Compositional Changes of Aromatic Steroid Hydrocarbons in Naturally Weathered Oil Residues in the Egyptian Western Desert

Naturally weathered oil residues from an arid dumpsite in Al-Alamein, Egypt were analyzed for monoaromatic and triaromatic steranes to demonstrate the utility of biomarker compounds in assessing the chemical composition changes during the degradation of the released oil residues in a terrestrial environment. The characterizations of individual aromatic compounds were based on gas chromatography/mass spectrometry (GC/MS) analyses. The results showed that triaromatic sterane distributions were similar in the oil residues of varying weathering degradation extents and correlated with a fresh crude oil sample of the Western Desert-sourced oil. Molecular ratios of triaromatic sterane compounds (ratios of C₂₈20R /C₂₈20S, C₂₇20R /C₂₈20R, and C₂₈20S /[C₂₆20R + C₂₇20S ]) were proved to be suitable for source identification. Major changes in chemical compositions during weathering of the oil residues were the depletion of short chain mono- and tri-aromatic steranes in samples that had undergone extensive degradation. The results of triaromatic sterane distribution are in good agreement with weathering classification based on the analyses of saturate and aromatic hydrocarbons and the ratios of n -alkanes, PAHs and saturate biomarker compounds.     

Stigmastane and hopanes as conserved biomarkers for estimating oil biodegradation in a former refinery plant-contaminated soil

In the last decade, a refinery plant located in Lido Adriano, East Ravenna (Italy) has been subject to mineral oil contamination. The mineral crude oil, extracted from the offshore in Adriatic sea, consisted of 78% aliphatics, cyclic alkanes and saturated polycyclic hydrocarbons, 9% aromatics, polycyclic aromatic hydrocarbons (PAHs) and alkylated derivatives, and 13% of tars/asphaltenes. Analysis of soil after 10 years of natural attenuation revealed a complete depletion of linear (n-C(9)-C(24)), light aromatics (C1-C3/benzenes) and PAHs (C2/naphthalene, C1/phenanthrene); besides a substantial degradation of isoprenoids prystane and phytane, branched and cyclic alkanes. The remaining contaminants which withstood to natural degradation was saturated polycyclic hydrocarbons (perhydro-PAH derivatives), unsaturated polycyclic hydrocarbons (tetrahydro, dihydro-PAH derivatives), terpanes, steranes and unidentified compounds. Such residues resulted in 80% reduction of its concentration after two months of laboratory treatment. Samples were extracted by organic solvents, separated by silica/alumina gel column chromatography and analyzed by gas chromatography-mass selective detector (GC-MSD). Identification and quantification of aliphatic, cyclic alkanes, typical PAHs, terpanes and steranes were carried out to chromatograms of M/Z=85, 83, individual M/Zs, M/Z=191 and 217, respectively. The present work shows that, among numerous biomarkers present in the source oil, stigmastane and two isomers of hopane showed invariable concentrations after laboratory experiments that mimic natural biodegradation in the field, so they can be used as conserved internal biomarkers. These are very useful tools to assess alterations in less stable classes of saturated compounds contained in petroleum. Marked degradation of perhydro, tetrahydro, dihydro-PAH derivatives in the laboratory treatment has been evidenced.     

Semipermeable membrane devices accumulate conserved ratios of sterane and hopane petroleum biomarkers

Semipermeable membrane devices (SPMDs) are commonly used as a time-integrated measure of aqueous concentrations of persistent hydrophobic chemicals, including PAH, pesticides, dioxins, and PCBs. Another class of persistent hydrophobic chemicals is petroleum biomarker compounds (hopanes and steranes) that are used for hydrocarbon source identification and allocation. In this study three different passive sampling device designs were exposed to a complex hydrocarbon mixture (Alaska North Slope crude) in a laboratory experiment to determine uptake rates of biomarkers into SPMDs. In addition to the standard triolein filled SPMDs, iso-octane filled SPMDs (ISPMDs) and unfilled low-density polyethylene strips (PESDs) were tested. Uptake rates and effective sampling rates were determined for 53 compounds. There was little variation in sampling rates among the individual biomarkers; average values (ld(-1)) for hopanes were 0.43+/-0.07 (PESD), 0.33+/-0.06 (SPMD), and 0.44+/-0.03 (ISPMD) and average sterane sampling rates were 0.57+/-0.04 (PESD), 0.42+/-0.03 (SPMD), and 0.53+/-0.03 (ISPMD). The primary reason for biomarker analysis is for source discrimination of petroleum. Nineteen different diagnostic ratios were measured, and were found to be remarkably well conserved between the oil, water, and all three devices. This indicates that SPMDs, PESDs, and ISPMDs should each be effective for source discrimination studies of petroleum contamination.     

Molecular markers, essential oils and diagnostic parameters as indicators for assessing the origin and constitution of organic matter in atmospheric aerosols

Atmospheric particulate matter was collected at two sites: an Abies boressi forest in central Greece and in Giesta, a coastal-rural site in the centre of Portugal. The extractable organic material consisted primarily of aliphatic hydrocarbons, acids, alcohols, and ketones, with a predominance of molecular weights >C20, derived from vascular plant waxes. Biomarkers for vegetation sources such as phytosterols, triterpenic compounds and essential oils were also detected. Microbial components (>C20), pyrogenic aromatic hydrocarbons and petroleum residues, including hopanes, steranes and cyclic and branched hydrocarbons, were present in the various aerosol extracts. In the apportionment of the various organic fractions to sources, vegetation waxes dominated, contributing to 50–60% of the solvent-extractable material in the aerosols. Petroleum residues and microbial components comprise 25–30% and 15–25% of the organic matter, respectively.     

Source diagnostic and weathering indicators of tar balls utilizing acyclic, polycyclic and S-heterocyclic components

This study represents a forensic chemical analysis to define the liability for the coastal bitumens polluting the beaches of the Mediterranean city of Alexandria. Six tar balls collected from several locations along the coast of the city were analyzed for their acyclic and polycyclic hydrocarbons as well as sulfur heterocycles using GC/FID, GC/AED and gas chromatography/mass spectrometry techniques. The analysis of one Egyptian crude oil is also included as a possible source oil. The tar ball samples were at early stages of weathering. Based on the GC traces and biomarker signatures, the tar balls could be genetically different. One sample collected from the Eastern Harbor region appears to be a Bunker C type fuel produced from Egyptian crudes. The refining process has removed the low molecular weight components. On the other hand, the wide n-alkane distribution together with the absence of an unresolved complex mixture suggests that crude oils probably from tank washings, ballast discharges or accident spills from tankers could have contributed significantly to the other tar ball samples. The distribution of source specific hopane and sterane markers revealed that the tar samples probably originate from different oil fields.     

Variations in speciated emissions from spark-ignition and compression-ignition motor vehicles in California's south coast air basin

The U.S. Department of Energy Gasoline/Diesel PM Split Study examined the sources of uncertainties in using an organic compound-based chemical mass balance receptor model to quantify the contributions of spark-ignition (SI) and compression-ignition (CI) engine exhaust to ambient fine particulate matter (PM2.5). This paper presents the chemical composition profiles of SI and CI engine exhaust from the vehicle-testing portion of the study. Chemical analysis of source samples consisted of gravimetric mass, elements, ions, organic carbon (OC), and elemental carbon (EC) by the Interagency Monitoring of Protected Visual Environments (IMPROVE) and Speciation Trends Network (STN) thermal/optical methods, polycyclic aromatic hydrocarbons (PAHs), hopanes, steranes, alkanes, and polar organic compounds. More than half of the mass of carbonaceous particles emitted by heavy-duty diesel trucks was EC (IMPROVE) and emissions from SI vehicles contained predominantly OC. Although total carbon (TC) by the IMPROVE and STN protocols agreed well for all of the samples, the STN/IMPROVE ratios for EC from SI exhaust decreased with decreasing sample loading. SI vehicles, whether low or high emitters, emitted greater amounts of high-molecular-weight particulate PAHs (benzo[ghi]perylene, indeno[1,2,3-cd]pyrene, and coronene) than did CI vehicles. Diesel emissions contained higher abundances of two- to four-ring semivolatile PAHs. Diacids were emitted by CI vehicles but are also prevalent in secondary organic aerosols, so they cannot be considered unique tracers. Hopanes and steranes were present in lubricating oil with similar composition for both gasoline and diesel vehicles and were negligible in gasoline or diesel fuels. CI vehicles emitted greater total amounts of hopanes and steranes on a mass per mile basis, but abundances were comparable to SI exhaust normalized to TC emissions within measurement uncertainty. The combustion-produced high-molecular-weight PAHs were found in used gasoline motor oil but not in fresh oil and are negligible in used diesel engine oil. The contributions of lubrication oils to abundances of these PAHs in the exhaust were large in some cases and were variable with the age and consumption rate of the oil. These factors contributed to the observed variations in their abundances to total carbon or PM2.5 among the SI composition profiles.     

Oil residues in Baltic sediment,mussel and fish. II. Study of the Finnish Archipelago 1979–1981

Aliphatic hydrocarbons and non-polar aromatic compounds have been analyzed after a spill of grude oil at Baltic Sea 1979 from drifting weathered oil glumps, sediment and sediment trap samples and bivalves at five areas of the Finnish Archipelago Sea. $\text{Mytilus}$ and flounder muscles and flounder livers were analyzed 1980 and 1981 from three of the previous five areas. The glumps were shown to be weathered grude oil. Sediment trap samples on the visibly contaminated area contained higher amounts of oil residues and in different ratios than the bottom sediments at the same areas. Extra local aliphatic hydrocarbon pollution was detected from the sediment traps at the reference area. Bivalves contained high amount of oil residues at visibly polluted but in some visibly non-polluted areas also 1979. Any significant decontamination was not detected at summer 1979 but next year the $\text{Mytilus}$ samples contained only traces if any and 1981 no detectable levels of aromatic oil residues. Aliphatic hydrocarbon residues in flounders showed no regional differences and no significant change of levels between 1980 and 1981. In contrary, the aromatic oil residues decreased to the non-detectable level in flounder muscles and to a significantly lower level in flouder livers 1981 related to 1980. Linear correlations of the analysis results are discussed.     

Microbial in situ degradation of aromatic hydrocarbons in a contaminated aquifer monitored by carbon isotope fractionation

We present an approach for characterizing in situ microbial degradation using the 13C/12C isotope fractionation of contaminants as an indicator of biodegradation. The 13C/12C isotope fractionation of aromatic hydrocarbons was studied in anoxic laboratory soil percolation columns with toluene or o-xylene as the sole carbon and electron source, and sulfate as electron acceptor. After approximately 2 months' of incubation, the soil microbial community degraded 32 mg toluene l(-1) and 44 mg o-xylene l(-1) to less than 0.05 mg l(-1), generating a stable concentration gradient in the column. The 13C/12C isotope ratio in the residual non-degraded fraction of toluene and o-xylene increased significantly, corresponding to isotope fractionation factors (alphaC) of 1.0015 and 1.0011, respectively. When the extent of biodegradation in the soil column was calculated based on the measured isotope ratios (R(t)) and an isotope fractionation factor (alphaC=1.0017) obtained from a sulfate-reducing batch culture the theoretical residual substrate concentrations (C(t)) matched the measured toluene concentrations in the column. This indicated that a calculation of biodegradation based on isotope fractionation could work in systems like soil columns. In a field study, a polluted, anoxic aquifer was analyzed for BTEX and PAH contaminants. These compounds were found to exhibit a significant concentration gradient along an 800-m groundwater flow path downstream of the source of contamination. A distinct increase in the carbon isotope ratio (delta13C) was observed for the residual non-degraded toluene (7.2 per thousand ), o-xylene (8.1 per thousand ) and naphthalene fractions (1.2 per thousand ). Based on the isotope values and the laboratory-derived isotope fractionation factors for toluene and o-xylene, the extent to which the residual substrate fraction in the monitoring wells had been degraded by microorganisms was calculated. The results revealed significant biodegradation along the groundwater flow path. In the wells at the end of the plume, the bioavailable toluene and o-xylene fractions had been almost completely reduced by in situ microbial degradation. Although indane and indene showed decreasing concentrations downstream of the groundwater flow path, suggesting microbial degradation, their carbon isotope ratios remained constant. As the physical properties of these compounds are similar to those of BTEX compounds, the constant isotope values of indane and indene indicated that microbial degradation did not lead to isotope fractionation of all aromatic hydrocarbons. In addition, physical interaction with the aquifer material during the groundwater passage did not significantly alter the carbon isotope composition of aromatic hydrocarbons.     

Fingerprinting of High Boiling Hydrocarbon Fuels,Asphalts and Lubricants

Fingerprinting of hydrocarbon products requires high resolution differentiation of individual hydrocarbon compounds in any mixture. This requires the applications of various measuring techniques. In this paper, we have chosen the heavy hydrocarbons in fuels, lubricants and paving material as examples to discuss the methods for chemical characterization and differentiation. In the category most frequently termed "semi-volatile hydrocarbons" with boiling points from about 500°F to 1200°F or higher, there are several families of hydrocarbons, both natural and refined that are not easily distinguished by conventional EPA tests. Among the groups which we will use as examples are asphalts, hydraulic fluid, transmission oil, motor lubricating oils, heating oils, crude oil and coal. These hydrocarbon families are best studied using combined gas chromatography-mass spectrometry in full scan mode and characterizing various homologous series of hydrocarbons at known fragment ions. The hydrocarbon series providing the best information are: (1) N -alkanes; (2) iso-alkanes; (3) steranes; (4) terpanes; (5) polynuclear aromatic hydrocarbons; (6) aromatic steranes; and (7) specific polycyclic compounds.     

Roadside measurements of size-segregated particulate organic compounds near gasoline and diesel-dominated freeways in Los Angeles,CA

Individual organic compounds such as hopanes and steranes (originating in lube oil) and selected polycyclic aromatic compounds (PAHs) (generated via combustion) found in particulate emissions from vehicles have proven useful in source apportionment of ambient particulate matter (PM). Detailed information on the size-segregated (ultrafine and accumulation mode) chemical characteristics of organic PM during the winter season originating from a pure gasoline traffic freeway (CA-110), and a mixed-fleet freeway with the highest fraction of heavy-duty diesel vehicles in the state of California (I-710) is reported in this study. Hopanes and steranes as well as high molecular weight PAHs such as benzo(ghi)perylene (BgP) and coronene levels are found comparable near these freeways, while elemental carbon (EC) and lighter molecular weight PAHs are found much elevated near I-710 compared to CA-110. The roadway organic speciation data presented here are compared with the emission factors (EFs) measured in the Caldecott tunnel, Berkeley, CA [Phuleria, H.C., Geller, M.D., Fine, P.M., Sioutas, C., 2006. Size-resolved emissions of organic tracers from light- and heavy-duty vehicles measured in a California roadway tunnel. Environmental Science and Technology 40, 4109–4118] for light-duty vehicles (LDVs) and heavy-duty vehicles. Very good agreement is observed between CA-110 measurements and LDV EFs as well as I-710 measurements and corresponding reconstructed EFs from Caldecott tunnel for hopanes and steranes as well as heavier PAHs such as BgP and coronene. Our results, therefore, suggest that the EFs for hopanes and steranes obtained in tunnel environments, where emissions are averaged over a large vehicle-fleet, enable reliable source apportionment of ambient PM, given the overall agreement between the roadway vs tunnel concentrations of these species.     

Fingerprinting of High Boiling Hydrocarbon Fuels,Asphalts and Lubricants

Fingerprinting of hydrocarbon products requires high resolution differentiation of individual hydrocarbon compounds in any mixture. This requires the applications of various measuring techniques. In this paper, we have chosen the heavy hydrocarbons in fuels, lubricants and paving material as examples to discuss the methods for chemical characterization and differentiation.In the category most frequently termed “semi-volatile hydrocarbons” with boiling points from about 500°F to 1200°F or higher, there are several families of hydrocarbons, both natural and refined that are not easily distinguished by conventional EPA tests. Among the groups which we will use as examples are asphalts, hydraulic fluid, transmission oil, motor lubricating oils, heating oils, crude oil and coal.These hydrocarbon families are best studied using combined gas chromatography-mass spectrometry in full scan mode and characterizing various homologous series of hydrocarbons at known fragment ions. The hydrocarbon series providing the best information are: (1)N -alkanes; (2) iso-alkanes; (3) steranes; (4) terpanes; (5) polynuclear aromatic hydrocarbons; (6) aromatic steranes; and (7) specific polycyclic compounds.     

Oil residues in Baltic sediment,mussel and fish. I. Development of the analysis methods

Sediment, sediment trap, Mytilus, Macoma and flounder samples from Northern Baltic (Finnish archipelago) have been analyzed for their contents of aliphatic and aromatic hydrocarbons. Androstane and hexaethylbenzene were used as internal standards. The analysis procedure consisted of alkaline degradation of fat, column fractionation of the two residue groups and final determination by glass capillary gas chromatography with FID for aliphatic hydrocarbon group and with mass spectrometry for non-polar aromatic residue group. The latter group was also determined by high pressure liquid chromatography. The residues due to oil pollution were distinguished from compounds of pure natural origin on the basis of statistical treatment of the determination results.     

Biodegradation of aromatic hydrocarbons by Haloarchaea and their use for the reduction of the chemical oxygen demand of hypersaline petroleum produced water

Ten halophilic Archaea (Haloarchaea) strains able to degrade aromatic compounds were isolated from five hypersaline locations; salt marshes in the Uyuni salt flats in Bolivia, crystallizer ponds in Chile and Cabo Rojo (Puerto Rico), and sabkhas (salt flats) in the Persian Gulf (Saudi Arabia) and the Dead Sea (Israel and Jordan). Phylogenetic identification of the isolates was determined by 16S rRNA gene sequence analysis. The isolated Haloarchaea strains were able to grow on a mixture of benzoic acid, p-hydroxybenzoic acid, and salicylic acid (1.5 mM each) and a mixture of the polycyclic aromatic hydrocarbons, naphthalene, anthracene, phenanthrene, pyrene and benzo[a]anthracene (0.3 mM each). Evaluation of the extent of degradation of the mixed aromatic hydrocarbons demonstrated that the isolates could degrade these compounds in hypersaline media containing 20% NaCl. The strains were shown to reduce the COD of hypersaline crude oil reservoir produced waters significantly beyond that achieved using standard hydrogen peroxide treatment alone.     

Bioremediation of diesel-contaminated soil with composting

The major objective of this research was to find the appropriate mix ratio of organic amendments for enhancing diesel oil degradation during contaminated soil composting. Sewage sludge or compost was added as an amendment for supplementing organic matter for composting of contaminated soil. The ratios of contaminated soil to organic amendments were 1:0.1, 1:0.3, 1:0.5, and 1:1 as wet weight basis. Target contaminant of this research was diesel oil, which was spiked at 10,000 mg/kg sample on a dry weight basis. The degradation of diesel oil was significantly enhanced by the addition of these organic amendments relative to straight soil. Degradation rates of total petroleum hydrocarbons (TPH) and n-alkanes were the greatest at the ratio of 1:0.5 of contaminated soil to organic amendments on wet weight basis. Preferential degradation of n-alkanes over TPH was observed regardless of the kind and the amount of organic amendments. The first order degradation constant of n-alkanes was about twice TPH degradation constant. Normal alkanes could be divided in two groups (C10-C15 versus C16-C20) based on the first order kinetic constant. Volatilization loss of TPH was only about 2% of initial TPH. Normal alkanes lost by volatilization were mainly by the compounds of C10 to C16. High correlations (r=0.80-0.86) were found among TPH degradation rate, amount of CO2 evolved, and dehydrogenase activity.     

Tracing the transformation of labelled [1-13C]phenanthrene in a soil bioreactor

[1-(13)C]-labelled phenanthrene was incubated in a closed bioreactor to study the flux and biotransformation of polycyclic aromatic hydrocarbon (PAH) in contaminated soils on a bulk and molecular level. The degradation of extractable phenanthrene was observed by GC-MS measurements and the mineralisation was monitored by (13)CO(2) production. The transformation of the (13)C-label into non-extractable soil-bound residues was determined by carbon isotopic measurements. With these data we were able to calculate a carbon budget of the (13)C-label. Moreover, the chemical structure of non-extractable bound residues was characterised by applying selective chemical degradation reactions to cleave xenobiotic subunits from the macromolecular organic soil matrix. The obtained low molecular weight products yielded (13)C-labelled compounds which were identified using IRM (isotope ratio monitoring)-GC-MS and structurally characterised with GC-MS. Most of the (13)C-labelled products obtained by chemical degradation of non-extractable bound residues are well-known metabolites of phenanthrene. Thus, metabolites of [1-(13)C]phenanthrene formed during biodegradation appear to be reactive components which are subsequently involved in the bound residue formation. Hydrolysable amino acids of the soil residues were significantly labelled with (13)C as confirmed by IRM-GC-MS measurements. Therefore, phenanthrene-derived carbon was transformed by anabolic microbial processes into typical biologically derived compounds. These substances are likely to be incorporated into humic-like material after cell death.     

Influence of combustion conditions on yields of solvent-extractable anhydrosugars and lignin phenols in chars: Implications for characterizations of biomass combustion residues

Anhydrosugars, such as levoglucosan and its isomers (mannosan, galactosan), as well as the solvent-extractable lignin phenols (methoxylated phenols) are thermal degradation products of cellulose/hemicellulose and lignin, respectively. These two groups of biomarkers are often used as unique tracers of combusted biomass inputs in diverse environmental media. However, detailed characterization of the relative proportion and signatures of these compounds in highly heterogeneous plant-derived chars are still scarce. Here we conducted a systematic study to investigate the yields of solvent-extractable anhydrosugars and lignin phenols in 25 lab-made chars produced from different plant materials under different combustion conditions. Solvent-extractable anhydrosugars and lignin phenols were only observed in chars formed below 350 °C and yields were variable across different combustion temperatures. The yields of mannosan (M) and galactosan (G) decreased more rapidly than those of levoglucosan (L) under increasing combustion severity (temperature and duration), resulting in variable L/M and L/(M + G) ratios, two diagnostic ratios often used for identification of combustion sources (e.g. hardwoods vs. softwoods vs. grasses). Our observations thus may provide an explanation for the wide ranges of values reported in the literature for these two ratios. On the other hand, the results of this study suggest that the ratios of the major solvent-extractable lignin phenols (vanillyls (V), syringyls (S), cinnamyls (C)) provide additional source reconstruction potential despite observed variations with combustion temperature. We thus propose using a property-property plot (L/M vs. S/V) as an improved means for source characterization of biomass combustion residues. The L/M-S/V plot has shown to be effective in environmental samples (soil organic matter, atmospheric aerosols) receiving substantial inputs of biomass combustion residues.     

Brazilian Oil Spills Chemical Characterization—Case Studies

In the last decade, PETROBRAS has experienced some significant oil spills cases and the PETROBRAS Research Center has played an important role in the company emergency response program by characterizing the spilled oil, monitoring the affected ecosystem, determining the fate of the oil in the environment, and, subsequently, helping the company in assessing the environmental damage. This paper presents the use of advanced chemical analytical techniques (GC/FID, P&T/GC/PID and GC/MS) in some Brazilian oil spill studies in order to determine fractions and individual petroleum hydrocarbons in different matrices such as water, groundwater, sediment, sand, fish and the spilled oil itself. The spill studies encompassed crude and fuel oil releases on land and coastal ecosystems, related to the incidents in Guanabara Bay (Rio de Janeiro), Barigui and Iguassu Rivers (Parana) and Sao Sebastiao Channel (Sao Paulo). Total petroleum hydrocarbons (TPH), n -alkanes, isoprenoids, unresolved complex mixtures (UCM), volatile monoaromatic compounds—benzene, toluene, ethylbenzene and xylenes (BTEX), parent and alkylated homologues polycyclic aromatic hydrocarbons (PAH), and terpanes and steranes were characterized for determining correlation to the spilled oil and other known oil sources and environmental assessment. Some of the acute ecotoxicity data for water and sediment samples is also presented.     

Crude oil and hydrocarbon-degrading strains of Rhodococcus rhodochrous isolated from soil and marine environments in Kuwait

Soil and marine samples collected from different localities in Kuwait were screened for microorganisms capable of oil degradation. Both fungi and bacteria were isolated. The fungal flora consisted of Aspergillus terreus, A. sulphureus, Mucor globosus, Fusarium sp. and Penicillum citrinum. Mucor globosus was the most active oil degrading fungus isolated. Bacterial isolates included Bacillus spp. Enterobacteriaceae, Pseudomonas spp., Nocardia spp., Streptomyces spp.,and Rhodococcus spp. Among these Rhodococcus strains were the most efficient in oil degradation and, relatively speaking, the most abundant. Bacterial and fungal isolates differed in their ability to degrade crude oil, with Rhodococcus isolates being more active that fungin in n-alkane biodegradation, particularly in the case of R. rhodochrous. In addition to medium chain n-alkanes, fungi utilized one or more of the aromatic hydrocarbons studied, while bacteria failed to do so. R. rhodochorous KUCC 8801 was shown by GLC and post-growth studies to be more efficient in oil degradation than isolates known to be active oil degraders.     

Biodegradation of phenol and phenol-related compounds by psychrophilic and cold-tolerant alpine yeasts

We characterized 32 cold-adapted, psychrophilic and cold-tolerant, yeast strains isolated from alpine habitats with regard to their taxonomy, growth temperature profile, and ability to degrade phenol and 18 phenol-related mono-aromatic compounds at 10 degrees C. Twenty of the strains were identified by sequencing of the ribosomal ITS region as seven species of the basidiomycota: Cryptococcus terreus (three strains), Cryptococcus terricola (one strain), Rhodosporidium lusitaniae (two strains), Rhodotorula creatinivora (10 strains), Rhodotorula ingeniosa (one strain), Mastigobasidium intermedium (one strain), and Sporobolomyces roseus (two strains). Twelve strains sharing closely related ITS sequences could not be identified to the species level; according to their ITS sequence they are included in the Microbotryomycetidae. These 12 strains were psychrophilic (no growth at temperatures above 20 degrees C); one-third of these strains did not grow above 15 degrees C. None of the 32 strains utilized any of the highly volatile mono-aromatic compounds (benzene, toluene, ethylbenzene, nitrobenzene, o-xylene, m-xylene, and p-xylene) as the sole carbon source. Non/low volatile aromatic compounds were degraded in the following order: phenol>hydroquinone>resorcinol>benzoate>catechol>salicylate>p-cresol>m-cresol. o-Cresol, guaiacol, p-nitrophenol, or p-nitrotoluene were not utilized for growth. R. creatinivora strains degraded up to seven compounds, whereas C. terricola and S. roseus strains degraded only two compounds. The toxicity of the compounds was determined via growth inhibition in the presence of toxicants and nutrients at 10 degrees C. R. creatinivora strains were characterized by higher IC50 values than other species, S. roseus was the most sensitive species. The most toxic compounds were the xylene isomers, ethylbenzene, p-nitrophenol, and m-cresol. There was a relation between the chemical structure of the compounds and their toxicity, whereas a relation between the toxicity of the compounds and the ability of the yeasts strains to utilize these compounds for growth was only detected in some cases.