Evaluation of a Gravity Flow Membrane Bioreactor for Treating Municipal Wastewater.

The biomass concentrator reactor (BCR), a gravity flow membrane bioreactor (MBR) design, was evaluated for use in treating a municipal wastewater stream. The BCR operates with less than 2.5 cm of pressure head and uses a 3 to 4 mm thick tortuous path membrane with pore size ranging from 18 to 28 µm to achieve solids separation. A two-stage, aerobic/anoxic reactor was evaluated for the removal of chemical oxygen demand (COD), ammonia, total nitrogen, and solids separation. The reactor was fed 72 L/day, with a hydraulic retention time of 9.3 hours, and had a solids retention time of 20 days. The influent COD was reduced by 93%, whereas, influent ammonia was reduced below 0.1 mg/L and total nitrogen was reduced by 53.7%. A lack of readily biodegradable COD limited denitrification and thus total nitrogen removal. The reactor solids were retained completely in the reactor by the membrane for the duration of testing.

Anaerobic biodegradation of soybean biodiesel and diesel blends under sulfate-reducing conditions.

Biotransformation of soybean biodiesel and its biodiesel/petrodiesel blends were investigated under sulfate-reducing conditions. Three blends of biodiesel, B100, B50, and B0, were treated using microbial cultures pre-acclimated to B100 (biodiesel only) and B80 (80% biodiesel and 20% petrodiesel). Results indicate that the biodiesel could be effectively biodegraded in the presence or absence of petrodiesel, whereas petrodiesel could not be biodegraded at all under sulfate-reducing conditions. The kinetics of biodegradation of individual Fatty Acid Methyl Ester (FAME) compounds and their accompanying sulfate-reduction rates were studied using a serum bottle test. As for the biodegradation of individual FAME compounds, the biodegradation rates for the saturated FAMEs decreased with increasing carbon chain length. For unsaturated FAMEs, biodegradation rates increased with increasing number of double bonds. The presence of petrodiesel had a greater effect on the rate of biodegradation of biodiesel than on the extent of removal.

Biodegradation and toxicity of vegetable oils in contaminated aquatic environments: Effect of antioxidants and oil composition.

Antioxidants may affect the oxidative rate of vegetable oils determining their fate and impact in contaminated aquatic media. In previous studies, we demonstrated the effectiveness of butylated hydroxytoluene (BHT), one of the most used antioxidants in edible oils, in enhancing the biodegradation of glyceryl trilinoleate, a pure triacylglycerol of cis,cis-9,12-octadecadienoic acid (C18:2 delta), through retarding its oxidative polymerization relatively to the oil with no added antioxidant. In this study, the effect of BHT on the biodegradation and toxicity of purified canola oil, a mixed-acid triacylglycerol with high C18:1 content, was investigated in respirometric microcosms and by use of the Microtox® assay. Investigations were carried out in the absence and presence (200 mg kg(-1)) of the antioxidant, and at an oil loading of 0.31 L m(-2) (333 gal acre(-1)). Substantial oil mineralization was achieved after 16 weeks of incubation (>77%) and was not significantly different (p>0.05) between the two BHT treatments, demonstrating an important role of the oil fatty acid composition in determining the potency of antioxidants and, consequently, the fate of spilled vegetable oils. Furthermore, for both treatments, toxicity was measured at early stages of the experiments and disappeared at a later stage of incubation. The observed transient toxicity was associated with the combined effect of toxic biodegradation intermediates and autoxidation products. These results were supported by the gradual disappearance of BHT in the microcosms initially supplemented with the antioxidant, reaching negligible amounts after only 2 weeks of incubation.

Effect of dispersants on the biodegradation of South Louisiana crude oil at 5 and 25 °C.

This article reports biodegradation rates for a commercial dispersant, JD-2000, South Louisiana crude oil (SLC) alone, and SLC dispersed with JD-2000 at 5 and 25 °C. Results from the biodegradation experiments revealed that Component X, a chemical marker for JD-2000, rapidly degraded at both temperatures. The application of JD-2000 decreased by half the overall biodegradation rate of aliphatic compounds at 25 °C. At 5 °C, a residual fraction consisting of iso- and n-alkanes (C29-C35) persisted after 56 d. The combination of dispersant and higher temperature resulted in faster removal rates for 2- and 3-ring polycyclic aromatic hydrocarbons. When compared with Corexit 9500, our results suggest that the chemistry of the surfactant (or surfactants) in JD-2000 might have favored oil dissolution (substrate transport to the aqueous phase) as an uptake mechanism over adhesion, which requires direct contact of the biomass with the oil.

Characterization of solidifiers used for oil spill remediation.

The physical characteristics and chemical composition of oil spill solidifiers were studied, and correlation of these properties with product effectiveness enabled determination of characteristics that are desirable in a good solidifier. The analyses revealed that the commercial products were primarily comprised of organic polymers and a few trace elements. A natural sorbent, which was composed entirely of plant based matter, was also evaluated, and it had the highest oil removal capacity, but it did not produce a solid mat-like final product. Generally, solidifiers with a carbonate group, pore size greater than 5 µm, and bulk densities lower than 0.3 g cm(-3) were found to have better efficiency and produced a cohesive rubbery final product that facilitated removal compared to sorbents. The importance of bulk density and pore size in the performance of the solidifier suggest that the primary mechanism of action was likely physical sorption.

Anaerobic biodegradation of soybean biodiesel and diesel blends under methanogenic conditions.

Biotransformation of soybean biodiesel and the inhibitory effect of petrodiesel were studied under methanogenic conditions. Biodiesel removal efficiency of more than 95% was achieved in a chemostat with influent biodiesel concentrations up to 2.45 g/L. The kinetics of anaerobic biodegradation of soybean biodiesel B100 (biodiesel only) with different petrodiesel loads was studied using biomass pre-acclimated to B100 and B80 (80% biodiesel and 20% petrodiesel). The results indicated that the biodiesel fraction of the blend could be effectively biodegraded, whereas petrodiesel was not biodegraded at all under methanogenic conditions. The presence of petrodiesel in blends with biodiesel had a greater inhibitory effect on the rate of biodegradation than the biodegradation efficiency (defined as the efficiency of methane production). Both the biodegradation rate coefficient and the methane production efficiency increased almost linearly with the increasing fraction of biodiesel. With the increasing fraction of petrodiesel, the biodegradation rate and efficiency were correlated with the concentration of soluble FAMEs in the water.

Parametric study to determine the effect of temperature on oil solidifier performance and the development of a new empirical correlation for predicting effectiveness.

Temperature can play a significant role in the efficacy of solidifiers in removing oil slicks on water. We studied and quantified the effect of temperature on the performance of several solidifiers using 5 different types of oils under a newly developed testing protocol by conducting experiments in constant temperature rooms set at 22°C and 5°C. The results indicated that solidifier efficiency decreased substantially at the lower temperature, especially at lower application rates. The removal efficiency of the solidifier was in general directly proportional to temperature, except for the heavier oils, where removal by attachment was observed. Solidifier products with lower powder bulk density exhibited the best removal effectiveness. Analysis of experimental data yielded empirical correlations involving certain operational variables such as application rate, temperature, solidifier property (bulk density), and oil property (viscosity). Regression analysis was used to fit a mathematical model to the experimental solidifier effectiveness data.

Aerobic biodegradation kinetics and mineralization of six petrodiesel/soybean-biodiesel blends.

The aerobic biodegradation kinetics and mineralization of six petrodiesel/soybean-biodiesel blends (B0, B20, B40, B60, B80, and B100), where B100 is 100% biodiesel, were investigated by acclimated cultures. The fatty acid methyl esters (FAMEs) of biodiesel were found to undergo rapid abiotic transformation in all experiments. The C10-C21 n-alkanes of petrodiesel were metabolized at significantly higher microbial utilization rates in the presence of biodiesel. The rates of mineralization of the blends were also enhanced in the presence of biodiesel; yet a similar enhancement in the extent of mineralization was not observed. Abiotic fuel-blends/aqueous-phase equilibration experiments revealed that the FAMEs of biodiesel were capable of cosolubilizing the n-alkanes of petrodiesel, a mechanism that fully explains the faster utilization and mineralization kinetics of petrodiesel in the presence of biodiesel without necessarily enhancing the extent of biomineralization. The biodegradation of six targeted aromatic compounds present in petrodiesel was also influenced by the amount of biodiesel in a blend. While toluene, o-xylene, and tetralin were not degraded in the B0 and B20 treatments, all of the targeted aromatic compounds were degraded to below detection limits in the B40 and B80 treatments. Biomass acclimated to B60, however, was unable to degrade most of the aromatic compounds. These results indicate that the amount of biodiesel in a blend significantly affects the absolute and relative abundance of the dissolved and bioavailable constituents of biodiesel and petrodiesel in a way that can considerably alter the biodegrading capacity of microbial cultures.

Key factors controlling the transport of silver nanoparticles in porous media.

The current study investigated the mobility of four silver nanoparticles (AgNPs) stabilized using different capping agents and represent the common stabilization mechanisms as well as surface charging scenarios in reactive and nonreactive porous media. The AgNPs were (1) uncoated H2-AgNPs (electrostatically stabilized) and (2) citrate coated AgNPs (Citrate-AgNPs) (electrostatically stabilized), (3) polyvinylpyrrolidone coated AgNPs (PVP-AgNPs) (sterically stabilized), and (4) branched polyethyleneimine coated AgNPs (BPEI-AgNPs) (electrosterically stabilized). The porous media were (1) quartz sand (QS), (2) ferrihydrite-coated sand (FcS), and (3) kaolin-coated sand (KcS). The H2-AgNPs and Citrate-AgNPs were readily mobile in QS but significantly retained in FcS and KcS with more deposition achieved in the KcS media. The deposition of the H2-AgNPs and Citrate-AgNPs followed the order of KcS > FcS > QS. The PVP-AgNPs breakthrough occurred more rapid as compared to the H2-AgNPs and Citrate-AgNPs but the deposition of PVP-AgNPs followed the same order of the electrostatically stabilized AgNPs (KcS > FcS > QS). The BPEI-AgNPs were readily mobile regardless of the porous media reactivity. Physicochemical interactions were the dominant filtration mechanism in the majority of the investigated cases but straining played the major role in the deposition of the electrostatically stabilized H2-AgNPs and Citrate-AgNPs in the KcS media. The results highlight the importance of both the stabilization mechanism and capping agent chemistry as key factors governing the transport of AgNPs in the environment.

Biodegradability of Corexit 9500 and dispersed South Louisiana crude oil at 5 and 25 °C.

The reported persistence of the dioctyl sodium sulfosuccinate (DOSS) surfactant in Corexit 9500 in the oil plumes formed during the Deepwater Horizon oil spill has contributed to concerns regarding the biodegradability and bioavailability of dispersed oil and dispersants used as an oil spill countermeasure in the Gulf of Mexico. We studied the biodegradation of DOSS and dispersed South Louisiana crude oil (SLC) in laboratory microcosms. Two oil-degrading cultures from the Gulf of Mexico were isolated, one from the surface (meso) and one from close to the area of the Macondo well (cryo). Each was enriched on SLC, the former at 25 °C, the latter at 5 °C. Results indicated that the meso culture rapidly and completely degraded DOSS, alkanes, and aromatics. The cryo culture metabolized the same compounds but with a lag of 28 d and a remaining residual of iso-alkanes, n-C(30-35), and the 4-ring PAHs.

Microbial kinetic model for the degradation of poorly soluble organic materials.

A novel mechanistic model is presented that describes the aerobic biodegradation kinetics of soybean biodiesel and petroleum diesel in batch experiments. The model was built on the assumptions that biodegradation takes place in the aqueous phase according to Monod kinetics, and that the substrate dissolution kinetics at the oil/water interface is intrinsically fast compared to biodegradation kinetics. Further, due to the very low aqueous solubility of these compounds, the change in the substrate aqueous-phase concentration over time was assumed to approaches zero, and that substrate aqueous concentration remains close to the saturation level while the non-aqueous phase liquid (NAPL) is still significant. No former knowledge of the saturation substrate concentration (S(sat)) and the Monod half-saturation constant (K(s)) was required, as the term S(sat)/(K(s) + S(sat)) in the Monod equation remained constant during this phase. The n-alkanes C10-C24 of petroleum diesel were all utilized at a relatively constant actual specific utilization rate of 0.01-0.02 mg-alkane/mg-biomass-hr, while the fatty acid methyl esters (FAMEs) of biodiesel were utilized at actual specific rates significantly higher with increasing carbon chain length and lower with increasing number of double bonds. The results were found to be in agreement with kinetic, genetic, and metabolic evidence reported in the literature pertaining to microbial decay rates, uptake mechanisms, and the metabolic pathway by which these compounds are assimilated into microorganisms. The presented model can be applied, without major modifications, to estimate meaningful kinetic parameters from batch experiments, as well as near source zone field application. We suggest the estimated actual microbial specific utilization rate (kC) of such materials to be a better measure of the degradation rate when compared to the maximum specific utilization rate (k), which might be orders of magnitude higher than kC and might never be observed in reality.

Biodegradation of alkylates under less agitated aquifer conditions.

The biodegradability of three alkylates (2,3-dimethylpentane, 2,4-dimethylpentane and 2,2,4-trimethylpentane) under less agitated aquifer conditions was investigated in this study. All three alkylates biodegraded completely under these conditions regardless of the presence or absence of ethanol or benzene, toluene, ethylbenzene, and xylenes (BTEX) in the feed. In the presence of ethanol, alkylates degradation was not inhibited by ethanol. However, alkylates degraded more slowly in the presence of BTEX suggesting competitive inhibition to microbial utilization of alkylates. In the sterile controls, alkylates concentrations remained unchanged throughout the experiments.

Biological nitrogen and carbon removal in a gravity flow biomass concentrator reactor for municipal sewage treatment.

A novel membrane system, the Biomass Concentrator Reactor (BCR), was evaluated as an alternative technology for the treatment of municipal wastewater. Because the BCR is equipped with a membrane whose average poresize is 20 µm (18-28 µm), the reactor requires low-pressure differential to operate (gravity). The effectiveness of this system was evaluated for the removal of carbon and nitrogen using two identical BCRs, identified as conventional and hybrid, that were operated in parallel. The conventional reactor was operated under full aerobic conditions (i.e., organic carbon and ammonia oxidation), while the hybrid reactor incorporated an anoxic zone for nitrate reduction as well as an aerobic zone for organic carbon and ammonia oxidation. Both reactors were fed synthetic wastewater at a flow rate of 71 L d(-1), which resulted in a hydraulic retention time of 9 h. In the case of the hybrid reactor, the recycle flow from the aerobic zone to the anoxic zone was twice the feed flow rate. Reactor performance was evaluated under two solids retention times (6 and 15 d). Under these conditions, the BCRs achieved nearly 100% mixed liquor solids separation with a hydraulic head differential of less than 2.5 cm. The COD removal efficiency was over 90%. Essentially complete nitrification was achieved in both systems, and nitrogen removal in the hybrid reactor was close to the expected value (67%).

Microtox aquatic toxicity of petrodiesel and biodiesel blends: the role of biodiesel's autoxidation products.

The acute Microtox toxicity of the water accommodated fraction (WAF) of six commercial soybean biodiesel/petrodiesel blends was investigated at different oil loads. We analyzed five fatty acid methyl esters (FAMEs), C10-C24 n-alkanes, four aromatics, methanol, and total organic carbon (TOC) content. At high oil loads, the WAFs' toxicity was significantly higher for blends containing biodiesel. At the lowest load, the WAFs' toxicity decreased almost linearly with decreasing biodiesel in the blend. At intermediate loads, the WAFs of all the blends appeared to have a similar toxicity. Analysis of WAFs confirmed the presence of autoxidation byproducts of FAMEs at high oil loads. Pure unsaturated FAMEs and n-alkanes were nontoxic when present in water at their reported solubility limits. However, 24-h equilibrated WAFs of pure FAMEs were highly toxic for C18:1 and C18:3, but not for C18:2. The authors concluded that at high oil loads, the acute toxicity of the WAFs was caused by FAMEs' autoxidation byproducts, whereas at low oil loads, the toxicity appeared to be caused primarily by the aromatic compounds present in petrodiesel. The addition of a synthetic antioxidant in biodiesel did not appear to affect the concentration of autoxidation byproducts in the WAF but resulted in a slight decrease in its toxicity. The major autoxidation byproducts identified in the WAF of commercial biodiesel were present neither in the WAFs of pure unsaturated FAMEs nor in the WAF of a different soybean biodiesel that was transesterified in our laboratory, which was nontoxic. We concluded that the process of transesterification of biodiesel might be a more critical factor in determining the aquatic toxicity of the fuel than the source of feedstock itself.

Assessing sustainability in real urban systems: the Greater Cincinnati Metropolitan Area in Ohio, Kentucky, and Indiana.

Urban systems have a number of factors (i.e., economic, social, and environmental) that can potentially impact growth, change, and transition. As such, assessing and managing these systems is a complex challenge. While, tracking trends of key variables may provide some insight, identifying the critical characteristics that truly impact the dynamic behavior of these systems is difficult. As an integrated approach to evaluate real urban systems, this work contributes to the research on scientific techniques for assessing sustainability. Specifically, it proposes a practical methodology based on the estimation of dynamic order, for identifying stable and unstable periods of sustainable or unsustainable trends with Fisher Information (FI) metric. As a test case, the dynamic behavior of the City, Suburbs, and Metropolitan Statistical Area (MSA) of Cincinnati was evaluated by using 29 social and 11 economic variables to characterize each system from 1970 to 2009. Air quality variables were also selected to describe the MSA's environmental component (1980-2009). Results indicate systems dynamic started to change from about 1995 for the social variables and about 2000 for the economic and environmental characteristics.

Partitioning behavior of petrodiesel/biodiesel blends in water.

The partitioning behavior of six petrodiesel/soybean-biodiesel blends (B0, B20, B40, B60, B80, and B100, where B100 is 100% unblended biodiesel) in water was investigated at various oil loads by the 10-fold dilution method. Five fatty acid methyl esters (FAMEs), C10-C20 n-alkanes, and four monoaromatic compounds were targeted for analysis. Only the aromatic compounds were partitioned according to Raoult's law at all oil loads. The partitioning of the FAMEs and n-alkanes at higher oil loads was found to be orders of magnitude higher than the reported aqueous solubilities of these compounds, and directly correlated with the amount of oil load applied. Depth filtration of the water-accommodated fractions (WAFs) significantly reduced the observed concentrations of the FAMEs and n-alkanes, but did not appreciably affect the aromatic compounds. The FAMEs and n-alkanes concentrations in the filtered WAFs agreed with the aqueous solubilities of those compounds reported in the literature, but the n-alkanes showed progressive deviations from those solubilities with the increase in the amount of biodiesel in the blends. Further dilution experiments on pure n-hexadecane confirmed the presence of a metastable colloidal phase that seems to be controlled by hydrophobic interactions and surface phenomena. The addition of biodiesel to the oil blend appeared to have a positive impact on the dissolved concentrations and the colloidal accommodation of the n-alkanes. Autoxidation of the biodiesel constituents was found to be significant, and increased with increasing oil loads. Chemical products such as hexanal, n-butyl acetate, diethylene glycol monobutyl ether, and diethylene glycol monobutyl ether acetate were positively identified among the FAMEs' autoxidation byproducts. Our data suggest a positive enhancement for biodiesel on the formation of the oil in water colloidal phase, possibly by forming a surfactant-cosurfactant-like pair of the FAMEs and their autoxidation byproducts.

Effect of butylated hydroxytoluene (BHT) on the aerobic biodegradation of a model vegetable oil in aquatic media.

Antioxidants added to vegetable oils to prevent lipid oxidation significantly affect their biodegradation in impacted aquatic environments. In this study, the effect of butylated-hydroxytoluene (BHT) on the biodegradation of glyceryl trilinoleate, a model vegetable oil highly susceptible to autoxidation, was determined. Biodegradation experiments were conducted in respirometric microcosms at an oil loading of 333 gal acre(-1) (0.31 L m(-2)) and BHT concentrations ranging from 0 to 800 mg kg(-1) (0, 50, 100, 200, 400, and 800 mg kg(-1)). Competition between polymerization and biodegradation of the oil was observed at all BHT concentrations and was significant in the microcosms not supplemented with the antioxidant. In all microcosms, intractable rigid polymers unavailable for bacterial degradation were formed. Infrared analysis evidenced the advanced stages of the oil autoxidation. After 19 weeks of incubation, only about 41% of the oil was mineralized in the microcosms with no BHT. However, mineralization exceeded 67% in the microcosms with added antioxidant and did not significantly increase with increasing BHT concentrations. Biodegradation rate constants were calculated by nonlinear regression and were not significantly different in the microcosms with added BHT (k = 0.001 h(-1)). Higher k values were measured in the microcosms lacking the antioxidant (k = 0.0023 h(-1)), most likely due to the increased oxygen consumption associated with the autoxidation process in this case. No toxicity was detected in all biotic microcosms at the end of the incubation period, while high toxicity (EC(50) = 4.78%) was measured in the abiotic blanks with no antioxidant and was attributed to the accumulation of autoxidation products.

The impact of stabilization mechanism on the aggregation kinetics of silver nanoparticles.

The use of silver nanoparticles (AgNPs) for various applications is growing drastically. The increase in use will eventually lead to their release into the environment. The tendency of AgNPs to aggregate and the kinetics of aggregation are major factors that govern their fate in the environment. Dynamic light scattering (DLS) was utilized to investigate the electrolyte-induced aggregation kinetics (NaNO3, NaCl and Ca(NO3)2) of coated and uncoated AgNPs which are electrostatically (H2-AgNPs and Citrate-AgNPs), sterically (polyvinylpyrrolidone (PVP)-AgNPs) and electrosterically (branched polyethyleneimine (BPEI)-AgNPs) stabilized. The aggregation kinetics of the electrostatically stabilized AgNPs was in agreement with the classical Derjaguin-Landau-Verwey-Overbeek (DLVO) theory and the AgNPs exhibited both reaction-limited and diffusion-limited regimes. The H2-AgNPs had critical coagulation concentrations (CCC) of 25, 30 and 3mM in the presence of NaNO3, NaCl and Ca(NO3)2 salts, respectively. The Citrate-AgNPs had CCC of 70, 70 and 5 mM in the presence of NaNO3, NaCl and Ca(NO3)2 salts, respectively. The values of the Hamaker constant for the electrostatically stabilized AgNPs were also determined and the values were in agreement with the reported values for metallic particles. The aggregation kinetics for both the sterically and electrosterically stabilized AgNPs (PVP-AgNPs and BPEI-AgNPs) was not in agreement with the DLVO theory and the particles were resistant to aggregation even at high ionic strength and electrolyte valence. The PVP-AgNPs and the BPEI-AgNPs had no critical aggregation concentration value at the investigated ionic strength values.

Assessment of the abiotic transformation of 17ß-estradiol in the presence of vegetable matter--II: the role of molecular oxygen.

This study characterizes the effect of oxygen in the abiotic transformation of estrogens when they are contacted with a surrogate of the vegetable wastes found in sewage. 17ß-Estradiol (E2) and 17ß-(14)C(4)-estradiol ((14)C-E2) were utilized as model compounds. Batch experiments were run under both oxic and anoxic conditions. In order to accomplish an accurate mass balance of the target estrogen, two analyses were performed simultaneously: first, radioactivity counting, and second, quantitation of E2 and (14)C-E2, as well as their transformation product estrone and (14)C(4)-estrone, by Liquid Chromatography tandem Mass Spectrometry. Under oxic conditions, the total concentration of (14)C-E2 was found to decrease by 78% in 72 h (15% and 7% remained in the liquid and solid phases, respectively). Conversely, when the estrogens were contacted with the synthetic influent under anoxic conditions, E2 was quantitatively recovered after 72 h (70% and 22% in aqueous and solid matrices, correspondingly). These results suggest that when the concentration of dissolved oxygen is null or limited, catalysis through an oxidative coupling mechanism is halted. Moreover, it was confirmed that the catalytic reaction occurred solely in the presence of the solid phase of the model vegetable matter.

Assessment of aquatic toxicity and oxygen depletion during aerobic biodegradation of vegetable oil: effect of oil loading and mixing regime.

The potential ecological impacts of aerobic biodegradation of vegetable oils on contaminated water columns was investigated in the laboratory at different oil loadings (100, 333, and 1,000 gal acre(-1)) and mixing regimes (fully, moderately, and nonmixed microcosms). The impacts were estimated by use of the Microtox assay and dissolved oxygen concentration measurements. The results of the Microtox assay showed no major toxicity at the 100 gal acre(-1) loading. Furthermore, oxygen was not completely depleted from the water column at this oil coverage. At higher oil loadings, oxygen was fully depleted from the mixed and nonmixed water columns. A transient toxicity in the aqueous phase was observed in the case of the moderately mixed microcosms at 333 gal acre(-1) and was maintained at moderate levels (EC(50) ∼ 30%) in the nonmixed microcosms. A substantial increase in toxicity (EC(50) ∼ 10%) was observed in both mixing conditions when the initial oil loading was increased to 1,000 gal acre(-1). At all oil loadings, significant toxicity (EC(50) < 2%) was found in the solid phase due to the strong partition of lipids to the biomass. Long and medium chains fatty acids associated with the measured toxicity were detected in both liquid and solid phases.