Biodegradation is one of the most effective and profitable methods for the elimination of toxic polychlorinated biphenyls (PCBs) and total petroleum hydrocarbons (TPH) from the environment. In this study, aerobic degradation of the mentioned pollutants by bacterial strains Mycolicibacterium frederiksbergense IN53, Rhodococcus erythropolis IN129, and Rhodococcus sp. IN306 and mixed culture M1 developed based on those strains at 1:1:1 ratio was analyzed. The effectiveness of individual strains and of the mixed culture was assessed based on carried out respirometric tests and chromatographic analyses. The Rhodococcus sp. IN306 turned out most effective in terms of 18 PCB congeners biodegradation (54.4%). The biodegradation index was decreasing with an increasing number of chlorine atoms in a molecule. Instead, the Mycolicobacterium frederiksbergense IN53 was the best TPH degrader (37.2%). In a sterile soil, contaminated with PCBs and TPH, the highest biodegradation effectiveness was obtained using inoculation with mixed culture M1, which allowed to reduce both the PCBs (51.8%) and TPH (34.6%) content. The PCBs and TPH biodegradation capacity of the defined mixed culture M1 was verified ex-situ with prism method in a non-sterile soil polluted with aged petroleum hydrocarbons (TPH) and spent transformer oil (PCBs). After inoculation with mixed culture M1, the PCBs were reduced during 6 months by 84.5% and TPH by 70.8% as well as soil toxicity was decreased.

Biological methods are currently the most commonly used methods for removing hazardous substances from land. This research work focuses on the remediation of oil-contaminated land. The biodegradation of aliphatic hydrocarbons and PAHs as a result of inoculation with biopreparations B1 and B2 was investigated. Biopreparation B1 was developed on the basis of autochthonous bacteria, consisting of strains Dietzia sp. IN118, Gordonia sp. IN101, Mycolicibacterium frederiksbergense IN53, Rhodococcus erythropolis IN119, Rhodococcus globerulus IN113 and Raoultella sp. IN109, whereas biopreparation B2 was enriched with fungi, such as Aspergillus sydowii, Aspergillus versicolor, Candida sp., Cladosporium halotolerans, Penicillium chrysogenum. As a result of biodegradation tests conducted under ex situ conditions for soil inoculated with biopreparation B1, the concentrations of TPH and PAH were reduced by 31.85% and 27.41%, respectively. Soil inoculation with biopreparation B2 turned out to be more effective, as a result of which the concentration of TPH was reduced by 41.67% and PAH by 34.73%. Another issue was the phytoremediation of the pre-treated G6-3B2 soil with the use of Zea mays. The tests were carried out in three systems (system 1—soil G6-3B2 + Zea mays; system 2—soil G6-3B2 + biopreparation B2 + Zea mays; system 3—soil G6-3B2 + biopreparation B2 with γ-PGA + Zea mays) for 6 months. The highest degree of TPH and PAH reduction was obtained in system 3, amounting to 65.35% and 60.80%, respectively. The lowest phytoremediation efficiency was recorded in the non-inoculated system 1, where the concentration of TPH was reduced by 22.80% and PAH by 18.48%. Toxicological tests carried out using Phytotoxkit^TM, Ostracodtoxkit^TM and Microtox^® Solid Phase tests confirmed the effectiveness of remediation procedures and showed a correlation between the concentration of petroleum hydrocarbons in the soil and its toxicity. The results obtained during the research indicate the great potential of bioremediation practices with the use of microbial biopreparations and Zea mays in the treatment of soils contaminated with petroleum hydrocarbons.

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.

Phytoremediation supported by bioaugmentation is a promising method considered for cleaning up areas polluted with petroleum hydrocarbons. In this study, phytoremediation was carried out using Echinacea purpurea as a phytoremediant on two types of soil: Soil DW—aged soil taken from an excavation pit, Soil OS—soil taken from an oil spill area. The tests for each soil were carried out in six test systems (non-inoculation, inoculation with the B1 microbial consortium, inoculation with the B2 microbial consortium, inoculation with the B1 microbial consortium with the addition of γ-PGA (γ-poly glutamic acid), inoculation with the B2 microbial consortium with the addition of γ-PGA and inoculation with the γ-PGA solution) for 6 months. The effectiveness of the remediation treatments used was assessed based on chromatographic analyses of soil and plant material (roots, shoots) and toxicological analyses using four types of toxicological tests (Phytotoxkit^TM (MicroBioTests Inc., Gent, Belgium), Ostracodtoxkit^TM (MicroBioTests Inc., Gent, Belgium), Microtox^® Solid Phase Test (Modern Water Inc., New Castle, DE, USA), MARA (NCIMB Ltd., Aberdeen, UK)). The research conducted showed that the most effective method of bioremediation of soils contaminated with petroleum hydrocarbons was phytoremediation supported by bioaugmentation with the microbial consortium B2 with γ-PGA, which allowed for reducing the concentration of total petroleum hydrocarbons (TPHs) and polycyclic aromatic hydrocarbons (PAHs) in the tested soils by 53.98% and 49.54% (Soil DW-5) and 60.47% and 37.55% (Soil OS-5), respectively. In turn, the lowest bioremediation efficiency was recorded in non-inoculated systems, for which the concentration of TPHs and PAHs at the end of the study decreased by 18.40% and 16.14% (Soil DW-1) and 21.87% and 18.20% (Soil OS-1), respectively. The results of toxicological analyses confirmed the relationship between the concentration of TPHs and PAHs in the soil and its toxicity level.

W artykule przedstawione zostały zagadnienia związane z opracowaniem metodyki chromatograficznego oznaczania wielopierścieniowych węglowodorów aromatycznych w próbkach gleb. Do tego celu stosowano chromatograf cieczowy serii Vanquish Core firmy Thermo Scientific wyposażony w detektor fluorescencyjny (FLD) i detektor UV-ViS. W ramach prowadzonych badań określono optymalne parametry pracy chromatografu, takie jak: objętość próbki, rodzaj eluentów, szybkość przepływu eluentów, gradient eluentów w trakcie analizy chromatograficznej i czas analizy, oraz dobrano kolumnę chromatograficzną umożliwiającą selektywny rozdział analitów. Następnie wykonano kalibrację układu chromatograficznego i walidację metody analitycznej oznaczania WWA, stosując roztwory kalibracyjne opracowane na podstawie roztworu wzorcowego PAH-Mix nr ref. 722393 firmy Macherey-Nagel. Walidacja metody analitycznej obejmowała wyznaczenie liniowości, odchylenia standardowego, względnego odchylenia standardowego, granicy wykrywalności (LOD) i granicy oznaczalności (LOQ) badanych WWA. Chromatograficzna metoda identyfikacji WWA w próbkach gleb wymaga przeprowadzenia analitów z matrycy stałej do matrycy ciekłej (etap izolacji), oczyszczenia próbki z substancji przeszkadzających oraz zagęszczenia ekstraktu (etap wzbogacania). W celu opracowania metody oznaczania WWA w próbkach gleb sprawdzono 3 metody przygotowania próbek do analizy chromatograficznej HPLC: metodę A (metoda QuEChERS), metodę B (ekstrakcja rozpuszczalnikiem w aparacie Soxhleta) i metodę C (ekstrakcja rozpuszczalnikiem wspomagana wytrząsaniem). Otrzymane ekstrakty były następnie oczyszczane na wybranym materiale sorpcyjnym: MgSO4 i PSA (metoda A), CN/SiOH (metoda B) i Al2O3 (metoda C). Do przetestowania wybranych metod przygotowania próbek gleby do analizy chromatograficznej HPLC wykorzystano glebę wzorcową PAHs by HPLC40g (SQC017-40G) firmy Sigma Aldrich, o znanych stężeniach analitów. Oczyszczone ekstrakty poddano analizie chromatograficznej HPLC, obejmującej identyfikację oraz ilościowe oznaczenie poszczególnych WWA, oraz obliczono stopnie odzysku analitów. Na podstawie analiz chromatograficznych i stopni odzysku wytypowano optymalną metodykę przygotowania próbek gleb do analizy chromatograficznej pod kątem oznaczania wielopierścieniowych węglowodorów aromatycznych. Opracowaną metodykę oznaczania WWA w próbkach gleb przetestowano na rzeczywistych próbkach gruntu pobranych z 2 otworów badawczych zlokalizowanych na terenie kopalni ropy naftowej usytuowanej w południowo-wschodniej Polsce.