Unlocking bioremediation potential for site restoration: A comprehensive approach for crude oil degradation in agricultural soil and phytotoxicity assessment.

Crude oil contamination has inflicted severe damage to soil ecosystems, necessitating effective remediation strategies. This study aimed to compare the efficacy of four different techniques (biostimulation, bioaugmentation, bioaugmentation + biostimulation, and natural attenuation) for remediating agricultural soil contaminated with crude oil using soil microcosms. A consortium of previously characterized bacteria Xanthomonas boreopolis, Microbacterium schleiferi, Pseudomonas aeruginosa, and Bacillus velezensis was constructed for bioaugmentation. The microbial count for the constructed consortium was recorded as 2.04 ± 0.11 × 108 CFU/g on 60 d in augmented and stimulated soil samples revealing their potential to thrive in chemically contaminated-stress conditions. The microbial consortium through bioaugmentation + biostimulation approach resulted in 79 ± 0.92% degradation of the total polyaromatic hydrocarbons (2 and 3 rings âˆ¼ 74%, 4 and 5 rings âˆ¼ 83% loss) whereas, 91 ± 0.56% degradation of total aliphatic hydrocarbons (C8-C16 ∼ 90%, C18-C28 ∼ 92%, C30 to C40 ∼ 88% loss) was observed in 60 d. Further, after 60 d of microcosm treatment, the treated soil samples were used for phytotoxicity assessment using wheat (Triticum aestivum), black chickpea (Cicer arietinum), and mustard (Brassica juncea). The germination rates for wheat (90%), black chickpea (100%), and mustard (100%) were observed in 7 d with improved shoot-root length and biomass in both bioaugmentation and biostimulation approaches. This study projects a comprehensive approach integrating bacterial consortium and nutrient augmentation strategies and underscores the vital role of innovative environmental management practices in fostering sustainable remediation of oil-contaminated soil ecosystems. The formulated bacterial consortium with a nutrient augmentation strategy can be utilized to restore agricultural lands towards reduced phytotoxicity and improved plant growth.

Holistic approach to waste mobil oil bioremediation: Valorizing waste through biosurfactant production for soil restoration.

The combustion of mobil oil leads to the emission of toxic compounds in the environment. In this study, the aromatic and aliphatic hydrocarbon fractions present in a waste mobil oil collected from automobile market were comprehensively identified and their toxicity was evaluated using wheat grain. Lysinibacillus sphaericus strain IITR51 isolated and characterized previously could degrade 30-80% of both aliphatic and aromatic hydrocarbons in liquid culture. Interestingly, the strain IITR51 produced 627 mg/L of rhamnolipid biosurfactant by utilizing 3% (v/v) of waste mobil oil in the presence of 1.5% glycerol as additional carbon source. In a soil microcosm study by employing strain IITR51, 50-86% of 3-6 ring aromatic hydrocarbons and 63-98% of aliphatic hydrocarbons (C8 to C22) were degraded. Addition of 60 µg/mL rhamnolipid biosurfactant enhanced the degradation of both aliphatic and aromatic hydrocarbons from 76.88% to 61.21%-94.11% and 78.27% respectively. The degradation of mobil oil components improved the soil physico-chemical properties and increased soil fertility to 64% as evident by the phytotoxicity assessments. The findings indicate that strain IITR51 with degradation capability coupled with biosurfactant production could be a candidate for restoring hydrocarbon contaminated soils.

Bioremediation of polycyclic aromatic hydrocarbons in crude oil by bacterial consortium in soil amended with Eisenia fetida and rhamnolipid.

The present study investigated the concerted effort of Eisenia fetida and rhamnolipid JBR-425 in combination with a five-member bacterial consortium exhibiting elevated degradation levels of low and high molecular weight polycyclic aromatic hydrocarbons (PAH) from soil contaminated with Digboi crude oil. Application of bacterial consortium (G2) degraded 30-89% of selected PAH from the artificial soil after a 45-day post-exposure, in which chrysene showed the highest level of degradation with 89% and benzo(a)pyrene is the lowest with 30%, respectively. Moreover, an acute exposure study observed that earthworm biomass decreased, and mortality rates increased with increasing crude oil concentrations (0.25 to 2%). Earthworms with a 100% survival rate at 1% crude oil exposure suggest the tolerance potential and its mutual involvement in the bioremediation of crude oil with selected bacterial consortia. Bacterial consortium assisted with E. fetida (G3) showed 98% chrysene degradation with a slight change in benzo(a)pyrene degradation (35%) in crude oil spiked soil. Besides, the most dominant PAH in crude oil found in the current work, fluoranthene, undergoes 93% and 70% degradation in G3 and G5 groups, respectively. However, rhamnolipid JBR-425 coupled with the bacterial consortium (G5) has resulted in 97% degradation of chrysene and 33% for benzo(a)pyrene. Overall, bacterial consortium assisted with earthworm group has shown better degradation of selected PAH than bacterial consortium with biosurfactant. Catalase (CAT), glutathione reductase (GST) activity and MDA content was found to be reduced in earthworms after sub-lethal exposure, suggesting oxidative stress prevalence via reactive oxygen species (ROS). Hence, the findings of the present work suggest that the application of a bacterial consortium, along with earthworm E. fetida, has huge potential for field restoration of contaminated soil with PAH and ecosystem sustainability.

Assessing the half-life and degradation kinetics of aliphatic and aromatic hydrocarbons by bacteria isolated from crude oil contaminated soil.

Pollution from the oil industries and refineries has worsened various environmental compartments. In this study, indigenous oil degrading bacteria were isolated from crude oil obtained from an Oil and Natural Gas Corporation (ONGC) asset in Ankleshwar, Gujarat, India. Based on 16S rRNA phylogeny, they were identified as Pseudomonas boreopolis IITR108, Microbacterium schleiferi IITR109, Pseudomonas aeruginosa IITR110, and Bacillus velezensis IITR111. The strain IITR108, IITR109, IITR110, and IITR111 showed 80-89% and 71-78% degradation of aliphatic (C8-C40) and aromatic (4-5 ring) hydrocarbons respectively in 45 d when supplemented with 3% (v/v) waste crude oil. When compared to individual bacteria, the consortium degrades 93.2% of aliphatic hydrocarbons and 85.5% of polyaromatic hydrocarbons. It was observed that the total aliphatic and aromatic content of crude oil 394,470 µg/mL and 47,050 µg/mL was reduced up to 9617.75 µg/mL and 4586 µg/mL respectively in 45 d when consortium was employed. The rate kinetics analysis revealed that the biodegradation isotherm followed first order kinetics, with a linear correlation between concentration (hydrocarbons) and time intervals. The half-life of aliphatic (C8-C40) and aromatic hydrocarbons ranged from 200 to 453 h and 459-714 h respectively. All the bacteria efficiently produced catabolic enzymes such as alkane monooxygenase, alcohol dehydrogenase, and lipase during the degradation of crude oil. These findings indicated that the bacterial consortium can be a better candidate for bioremediation and reclamation of aliphatic and aromatics hydrocarbon contaminated sites.

Characterization and properties of the biosurfactant produced by PAH-degrading bacteria isolated from contaminated oily sludge environment.

The aim of the present study was to investigate biosurfactant production ability of five different polyaromatic hydrocarbon (PAH)-metabolizing bacteria, such as Ochrobactrum anthropi IITR07, Pseudomonas mendocina IITR46, Microbacterium esteraromaticum IITR47, Pseudomonas aeruginosa IITR48, and Stenotrophomonas maltophilia IITR87. These bacteria showed biosurfactant production using 2% glucose as rich substrate; strain IITR47 yielded the highest with 906 and 534 mg/L biosurfactant in the presence of naphthalene and crude oil as the unique carbon sources. P. aeruginosa IITR48 showed the least surface tension at 29 N/m and the highest emulsification index at 63%. The biosurfactants produced were identified as glycolipid and rhamnolipid based on Fourier transform infrared spectroscopy analysis. In particular, the biosurfactant produced by bacteria S. maltophilia IITR87 efficiently emulsified mustard oil with an E24 value of 56%. It was observed that, all five biosurfactants from these degrader strains removed 2.4-, 1.7-, 0.9-, 3.8-, and 8.3-fold, respectively, crude oil from contaminated cotton cloth. Rhamnolipid derived from IITR87 was most efficient, exhibiting highest desorption of crude oil. These biosurfactants exhibited good stability without significantly losing its emulsification ability under extreme conditions, thus can be employed for bioremediation of PAHs from diverse contaminated ecosystem. Graphical Abstract.

Improved polycyclic aromatic hydrocarbon degradation in a crude oil by individual and a consortium of bacteria.

In this study, we report the ability of Stenotrophomonas maltophilia, Ochrobactrum anthropi, Pseudomonas mendocina, Microbacterium esteraromaticum and Pseudomonas aeruginosa to degrade multiple polycyclic aromatic hydrocarbons (PAHs) present in crude oil. The PAHs in the crude oil sample obtained from Digboi oil refinery, India were estimated to be naphthalene (10.0 mg L-1), fluorene (1.9 mg L-1), phenanthrene (3.5 mg L-1) and benzo(b)fluoranthene (6.5 mg L-1). Exposure of individual bacteria to crude oil showed high rate of biodegradation of specific PAHs by M. esteraromaticum, 81.4%-naphthalene; P. aeruginosa, 67.1%-phenanthrene and 61.0%-benzo(b)fluoranthene; S. maltophilia, 47.9%-fluorene in 45 days. However, consortium of these bacteria showed enhanced biodegradation of 89.1%-naphthalene, 63.8%-fluorene, 81% of phenanthrene and 72.8% benzo(b)fluoranthene in the crude oil. The degradation was further improved up to 10% by consortium on addition of 40 µg mL-1 rhamnolipid JBR-425 biosurfactant. These results suggest that the developed bacterial consortium has significant potential in PAH remediation.