Microbial sulfate-reducing activities in anoxic sediment from Marine Lake Grevelingen: screening of electron donors and acceptors

Sulfate-reducing bacteria in marine sediments mainly utilize sulfate as a terminal electron acceptor with different organic compounds as electron donors. This study investigated microbial sulfate-reducing activity of coastal sediment from Marine Lake Grevelingen (MLG), the Netherlands using different electron donors and electron acceptors. All four electron donors (ethanol, lactate, acetate and methane) showed sulfate-reducing activity with sulfate as electron acceptor, suggesting the presence of an active sulfate-reducing bacterial population in the sediment, even at dissolved sulfide concentrations exceeding 12 mM. Ethanol showed the highest sulfate reduction rate of 55 µmol g _VSS ^?1 day^?1 compared to lactate (32 µmol g _VSS ^?1 day^?1), acetate (26 µmol g _VSS ^?1 day^?1) and methane (4.7 µmol g _VSS ^?1 day^?1). Sulfide production using thiosulfate and elemental sulfur as electron acceptors and methane as the electron donor was observed, however, mainly by disproportionation rather than by anaerobic oxidation of methane coupled to sulfate reduction. This study showed that the MLG sediment is capable of performing sulfate reduction by using diverse electron donors, including the gaseous and cheap electron donor methane.     

Coupling hydrocarbon degradation to anaerobic respiration and mineral diagenesis: theoretical constraints

The diagenetic mineral assemblages in petroleum reservoirs control the formation fluid pH and pCO₂. Anaerobic biodegradation of petroleum is controlled by the transfer of electrons from reduced organic species to inorganic, redox sensitive, aqueous and mineral species in many cases through intermediates such as H₂ and CH₃COO^?. The terminal electron accepting reactions induce the dissolution or precipitation of the same minerals that control the ambient pH and pCO₂ in petroleum reservoirs. In this study, we develop a model for anaerobic biodegradation of petroleum that couples the production of acetate and H₂ to ‘late stage’ diagenetic reactions. The model reveals that the principal terminal electron accepting process and electron donor control the type of diagenetic reaction, and that the petroleum biodegradation rate is controlled through thermodynamic restriction by the minimum ΔG required to support a specific microbial metabolism, the fluid flux and the mineral assemblage. These relationships are illustrated by modeling coupled microbial diagenesis and biodegradation of the Gullfaks oil reservoir. The results indicate that the complete dissolution of albite by acids generated during oil biodegradation and the corresponding elevated pCO₂ seen in the Gullfaks field are best explained by methanogenic respiration coupled to hydrocarbon degradation and that the biodegradation rate is likely controlled by the pCH₄. Biodegradation of Gullfaks oil by a consortium that includes either Fe³⁺‐reducing or ‐reducing bacteria cannot explain the observed diagenetic mineral assemblage or pCO₂. For octane, biodegradation, not water washing, was the principal agent for removal at fluid velocities <20 m Myr^?1.     

Microbial activity and community structure in a lake sediment used for psychrophilic anaerobic wastewater treatment

Aims: The aim of the study was to investigate the feasibility of a continuous reactor for psychrophilic anaerobic wastewater treatment by using the sludge from cold natural environment. Methods and Results: Six sludge samples (S1–S6) were collected from different cold natural locations to select sludge with high anaerobic microbial activity under low temperatures. After a 225‐day incubation, the maximum specific methane production rate of a waterfowl lake sediment (S1) at 15°C (70·5 mLCH₄ gVSS^?1 day^?1) was much higher than all other samples. S1 was thus chosen as the seed sludge for the reactor treating synthetic brewery wastewater at 15°C, by immobilizing the micro‐organisms on polyurethane foam carriers. The chemical oxygen demand (COD) removal efficiency reached over 80% after 240‐day operation at an organic loading rate of 5·3 kg m^?3 day^?1, and significant enrichment of biomass was observed. Clone libraries of the microbial communities in the inoculum had high diversities for both archaea and bacteria. Along with a decrease in microbial community diversities, the dominant bacteria (79·5%) at the end of the operation represented the phylum Firmicutes, while the dominant archaeon (41·5%) showed a similarity of 98% with the psychrotolerant methanogen Methanosarcina lacustris. Conclusions: The possibility of using anaerobic micro‐organisms from cold environments in anaerobic wastewater treatment under psychrophilic conditions is supported by these findings. Significance and Impact of the Study: This study enriches the theory on microbial community and the application on anaerobic treatment of sludge from cold natural environments.     

High‐pressure systems for gas‐phase free continuous incubation of enriched marine microbial communities performing anaerobic oxidation of methane

Novel high‐pressure biotechnical systems that were developed and applied for the study of anaerobic oxidation of methane (AOM) are described. The systems, referred to as high‐pressure continuous incubation system (HP‐CI system) and high‐pressure manifold‐incubation system (HP‐MI system), allow for batch, fed‐batch, and continuous gas‐phase free incubation at high concentrations of dissolved methane and were designed to meet specific demands for studying environmental regulation and kinetics as well as for enriching microbial biomass in long‐term incubation. Anoxic medium is saturated with methane in the first technical stage, and the saturated medium is supplied for biomass incubation in the second stage. Methane can be provided in continuous operation up to 20 MPa and the incubation systems can be operated during constant supply of gas‐enriched medium at a hydrostatic pressure up to 45 MPa. To validate the suitability of the high‐pressure systems, we present data from continuous and fed‐batch incubation of highly active samples prepared from microbial mats from the Black Sea collected at a water depth of 213 m. In continuous operation in the HP‐CI system initial methane‐dependent sulfide production was enhanced 10‐ to 15‐fold after increasing the methane partial pressure from near ambient pressure of 0.2 to 10.0 MPa at a hydrostatic pressure of 16.0 MPa in the incubation stage. With a hydraulic retention time of 14 h a stable effluent sulfide concentration was reached within less than 3 days and a continuing increase of the volumetric AOM rate from 1.2 to 1.7 mmol L^?1 day^?1 was observed over 14 days. In fed‐batch incubation the AOM rate increased from 1.5 to 2.7 and 3.6 mmol L^?1 day^?1 when the concentration of aqueous methane was stepwise increased from 5 to 15 mmol L^?1 and 45 mmol L^?1. A methane partial pressure of 6 MPa and a hydrostatic pressure of 12 MPa in manifold fed‐batch incubation in the HP‐MI system yielded a sixfold increase in the volumetric AOM rate. Over subsequent incubation periods AOM rates increased from 0.6 to 1.2 mmol L^?1 day^?1 within 26 days of incubation. No inhibition of biomass activity was observed in all continuous and fed‐batch incubation experiments. The organisms were able to tolerate high sulfide concentrations and extended starvation periods. Biotechnol. Bioeng. 2010; 105: 524–533. © 2009 Wiley Periodicals, Inc.     

Microbiological investigations of high-temperature horizons of the Kongdian petroleum reservoir in connection with field trial of a biotechnology for enhancement of oil recovery

The physicochemical conditions and microbiological characteristics of the formation waters of the Kongdian oilfield of the Dagang oilfield (China) were studied. It was demonstrated that this oilfield is a high-temperature ecosystem with formation waters characterized by low mineralization. The concentrations of nitrogen and phosphorus compounds, as well as of electron acceptors, are low. Oil and oil gas are the main organic matter sources. The oilfield is exploited with water-flooding. The oil stratum was inhabited mostly by anaerobic thermophilic microorganisms, including fermentative (10²–10⁵ cells/ml), sulfate-reducing (0–10² cells/ml), and methanogenic (0–10³ cells/ml) microorganisms. Aerobic bacteria were detected mainly in the near-bottom zone of injection wells. The rate of sulfate reduction varied from 0.002 to 18.940 μg S^2? l^?1 day^?1 and the rate of methanogenesis from 0.012 to 16.235 μg CH₄ l^?1 day^?1. Microorganisms with great biotechnological potential inhabited the oilfield. Aerobic thermophilic bacteria were capable of oxidizing oil with formation of biomass, the products of partial oxidation of oil (volatile acids), and surfactants. During growth on the culture liquid of oil-oxidizing bacteria, methanogenic communities produced methane and carbon dioxide, which also had oil-releasing capabilities. Using various labeled tracers, the primary filtration flows of injected solutions at the test site were studied. Our comprehensive investigations allowed us to conclude that the method for microbial enhancement of oil recovery based on the activation of the stratal microflora can be applied in the Kongdian oilfield.     

Microbial diversity and community structure of a highly active anaerobic methane‐oxidizing sulfate‐reducing enrichment

Anaerobic oxidation of methane (AOM) is an important methane sink in the ocean but the microbes responsible for AOM are as yet resilient to cultivation. Here we describe the microbial analysis of an enrichment obtained in a novel submerged‐membrane bioreactor system and capable of high‐rate AOM (286 μmol g_{dry weight}^?1 day^?1) coupled to sulfate reduction. By constructing a clone library with subsequent sequencing and fluorescent in situ hybridization, we showed that the responsible methanotrophs belong to the ANME‐2a subgroup of anaerobic methanotrophic archaea, and that sulfate reduction is most likely performed by sulfate‐reducing bacteria commonly found in association with other ANME‐related archaea in marine sediments. Another relevant portion of the bacterial sequences can be clustered within the order of Flavobacteriales but their role remains to be elucidated. Fluorescent in situ hybridization analyses showed that the ANME‐2a cells occur as single cells without close contact to the bacterial syntrophic partner. Incubation with ¹³C‐labelled methane showed substantial incorporation of ¹³C label in the bacterial C₁₆ fatty acids (bacterial; 20%, 44% and 49%) and in archaeal lipids, archaeol and hydroxyl‐archaeol (21% and 20% respectively). The obtained data confirm that both archaea and bacteria are responsible for the anaerobic methane oxidation in a bioreactor enrichment inoculated with Eckernförde bay sediment.     

Identification of functionally active aerobic methanotrophs in sediments from an arctic lake using stable isotope probing

Arctic lakes are a significant source of the greenhouse gas methane (CH₄), but the role that methane oxidizing bacteria (methanotrophs) play in limiting the overall CH₄ flux is poorly understood. Here, we used stable isotope probing (SIP) techniques to identify the metabolically active aerobic methanotrophs in upper sediments (0–1 cm) from an arctic lake in northern Alaska sampled during ice‐free summer conditions. The highest CH₄ oxidation potential was observed in the upper sediment (0–1 cm depth) with 1.59 µmol g wet weight^?1 day^?1 compared with the deeper sediment samples (1–3 cm, 3–5 cm and 5–10 cm), which exhibited CH₄ oxidation potentials below 0.4 µmol g wet weight^?1 day^?1. Both type I and type II methanotrophs were directly detected in the upper sediment total communities using targeted primer sets based on 16S rRNA genes. Sequencing of 16S rRNA genes and functional genes (pmoA and mxaF) in the ¹³C‐DNA from the upper sediment indicated that type I methanotrophs, mainly Methylobacter, Methylosoma, Methylomonas and Methylovulum miyakonense, dominated the assimilation of CH₄. Methylotrophs, including the genera Methylophilus and/or Methylotenera, were also abundant in the ¹³C‐DNA. Our results show that a diverse microbial consortium acquired carbon from CH₄ in the sediments of this arctic lake.     

Simultaneous hydrocarbon biodegradation and biosurfactant production by oilfield-selected bacteria

Aims: To study the bacterial diversity associated with hydrocarbon biodegradation potentiality and biosurfactant production of Tunisian oilfields bacteria. Methods and Results: Eight Tunisian hydrocarbonoclastic oilfields bacteria have been isolated and selected for further characterization studies. Phylogenetic analysis revealed that three thermophilic strains belonged to the genera Geobacillus, Bacillus and Brevibacillus, and that five mesophilic strains belonged to the genera Pseudomonas, Lysinibacillus, Achromobacter and Halomonas. The bacterial strains were cultivated on crude oil as sole carbon and energy sources, in the presence of different NaCl concentrations (1, 5 and 10%, w/v), and at 37 or 55°C. The hydrocarbon biodegradation potential of each strain was quantified by GC–MS. Strain C450R, phylogenetically related to the species Pseudomonas aeruginosa, showed the maximum crude oil degradation potentiality. During the growth of strain C450R on crude oil (2%, v/v), the emulsifying activity (E24) and glycoside content increased and reached values of 77 and 1·33 g l^?1, respectively. In addition, the surface tension (ST) decreased from 68 to 35·1 mN m^?1, suggesting the production of a rhamnolipid biosurfactant. Crude biosurfactant had been partially purified and characterized. It showed interest stability against temperature and salinity increasing and important emulsifying activity against oils and hydrocarbons. Conclusions: The results of this study showed the presence of diverse aerobic bacteria in Tunisian oilfields including mesophilic, thermophilic and halotolerant strains with interesting aliphatic hydrocarbon degradation potentiality, mainly for the most biosurfactant produced strains. Significance and Impact of the Study: It may be suggested that the bacterial isolates are suitable candidates for practical field application for effective in situ bioremediation of hydrocarbon‐contaminated sites.     

Microbial processes in the Athabasca Oil Sands and their potential applications in microbial enhanced oil recovery

The Athabasca Oil Sands are located within the Western Canadian Sedimentary Basin, which covers over 140,200 km² of land in Alberta, Canada. The oil sands provide a unique environment for bacteria as a result of the stressors of low water availability and high hydrocarbon concentrations. Understanding the mechanisms bacteria use to tolerate these stresses may aid in our understanding of how hydrocarbon degradation has occurred over geological time, and how these processes and related tolerance mechanisms may be used in biotechnology applications such as microbial enhanced oil recovery (MEOR). The majority of research has focused on microbiology processes in oil reservoirs and oilfields; as such there is a paucity of information specific to oil sands. By studying microbial processes in oil sands there is the potential to use microbes in MEOR applications. This article reviews the microbiology of the Athabasca Oil Sands and the mechanisms bacteria use to tolerate low water and high hydrocarbon availability in oil reservoirs and oilfields, and potential applications in MEOR.     

Microbial processes of the carbon and sulfur cycles in the Chukchi Sea

The research performed in August 2004 within the framework of the Russian-American Long-term Census of the Arctic (RUSALCA) resulted in the first data concerning the rates of the key microbial processes in the water column and bottom sediments of the Bering strait and the Chukchi Sea. The total bacterial counts in the water column varied from 30 × 10³ cells ml^?1 in the northern and eastern parts to 245 × 10³ cells ml^?1 in the southern part. The methane content in the water column of the Chukchi sea varied from 8 nmol CH₄l^?1 in the eastern part of the sea to 31 nmol CH₄l^?1 in the northern part of the Herald Canyon. Microbial activity occurred in the upper 0–3 cm of the bottom sediments; the methane formation rate varied from 0.25 to 16 nmol CH₄dm^?3 day^?1. The rates of methane oxidation varied from 1.61 to 14.7 nmol CH₄dm^?3 day^?1. The rates of sulfate reduction varied from 1.35 to 16.2 μmol SO ₄ ^2? dm^?1 day^?1. The rate of methane formation in the sediments increased with depth, while sulfate reduction rates decreased (less than 1 μmol SO ₄ ^2? dm^?3 day^?1). These high concentrations of biogenic elements and high rates of microbial processes in the upper sediment layers suggest a specific type of trophic chain in the Chukchi Sea. The approximate calculated balance of methane emission from the water column into the atmosphere is from 5.4 to 57.3 μmol CH₄m^?2 day^?1.     

The effect of nutrients and environmental conditions on biomass and oil production in Botryococcus braunii Race B strains

The green alga Botryococcus braunii is widely recognized as a source of non-fossil oil. However, limitations in Botryococcus biomass production hamper its commercial exploitation. This study examines the effects of nutrients (nitrogen and iron) and environmental conditions (temperature, light intensity and photoperiod) on biomass and oil production in two B. braunii Race B strains, Kossou-4 and Overjuyo-3. The highest biomass and oil production were obtained at a nitrogen concentration of 750 mg l^?1, iron concentration of 6 mg l^?1, at 25°C and at 135 µmol photons m^?2 s^?1 with a photoperiod of 16 h light:8 h darkness. Culturing the strains in Blue-green (BG11) medium containing optimized nutrients under optimal conditions resulted in an up to ~10.6-fold increase in biomass. In Kossou-4 and Overjuyo-3 strains, biomass increased from 1.647 g 10 l^?1 and 3.137 g 10 l^?1 respectively in normal BG11 medium to 17.390 g 10 l^?1 and 21.721 g 10 l^?1 in optimized BG11 media and growth conditions. This was accompanied by ~8–10.5-fold increase in oil production compared with that in normal BG11 medium. Oil (0.324 g 10 l^?1 and 0.211 g 10 l^?1) was produced in normal BG11 medium in Kossou-4 and Overjuyo-3 strains respectively, compared with 2.642 g 10 l^?1 (Kossou-4) and 2.206 g 10 l^?1 (Overjuyo-3) in modified BG11 media under optimized conditions. Therefore, optimization of nutrients and environmental conditions can increase biomass and oil production in the two strains of B. braunii.     

Low concentrations of glyphosate‐based herbicide cause complete loss of sperm motility of yellowtail tetra fish Astyanax lacustris

Environmental relevant concentrations of glyphosate‐based herbicide as 50 µg l^?1, 300 µg l^?1 and 1800 µg l^?1 can affect sperm quality of yellowtail tetra fish Astyanax lacustris . Viability of sperm cells was impaired at 300 µg l^?1, a concentration that is within legal limits in U.S.A. waterbodies, while motility was impaired at 50 µg l^?1, which is the more stringent limit set in Brazilian law. Therefore, environment protection agencies must review regulations of glyphosate‐based herbicides on water bodies.     

Complex nitrogen cycling in the sponge Geodia barretti

Marine sponges constitute major parts of coral reefs and deep‐water communities. They often harbour high amounts of phylogenetically and physiologically diverse microbes, which are so far poorly characterized. Many of these sponges regulate their internal oxygen concentration by modulating their ventilation behaviour providing a suitable habitat for both aerobic and anaerobic microbes. In the present study, both aerobic (nitrification) and anaerobic (denitrification, anammox) microbial processes of the nitrogen cycle were quantified in the sponge Geodia barretti and possible involved microbes were identified by molecular techniques. Nitrification rates of 566 nmol N cm^?3 sponge day^?1 were obtained when monitoring the production of nitrite and nitrate. In support of this finding, ammonia‐oxidizing Archaea (crenarchaeotes) were found by amplification of the amoA gene, and nitrite‐oxidizing bacteria of the genus Nitrospira were detected based on rRNA gene analyses. Incubation experiments with stable isotopes (¹⁵NO₃^‐ and ¹⁵NH₄⁺) revealed denitrification and anaerobic ammonium oxidation (anammox) rates of 92 nmol N cm^?3 sponge day^?1 and 3 nmol N cm^?3 sponge day^?1 respectively. Accordingly, sequences closely related to ‘Candidatus Scalindua sorokinii’ and ‘Candidatus Scalindua brodae’ were detected in 16S rRNA gene libraries. The amplification of the nirS gene revealed the presence of denitrifiers, likely belonging to the Betaproteobacteria. This is the first proof of anammox and denitrification in the same animal host, and the first proof of anammox and denitrification in sponges. The close and complex interactions of aerobic, anaerobic, autotrophic and heterotrophic microbial processes are fuelled by metabolic waste products of the sponge host, and enable efficient utilization and recirculation of nutrients within the sponge–microbe system. Since denitrification and anammox remove inorganic nitrogen from the environment, sponges may function as so far unrecognized nitrogen sinks in the ocean. In certain marine environments with high sponge cover, sponge‐mediated nitrogen mineralization processes might even be more important than sediment processes.     

Arthrospira platensis biomass with high protein content cultivated in continuous process using urea as nitrogen source

Aims: Arthrospira platensis has been studied for single‐cell protein production because of its biomass composition and its ability of growing in alternative media. This work evaluated the effects of different dilution rates (D) and urea concentrations (N₀) on A. platensis continuous culture, in terms of growth, kinetic parameters, biomass composition and nitrogen removal. Methods and results: Arthrospira platensis was continuously cultivated in a glass‐made vertical column photobioreactor agitated with Rushton turbines. There were used different dilution rates (0·04–0·44 day^?1) and urea concentrations (0·5 and 5 mmol l^?1). With N₀ = 5 mmol l^?1, the maximum steady‐state biomass concentration was1415 mg l^?1, achieved with D = 0·04 day^?1, but the highest protein content (71·9%) was obtained by applying D = 0·12 day^?1, attaining a protein productivity of 106·41 mg l^?1 day^?1. Nitrogen removal reached 99% on steady‐state conditions. Conclusions: The best results were achieved by applying N₀ = 5 mmol l^?1; however, urea led to inhibitory conditions at D ≥ 0·16 day^?1, inducing the system wash‐out. The agitation afforded satisfactory mixture and did not harm the trichomes structure. Significance and Impact of the Study: These results can enhance the basis for the continuous removal of nitrogenous wastewater pollutants using cyanobacteria, with an easily assembled photobioreactor.     

Uptake modes of octadecane by Pseudomonas sp. DG17 and synthesis of biosurfactant

Aims: In order to gain more insight into the uptake modes of octadecane by bacteria. Methods and Results: A strain that could utilize octadecane well was isolated from crude oil contaminated soil, and named as Pseudomonas sp. DG17 by 16S rDNA analysis. Culture growth result showed that Pseudomonas sp. DG17 grew well in the addition of 200 and 400 mg l^?1 of octadecane, which showed that physical contact between substrate and bacteria was important in the substrate biodegradation. Meanwhile, Pseudomonas sp. DG17 produced rhamnolipids biosurfactant that contains 10 congeners, thus causing the surface tension of the culture medium decline and facilitating the contact between hydrocarbon and bacteria. Scanning‐electron‐microscopy results showed that a disruption of the surface membranes in certain zones was observed in some of the cells grown in 400 mg l^?1 octadecane at 176 h compared with the cells in exponential phase at 72 h due to the production of biosurfactant‐rhamnolipid. Conclusions: These results indicated the possibility that the direct contact with insoluble octadecane droplets occurred before the contact with pseudosolubilization smaller oil droplets. Significance: This report throws more light on the uptake mechanisms of octadecane by bacteria, and proposes the possibility that role of biosurfactant is to increase the contact between hydrocarbon and bacteria by changing the cell membrane structure which needs studied in depth. Impact of Study: Results of this study are useful in the bioremediation of petroleum polluted soil.     

Microbial processes of the carbon and sulfur cycles in an ice‐covered,iron‐rich meromictic lake Svetloe (Arkhangelsk region,Russia)

Biogeochemical, isotope geochemical and microbiological investigation of Lake Svetloe (White Sea basin), a meromictic freshwater was carried out in April 2014, when ice thickness was ～0.5 m, and the ice‐covered water column contained oxygen to 23 m depth. Below, the anoxic water column contained ferrous iron (up to 240 μμM), manganese (60 μM), sulfide (up to 2 μM) and dissolved methane (960 μM). The highest abundance of microbial cells revealed by epifluorescence microscopy was found in the chemocline (redox zone) at 23–24.5 m. Oxygenic photosynthesis exhibited two peaks: the major one (0.43 μmol C L^?1 day^?1) below the ice and the minor one in the chemocline zone, where cyanobacteria related to Synechococcus rubescens were detected. The maximum of anoxygenic photosynthesis (0.69 μmol C L^?1 day^?1) at the oxic/anoxic interface, for which green sulfur bacteria Chlorobium phaeoclathratiforme were probably responsible, exceeded the value for oxygenic photosynthesis. Bacterial sulfate reduction peaked (1.5 μmol S L^?1 day^?1) below the chemocline zone. The rates of methane oxidation were as high as 1.8 μmol CH₄ L^?1 day^?1 at the oxi/anoxic interface and much lower in the oxic zone. Small phycoerythrin‐containing Synechococcus‐related cyanobacteria were probably involved in accumulation of metal oxides in the redox zone.     

Use of Rice Husk as Bulking Agent in Bioremediation of Automobile Gas Oil Impinged Agricultural Soil

Evaluation of rice husk (RH) as bulking agent in bioremediation of automobile gas oil (AGO) hydrocarbon polluted agricultural soil using renewal by enhanced natural attenuation (RENA) as control was the subject of the present investigation. The effect of different parameters such as total petroleum hydrocarbon (TPH), dehydrogenase activity (DHA), optical density and pH on bioremediation performance were evaluated. The studied parameters such as microbial dynamics, percentage degradation and DHA were found to be higher in RH-amended system and differed significantly with control at P < 0.05. RH resulted in high removal efficiency of 97.85 ± 0.93% under a two-month incubation period, while RENA had lesser removal efficiency of 53.15 ± 3.81%. Overall hydrocarbon biodegradation proceeded very slowly in the RENA particularly from week 0 to 4. Experimental data perfectly fitted into the first-order kinetic and generated high r² values (0.945), first-order degradation constant (0.47 day^?1), and shorter degradation half-life (1.50 d)—t_1/2 = Ln2/K and Ln2 numerically equals to 0.693 and hence written as 0.693/K. Micrococcus luteus and Rhizopus arrhizus were isolated in the present study, which displayed extreme AGO hydrocarbon biodegradative abilities. The use of RH in hydrocarbon-polluted soil significantly increased biodegradation rate and resulted in effective AGO cleanup within 2 months period. Therefore, RH provides an alternative source of bioremediation material in field application for abundant petroleum hydrocarbon soil pollution.     

Characterization of an alkane-degrading methanogenic enrichment culture from production water of an oil reservoir after 274 days of incubation

Oil reservoirs represent special habitats for the activity of anaerobic microbial communities in the transformation of organic compounds. To understand the function of microbial communities in oil reservoirs under anaerobic conditions, an alkane-degrading methanogenic enrichment culture was established and analyzed. Results showed that a net 538 ??mol of methane higher than the controls were produced over 274 days of incubation in microcosms amended with alkanes and a decrease in the alkanes profile was also observed. Phylogenetic analysis of 16S rRNA gene sequences retrieved from the enrichment microcosms indicated that the archaeal phylotypes were mostly related to members of the orders Methanobacteriales and Methanosarcinales. The bacterial clone library was composed of sequences affiliated with the Firmicutes, Proteobacteria, Deferribacteres, and Bacteroidetes. However, most of the bacterial clones retrieved from the enrichment cultures showed low similarity to 16S rRNA gene sequences of the cultured members, indicating that the enrichment cultures contained novel bacterial species. Though alkane-degrading methanogenic enrichment consortium has rarely been reported from petroleum reservoirs, our results indicated that oilfield production water harbors a microbial community capable of syntrophic conversion of n-alkanes to methane, which sheds light on the bio-utilization of marginal oil reservoirs for enhanced energy recovery.     

石油烃的厌氧生物降解对油藏残余油气化开采的启示

利用微生物将油藏中难以动用的原油就地转化为甲烷,以天然气的形式开采、或作为战略资源就地储备,从而大幅度提高油气资源的利用率,是当前国际上研究的前沿课题。本文综述了石油烃厌氧生物降解转化为甲烷的菌群结构、反应热力学和反应动力学等基础科学问题的最新研究进展,讨论了油藏残余油气化开采技术的可行性及开发潜力,提出了该技术进一步研究的方向。     

Calcium‐dependent behavioural responses to acute copper exposure in Oncorhynchus mykiss

Using rainbow trout Oncorhynchus mykiss, the present study demonstrated that: (1) calcium (Ca) increased the range of copper (Cu) concentrations that O. mykiss avoided; (2) Ca conserved the maintenance of pre‐exposure swimming activity during inescapable acute (10 min) Cu exposure. Data showed that when presented with a choice of Cu‐contaminated water (ranging from 0 to 454 µg Cu l^?1) and uncontaminated water in a choice tank, O. mykiss acclimated and tested at low Ca concentration (3 mg Ca l^?1) avoided the 10 µg Cu l^?1 only. By contrast, O. mykiss acclimated and tested at high Ca concentration (158 mg Ca l^?1) avoided all the Cu concentrations ≥37 µg l^?1. The Cu avoidance was connected with increased spontaneous swimming speed in the Cu‐contaminated water. When subjected to inescapable Cu exposure (35 µg Cu l^?1), O. mykiss acclimated and tested at low Ca concentration reduced their spontaneous swimming speed, whereas no response was observed in O. mykiss acclimated and tested at high Ca concentration. Collectively, the data support the conclusion that in O. mykiss the behavioural responses to acute Cu exposure are Ca‐dependent.