Influence of bioremediation conditions on biodegradation of saturated and aromatic hydrocarbons in oil-type pollutants

Cilj ovog rada je bio da se na osnovu rezultata četiri različita eksperimenta objasni uticaj bioremedijacionih uslova na biodegradaciju zasićenih i aromatičnih ugljovodonika u zagađivačima naftnog tipa. Primenjen je ogled simulirane biodegradacije u laboratoriji, ogled ex situ bioremedijacije koji je izvođen u rafineriji nafte Pančevo od maja do novembra 2006. godine, ogled ex situ bioremedijacije zemljišta zagađenog mazutom koji je trajao od septembra 2009. do marta 2010. godine i ogled višestepene in situ bioremedijacije izdani kontaminirane naftnim derivatima koji je trajao od maja do jula 2012. godine. U prve tri ekesperimentalne studije za karakterizaciju supstrata korišćene su iste analitičke tehnike. Nakon ekstrakcije organske supstance, dobijeni ekstrakti su razdvojeni hromatografijom na koloni u četiri različite frakcije: frakciju zasićenih ugljovodonika, frakciju aromata, frakciju alkohola i frakciju masnih kiselina. U četvrtoj studiji nakon ekstrakcije dobijeni ekstrakti su razdvojeni hromatografijom na koloni u tri različite frakcije: frakciju zasićenih ugljovodonika, frakciju aromata i frakciju NSO jedinjenja. Pojedine frakcije u svim studijama su analizirane gasnom hromatografijom-masenom spektrometrijom (GC-MS). Tokom ogleda simulirane bioedgradacije u laboratoriji isptitivan je bioremedijacioni potencijal aerobnih zimogenih mikroorganizma iz zemljišta. Korišćeni mikroorganizmi su pokazali najviši bioremedijacioni potencijal u razgradnji n-alkana i izoprenoida, visok bioedegradacioni potencijal pri degradaciji fenantrena i metil-fenantrena i nizak pri degradaciji dimetil-fenatrena, sterana i terpana. Rezultati ovog ogleda simulirane biodegradacije u laboratoriji originalni su rezultati doktorske teze dr Mile Ilić111. U drugoj studiji su izvođeni eksperimenti ex situ bioremedijacije kontaminiranog zemljišta na lokalitetu rafinerije nafte Pančevo. Analizom uzoraka su ustanovljene promene u sastavu naftnog zagađivača samo u poslednjoj fazi ekesperimenta. Aktivnost mikroorganizma ogledala se u smanjenju teških komponenti nafte, čime je naftna zagađujuća supstanca dovedena u oblik koji se efikasnije može ukloniti iz zemljišta. Rezultati ovog ogleda ex situ bioremedijacije originalni su rezultati doktorske teze dr Vladimira Beškoskog 105. Milan D. Novaković Doktorska disertacija Rezime VI U trećoj studiji su izvođeni eksperimenti ex situ bioremedijacije zemljišta zagađenog mazutom. Deo „iskopanog“ zemljišta je odvojen u kontrolnu gomilu, koja je bila izložena prirodnim biodegradacijonim procesima. Tokom biodegradacionog ogleda uočena je preferencijalna i brža degradacija fenantrena u odnosu na metil supstituisane fenentrene. U poslednjoj studiji su izvođeni ogledi višestepene in situ bioremedijacije izdani kontaminirane naftnim zagađivačem. Na kraju eksperimenta uočena je kompletna degradacija svih ispitivanih jedinjenja, n-alakana i izoprenoida, sterana, terpana, fenantrena, metilfenantrena, dimetil-fenatrena i trimetil-fenatrena. Rezultati izloženi u ovoj doktorskoj tezi deo su velikog istraživanja tokom kojeg su primenom četiri različita pristupa detaljno i sveobuhvatno analizirani različiti aspekti bioremedijacije životne sredine zagađene naftnim zagađujućim supstancama. Kao sto je već naglašeno, rezultati ogleda simulirane biodegradacije u laboratoriji i ogleda ex situ bioremedijacije koji je izvođen u rafineriji nafte Pančevo od maja do novembra 2006 originalni su rezultati dve doktorske teze koje su odbranjene na Hemijskom fakultetu Univerziteta u Beogradu 111, 105. U ovoj doktorskoj tezi originalno su izloženi rezultati eksperimenta ex situ bioremedijacije zemljišta zagađenog mazutom i ogleda višestepene in situ bioremedijacije izdani kontaminirane naftnim zagađivačem. Ovi rezultati su prvo nezavisno prodiskutovani, a zatim i upoređeni sa prethodno publikovanim rezultatima celokupnog istraživanjaThe aim of this paper is to explain influence of bioremediation conditions on the biodegradation of saturated and aromatic hydrocarbons of oil-type pollutants based on the results of four different experiments. Experiment of simulated biodegradation under laboratory conditions, experiment of ex situ bioremediation which was performed at the locality of the Oil Refinery Pančevo from May to November 2006, experiment of ex situ bioremediation of soil contaminated with heavy residual fuel oil which was performed during the period from September 2009 to March 2010 and experiment of multi stage in situ bioremediation of aquifer contaminated with petroleum derivatives which was performed from May to July 2012 were done. In the first three experiments for substrate characterizations the same analytical procedure was used. After extraction of organic substance, extracts were separated by column chromatography on fraction of saturated hydrocarbons, aromatics, alcohols and fraction of fatty acids. In the fourth experiment obtained extracts after extractions were separated by column chromatography on fraction of saturated hydrocarbons, aromatics and NSO compounds. Individual fractions form all experiments were analyzed by gas chromatography-mass spectrometry (GC-MS). In the experiment of simulated biodegradation under laboratory conditions, bioremediation potential of the aerobic zymogenous microorganisms from soil for crude oil biodegradation was investigated. Microorganisms showed the highest bioremediation potential in degradadtion of n-alkanes and isoprenoids, a high potential was confirmed in the biodegradation of phenanthrene and methyl phenanthrenes and low potential was noted in degradation of dimethyl phenantrenes, steranes and triterpanes. The results of this experiment of simulated biodegradation under the laboratory conditions are original results of a doctoral thesis of Dr. Mila Ilić111. In the experiment of ex situ bioremediation at the locality of the Oil Refinery Pančevo, significant changes in the composition of the oil pollutant determined by analysis of the samples Milan D. Novaković Doktorska disertacija Abstract VIII occured only during the last experiment phases. The activity of microorganisms was reflected in the decrease of the heavy compounds, wherby the oil pollutant was transformed to a form that could be removed more efficiently. The results of this experiment of ex situ bioremediation are original results of a doctoral thesis of Dr. Vladimir Beškoski105. In the third study, experiment of ex situ bioremediation of soil contaminated with heavy residual fuel oil was performed. Part of excavated soil was separated into a control pile, which was exposed to natural biodegradation processes. A preferential and accelerated degradation of phenathrene versus methyl-phenantrenes was observed. In the last study, experiment of multi stage in situ bioremediation of aquifer contaminated with petroleum derivatives was performed. At the end of experiment complete degradation of all examined compounds, n-alkanes and isoprenoides, steranes, terpanes, phenanhtrene, methylphenantrenes, dimethyl-pehanthrenes and trimethyl-phenthrenes was observed. The results presented in this doctoral thesis are part of a large research in which by application of four different approaches thoroughly were analyzed various aspects of bioremediation of environment contaminated by oil pollutants. As already pointed out, the results of experiments of simulated biodegradation under laboratory conditions and ex situ bioremediation which was performed at the Oil Refinery Pančevo from May to November 2006, are the original results of two doctoral theses which were defended at the Faculty of Chemistry, University of Belgrade111, 105. This doctoral thesis originally presents the results of experiments of ex situ bioremediation of soil contaminated with heavy residual fuel oil and experiment of multistage in situ bioremediation of aquifers contaminated with petroleum pollutants. These results are firstly independently discussed, and then compared with previously published results of this whole researc