The article presents issues related to the possibility of using toxicological tests as a tool to monitor the progress of soil treatment contaminated with petroleum substances (TPH, PAH), Zn, Pb and Cd in bio-phytoremediation processes. In order to reduce the high content of petroleum pollutants (TPH = 56,371 mg kg⁻¹ dry mass, PAH = 139.3 mg kg⁻¹ dry mass), the technology of stepwise soil treatment was applied, including basic bioremediation and inoculation with biopreparations based of indigenous non-pathogenic species of bacteria, fungi and yeasts. As a result of basic bioremediation in laboratory conditions (ex-situ method), the reduction of petroleum pollutants TPH by 33.9% and PAH by 9.5% was achieved. The introduction of inoculation with biopraparation-1 prepared on the basis of non-pathogenic species of indigenous bacteria made it possible to reduce the TPH content by 86.3%, PAH by 40.3%. The use of a biopreparation-1 enriched with indigenous non-pathogenic species of fungi and yeasts in the third series of inoculation increased to an increase in the degree of biodegradation of aliphatic hydrocarbons with long carbon chains and PAH by a further 28.9%. In the next stage of soil treatment after biodegradation processes, which was characterized by an increased content of heavy metals (Zn, Pb, Cd) and naphthalene, chrysene, benzo(a)anthracene and benzo(ghi)perylene belonging to polycyclic aromatic hydrocarbons, phytoremediation with the use of Melilotus officinalis was applied. After the six-month phytoremediation process, the following was achieved: Zn content by 25.1%, Pb by 27.9%, Cd by 23.2% and TPH by 42.2% and PAH by 49.9%. The rate of removal of individual groups of hydrocarbons was in the decreasing order: C₁₂–C₁₈ > C₆–C₁₂ > C₁₈–C₂₅ > C₂₅–C₃₆. PAHs tended to be removed in the following order: chrysene > naphthalene > benzo(a)anthracene > benzo(ghi)perylene. The TF and BCF coefficients were calculated to assess the capacity of M. officinalis to accumulate metal in tissues, uptake from soil and transfer from roots to shoots. The values of TF translocation coefficients were, respectively, for Zn (0.44), Pb (0.12), Cd (0.40). The calculated BCF concentration factors (BCF_roots > BCF_shoots) show that heavy metals taken up by M. officinalis are mainly accumulated in the root tissues in the following order Zn > Pb > Cd, revealing a poor metal translocation from the root to the shoots. This process was carried out in laboratory conditions for a period of 6 months. The process of phytoremediation of contaminated soil using M. officinalis assisted with fertilization was monitored by means of toxicological tests: Microtox, Ostracodtoxkit F^TM, MARA and Phytotoxkit^TM. The performed phytotoxicity tests have indicated variable sensitivity of the tested plants on contaminants occurring in the studied soils, following the sequence: Lepidium sativum < Sorghum saccharatum < Sinapis alba. The sensitivity of toxicological tests was comparable and increased in the order: MARA < Ostracodtoxkit F^TM < Microtox. The results of the toxicological monitoring as a function of the time of soil treatment, together with chemical analyses determining the content of toxicants in soil and biomass M. officinalis, clearly confirmed the effectiveness of the applied concept of bioremediation of soils contaminated with zinc, lead and cadmium in the presence of petroleum hydrocarbons.

The article discusses the results of biomonitoring research at the Underground Gas Storage (UGS). Hydrogen sulphide, as one of the products of microbiological reaction and transformation, as well as a product of chemical reactions in rocks, is a subject of interest for global petroleum companies. The materials used in this research work were formation waters and stored natural gas. The biomonitoring of reservoir waters and cyclical analyses of the composition of gas stored at UGS Wierzchowice enabled the assessment of the microbiological condition of the reservoir environment and individual storage wells in subsequent years of operation. Investigations of the formation water from individual wells of the UGS Wierzchowice showed the presence of sulphate reducing bacteria bacteria (SRB), such as Desulfovibrio and Desulfotomaculum genera and bacteria that oxidize sulphur compounds. In the last cycles of UGS Wierzchowice, the content of hydrogen sulphide and sulphides in the reservoir waters ranged from 1.22 to 15.5 mg/dm³. The monitoring of natural gas received from UGS production wells and observation wells, which was carried out in terms of the determination of hydrogen sulphide and organic sulphur compounds, made it possible to observe changes in their content in natural gas in individual storage cycles. In the last cycles of UGS Wierzchowice, the content of hydrogen sulphide in natural gas from production wells ranged from 0.69 to 2.89 mg/dm³, and the content of organic sulphur compounds converted to elemental sulphur ranged from 0.055 to 0.130 mg S_el./Nm³. A higher hydrogen sulphide content was recorded in natural gas from observation wells in the range of 2.02–25.15 mg/Nm³. In order to explain the causes of hydrogen sulphide formation at UGS Wierzchowice, isotopic analyses were performed to determine the isotope composition of δ³⁴S_H2S, δ³⁴S_SO4, δ¹⁸O_SO4 in natural gas samples (production and observation wells) and in the deep sample of reservoir water. The results of isotope tests in connection with microbiological tests, chromatographic analyses of sulphur compounds in natural gas collected from UGS Wierzchowice and an analysis of the geological structure of the Wierzchowice deposit allow us to conclude that the dominant processes responsible for the formation of hydrogen sulphide at UGS Wierzchowice are microbiological, consisting of microbial sulphate reduction (MSR). The presented tests allow for the control and maintenance of hydrogen sulphide at a low level in the natural gas received from the Wierzchowice Underground Gas Storage facility.

An analysis of environmental threats showed that the complete influence of shale gas production on the environment is bigger than in the case of gas production from a traditional oilfield. Hydraulic fracturing of shale, done on a much larger scale, results in huge amounts of liquid waste that must be managed in a rational way. An optimum solution to this problem is reusing flowback water to develop fluids for later fracturing. This article discusses the composition of the fluids used in hydraulic fracturing of non-conventional deposits as well as the flowback water. In the case of the examined liquids, toxicological tests have been carried out using microbiotests of ToxKit types Microtox, MARA, Daphtoxkit F magna, and Thamnotoxkit F. The research was done on a sample of fluids for hydraulic fracturing, and on flowback water obtained in hydraulic fracturing of shale formations in Poland. The tests are essential for correctly managing flowback waters after fracturing.