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 uticajbioremedijacionih uslova na biodegradaciju zasićenih i aromatičnih ugljovodonika uzagađivačima naftnog tipa. Primenjen je ogled simulirane biodegradacije u laboratoriji, ogled exsitu 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 septembra2009. do marta 2010. godine i ogled višestepene in situ bioremedijacije izdani kontaminiranenaftnim derivatima koji je trajao od maja do jula 2012. godine.U prve tri ekesperimentalne studije za karakterizaciju supstrata korišćene su isteanalitičke tehnike. Nakon ekstrakcije organske supstance, dobijeni ekstrakti su razdvojenihromatografijom na koloni u četiri različite frakcije: frakciju zasićenih ugljovodonika, frakcijuaromata, frakciju alkohola i frakciju masnih kiselina. U četvrtoj studiji nakon ekstrakcije dobijeniekstrakti su razdvojeni hromatografijom na koloni u tri različite frakcije: frakciju zasićenihugljovodonika, frakciju aromata i frakciju NSO jedinjenja. Pojedine frakcije u svim studijama suanalizirane gasnom hromatografijom-masenom spektrometrijom (GC-MS).Tokom ogleda simulirane bioedgradacije u laboratoriji isptitivan je bioremedijacionipotencijal aerobnih zimogenih mikroorganizma iz zemljišta. Korišćeni mikroorganizmi supokazali najviši bioremedijacioni potencijal u razgradnji n-alkana i izoprenoida, visokbioedegradacioni potencijal pri degradaciji fenantrena i metil-fenantrena i nizak pri degradacijidimetil-fenatrena, sterana i terpana. Rezultati ovog ogleda simulirane biodegradacije ulaboratoriji originalni su rezultati doktorske teze dr Mile Ilić111.U drugoj studiji su izvođeni eksperimenti ex situ bioremedijacije kontaminiranogzemljišta na lokalitetu rafinerije nafte Pančevo. Analizom uzoraka su ustanovljene promene usastavu naftnog zagađivača samo u poslednjoj fazi ekesperimenta. Aktivnost mikroorganizmaogledala se u smanjenju teških komponenti nafte, čime je naftna zagađujuća supstanca dovedenau oblik koji se efikasnije može ukloniti iz zemljišta. Rezultati ovog ogleda ex situbioremedijacije originalni su rezultati doktorske teze dr Vladimira Beškoskog 105.Milan D. Novaković Doktorska disertacijaRezime VIU trećoj studiji su izvođeni eksperimenti ex situ bioremedijacije zemljišta zagađenogmazutom. Deo „iskopanog“ zemljišta je odvojen u kontrolnu gomilu, koja je bila izloženaprirodnim biodegradacijonim procesima. Tokom biodegradacionog ogleda uočena jepreferencijalna i brža degradacija fenantrena u odnosu na metil supstituisane fenentrene.U poslednjoj studiji su izvođeni ogledi višestepene in situ bioremedijacije izdanikontaminirane naftnim zagađivačem. Na kraju eksperimenta uočena je kompletna degradacijasvih 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 suprimenom četiri različita pristupa detaljno i sveobuhvatno analizirani različiti aspektibioremedijacije životne sredine zagađene naftnim zagađujućim supstancama. Kao sto je većnaglašeno, rezultati ogleda simulirane biodegradacije u laboratoriji i ogleda ex situbioremedijacije koji je izvođen u rafineriji nafte Pančevo od maja do novembra 2006 originalnisu rezultati dve doktorske teze koje su odbranjene na Hemijskom fakultetu Univerziteta uBeogradu 111, 105.U ovoj doktorskoj tezi originalno su izloženi rezultati eksperimenta ex situbioremedijacije zemljišta zagađenog mazutom i ogleda višestepene in situ bioremedijacije izdanikontaminirane naftnim zagađivačem. Ovi rezultati su prvo nezavisno prodiskutovani, a zatim iupoređeni sa prethodno publikovanim rezultatima celokupnog istraživanjaThe aim of this paper is to explain influence of bioremediation conditions on thebiodegradation of saturated and aromatic hydrocarbons of oil-type pollutants based on the resultsof four different experiments. Experiment of simulated biodegradation under laboratoryconditions, experiment of ex situ bioremediation which was performed at the locality of the OilRefinery Pančevo from May to November 2006, experiment of ex situ bioremediation of soilcontaminated with heavy residual fuel oil which was performed during the period fromSeptember 2009 to March 2010 and experiment of multi stage in situ bioremediation of aquifercontaminated with petroleum derivatives which was performed from May to July 2012 weredone.In the first three experiments for substrate characterizations the same analytical procedurewas used. After extraction of organic substance, extracts were separated by columnchromatography on fraction of saturated hydrocarbons, aromatics, alcohols and fraction of fattyacids. In the fourth experiment obtained extracts after extractions were separated by columnchromatography on fraction of saturated hydrocarbons, aromatics and NSO compounds.Individual fractions form all experiments were analyzed by gas chromatography-massspectrometry (GC-MS).In the experiment of simulated biodegradation under laboratory conditions,bioremediation potential of the aerobic zymogenous microorganisms from soil for crude oilbiodegradation was investigated. Microorganisms showed the highest bioremediation potential indegradadtion of n-alkanes and isoprenoids, a high potential was confirmed in the biodegradationof phenanthrene and methyl phenanthrenes and low potential was noted in degradation ofdimethyl phenantrenes, steranes and triterpanes. The results of this experiment of simulatedbiodegradation 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 samplesMilan D. Novaković Doktorska disertacijaAbstract VIIIoccured only during the last experiment phases. The activity of microorganisms was reflected inthe decrease of the heavy compounds, wherby the oil pollutant was transformed to a form thatcould be removed more efficiently. The results of this experiment of ex situ bioremediation areoriginal results of a doctoral thesis of Dr. Vladimir Beškoski105.In the third study, experiment of ex situ bioremediation of soil contaminated with heavyresidual fuel oil was performed. Part of excavated soil was separated into a control pile, whichwas exposed to natural biodegradation processes. A preferential and accelerated degradation ofphenathrene versus methyl-phenantrenes was observed.In the last study, experiment of multi stage in situ bioremediation of aquifer contaminatedwith petroleum derivatives was performed. At the end of experiment complete degradation of allexamined 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 byapplication of four different approaches thoroughly were analyzed various aspects ofbioremediation of environment contaminated by oil pollutants. As already pointed out, the resultsof experiments of simulated biodegradation under laboratory conditions and ex situbioremediation 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 situbioremediation of soil contaminated with heavy residual fuel oil and experiment of multistage insitu bioremediation of aquifers contaminated with petroleum pollutants. These results are firstlyindependently discussed, and then compared with previously published results of this wholeresearc