Petroleum pollutant degradation by surface water microorganisms.

BACKGROUND, AIMS AND SCOPE: It is well known that the composition of petroleum or some of its processing products changes in the environment mostly under the influence of microorganisms. A series of experiments was conducted in order to define the optimum conditions for an efficient biodegradation of petroleum pollutant, or bioremediation of different segments of the environment. The aim of these investigations was to show to what extent the hydrocarbons of a petroleum pollutant are degraded by microbial cultures which were isolated as dominant microorganisms from a surface water of a wastewater canal of an oil refinery and a nitrogen plant. Biodegradation experiments were conducted on one paraffinic, and one naphthenic type of petroleum during a three month period under aerobic conditions, varying the following parameters: Inorganic (Kp) or an organic medium (Bh) with or without exposition to light. METHODS: Microorganisms were analyzed in a surface water sample from a canal (Pancevo, Serbia), into which wastewater from an oil refinery and a nitrogen plant is released. The consortia of microorganisms were isolated from the water sample (most abundant species: Phormidium foveolarum--filamentous Cyanobacteria, blue-green algae and Achanthes minutissima, diatoms, algae). The simulation experiments of biodegradation were conducted with the biomass suspension and crude oils Sirakovo (Sir, paraffinic type) and Velebit (Ve, naphthenic type). After a three month period, organic substance was extracted by means of chloroform. In the extracts, the content of saturated hydrocarbons, aromatic hydrocarbons, alcohols and fatty acids was determined (the group composition). n-Alkanes and isoprenoid aliphatic alkanes, pristane and phytane, in the aliphatic fractions, were analyzed using gas chromatography (GC). Total isoprenoid aliphatic alkanes and polycyclic alkanes of sterane and triterpane types were analyzed by GC-MS. RESULTS AND DISCUSSION: Paraffinic type petroleums have a significant loss of saturated hydrocarbons. For naphthenic type petroleum, such a trend has not been observed. The most intensive degradation of n-alkanes and isoprenoid aliphatic alkanes (in paraffinic oil) and isoprenoids (in naphthenic oil) was observed using the inorganic medium Kp in the light; the microbial conversion is somewhat lower with Kp in the dark; with organic medium Bh in the light the degradation is of low intensity; with the same medium in the dark the degradation is hardly to be seen. Steranes and triterpanes were not affected by microbial degradation under the conditions used in our experiments. Obviously, the petroleum biodegradation was restricted to the acyclic aliphatics (n-alkanes and isoprenoids). CONCLUSION: Phormidium foveolarum (filamentous Cyanobacteria--blue-green algae) and Achanthes minutissima (diatoms, algae), microbial cultures isolated as dominant algae from a surface water in a wastewater canal of an oil refinery and a nitrogen plant, have degradable effects dominantly involving petroleum hydocarbons. Petroleum microbiological degradation is more intensive when inorganic medium (in the light) is applied. Having in mind that the inorganic pollutants have been released into the canal as well, this medium reflects more the natural environmental conditions. Polycyclic alkanes of sterane and triterpane type, in spite of the fact that these compounds could be degraded, have remained unchanged regarding abundance and distribution. Since this is the case even for naphthenic type petroleum (which is depleted in n-alkanes), it can be concluded that the biodegradation of petroleum type pollutants, under natural conditions, will be restrained to the n-alkane and isoprenoid degradation. RECOMMENDATION AND OUTLOOK: Performed experiments and simulations of petroleum microbiological degradation may serve for the prediction of the fate of petroleum type pollutants, as well as for definition of conditions for bioremediation of some environmental segments.

Investigation of interactions between surface water and petroleum-type pollutants.

BACKGROUND, AIMS AND SCOPE: In oil spill investigations, one of the most important steps is a proper choice of approaches that imply an investigation of samples taken from different sedimentary environments, samples of oil contaminants taken in different periods of time and samples taken at different distances from the oil spill. In all these cases, conclusion on the influence of the environment, microorganisms or migration on the oil contaminants' composition can be drawn from the comparison of chemical compositions of the investigated contaminants. However, in case of water contaminants, it is very important to define which part of organic matter has been analyzed. Namely, previous investigations showed that there were some differences in chemical composition of the same oil contaminant depending on the intensity of its contact with ground water. The aim of this work is to define more precisely the interactions between oil contaminant and water, i.e. the influence of the intensity of interaction between the oil contaminant and water on its chemical composition. The study was based on a comparison of four fractionated extracts of an oil pollutant, after they had been analyzed in details. METHODS: Oil polluted surface water (wastewater canal, Pancevo, Serbia) was investigated. The study was based on a comparison of four extracts of an oil contaminant: extract 1 (decanted part), and extracts 2, 3 and 4 (extracted by shaking for 1 minute, 5 minutes and 24 hours, respectively). The fractionated extracts were saponified with a solution of KOH in methanol, and neutralized with 10% hydrochloric acid. The products were dissolved in a mixture of dichloromethane and hexane, and individually fractionated by column chromatography on alumina and silica gel (saturated hydrocarbon, aromatic, alcohol and fatty acid fractions). n-Alkanes and isoprenoid aliphatic alkanes, polycyclic alkanes of sterane and triterpane types, alcohols and fatty acids were analyzed using gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). delta 13C(PDB) values of individual n-alkanes in the aliphatic fractions were determined using gas chromatography-isotope ratio monitoring-mass spectrometry (GC-irmMS). RESULTS AND DISCUSSION: Extracts 1 and 2 are characterized by uniform distribution of n-alkanes, whereas extract 3 is characterized by an even-numbered members dominating the odd-ones, and extract 4 showed a bimodal distribution. Extract 1 is characterized by the least negative delta 13C(PDB) values of C19-C26 n-alkanes. Sterane and triterpane analysis confirmed that all extracts originated from the same oil contaminant. n-Fatty acids, C19-C24, in all extracts are very low, being somewhat higher in extract 4. Even-numbered n-alcohols, C12-C16, were identified in the highest concentration in extract 3. It was assumed that algae were responsible for the composition of extract 3. Furthermore, a possible reason for higher concentrations of C19-C26 n-alkanes and C19-C24 fatty acids in extract 4 is the formation of inclusion compounds with colloidal micelles formed between the oil contaminant's NSO-compounds and water. CONCLUSION: It was undoubtedly confirmed that there were specific differences in the compositions of the different extracts depending on the intensity of the interaction between the oil contaminant and the surface water. RECOMMENDATION AND OUTLOOK: When comparing the composition of oil contaminants from different water samples (regardless of the ultimate investigation goal) it is necessary to compare the extracts isolated under the same conditions, in other words, extracts that were in the same or very similar interaction with water.