Alkanes In Crude Oils Research Articles

Differential biodegradation of alkanes in crude oil by three oleophilic strains

Ex situ bioremediation of crude oil in bioreactors by oleophilic bacteria permits to regulate temperature, nutrients and oxygen and the results in petroleum removal are much more effective than in situ bioremediation. This article quantify the biodegradation of alkanes which conform crude oil in bioreactors by three oleophilic strains. Samples were collected from the bioreactors and extracted in dichloromethane (10:1 aqueous/organic v/v) for amplification in GC-FID analysis of individual alkanes. 94% of crude oil (initial concentration 8.6 g l−1) is removed in 24 days by Bacillus licheniformis and in 38 days by Pseudomonas putida and Paenibacillus glucanolyticus. Higher biodegradation rates (9.0–26.5 mg l−1d−1) are reached for the lightest fraction of crude oil by Bacillus licheniformis (C10–C16), and much lower for C22–C25 alkanes (4.4–8.9 mg l−1d−1). Pseudomonas putida and Paenibacillus glucanolyticus degrade a wide range of alkanes at 1.6–9.6 mg l−1d−1, with preference over C8–C16 hydrocarbons. Heptane and octane (alkanes cited to be toxic) are difficult for biodegradation in two strains (B. licheniformis and P. putida) and a recalcitrant “window” C18–C21 is observed for the three oleophilic strains in the alkane range studied.

Understanding the phase behavior during CO2 flooding by dissipative particle dynamics

Understanding the phase behavior of a CO2-oil–water surfactant system is critical for carbon dioxide flooding engineering. Using dissipative particle dynamics, we investigate how the alkanes in crude oil are extracted after the injection of supercritical carbon dioxide into the formation, and what effect this extraction has on the miscible and immiscible displacement of carbon dioxide and crude oil. The images of the extraction are exhibited at the molecular level, and the effects of temperature and pressure on the extraction are investigated. The phase distribution of carbon dioxide, oil, and water under immiscible phase is simulated using actual reservoir fluid data based on a comprehensive understanding of extraction. Oil tends to be spread at the interface of carbon dioxide and water when it is immiscible. The miscible pictures were generated under conditions of reduced CO2-oil–water miscible pressure by adding surfactants to the CO2-oil–water system, and the fluid distribution characteristics under immiscible and miscible phases were examined. dissipative particle dynamics (DPD) simulation demonstrates that when the molar proportion of carbon dioxide is less than 0.4, the solid phase surface can always create a water film of varied thickness, regardless of whether the surface charge of the solid phase is positive or negative. As the molar fraction of carbon dioxide increases, the water film’s thickness decreases.

Identification of long-chain alkane-degrading (LadA) monooxygenases in Aspergillus flavus via in silico analysis.

Efficient degradation of alkanes in crude oil by the isolated Aspergillus flavus MM1 alluded to the presence of highly active alkane-degrading enzymes in this fungus. A long-chain alkane-degrading, LadA-like enzyme family in A. flavus was identified, and possible substrate-binding modes were analyzed using a computational approach. By analyzing publicly available protein databases, we identified six uncharacterized proteins in A. flavus NRRL 3357, of which five were identified as class LadAα and one as class LadAβ, which are eukaryotic homologs of bacterial long-chain alkane monooxygenase (LadA). Computational models of A. flavus LadAα homologs (Af1-Af5) showed overall structural similarity to the bacterial LadA and the unique sequence and structural elements that bind the cofactor Flavin mononucleotide (FMN). A receptor-cofactor-substrate docking protocol was established and validated to demonstrate the substrate binding in the A. flavus LadAα homologs. The modeled Af1, Af3, Af4, and Af5 captured long-chain n-alkanes inside the active pocket, above the bound FMN. Isoalloxazine ring of reduced FMN formed a π–alkyl interaction with the terminal carbon atom of captured alkanes, C16–C30, in Af3–Af5 and C16–C24 in Af1. Our results confirmed the ability of identified A. flavus LadAα monooxygenases to bind long-chain alkanes inside the active pocket. Hence A. flavus LadAα monooxygenases potentially initiate the degradation of long-chain alkanes by oxidizing bound long-chain alkanes into their corresponding alcohol.

Superimposed secondary alteration of oil reservoirs. Part II: The characteristics of biomarkers under the superimposed influences of biodegradation and thermal alteration

Petroleum biomarkers could be altered to varying extents under the influence of biodegradation and thermal alteration in reservoir, and their respective influence has been well documented. However, the superimposed influence of successive biodegradation and thermal alteration has not been systematically studied so far. In this study, a sequence of crude oils and oil sands that are of identical origin but at different biodegradation levels from the Liaohe Basin of China were artificially matured via closed-system pyrolysis. The distributions of biomarkers such as steranes and hopanes in the variably biodegraded oil samples and in their pyrolyzates at varying artificial thermal maturities were compared. The results showed that the presence of normal (n-) alkanes in crude oil does not necessarily excludes the possibility of previous moderate–severe biodegradation as n-alkanes can be further generated under advanced thermal stress. Besides, biomarkers in the oils that have suffered superimposed alterations could be derived not only from the modifications of free biomarkers in the oils, but also from the release of biomarkers bound in the asphaltenes. The mix of these two sources complicated the biomarker profiles in this type of oils. Most source related parameters based on steranes and tri-, tetra- and penta-cyclic terpanes seem to be reliable only when the oil reservoir has suffered biodegradation not beyond moderate level and the maturity of the crude oil is below 0.91 Easy%Ro. The TT23/(TT23 + TT24) ratio, however, was found to remain constant even after severe biodegradation and thermal alteration, making it applicable in oil-source correlation in superimposed oil reservoirs. In addition, isomerization-based maturity parameters such as methylphenanthrene index of crude oils are reliable if subsequent thermal alteration reached above 0.91 Easy%Ro despite prior moderate–severe biodegradation, because the related isomerization reactions would approach the same level observed for the non-biodegraded oil after subsequent thermal alteration.

Isolation, screening, and crude oil degradation characteristics of hydrocarbons-degrading bacteria for treatment of oily wastewater.

The aim of this study was to isolate hydrocarbons-degrading bacteria for treatment of oily wastewater from long-standing petroleum-polluted sediments in Bohai Bay, China. Six hydrocarbons-degrading bacteria were screened and identified as Pseudomonas sp. and Bacillus sp. A new approach using a combination of various bacterial species in petroleum biodegradation was proposed and evaluated for its degradation characteristics. Gas chromatography-flame ionization detection (GC-FID) analysis showed that mixed bacterial agents (YJ01) degraded 80.64% of crude oil and 76.30% of crude oil alkanes, exhibiting good biodegradation effect. Besides, after 14 days of culture, the biodegradation assessment markers, pristane and phytane, showed significant degradation rates of 46.75% and 78.23%, respectively. Kinetic analysis indicated that the degradation trends followed a single first-order kinetics model and the degradation half-life (t1/2) of 15 g/L crude oil was significantly shorter (5.48 days). These results indicated that YJ01 could degrade a wider range of hydrocarbons as well as some recalcitrant hydrocarbon components, and can be applied for bioremediation and treatment of oil-contaminated environment.

Enrichment and Characterization of a Psychrotolerant Consortium Degrading Crude Oil Alkanes Under Methanogenic Conditions.

Anaerobic alkane degradation via methanogenesis has been intensively studied under mesophilic and thermophilic conditions. While there is a paucity of information on the ability and composition of anaerobic alkane-degrading microbial communities under low temperature conditions. In this study, we investigated the ability of consortium Y15, enriched from Shengli oilfield, to degrade hydrocarbons under different temperature conditions (5-35 °C). The consortium could use hexadecane over a low temperature range (15-30 °C). No growth was detected below 10 °C and above 35 °C, indicating the presence of cold-tolerant species capable of alkane degradation. The preferential degradation of short chain n-alkanes from crude oil was observed by this consortium. The structure and dynamics of the microbial communities were examined using terminal restriction fragment length polymorphism (T-RFLP) fingerprinting and Sanger sequencing of 16S rRNA genes. The core archaeal communities were mainly composed of aceticlastic Methanosaeta spp. Syntrophaceae-related microorganisms were always detected during consecutive transfers and dominated the bacterial communities, sharing 94-96 % sequence similarity with Smithella propionica strain LYP(T). Phylogenetic analysis of Syntrophaceae-related clones in diverse methanogenic alkane-degrading cultures revealed that most of them were clustered into three sublineages. Syntrophaceae clones retrieved from this study were mainly clustered into sublineage I, which may represent psychrotolerant, syntrophic alkane degraders. These results indicate the wide geographic distribution and ecological function of syntrophic alkane degraders.

Progressive degradation of crude oil n-alkanes coupled to methane production under mesophilic and thermophilic conditions.

Although methanogenic degradation of hydrocarbons has become a well-known process, little is known about which crude oil tend to be degraded at different temperatures and how the microbial community is responded. In this study, we assessed the methanogenic crude oil degradation capacity of oily sludge microbes enriched from the Shengli oilfield under mesophilic and thermophilic conditions. The microbial communities were investigated by terminal restriction fragment length polymorphism (T-RFLP) analysis of 16S rRNA genes combined with cloning and sequencing. Enrichment incubation demonstrated the microbial oxidation of crude oil coupled to methane production at 35 and 55°C, which generated 3.7±0.3 and 2.8±0.3 mmol of methane per gram oil, respectively. Gas chromatography-mass spectrometry (GC-MS) analysis revealed that crude oil n-alkanes were obviously degraded, and high molecular weight n-alkanes were preferentially removed over relatively shorter-chain n-alkanes. Phylogenetic analysis revealed the concurrence of acetoclastic Methanosaeta and hydrogenotrophic methanogens but different methanogenic community structures under the two temperature conditions. Candidate divisions of JS1 and WWE 1, Proteobacteria (mainly consisting of Syntrophaceae, Desulfobacteraceae and Syntrophorhabdus) and Firmicutes (mainly consisting of Desulfotomaculum) were supposed to be involved with n-alkane degradation in the mesophilic conditions. By contrast, the different bacterial phylotypes affiliated with Caldisericales, “Shengli Cluster” and Synergistetes dominated the thermophilic consortium, which was most likely to be associated with thermophilic crude oil degradation. This study revealed that the oily sludge in Shengli oilfield harbors diverse uncultured microbes with great potential in methanogenic crude oil degradation over a wide temperature range, which extend our previous understanding of methanogenic degradation of crude oil alkanes.

Isolation and characterization of crude oil degrading bacteria from the Persian Gulf (Khorramshahr provenance)

Fifteen crude oil degrading bacteria were isolated from oil contaminated sites in the Persian Gulf at Khorramshahr provenance. These bacteria were screened with two important factors such as growth rate on crude oil and hydrocarbon biodegradation, and then three strains were selected from 15 isolated strains for further study. One strain (PG-Z) that show the best crude oil biodegradation was selected between all isolates. Nucleotides sequencing of the gene encoding for 16S rRNA show that strain PG-Z belong to Corynebacterium variabile genus. This strain was efficient in degrading of crude oil. This strain was capable to degraded 82% of crude-oil after one week incubation in ONR7a medium. The PG-Z strain had high emulsification activity and biosurfactant production between all isolates. GC–MS analysis shows that C. variabile strain PG-Z can degrade different alkanes in crude oil.

The detection of high‐mass aliphatics in petroleum by matrix‐assisted laser desorption/ionisation mass spectrometry

The development of simplified procedures for isolating high-mass alkanes present in crude oils is described. The new procedures, which bypass the sample recovery step with hot toluene in the conventional alkane-isolation procedure, also provide an effective sample preparation route, prior to analysis by matrix-assisted laser desorption/ionisation mass spectrometry (MALDI-MS). Urea-alkane adducts are formed by mixing sample and urea solutions on chromatographic paper or silica-coated plates. Unreacted hydrocarbons are removed by developing the plates with chloroform. In a second development with water, adducts are broken up in situ and the liberated urea removed, leaving bands of isolated alkanes behind. For MALDI-MS, strips of paper/plates, carrying the isolated alkanes, are fixed on metal target plates. The samples are treated with matrix (AgNO(3)) and analysed by MALDI-MS. The observed signal represents silver ion adducts of the isolated alkanes. Silver appears to work without much fragmentation and to generate whole silver adduct ions. Much improved MALDI-MS detection sensitivity and a wider range of masses was observed when samples were ablated from paper/plate surfaces, than by ablation from bulk samples spread over a smooth surface--the conventional method. Chromatographic paper gave better resolution and a broader range of masses than silica-coated plates. The analytical sequences have been confirmed using standard alkanes (C(20)-C(60)) and Polywax. The proposed procedures enhanced the sensitivity and detection range of the MS analysis. The method was useful in detecting n-alkanes to m/z 1500 (C(100)) and required relatively small quantities of sample and reagents. It provides a promising qualitative analysis route for the rapid isolation and reliable determination of alkanes in crude oils.

Isolation and characterization of crude-oil-degrading bacteria from the Persian Gulf and the Caspian Sea

Twenty-five crude-oil-degrading bacteria were isolated from oil-contaminated sites in the Persian Gulf and the Caspian Sea. Based on a high growth rate on crude oil and on hydrocarbon degradation ability, 11 strains were selected from the 25 isolated strains for further study. Determination of the nucleotide sequence of the 16S rRNA gene showed that these isolated strains belonged to genera Acinetobacter, Pseudomonas, Gordonia, Rhodococcus, Cobetia, Halomonas, Alcanivorax, Marinobacter and Microbacterium. Among the 11 isolates, strains BS (Acinetobacter calcoaceticus, 98%) and PG-12 (Alcanivorax dieselolei, 98%) were the most effective in degrading crude oil. Rate of crude-oil degradation of 82% (isolate BS) and 71% (isolate PG-12) were observed after 1week of cultivation in mineral medium. These strains had high emulsification activity and biosurfactant production. GC–MS analysis showed that A. dieselolei PG-12 can degrade different alkanes in crude oil. Screening of the distribution of the alkane hydroxylase gene in 25 isolates in relation to the source of isolation indicated that the group (II) alkane hydroxylase is prevalent in the Caspian Sea, but in the Persian Gulf, the frequency of the group (III) alkane hydroxylase gene is greater than that of the group (II) alkane hydroxylase gene.

The quantitative significance of Syntrophaceae and syntrophic partnerships in methanogenic degradation of crude oil alkanes

Libraries of 16S rRNA genes cloned from methanogenic oil degrading microcosms amended with North Sea crude oil and inoculated with estuarine sediment indicated that bacteria from the genera Smithella (Deltaproteobacteria, Syntrophaceace) and Marinobacter sp. (Gammaproteobacteria) were enriched during degradation. Growth yields and doubling times (36 days for both Smithella and Marinobacter) were determined using qPCR and quantitative data on alkanes, which were the predominant hydrocarbons degraded. The growth yield of the Smithella sp. [0.020 g(cell-C)/g(alkane-C)], assuming it utilized all alkanes removed was consistent with yields of bacteria that degrade hydrocarbons and other organic compounds in methanogenic consortia. Over 450 days of incubation predominance and exponential growth of Smithella was coincident with alkane removal and exponential accumulation of methane. This growth is consistent with Smithella's occurrence in near surface anoxic hydrocarbon degrading systems and their complete oxidation of crude oil alkanes to acetate and/or hydrogen in syntrophic partnership with methanogens in such systems. The calculated growth yield of the Marinobacter sp., assuming it grew on alkanes, was [0.0005 g(cell-C)/g(alkane-C)] suggesting that it played a minor role in alkane degradation. The dominant methanogens were hydrogenotrophs (Methanocalculus spp. from the Methanomicrobiales). Enrichment of hydrogen-oxidizing methanogens relative to acetoclastic methanogens was consistent with syntrophic acetate oxidation measured in methanogenic crude oil degrading enrichment cultures. qPCR of the Methanomicrobiales indicated growth characteristics consistent with measured rates of methane production and growth in partnership with Smithella.

Biodegradation of Crude Oil by Thermophilic Bacteria Isolated from a Volcano Island

One-hundred and fifty different thermophilic bacteria isolated from a volcanic island were screened for detection of an alkane hydroxylase gene using degenerated primers developed to amplify genes related to the Pseudomonas putida and Pseudomonas oleovorans alkane hydroxylases. Ten isolates carrying the alkJ gene were further characterized by 16s rDNA gene sequencing. Nine out of ten isolates were phylogenetically affiliated with Geobacillus species and one isolate with Bacillus species. These isolates were able to grow in liquid cultures with crude oil as the sole carbon source and were found to degrade long chain crude oil alkanes in a range between 46.64% and 87.68%. Results indicated that indigenous thermophilic hydrocarbon degraders of Bacillus and Geobacillus species are of special significance as they could be efficiently used for bioremediation of oil-polluted soil and composting processes.

Hydrogen isotopic compositions, distributions and source signals of individual n-alkanes for some typical crude oils in Lunnan Oilfield, Tarim Basin, NW China

Isotopic compositions of carbon-bound hydrogen in individual n-alkanes from several typical crude oil samples from Lunnan Oilfield, Tarim Basin, NW China, were firstly measured using newly developed gas chromatography-thermal conversion-isotope ratio mass spectrometry. The similar range of δ D of individual n -alkanes of crude oils among reservoirs of different geological times reflects that hydrocarbons are all derived from the same marine depositional environment. Compared to the theoretic value (-150‰) and the reported δ D values ( n C13― n C27, -160‰―-90‰) of individual n -alkanes for Ordovician-sourced crude oils in the Canadian Williston Basin, the hydrogen isotopic composition of individual n -alkanes in crude oils from Lunnan Oilfield is characterized by heavy hydrogen isotopic values ( n C12― n C27, -120‰―-60‰). In terms of the factors that control the fractionation of hydrogen isotopes, relatively saline depositional environment and higher thermal maturation were attributed to the heavy d D values of individual n-alkanes in crude oils from Lunnan Oilfield.

04/02236 Macrocyclic alkanes in crude oils and sedimentextracts: enrichment using molecular sieves: Audino, M. et al. Organic Geochemistry, 2004, 35, (5), 661–663

04/02236 Macrocyclic alkanes in crude oils and sedimentextracts: enrichment using molecular sieves: Audino, M. et al. Organic Geochemistry, 2004, 35, (5), 661–663

Macrocyclic alkanes in crude oils and sediment extracts: enrichment using molecular sieves

A molecular sieve method using NaX (13X) has been used to separate macrocyclic alkanes (in the carbon number range C 14–C 34), markers of the freshwater alga Botryococcus braunii, from the branched/cyclic hydrocarbon fractions of sediments and crude oils. The macrocyclic alkanes were included in the 13X sieve and were recovered by HF digestion.

Macrocyclic alkanes in crude oils from the algaenan of Botryococcus braunii

New classes of biomarkers, macrocyclic alkanes, have been previously identified in high grade oil shales (torbanites, Late Carboniferous-Late Permian) containing remains of the freshwater alga Botryococcus braunii ( B. braunii). In this study we report the occurrence of these compounds in two Indonesian crude oils (Minas and Duri, Tertiary) utilising metastable reaction monitoring GC–MS experiments. Based on δ 13C data and the distributions of the macrocyclic alkanes we suggest that the macrocyclic alkanes are likely formed by an olefin metathesis reaction of a structurally-specific polyunsaturated precursor i.e. the algaenan of B. braunii. The macrocyclic alkanes appear to be novel markers of B. braunii and add to the catalogue of the characteristic hydrocarbons derived from this alga. More importantly, these components could be original markers specific to the highly resistant algaenan of B. braunii in sediments and crude oils, unlike n-alkanes which can have multiple origins.

Hydrogen isotopic compositions of individual alkanes as a new approach to petroleum correlation: case studies from the Western Canada Sedimentary Basin

Isotopic compositions of carbon-bound hydrogen in individual n-alkanes and acyclic isoprenoid alkanes, from a number of crude oil samples, were measured using gas chromatography-thermal conversion-isotope ratio mass spectrometry. The precision of this technique is better than 3‰ for most alkanes, compared to the large range of δD variation among the samples (up to 160‰). The oils were selected from major genetic oil families in the Western Canada Sedimentary Basin, with source rocks ranging in age from Ordovician (and possibly Cambrian) to Cretaceous. The hydrogen isotopic composition of alkanes in crude oils is controlled by three factors: isotopic compositions of biosynthetic precursors, source water δD values, and postdepositional processes. The inherited difference in the lipid's biosynthetic origins and/or pathways is reflected by a small hydrogen isotopic variability within n-alkanes, but much larger differences in the δD values between n-alkanes and pristane/phytane. The shift toward lighter hydrogen isotopic compositions from Paleozoic to Upper Cretaceous oils in the WCSB reflects a special depositional setting and/or a minor contribution of terrestrial organic matter. The strong influence of source water δD values is demonstrated by the distinctively lower δD values of lacustrine oils than marine oils, and also by the high values for oils with source rocks deposited in evaporative environments. Thermal maturation may alter the δD values of the alkanes in the oil to some extent, but secondary oil migration does not appear to have had any significant impact. The fact that oils derived from source rocks that could be of Cambrian age still retain a strong signature of the hydrogen isotopic compositions of source organic matter, and source water, indicates that δD values are very useful for oil-source correlation and for paleoenvironmental reconstructions.

Isolation and characterization of a novel hydrocarbon-degrading, Gram-positive bacterium, isolated from intertidal beach sediment, and description of Planococcus alkanoclasticus sp. nov.

Characterization of a bacterial isolate (strain MAE2) from intertidal beach sediment capable of degrading linear and branched alkanes. A Gram-positive, aerobic, heterotrophic bacterium (strain MAE2), that was capable of extensive degradation of alkanes in crude oil but had a limited capacity for the utilization of other organic compounds, was isolated from intertidal beach sediment. MAE2 had an obligate requirement for NaCl but could not tolerate high salt concentrations. It was capable of degrading branched and n-alkanes in crude oil from C11 to C33, but was unable to degrade aromatic hydrocarbons. Comparative 16S rRNA sequence analysis placed the isolate with members of the genus Planococcus. That finding was corroborated by chemotaxonomic and physiological data. The fatty acid composition of strain MAE2 was very similar to the type species of the genus Planococcus, P. citreus (NCIMB 1493T) and P. kocurii (NCIMB 629T), and was dominated by branched acids, mainly a15:0. However, the 16S rRNA of strain MAE2 had less than 97% sequence identity with the type strains of P. citreus (NCIMB 1439T), P. kocurii (NCIMB 629T) and two Planococcus spp. (strain MB6-16 and strain ICO24) isolated from Antarctic sea ice. This indicated that strain MAE2 represented a separate species from these planococci. Morphologically, the isolate resembled P. okeanokoites (NCIMB 561T) and P. mcmeekinii S23F2 (ATCC 700539T). The cellular fatty acid composition of P. okeanokoites and P. mcmeekinii was considerably different from strain MAE2, and the mol % G + C content of P. mcmeekinii was far lower than that of MAE2. On the basis of phenotypic and genotypic data, it is proposed that strain MAE2 is a new species of Planococcus, Planococcus alkanoclasticus sp. nov., for which the type strain is P. alkanoclasticus MAE2 (NCIMB 13489T). Planococcus species are abundant members of the bacterial community in a variety of marine environments, including some in sensitive Antarctic ecosystems. The occurrence of hydrocarbon-degrading Planococcus spp. is potentially of importance in controlling the impact of hydrocarbon contamination in sensitive marine environments.

Time-of-flight mass spectrometric analysis of high-molecular-weight alkanes in crude oil by silver nitrate chemical ionization after laser desorption.

Time-of-flight mass spectrometry was used for the first time to analyze the hydrocarbons in crude oil. Alkanes in the saturated fraction of a crude oil sample were chemically ionized by the laser desorption of silver nitrate, and the silver-attached C24-C60 alkanes were resolved with mass accuracy below 7 ppm. This technique was used to evaluate the biodegradation of aliphatic hydrocarbons and cycloalkanes by oil-degrading microorganisms resident in seawater. It is shown that the aliphatic hydrocarbons were degraded in the range of 60-80%, while the mono-, di-, tri-, tetra-, and pentacycloalkanes were degraded in the range of 40-55, 20-30, 10-16, 5-9, and <5%, respectively. Its high sensitivity and speed of application could result in an analysis by laser desorption silver chemical ionization time-of-flight mass spectrometry being the method of choice for determining high-molecular-weight hydrocarbons in various petroleum products.

Effects of surfactant mixtures, including Corexit 9527, on bacterial oxidation of acetate and alkanes in crude oil.

Mixtures of nonionic and anionic surfactants, including Corexit 9527, were tested to determine their effects on bacterial oxidation of acetate and alkanes in crude oil by cells pregrown on these substrates. Corexit 9527 inhibited oxidation of the alkanes in crude oil by Acinetobacter calcoaceticus ATCC 31012, while Span 80, a Corexit 9527 constituent, markedly increased the oil oxidation rate. Another Corexit 9527 constituent, the negatively charged dioctyl sulfosuccinate (AOT), strongly reduced the oxidation rate. The combination of Span 80 and AOT increased the rate, but not as much as Span 80 alone increased it, which tentatively explained the negative effect of Corexit 9527. The results of acetate uptake and oxidation experiments indicated that the nonionic surfactants interacted with the acetate uptake system while the anionic surfactant interacted with the oxidation system of the bacteria. The overall effect of Corexit 9527 on alkane oxidation by A. calcoaceticus ATCC 31012 thus seems to be the sum of the independent effects of the individual surfactants in the surfactant mixture. When Rhodococcus sp. strain 094 was used, the alkane oxidation rate decreased to almost zero in the presence of a mixture of Tergitol 15-S-7 and AOT even though the Tergitol 15-S-7 surfactant increased the alkane oxidation rate and AOT did not affect it. This indicated that there was synergism between the two surfactants rather than an additive effect like that observed for A. calcoaceticus ATCC 31012.