic pollutants.

Total petroleum hydrocarbons and influencing factors in co-composting of rural sewage sludge and organic solid wastes

Co-composting is an efficient strategy for collaborative disposal of multiple organic wastes in rural areas. In this study, we explored the co-composting of rural sewage sludge and other organic solid wastes (corn stalks and kitchen waste), with a focus on the variation of total petroleum hydrocarbons (TPH) during this process. 12% corn-derived biochar was applied in the composting (BC), with no additives applied as the control treatment (CK). The TPH contents of piles after composting ranged from 0.70 to 0.74 mg/g, with overall removal efficiencies of 35.6% and 61.1% for CK and BC, respectively. The results indicate that the addition of 12% biochar increased the rate of TPH degradation and accelerated the degradation process. 16s rDNA high-throughput sequencing was applied to investigate the biodiversity and bacterial community succession during the composting process. Diverse bacterial communities with TPH degradation functions were observed in the composting process, including Acinetobacter, Flavobacterium, Paenibacillus, Pseudomonas, and Bacillus spp. These functional bacteria synergistically degraded TPH, with cooperative behavior dominating during composting. Biochar amendment enhanced the microbial activity and effectively promoted the biodegradation of TPH. The physicochemical properties of the compost piles, including environmental factors (pH and temperature), nutrients (nitrogen, phosphorus, potassium), and humic substances produced in composting (humic acids and fulvic acids), directly and indirectly affected the variation in TPH contents. In conclusion, this work illustrates the variation in TPH content and associated influencing factors during co-composting of rural organic solid wastes, providing valuable guidance toward the further optimization of rural organic waste management.

Monitoring TPH biodegradation in soil around Ray oil refinery by natural attenuation, biostimulation and bioaugmentation treatments

In this study, total petroleum hydrocarbons (TPHs) decontaminating mechanisms for soils around the Rey refinery complex (South of Tehran, Iran) was investigated. Natural attenuation, biostimulation and bioaugmentation (separately and in combination) methods were evaluated TPHs and soil microbial respiration in 210 days, using laboratory treatments The modified methods were applied through 13 different treatments, including improving the environmental conditions for native bacteria (natural attenuation for treatments 1-8), adding non-native bacterial complex (bioaugmentation for treatment 9) and intensifying and stimulating growth while adding non-native bacterial complex (biostimulation-bioaugmentation for treatments 10-13). Although, overall of the treatments, a significant decreasing TPHs concentration were observed over the time, biostimulation-bioaugmentation treatments had the highest amount of TPHs decomposition, the highest rate of bio-respiration, the lowest half-life times (t1/2), and the highest remediation efficiency and biodegradation constants rate. Among natural attenuation treatments, modifiers with manure and sawdust had the greatest effect on reducing the TPHs concentration and the highest rate of bio-respiration. The first-order kinetic model was fitted to the data related to biodegradation in a satisfactory manner. The results showed that there was a strong and positive linear correlation between decreasing TPHs concentration and microbial respiration in all modifiers. Although for the bacterial treatments and at the early stages of inoculation, the rate of total respiration was low, but as the time passed and with adaptation of the effective inoculated bacteria to contaminated soil, the respiration rate gradually was increased. Due to its low cost and low environmental risk, the proposed bioremediation technique for oil contaminated soil can be recommended to the region.

Plant-derived saponin enhances biodegradation of petroleum hydrocarbons in the rhizosphere of native wild plants

Plant-derived saponins are bioactive surfactant compounds that can solubilize organic pollutants in environmental matrices, thereby facilitating pollutant remediation. Externally applied saponin has potential to enhance total petroleum hydrocarbon (TPH) biodegradation in the root zone (rhizosphere) of wild plants, but the associated mechanisms are not well understood. For the first time, this study evaluated a triterpenoid saponin (from red ash leaves, Alphitonia excelsa) in comparison to a synthetic surfactant (Triton X-100) for their effects on plant growth and biodegradation of TPH in the rhizosphere of two native wild species (a grass, Chloris truncata, and a shrub, Hakea prostrata). The addition of Triton X-100 at the highest level (1000 mg/kg) in the polluted soil significantly hindered the plant growth (reduced plant biomass and photosynthesis) and associated rhizosphere microbial activity in both the studied plants. Therefore, TPH removal in the rhizosphere of both plant species treated with the synthetic surfactant was not enhanced (at the lower level, 500 mg/kg soil) and even slightly decreased (at the highest level) compared to that in the surfactant-free (control) treatment. By contrast, TPH removal was significantly increased with saponin application (up to 60% in C. truncata at 1000 mg/kg due to enhanced plant growth and associated rhizosphere microbial activity). No significant difference was observed between the two saponin application levels. Dehydrogenase activity positively correlated with TPH removal (p < 0.001) and thus this parameter could be used as an indicator to predict the rhizoremediation efficiency. This work indicates that saponin-amended rhizoremediation could be an environmentally friendly and effective biological approach to remediate TPH-polluted soils. It was clear that the enhanced plant growth and rhizosphere microbial activity played a crucial role in TPH rhizoremediation efficiency. The saponin-induced molecular processes that promoted plant growth and soil microbial activity in the rhizosphere warrant further studies.

Efficient removal of heavily oil-contaminated soil using a combination of fenton pre-oxidation with biostimulated iron and bioremediation

Crude oil contamination severely deteriorates soils quality. Bioremediation utilizing soil indigenous organisms could be employed to decompose petroleum hydrocarbons thanks to its low cost and minor environmental disturbance. However, slow kinetics limit the successful application of this biotechnique. Pretreating oil-contaminated soils with Fenton pre-oxidation could accelerate the subsequent bioremediation process. This study was to explore the mechanisms behind the rapid propagation of indigenous petroleum-degrading bacteria (IPDB) and the efficient degradation of total petroleum hydrocarbons (TPH) in soil after Fenton pre-oxidation with biostimulated iron. Biostimulated iron and non-biostimulated iron were used in the experiments, where Fenton pre-oxidation was combined with the bioremediation of oil-contaminated soil (TPH = 13221 mg/kg). Although the amount of Fenton pre-oxidized TPH (3331–3775 mg/kg) was similar with biostimulated and non-biostimulated irons, the biodegradation of TPH after Fenton pre-oxidation with biostimulated iron (5840 mg/kg) was much higher than that with non-biostimulated iron (3034–4034 mg/kg). Moreover, abundant nutrients and a high population of residual IPDB were found after Fenton pre-oxidation with biostimulated iron, which benefited stable consumption of NH3–N and dissolved organic carbon (DOC) by IPDB during the subsequent bioremediation. However, Fenton pre-oxidation with non-biostimulated iron either resulted in greater damage to IPDB or produced fewer nutrients, thereby failing to ensure the continuous propagation of IPDB during the subsequent bioremediation. Therefore, we propose that Fenton pre-oxidation with biostimulated iron should be applied to heavily oil-contaminated soils prior to bioremediation.

Field Test of In Situ Groundwater Treatment Applying Oxygen Diffusion and Bioaugmentation Methods in an Area with Sustained Total Petroleum Hydrocarbon (TPH) Contaminant Flow

Contamination of groundwater by petroleum hydrocarbons is a widespread environmental problem in many regions. Contamination of unsaturated and saturated zones could also pose a significant risk to human health. The main purpose of the study was to assess the efficiency of biodegradation of total petroleum hydrocarbon (TPH) in situ, in an area with loam and sandy loam soils, and to identify features and characteristics related to groundwater treatment in an area with a persistent flow of pollutants. We used methods of biostimulation (oxygen as stimulatory supplement) and bioaugmentation to improve water quality. Oxygen was added to the groundwater by diffusion through silicone tubing. The efficiency of groundwater treatment was determined by detailed monitoring. Implementation of the applied measure resulted in an average reduction in TPH concentration of 73.1% compared with the initial average concentration (4.33 mg/L), and in the local area, TPH content was reduced by 95.5%. The authors hope that this paper will contribute to a better understanding of the topic of groundwater treatment by in situ biodegradation of TPH. Further studies on this topic are particularly needed to provide more data and details on the efficiency of groundwater treatment under adverse geological conditions.

Reduction of the soil environmental impact caused by the presence of total petroleum hydrocarbons (TPH) by using Pseudomonas sp.

This research focuses on the bioaugmentation with Pseudomonas sp. (native) and Pseudomonas aeruginosa (commercial) for the biodegradation of total petroleum hydrocarbons (TPH) of the environmental soil samples of the AqLab laboratory in Orellana, Ecuador. Two treatments of sterilized soil (one inoculated with the native strain and the other inoculated with the commercial strain), where used for physical- -chemical analyzes as well as the degradation of TPH. They were evaluated every 20 days for a total period of 80–100 days. The native bacterium was isolated from the laboratory agglomerates in a selective culture medium specific for Pseudomonas sp. The biodegradation of the TPH exhibited a positive result after 80 and 100 days of treatment, with a reduction of 84 and 96% of initial TPH after the bacterial inoculation. The comparison between the two strains evaluated, commercial and native, showed a greater efficiency of biodegradation by the native strain isolated directly from the agglomerates, suggesting working with native strains of the place that have a greater adaptability to the contaminated environment that would ensure bioremediation processes faster and more efficient, low cost and environmentally friendly.

Evaluation of the Effectiveness of the Biopreparation in Combination with the Polymer γ-PGA for the Biodegradation of Petroleum Contaminants in Soil.

Biodegradation is a method of effectively removing petroleum hydrocarbons from the natural environment. This research focuses on the biodegradation of aliphatic hydrocarbons, monoaromatic hydrocarbons such as benzene, toluene, ethylbenzene, and all three xylene isomers (BTEX) and polycyclic aromatic hydrocarbons (PAHs) as a result of soil inoculation with a biopreparation A1 based on autochthonous microorganisms and a biopreparation A1 with the addition of γ-PGA. The research used biopreparation A1 made of the following strains: Dietzia sp. IN133, Gordonia sp. IN138 Mycolicibacterium frederiksbergense IN53, Rhodococcus erythropolis IN119, Rhodococcus sp. IN136 and Pseudomonas sp. IN132. The experiments were carried out in laboratory conditions (microbiological tests, respirometric tests, and in semi-technical conditions (ex-situ prism method). The biodegradation efficiency was assessed on the basis of respirometric tests, chromatographic analyses and toxicological tests. As a result of inoculation of AB soil with the biopreparation A1 within 6 months, a reduction of total petroleum hydrocarbons (TPH) (66.03%), BTEX (80.08%) and PAHs (38.86%) was achieved and its toxicity was reduced. Inoculation of AB soil with the biopreparation A1 with the addition of γ-PGA reduced the concentration of TPH, BTEX and PAHs by 79.21%, 90.19%, and 51.18%, respectively, and reduced its toxicity. The conducted research has shown that the addition of γ-PGA affects the efficiency of the biodegradation process of petroleum pollutants, increasing the degree of TPH biodegradation by 13.18%, BTEX by 10.11% and PAHs by 12.32% compared to pure biopreparation A1.

A highly efficient method with low energy and water consumption in biodegradation of total petroleum hydrocarbons of oily sludge

The hydrocarbons in petroleum sludge are environmental pollutants. It is crucial to eliminate this type of pollution. In this study, a comprehensive and operational study has been conducted on the total petroleum hydrocarbons (TPH) biodegradation in oily sludge. The experiments were performed in a semi-solid phase by two degrader bacterium, Arthrobacter citreus and Rhodococcus jostii, and they were compared with the slurry phase. Solid samples were prepared in three mixing modes of oily sludge with clay. Experiments were conducted by semi-solid bioreactors and other methods on samples contaminated with petroleum hydrocarbons. The performance of the semi-solid bioreactor for the removal of clay-free oily sludge samples showed the best results, in which biodegradation of TPH was 90.33%. GC analyses were conducted on samples before and after biodegradation. It was observed that the contaminants were decomposed uniformly by the microorganisms, except for a combination with a large peak in 12 min. For the best case, the GC-MS test was performed before and after biodegradation. Compounds with a high concentration in the sludge were significantly reduced. Only one heavy aromatic compound was detected in 51.628 min, which decomposed quite slowly and produced a large peak. It was found that 78.2% of the compounds were removed completely.