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.

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.

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.

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.

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.

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.

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 environmental factors on the effectiveness of ammoniated bagasse in wicking oil from contaminated wetlands.

Ammoniated bagasse is a plant-derived organic sorbent that can be used for capturing oil and for supplying slow-release nutrients to oil-degrading microorganisms. We investigated the oil-wicking behavior of this sorbent under various conditions for its effectiveness in remediating oil-contaminated wetlands. Abiotic microcosms simulating a wetland environment were used to assess the influence of sand particle sizes (20 x 30 and 60 x 80 U.S. mesh), degrees of oil saturation (25% and 75%), water table levels (on top of the clean sand layer, oiled-sand layer, and sorbent layer), and the presence of sorbent. Results indicated that oil wicking favors higher oil contamination, larger sand particle size, and low water coverage. Water coverage was the predominant factor limiting the effectiveness of sorbent. The most plausible explanation for this limitation was that sorbent captured more water than oil at higher water coverage.

Biodegradability of lingering crude oil 19 years after the Exxon Valdez oil spill.

In 2001 and 2003, geospatial surveys of lingering oil were conducted in Prince William Sound (PWS) resulting in a prediction of significant acreage being contaminated with substantial subsurface oil from the 1989 Exxon Valdez oil spill (EVOS). In 2007, other researchers developed a mass weathering index (MWI) based on the degree of weathering of PAHs normalized to conserved biomarkers: if the degree of weathering of oil is 70% or more, further attempts at bioremediation would be unjustified. The objective of our study was to measure the biodegradability of the 19-year lingering oil in laboratory microcosms. Samples of beach substrate were collected from representative sites in PWS contaminated with oil residues of varying weathering states according to the MWI model. Enough sacrificial microcosms were set up to accommodate two treatments for each site (natural attenuation and biostimulation). Results indicated that lingering oil is biodegradable. Nutrient addition stimulated biodegradation compared to natural attenuation in all treatments regardless of the degree of weathering. The most weathered oil according to the MWI was the most biodegradable. Substantial biodegradation occurred in the natural attenuation microcosms due to the high sediment Total Kjeldahl Nitrogen (TKN), which served as a nitrogen source for biodegradation. Most of the observed biodegradation was due to the presence of dissolved oxygen. Nitrogen was a limiting factor but oxygen was the predominant one.

Efficient degradation of MtBE and other gasoline-originated compounds by means of a biological reactor of novel conception: two case studies in Italy and the USA.

The presence of MtBE in groundwaters, due to its past use as an oxygenate additive to "green" gasoline, poses potential threats to human health as a possible carcinogen. As its chemical properties render it very mobile and poorly treatable by means of bioremediation, chemical oxidation, stripping, or carbon adsorption, a new, ex-situ treatment process was recently developed by the US EPA and the University of Cincinnati. The objective of the study was to evaluate the effectiveness of this pump-and-treat technology in actual contaminated groundwater plumes in the field in two different conditions. The technology based on the combination of aerobic degradation and a special filtration medium is simple to operate, easily meets the challenge of producing a highly polished effluent in a cost-effective fashion, compared to other processes.

Assessment of chemical dispersant effectiveness in a wave tank under regular non-breaking and breaking wave conditions.

Current chemical dispersant effectiveness tests for product selection are commonly performed with bench-scale testing apparatus. However, for the assessment of oil dispersant effectiveness under real sea state conditions, test protocols are required to have hydrodynamic conditions closer to the natural environment, including transport and dilution effects. To achieve this goal, Fisheries and Oceans Canada and the US Environmental Protection Agency (EPA) designed and constructed a wave tank system to study chemical dispersant effectiveness under controlled mixing energy conditions (regular non-breaking, spilling breaking, and plunging breaking waves). Quantification of oil dispersant effectiveness was based on observed changes in dispersed oil concentrations and oil-droplet size distribution. The study results quantitatively demonstrated that total dispersed oil concentration and breakup kinetics of oil droplets in the water column were strongly dependent on the presence of chemical dispersants and the influence of breaking waves. These data on the effectiveness of dispersants as a function of sea state will have significant implications in the drafting of future operational guidelines for dispersant use at sea.

Biodegradability of alkylates as a sole carbon source in the presence of ethanol or BTEX.

The biodegradability of alkylate compounds in serum bottles was investigated in the presence and absence of ethanol or benzene, toluene, ethylbenzene, and p-xylene (BTEX). The biomass was acclimated to three different alkylates, 2,3-dimethylpentane, 2,4-dimethylpentane and 2,2,4-trimethylpentane in porous pot reactors. The alkylates were completely mineralized in all three sets of experiments. They degraded more slowly in the presence of BTEX than in their absence because BTEX inhibited the microbial utilization of alkylates. However, in the presence of ethanol, their slower biodegradation was not related to inhibition by the ethanol. Throughout the experiments alkylates, ethanol, and BTEX concentrations did not change in the sterile controls.

Biodegradation kinetics and toxicity of vegetable oil triacylglycerols under aerobic conditions.

The aerobic biodegradation of five triacylglycerols (TAGs), three liquids [triolein (OOO), trilinolein (LLL), and trilinolenin (LnLnLn)] and two solids [tripalmitin (PPP) and tristearin (SSS)] was studied in water. Respirometry tests were designed and conducted to determine the biochemical oxygen demand (BOD) parameters of the compounds. In the case of the solid lipids, the degradation process was limited by their extremely non-polar nature. When added to water, PPP and SSS formed irregular clumps or gumballs, not a fine and uniform suspension required for the lipase activity. After 30 days, appreciable mineralization was not achieved; therefore, first-order biodegradation coefficients could not be determined. The bioavailability of the liquid TAGs was restricted due to the presence of double bonds in the fatty acids (FAs). An autoxidation process occurred in the allylic chains, resulting in the production of hydroperoxides. These compounds polymerized and became non-biodegradable. Nevertheless, the non-oxidized fractions were readily mineralized, and BOD rate constants were estimated by non-linear regression: LLL (k=0.0061h(-1)) and LnLnLn (k=0.0071h(-1)) were degraded more rapidly than OOO (k=0.0025h(-1)). Lipids strongly partitioned to the biomass and, therefore, Microtox toxicity was not observed in the water column. However, EC(50) values (<15% sample volume) were measured in the solid phase.

A universal nutrient application strategy for the bioremediation of oil-polluted beaches.

Biostimulation by nutrient application is a viable technology for restoring oil-contaminated beaches. Maximizing the nutrient residence time is key for achieving a rapid cost-effective cleanup. We considered the nutrient injection strategy through a perforated pipe at the high tide line and we simulated numerically beach hydraulics, which allowed us to estimate the optimal injection flow rate of nutrient solution. Our results indicate that the optimal application is one that starts following the falling high tide and lasts for half tidal cycle. The saturated wet-front of the nutrient solution on the beach surface would move seaward with the same speed of the falling tide keeping a constant distance with the tide line. The numerical results were generalized to beaches of wide ranges of hydraulic and tidal properties using a novel dimensionless formulation for water flow and solute transport in porous media. Nomographs were presented to provide the flow rate based on four parameters: The beach slope and hydraulic conductivity, and tidal amplitude and period.

Effects of chemical dispersants and mineral fines on crude oil dispersion in a wave tank under breaking waves.

The interaction of chemical dispersants and suspended sediments with crude oil influences the fate and transport of oil spills in coastal waters. A wave tank study was conducted to investigate the effects of chemical dispersants and mineral fines on the dispersion of oil and the formation of oil-mineral-aggregates (OMAs) in natural seawater. Results of ultraviolet spectrofluorometry and gas chromatography flame ionized detection analysis indicated that dispersants and mineral fines, alone and in combination, enhanced the dispersion of oil into the water column. Measurements taken with a laser in situ scattering and transmissometer (LISST-100X) showed that the presence of mineral fines increased the total concentration of the suspended particles from 4 to 10microl l(-1), whereas the presence of dispersants decreased the particle size (mass mean diameter) of OMAs from 50 to 10microm. Observation with an epifluorescence microscope indicated that the presence of dispersants, mineral fines, or both in combination significantly increased the number of particles dispersed into the water.

Effects of ferric hydroxide on methanogenesis from lipids and long-chain fatty acids in anaerobic digestion.

The addition of ferric hydroxide to sludge from a municipal anaerobic digester stimulated the rate of methanogenesis from canola oil when the initial oil concentration was high (4600 mg/L; P < 0.002), but not when it was low (920 mg/L; P > 0.05). Similar trends were observed when oleic acid, a fatty acid that is a major component of canola oil triglycerides, was provided, but the effects were statistically significant only when the initial concentration of ferric hydroxide was also high (18 g/L; P = 0.015). Iron reduction occurred when ferric hydroxide was added to microcosms containing anaerobic digester sludge, but the extent of ferrous iron production was much less in acetate-amended microcosms than in those that were provided with canola oil or oleic acid. Methanogenesis and acetate consumption were completely inhibited when the initial acetate concentration was approximately 5000 mg/L, regardless of the initial ferric hydroxide concentration. The main effect of ferric hydroxide in this system appears to have been a result of stimulation of the rate of fatty acid oxidation.

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.

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.

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.

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.