20.500.12556/DKUM-92212 Določevanje metilnih estrov maščobnih kislin med procesi bioremediacije z dizelskim gorivom onesnaženih zemljin doktorska disertacija Determination of fatty acids methyl esters during bioremediation of diesel fuel-contaminated soil Naftni derivati prehajajo v tla predvsem v bližini industrijskih obratov in so zaradi široke uporabe eno najpogostejših onesnaževal tal. V skladu z Zakonom o varstvu okolja (Uradni list RS, št. 44/22, 18/23 – ZDU-1O, 78/23 – ZUNPEOVE in 23/24) mora v primeru okoljske nesreče povzročitelj nemudoma izvesti vse potrebne ukrepe za omejitev okoljske škode. V tem delu se osredotočamo na dva zemeljska izkopa (izkop iz Skladišča naftnih derivatov v Račah in izkop iz Terminala Instalacija Sermin), ki sta nastala v sklopu interventne sanacije tal po razlitju dizelskega goriva v industrijskih obratih podjetja Petrol d. d. kot intervencijska izkopa z namenom reševanja okoljske škode. Oba zemeljska izkopa sta bila bioremediirana z uporabo tehnike fitoremediacije znotraj industrijskih obratov na obeh lokacijah razlitja. Za identifikacijo in spremljanje mikroorganizmov med procesom bioremediacije smo uporabili metodologijo analiz fosfolipidnih maščobnih kislin, ki se jih določuje kot metilne estre maščobnih kislin s plinsko kromatografijo z masno spektrometrijo. Namen je bil kvantitativno določanje posameznih mikrobioloških komponent tal kot ekosistemske enote. Zaradi uporabe tehnike fitoremediacije smo raziskave osredotočili tudi na vpliv dizelskega goriva na samo rastlinsko komponento. Analize foliarnega tkiva dizelskemu gorivu izpostavljenih rastlin so vključevale rastline, ki so uspevale tako v industrijskem obratu na remediacijskem polju kot na neonesnaženih referenčnih tleh in pod nadzorovanimi pogoji. Izvedli smo analize metilnih estrov maščobnih kislin po direktni ekstrakciji lipidov iz fotosintetskih tkiv izpostavljenih rastlin. Namen vpeljave metodologije, ki določuje lipidno sestavo foliarnega tkiva, je bil identificirati učinke dizelskega goriva kot onesnaževala na fiziološke spremembe pri izpostavljenih rastlinah. Raziskave na zemeljskem izkopu, ki je izviral iz Terminala Instalacija Sermin, so vključevale preverjanje vpliva podnebja na razvoj mikrobiološke združbe med procesom bioremediacije. S tem razlogom smo del zemljine iz Instalacije Sermin (z značilnim submediteranskim podnebjem) prenesli v Lovrenc na Dravskem polju (z zmernim kontinentalnim podnebjem). Pri raziskavah na zemeljskem izkopu iz skladišča naftnih derivatov v Račah pa smo se osredotočili na preiskovanje vplivov biostimulatorjev (zeolit in komercialni pripravek iz alg), ter vplivov različnih tipov vegetacije na razvoj mikrobiološke združbe. Vpliv podnebja na hitrost razgradnje je bil v prid milejši klimi zmernega kontinentalnega podnebja, kjer je bila po enem letu dosežena 75-odstotna stopnja razgradnje dizelskega goriva, medtem ko je bila v submediteranskem podnebju po enem letu dosežena 60-odstotna stopnja razgradnje dizelskega goriva. S spremljanjem mikrobiološke združbe med procesi bioremediacije pa smo potrdili, da vegetacija vpliva na samo dinamiko mikrobne populacije, ne pa tudi na stopnjo razgradnje ogljikovodikov v tleh. Uporabljena biostimulatorja nista imela vpliva ne na hitrost razgradnje dizelskega goriva in ne na spremembe mikrobiološke združbe, kar pa je potrdilo relativno stabilno in raznoliko avtohtono mikrobno skupnost. Po enoletnem spremljanju mikrobiološke združbe med remediacijo zemeljskega izkopa v Serminu smo raziskave nadaljevali na rastlinski komponenti. S preiskavami na foliarnem tkivu smo potrdili, da se relativna koncentracija linolenske kisline zmanjšuje, medtem ko se koncentraciji palmitinske in linolne kisline povečujeta z naraščajočo koncentracijo dizelskega goriva v zemljinah. Oba zemeljska izkopa sta bila še pred zaključkom tega dela uspešno remediirana in ponovno uporabljena na omenjenih lokacijah. Petroleum products are because of their ubiquitous use among the most common pollutants found in soil. According to the Environmental Protection Act (Official Gazette of the Republic of Slovenia, No. 44/22, 18/23 – ZDU-1O, 78/23 – ZUNPEOVE, and 23/24), in the event of environmental pollution, the responsible party must immediately take all necessary measures to limit environmental damage. In this work, we focus on two excavated soils (from the Fuel Storage Facility in Rače and from the Terminal Instalacija Sermin), which were created by excavating the soil following a diesel spill at the industrial facilities of Petrol d. d. company, to suppress potential environmental damage. Both excavated soils were bioremediated by introducing phytoremediation techniques at the industrial facilities of both spill sites. To identify and monitor microorganisms during the bioremediation process, phospholipid fatty acid analysis were performed, which were determined as fatty acid methyl esters using gas chromatography-mass spectrometry. The purpose of this work was to quantitatively evaluate the individual microbial components of the soil as an ecosystemic unit. However, owing to the introduction of the phytoremediation technique, we also focused our research on the effects of diesel on the plant component itself. The analyses of the foliar tissue of diesel fuel-exposed plants included plants that grew in the land farming unit in the industrial facility, in uncontaminated reference soil, and under fully controlled conditions. The latter was carried out as fatty acid methyl ester analyses after direct lipid extraction from the photosynthetic tissues of the exposed plants. The purpose of introducing a methodology to determine the lipid composition of foliar tissue was to identify the actual effects of diesel as a pollutant on physiological changes in the exposed plants. Research on the soil originating from Terminal Instalacija Sermin included assessing the influence of climatic conditions on the microbial community during the bioremediation process. Part of the soil from the Terminal Sermin (with a typical sub-Mediterranean climate) was transferred to Lovrenc na Dravskem polju (with a moderate continental climate). Research on excavated soil from the Rače fuel storage has focused on investigating the effects of biostimulants, such as zeolite and commercial algae preparation, and the influence of different vegetation types on the microbial community dynamics during the bioremediation process. The influence of climate on the degradation rate favored the milder temperate continental climate, where 75% diesel degradation was achieved after one year, compared to the sub-Mediterranean climate, where 60% diesel degradation was achieved after one year. By monitoring the microbial community during both bioremediation processes, we confirmed that vegetation influences microbial population dynamics but not the rate of hydrocarbon degradation. In the case of excavated soil from Rače fuel storage, neither the diesel degradation rate nor the changes in the microbial community were influenced by the biostimulants used, confirming a relatively stable and diverse indigenous microbial community. After one year of monitoring the microbial community during the bioremediation of the Sermin excavated soil, we focused our investigations on the plant components, confirming that the relative concentration of linolenic acid decreased, whereas the concentrations of palmitic acid and linoleic acid increased with increasing concentrations of diesel in the soils. Both excavated soils were successfully remediated and reused at the aforementioned industrial facilities prior to the completion of this thesis. analiza fosfolipidnih maščobnih kislin foliarna lipidna sestava bioremediacija fitoremediacija onesnaženje tal z dizelskim gorivom analysis of phospholipid fatty acids foliar lipid composition bioremediation phytoremediation diesel fuel soil-pollution true true false [T. Cenčič Predikaka] Slovenski jezik Angleški jezik Doktorsko delo/naloga 2025-03-20 15:15:12 2025-12-19 14:17:49 2025-12-23 03:18:32 0000-00-00 00:00:00 2025 0 Maribor 0 Maribor X , 99 str. 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 543.635.4:502.521:[504.5:665.75](043.3) 262595331 DOK_Cencic_Predikaka_Tjasa_2025.pdf DOK_Cencic_Predikaka_Tjasa_2025.pdf 1 1CDDD911E7209A2B60FC46F38E596B1F 9ee9f417e9b20e090c60690f1d896ab417dca31580dcc9041f139a47a4b439f2 640e8868-dcd5-11f0-a44c-00155d580e02 https://dk.um.si/Dokument.php?lang=slv&id=201570 Fakulteta za kemijo in kemijsko tehnologijo 0 0 0