Biodegradation of Crude Oil by Individual Bacterial Strains and a Mixed Bacterial Consortium Isolated from Hydrocarbon Contaminated Areas

A preliminary study was undertaken to determine the optimal conditions for the biodegradation of a crude oil. Among 57 oil‐degrading bacterial cultures isolated from oil‐contaminated soil samples, Bacillus sp. IOS1‐7, Corynebacterium sp. BPS2‐6, Pseudomonas sp. HPS2‐5, and Pseudomonas sp. BPS1‐8 were selected for the study based on the efficiency of crude oil utilization. Along with the selected individual strains, a mixed bacterial consortium prepared using the above strains was also used for degradation studies. The mixed bacterial consortium showed more growth and degradation than did individual strains. At 1% crude oil concentration, the mixed bacterial consortium degraded a maximum of 77% of the crude oil. This was followed by 69% by Pseudomonas sp. BPS1‐8, 64% by Bacillus sp. IOS1‐7, 45% by Pseudomonas sp. HPS2‐5, and 41% by Corynebacterium sp. BPS2‐6. The percentage of degradation by the mixed bacterial consortium decreased from 77 to 45% as the concentration of crude oil was increased from 1 to 12%. Temperature of 35°C and pH 7 were found to be optimum for maximum degradation.     

Biodegradation by Co‐inoculated Bacteria and Fungi Alleviates Adverse Effects of Red‐S3B on Growth and Nitrogen Uptake of Wheat

Textile wastewater contains huge quantities of nitrogen (N)‐containing azo‐dyes. Irrigation of crops with such wastewater adds toxic dyes into our healthy soils. One of the ways to prevent their entry to soils could be these waters after the dyes' biodegradation. Therefore, the present study was conducted to evaluate the impact of textile dyes on wheat growth, dye degradation efficiency of bacteria‐fungi consortium, and alleviation of dye toxicity in wheat by treatment with microbial consortium. Among dyes, Red‐S3B (3.19% N) was found to be the most toxic to germination and growth of seven‐day‐old wheat seedlings. Shewanella sp. NIAB‐BM15 and Aspergillus terreus NIAB‐FM10 were found to be efficient degraders of Red‐S3B. Their consortium completely decolorized 500 mg L^?1 Red‐S3B within 4 h. Irrigation with Red‐S3B‐contaminated water after treatment with developed consortium increased root length, shoot length, root biomass, and shoot biomass of 30‐day‐old wheat seedlings by 47, 18, 6, and 25%, respectively, than untreated water. Moreover, irrigation after microbial treatment of dye‐contaminated water resulted in 20 and 51% increase in shoot N content and N uptake, respectively, than untreated water. Thus, co‐inoculation of bacteria and fungi could be a useful bioremediation strategy for the treatment of azo‐dye‐polluted water.     

Biodegradation of petroleum hydrocarbons at low temperature in the presence of the dispersant Corexit 9500

Our study examined the effects of Corexit 9500 and sediment on microbial mineralization of specific aliphatic and aromatic hydrocarbons found in crude oil. We also measured gross mineralization of crude oil, dispersed crude oil and dispersant by a marine microbial consortium in the absence of sediment. When provided as carbon sources, our consortium mineralized Corexit 9500 the most rapidly, followed by fresh oil, and finally weathered oil or dispersed oil. However, mineralization in short term assays favored particular components of crude oil (2-methyl-naphthalene > dodecane > phenanthrene > hexadecane > pyrene) and was not affected by addition of nutrients or sediment (high sand, low organic carbon). Adding dispersant inhibited hexadecane and phenanthrene mineralization but did not affect dodecane and 2-methyl-naphthalene mineralization. Thus, the effect of dispersant on biodegradation of a specific hydrocarbon was not predictable by class. The results were consistent for both high and low oiling experiments and for both fresh and weathered oil. Overall, our results indicate that environmental use of Corexit 9500 could result in either increases or decreases in the toxicity of residual oil through selective microbial mineralization of hydrocarbons.     

An assessment of a mesocosm approach to the study of microbial respiration in a sandy unsaturated zone

Microbial respiration rates were determined through a 3.2 m thick, sandy unsaturated zone in a 2.4 m diameter x 4.6 m high mesocosm. The mesocosm was maintained under near constant temperature (18 degrees to 23 degrees C) and reached steady moisture content conditions after several hundred days. Soil-gas CO2 concentrations in the mesocosm ranged from 0.09% to 3.31% and increased with depth. Respiration rates within the mesocosm were quantified over a 342-day period using measured CO2 concentrations and a transient, one-dimensional finite-element model. Microbial respiration rates were 2 x 10(-1) micrograms C.g-1.d-1 throughout most of the system, but decreased to 10(-4) to 10(-3) micrograms C.g-1.d-1 within the capillary fringe. Microbial respiration rates were also determined in minicosms (500 g sample mass) over a range in temperatures (4 degrees to 30 degrees C) and volumetric moisture contents (0.044 to 0.37). The functional dependence of CO2 production on temperature and soil-moisture content was similar for the two scales of laboratory observation. Respiration rates in the minicosms, for temperatures and moisture contents in the mesocosm, were up to an order of magnitude greater than those determined for the mesocosm. The higher respiration rates in the minicosms, compared to the mesocosm, were attributed to greater disturbance of the samples and to shorter acclimation time in the minicosms. Extrapolating the laboratory respiration rates to field conditions yielded rates that were two to three orders of magnitude greater than rates previously determined in situ for C-horizon material. Results show that in situ microbial reaction rates determined using disturbed samples in minicosms and mesocosms yielded respiration rates that greatly exceeded field conditions. Mesocosms can, however, provide a useful environment for conducting process-related research in unsaturated environments.     

Analysis of community structure of a microbial consortium capable of degrading benzo(a)pyrene by DGGE

A microbial consortium was obtained by enrichment culture of sea water samples collected from Botan oil port in Xiamen, China, using the persistent high concentration of a mixture of polycyclic aromatic hydrocarbons enrichment strategy. Denaturing gradient gel electrophoresis (DGGE) was used to investigate the bacterial composition and community dynamic changes based on PCR amplification of 16S rRNA genes during batch culture enrichment. Using the spray-plate method, three bacteria, designated as BL01, BL02 and BL03, which corresponded to the dominant bands in the DGGE profiles, were isolated from the consortium. Sequence analysis showed that BL01, BL02 and BL03 were phylogenetically close to Ochrobactrum sp., Stenotrophomonas maltophilia and Pseudomonas fluorescens, respectively. The degradation of benzo(a)pyrene (BaP), a model high-molecular-weight polycyclic aromatic hydrocarbon (HMW PAH) compound was investigated using individual isolates, a mixture of the three isolates, and the microbial consortium (BL) originally isolated from the oil port sea water. Results showed that the order of degradative ability was BL > the mixture of the three isolates > individual isolates. BL degraded 44.07% of the 10 ppm BaP after 14 days incubation, which showed the highest capability for HMW PAH compound degradation.Our results revealed that this high selective pressure strategy was feasible and effective in enriching the HMW PAH-degraders from the original sea water samples.     

Role of a moderately halophilic bacterial consortium in the biodegradation of polyaromatic hydrocarbons

Polycyclic aromatic hydrocarbons are ubiquitous pollutants in the environment, and most high molecular weight PAHs cause mutagenic, teratogenic and potentially carcinogenic effects. While several strains have been identified that degrade PAHs, the present study is focused on the degradation of PAHs in a marine environment by a moderately halophilic bacterial consortium. The bacterial consortium was isolated from a mixture of marine water samples collected from seven different sites in Chennai, India. The low molecular weight (LMW) PAHs phenanthrene and fluorine, and the high molecular weight (HMW) PAHs pyrene and benzo(e)pyrene were selected for the degradation study. The consortium metabolized both LMW and HMW PAHs. The consortium was also able to degrade PAHs present in crude oil-contaminated saline wastewater. The bacterial consortium was able to degrade 80% of HMW PAHs and 100% of LMW PAHs in the saline wastewater. The strains present in the consortium were identified as Ochrobactrum sp., Enterobacter cloacae and Stenotrophomonas maltophilia. This study reveals that these bacteria have the potential to degrade different PAHs in saline wastewater.     

Biodegradation of polycyclic aromatic hydrocarbons by a halotolerant bacterial strain Ochrobactrum sp. VA1

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous pollutants in the environment and are derived from both man-made and natural resources. The present study is focused on the degradation of PAHs by a halotolerant bacterial strain under saline conditions. The bacterial strain VA1 was isolated from a PAH-degrading consortium that was enriched from marine water samples that were collected from different sites at Chennai, India. In the present study, a clearing zone formed on PAH-amended mineral salt agar media confirmed the utilization of PAH by the bacterial strain VA1. The results show that the strain VA1 was able to degrade anthracene (88%), phenanthrene (98%), naphthalene (90%), fluorene (97%), pyrene (84%), benzo(k)fluoranthene (57%) and benzo(e)pyrene (50%) at a 30 g/L NaCl concentration. The present study reveals that the VA1 strain was able to degrade PAHs in petroleum wastewater under saline conditions. The promising PAH-degrading halotolerant bacterial strain, VA1, was identified as Ochrobactrum sp. using biochemical and molecular techniques.     

Long-term effects of crude oil on microbial processes in subarctic marine sediments: Studies on sediments amended with organic nutrients

Subarctic marine sediments amended with various organic compounds were exposed to fresh Cook Inlet crude oil at a concentration of 50 ppt for either 6 or 8 months. After the sediments were initially treated, they were returned to the approximate location where they were collected and left to be exposed to natural environmental conditions until they were retrieved for analysis. As a result of crude oil treatment, the activities of the enzymes that hydrolyse structural polysaccharides were reduced and the activities of the enzymes that hydrolyse storage polysaccharides were stimulated. In addition to these changes, we observed changes in phosphatase activity, nitrogen fixation rates, potential denitrification rates, methane concentrations, CO₂ production rates, and the glucose uptake and mineralization rates. The effect of the crude oil perturbation was different depending on the organic compound used in the amended sediments. Many of these differences could be explained by the effect of crude oil on the hydrolases which were responsible for degrading the compound in question. The results of this study suggest the effect of crude oil on microbial processes may be affected by the type of organic material present in the impacted marine sediment. This study also illustrates the value of measuring hydrolase activity in studies designed to determine the effects of this or any other pollutant on microbial processes in marine sediments.     

Cracking conditions of crude oil under different geological environments

There are mainly 3 kinds of existing states of oil generating from source rocks, that is, dispersive liquid hydrocarbon inside of source rock, dispersive liquid hydrocarbon outside of source rock and concentrated liquid hydrocarbon outside of source rock. Because of the differences in thermal history and medium conditions around, and the interaction of organic and inorganic matter, the liquid hydrocarbon with 3 kinds of existing state has different cracking conditions. The gas generation dynamics experiments of crude oil matching different mediums indicate that the distribution of activation energy of methane changes a lot according to medium difference. The carbonate has a main influence on oil cracking conditions and can largely reduce its activation energy, which reflects the lower cracking temperature of crude oil. The mudstone takes a second place and the sandstone is the smallest. The catalytic cracking function to the oil of the carbonate, of the mudstone and of the sandstone changes weaken in turn. The corresponding R _o values of main gas generation period in different mediums are as follows: 1.5%–3.8% with pure crude oil, 1.2%–3.2% with dispersive crude oil in carbonate, 1.3%～3.4% with dispersive crude oil in mudstone and 1.4%–3.6% with dispersive crude oil in sandstone. The influence of pressure to crude oil cracking is relatively complicated. In the low heating speed condition, pressure restrains the oil cracking and gas generation, but in the high heating speed condition, pressure has an indistinctive influence to the oil cracking and gas generation. Pressure also makes a different effort in different evolvement stage. Taking the middle and lower Cambrian source rocks in the Tarim Basin as an example, primary oil generating quantity is 2232.24×10⁸t, residual oil and oil cracking gas quantity is 806.21×10⁸t and 106.95×10¹²m³ respectively.     

Evaluating crude oil chemical dispersion efficacy in a flow-through wave tank under regular non-breaking wave and breaking wave conditions

Testing dispersant effectiveness under conditions similar to that of the open environment is required for improvements in operational procedures and the formulation of regulatory guidelines. To this end, a novel wave tank facility was fabricated to study the dispersion of crude oil under regular non-breaking and irregular breaking wave conditions. This wave tank facility was designed for operation in a flow-through mode to simulate both wave- and current-driven hydrodynamic conditions. We report here an evaluation of the effectiveness of chemical dispersants (Corexit^® EC9500A and SPC 1000^™) on two crude oils (Medium South American [MESA] and Alaska North Slope [ANS]) under two different wave conditions (regular non-breaking and plunging breaking waves) in this wave tank. The dispersant effectiveness was assessed by measuring the water column oil concentration and dispersed oil droplet size distribution. In the absence of dispersants, nearly 8-19% of the test crude oils were dispersed and diluted under regular wave and breaking wave conditions. In the presence of dispersants, about 21-36% of the crude oils were dispersed and diluted under regular waves, and 42-62% under breaking waves. Consistently, physical dispersion under regular waves produced large oil droplets (volumetric mean diameter or VMD ? 300 μm), whereas chemical dispersion under breaking waves created small droplets (VMD ? 50 μm). The data can provide useful information for developing better operational guidelines for dispersant use and improved predictive models on dispersant effectiveness in the field.     

Mesocosm experiments identifying hotspots of groundwater upwelling in a water column by fibre optic distributed temperature sensing

Lacustrine groundwater discharge (LGD) can substantially impact ecosystem characteristics and functions. Fibre optic distributed temperature sensing (FO‐DTS) has been successfully used to locate groundwater discharge into lakes and rivers at the sediment–water interface, but locating groundwater discharge would be easier if it could be detected from the more accessible water surface. So far, it is not clear if how and under which conditions the LGD signal propagates through the water column to the water surface–atmosphere interface, and what perturbations and signal losses occur along this pathway. In the present study, LGD was simulated in a mesocosm experiment. Under winter conditions, water with temperatures of 14 to 16 °C was discharged at the bottom of a 10 × 2.8‐m mesocosm. Water within this mesocosm ranged from 4.0 to 7.4 °C. Four layers (20, 40, 60, and 80 cm above the sediment) of the 82 cm deep mesocosm were equipped with FO‐DTS for tracing thermal patterns in the mesocosm. Aims are (a) to test whether the positive buoyancy of relatively warm groundwater imported by LGD into shallow water bodies allows detection of LGD at the lake's water surface–atmosphere interface by FO‐DTS, (b) to analyse the propagation of the temperature signal from the sediment‐water interface through the water column, and (c) to learn more about detectability of the signal under different discharge rates and weather conditions. The experiments supported the benchmarking of scale dependencies and robustness of FO‐DTS applications for measuring upwelling into aquatic environments and revealed that weather conditions can have important impacts on the detection of upwelling at water surface–atmosphere interfaces at larger scales.     

Gladstone, Australia Field Studies: Weathering and Degradation of Hydrocarbons in Oiled Mangrove and Salt Marsh Sediments With and Without the Application of an Experimental Bioremediation Protocol

This field study was a combined chemical and biological investigation of the relative rates of weathering and biodegradation of oil spilled in sediments and testing the influence of a bioremediation protocol. The aim of the chemistry work presented here was to determine whether the bioremediation protocol affected the rate of penetration, dissipation or long-term retention of a medium range crude oil (Gippsland) and a Bunker C oil stranded in tropical Rhizophora sp. mangrove and Halosarcia sp. salt marsh environments. Permission for the planned oil spills was granted in the Port Authority area of Gladstone, Queensland (Australia). Sediment cores from three replicate plots of each treatment for mangroves and four replicate plots for the salt marsh (oil only and oil plus bioremediation) were analysed for total hydrocarbons (THC) and for individual alkane markers using gas chromatography with flame ionization detection (GC–FID). Sediments were collected at day 2, then 1, 2, 5 or 6 and 12 or 13 months post-spill for mangroves and day 2, 1, 3 and 9 months post-spill for salt marshes. Over this time, hydrocarbons in all of the oil treated plots decreased exponentially. There was no statistical difference in initial oil concentrations, penetration of oil to depth, or in the rates of oil dissipation between untreated oil and bioremediated oil in the mangrove plots. The salt marsh plots treated with the waxy Gippsland oil showed a faster rate of biodegradation of the oil in the bioremediated plots. In this case only, the degradation rate significantly impacted the mass balance of remaining oil. The Bunker C oil contained only minor amounts of highly degradable n-alkanes and bioremediation did not significantly impact its rate of loss in the salt marsh sediments. At the end of each experiment, there were still n-alkanes visible in the gas chromatograms of residual oils. Thus it was concluded that there was unlikely to be any change in the stable internal biomarkers of the oils over this time period. The predominant removal processes in both habitats were evaporation and dissolution, with a lag-phase of 1–2 months before the start of microbial degradation.     

Microbial Degradation of Tar Oil Compounds under Different Redox Conditions

The microbial transformation of typical tar oil compounds such as acridine, benzo(b)thiophene, dibenzofuran, indane, and indene under different redox conditions was investigated in microcosm studies. Under aerobic conditions the inherent contamination in polluted soil as well as the added N‐, S‐, O‐ heterocyclic and homocyclic compounds were transformed predominantly apart from thiophene. 1‐Indanone was detected by RP‐HPLC‐DAD and GC‐MS as an intermediate metabolite from indane and indene. Under nitrate and sulfate reducing conditions indane, benzo(b)thiophene, and dibenzofuran were transformed in assays with the polluted soil samples from well B 65 and B 66 within 426 days, whereas they were persistent in assays with the highly polluted soil B 67. All heterocyclic and homocyclic compounds added to the non‐contaminated soil from well B 85 were not degraded under nitrate and sulfate reducing conditions, too. The results indicate that for the decision, wether natural attenuation can be implemented in the remediation of contaminated site, in addition to BTEX and PAHs especially the fate of further tar oil compounds in anoxic aquifers has to be considered.     

Isolation and characterization of two crude oil-degrading yeast strains, Yarrowia lipolytica PG-20 and PG-32, from the Persian Gulf

Among six crude oil-degrading yeasts that were isolated from an oil-polluted area in the Persian Gulf, two yeast strains showed high degradation activity of aliphatic hydrocarbons. From an analysis of 18S rRNA sequences and biochemical characteristics, these strains were identified as Yarrowia lipolytica strains PG-20 and PG-32. Gas Chromatography (GC) analysis of the crude oil remaining in the culture medium after 1 week at 30°C showed that the strains PG-20 and PG-32 degraded 68% and 58% of crude oil, respectively. The optimal growth condition and biodegradation of hydrocarbons was in ONR medium with an acidic pH (pH5). These two strains may degrade aliphatic hydrocarbons more efficiently than aromatic hydrocarbons, although strain PG-20 had better degradation than strain PG-32. The two Y. lipolytica strains reduce surface tension when cultured on hydrocarbon substrates (1% v/v). These strains showed a cell surface hydrophobicity higher than 70%. These results suggested that Y. lipolytica strains PG-20 and PG-32 have high crude oil degrading activity due to their high emulsifying activity and cell hydrophobicity. In conclusion, these yeast strains can be useful for the bioremediation process in the Persian Gulf and decreasing oil pollution in this marine ecosystem.     

Nutrient fluxes induced by disturbance in Meiliang Bay of Lake Taihu

A wave flume experiment was conducted to study nutrient fluxes at water-sediment interface of Meiliang Bay under different hydrodynamic conditions. The results reveal that hydrodynamics has remarkable effects on nutrient fluxes in this area. With a bottom wave stress of 0.019 N m-2 (equivalent to disturbance caused by wind SE 5-7 m s-1 at the sediment sample site of Meiliang Bay), the fluxes of TN, TDN and NH4+-N were separately 1.92× 10-3, -1.81 × 10-4 and 5.28× 10-4 mg m-2 s-1(positive for upward and negative for downward), but for TP, TDP and SRP, the fluxes were 5.69 × 10-4, 1.68 × 10-4 and -1.29 × 10-4 mgm-2 s-1. In order to calculate the released amount of nutrients based on these results, statistic analysis on the long-term meteorological data was conducted.The result shows that the maximum lasting time for wind SE 5-7 m s-1 in this area is about 15 h in summer. Further calculation shows that 111 t TN, 32 t NH4+-N, 34 t TP and 10 t TDP can be released into water (the sediment area was 47.45% of the whole surface area), resulting in concentration increase of 0.025, 0.007, 0.007 and 0.002 mg L-1 separately. With stronger disturbance (bottom wave stress is 0.217 N m-2 which is equivalent to disturbance caused by wind SE 10-11 m s-1 at the same site), there has been significant increase of nutrient fluxes (1.16× 10-2, 6.76×10-3, 1.14× 10-2 and 2.14× 10-3 mgm-2 s-1 for TN, DTN and NH4+-N and TP). The exceptions were TDP with flux having a decrease (measured to be 9.54× 10-5 mgm-2 s-1 ) and SRP with flux having a small increase (measured to be 5.42 × 10-5 mgm-2 s-1). The same statistic analysis on meteorological data reveal that the maximum lasting time for wind SE 10-11 m s-1 is no more than 5 h. Based on the nutrient fluxes and the wind lasting-time, similar calculations were also made suggesting that 232 t TN, 134.9 t TDN, 228 t NH4+-N, 42.7 t TP, 2.0 t TDP and 1.1 t SRP will be released from sediment at this hydrodynamic condition resulting in the concentration increases of 0.050, 0.029, 0.049, 0.009, 0.0004 and 0.0002 mg L-1. Therefore in shallow lakes, surface disturbance can lead to significant increase of nutrient concentrations although some components in water column had negative flux with weak disturbance (e.g. TDN and SRP in this experiment). In this case, sediment looks to be a source of nutrients. These nutrients deposited in sediment can be carried or released into water with sediment resuspension or changes of environmental conditions at water-sediment interface, which can have great effects on aquatic ecosystem and is also the characteristics of shallow lakes.     

Natural attenuation, biostimulation and bioaugmentation on biodegradation of polycyclic aromatic hydrocarbons (PAHs) in mangrove sediments

The biodegradability of a mixture of PAHs, namely fluorene (Fl), phenanthrene (Phe) and pyrene (Pyr), in mangrove sediment slurry was investigated. At the end of week 4, natural attenuation based on the presence of autochthonous microorganisms degraded more than 99% Fl and Phe but only around 30% of Pyr were degraded. Biostimulation with addition of mineral salt medium degraded over 97% of all three PAHs, showing that nutrient amendment could enhance Pyr degradation. Bioaugmentation with inoculation of a PAH-degrading bacterial consortium enriched from mangrove sediments did not show any promotion effect and the degradation percentages of three PAHs were similar to that by natural attenuation. Some inhibitory effect was observed in bioaugmentation treatment in week 1 with only 50% Fl and 70% Phe degraded. These results indicate that autochthonous microbes may interact and even compete with the enriched consortium during PAH biodegradation. Natural attenuation appeared to be the most appropriate way to remedy Fl- and Phe-contaminated mangrove sediments while biostimulation was more capable to degrade Pyr-contaminated sediments. The study also shows that although a large portion of the added PAHs (more than 95%) was adsorbed onto the sediments at the beginning of the experiment, most PAHs were degraded in 4 weeks, suggesting that the degraders could utilize the adsorbed PAHs efficiently.     

Capability of Paraguaçu estuary (Todos os Santos Bay,Brazil) to form oil–SPM aggregates (OSA) and their ecotoxicological effects on pelagic and benthic organisms

For experiments concerning the formation of oil–suspended particulate matter (SPM) aggregates (OSA), oil and sediment samples were collected from Campos Basin and six stations of Paraguaçu estuary, Todos os Santos Bay, Brazil, respectively. The sediments samples were analyzed for organic matter determined by the EMBRAPA method, nitrogen determined by the Kjeldahl method, and phosphorus determined by the method described by Aspila. The oil trapped in OSA was extracted following the method described by Moreira. The experiment showed a relationship between the amount of organic matter and OSA formation and consequently the dispersion of the studied oil. On the basis of the buoyancy of OSA and the ecotoxicological effects on pelagic and benthic community, the priority areas for application of remediation techniques are Cachoeira, Maragogipe, and Salinas da Margarida because of the large amount of oil that accumulated at the bottom of the experiment flask (5.85%, 27.95%, and 38,98%; 4.2%, 17.66%, and 32.64%; and 11.82%, 8.07%, and 10.91% respectively).     

Effect of bioemulsificant exopolysaccharide (EPS2003) on microbial community dynamics during assays of oil spill bioremediation: A microcosm study

Microcosms experiments were carried out to evaluate the effect of bioemulsificant exopolysaccharide (EPS₂₀₀₃) on microbial community dynamics. An experimental seawater microcosm, supplemented with crude oil and EPS₂₀₀₃ (SW + OIL + EPS₂₀₀₃), was monitored for 15 days and compared to control microcosm (only oil-polluted seawater, SW + OIL). Determination of bacterial abundance, heterotrophic cultivable and hydrocarbon-degrading bacteria were carried out during all experimentation period. The microbial community dynamic was monitored by isolation of total RNA, RT-PCR amplification of 16S rRNA, cloning and sequencing. Oil degradation was monitored by GC–MS analysis. Bioemulsificant addition stimulated an increase of the total bacterial abundance, change in the community structure and activity. The bioemulsificant also increased of 5 times the oil biodegradation rate.The data obtained from microcosm experiment indicated that EPS₂₀₀₃ could be used for the dispersion of oil slicks and could stimulate the selection of marine hydrocarbon degraders thus increasing bioremediation process.     

Relationship between structures of substituted indolic compounds and their degradation by marine anaerobic microorganisms

Degradation of selected indolic compounds including indole, 1-methylindole, 2-methylindole, and 3-methylindole was assessed under methanogenic and sulfate-reducing conditions using the serum-bottle anaerobic technique and marine sediment from Victoria Harbour, Hong Kong as an inoculum. Our results showed that indole degradation was achieved in 28 days by a methanogenic consortium and 35 days by a sulfate-reducing consortium. During degradation under both conditions, two intermediates were isolated, purified and identified as oxindole and isatin (indole-2,3-dione) suggesting that both methanogenic and sulfate-reducing bacteria use an identical degradation pathway. Degradation processes followed two steps of oxidation accomplished by hydroxylation and then dehydrogenation at 2- and then 3-position sequentially prior to the cleavage of the pyrrole ring between 2- and 3-positions. However, none of 1-methylindole or 2-methylindole was degraded under any conditions. 3-Methylindole (3-methyl-1H-indole, skatole) was transformed under methanogenic conditions and mineralized only under sulfate-reducing conditions. It is clear that methyl substitution on 1- or 2-position inhibits the initial attack by hydroxylation enzymes making them more persistent in the environment and posing longer toxic impact.     

Chemical evolution of Macondo crude oil during laboratory degradation as characterized by fluorescence EEMs and hydrocarbon composition

The fluorescence EEM technique, PARAFAC modeling, and hydrocarbon composition were used to characterize oil components and to examine the chemical evolution and degradation pathways of Macondo crude oil under controlled laboratory conditions. Three major fluorescent oil components were identified, with Ex/Em maxima at 226/328, 262/315, and 244/366 nm, respectively. An average degradation half-life of ～20 d was determined for the oil components based on fluorescence EEM and hydrocarbon composition measurements, showing a dynamic chemical evolution and transformation of the oil during degradation. Dispersants appeared to change the chemical characteristics of oil, to shift the fluorescence EEM spectra, and to enhance the degradation of low-molecular-weight hydrocarbons. Photochemical degradation played a dominant role in the transformation of oil components, likely an effective degradation pathway of oil in the water column. Results from laboratory experiments should facilitate the interpretation of field-data and provide insights for understanding the fate and transport of oil components in the Gulf of Mexico.