The influence of structural components of alkyl esters on their anaerobic biodegradation in marine sediment

Ester-based organic compounds are one type of synthetic base fluid added to drilling mud used during off-shore oil-drilling operations in the Gulf of Mexico. Concern over the environmental impact of synthetic base fluid (SBF) contaminated rock cuttings discharged into the Gulf of Mexico has prompted the promulgation of EPA regulations requiring that all SBF be tested for biodegradability in marine sediment prior to their use in the Gulf. In order to allow the design or selection of suitably biodegradable esters, the anaerobic biodegradability of a variety of ester compounds was tested using a marine sediment inoculum to reveal the effect of: (a) increasing the chain length of the acid moiety, (b) increasing the chain length of the alcohol moiety; (c) alternating the relative size of the alcohol and acid moieties, (d) branching in the alcohol moiety, and (e) the presence of an unsaturated bond in the acidic moiety. The chemical structure of esters was found to affect the completeness and rate of anaerobic biodegradation, and would affect their ability to be certified for use as an SBF in the Gulf of Mexico. Recommendations for ester usage include using esters that have a total carbon number of between 12 and 18 and avoiding the use of branched alcohols (or acids by inference). The presence of an unsaturated bond in the acid (or alcohol by inference) increased biodegradability of the ester.     

Hydrogen as an electron donor for sulfate-reducing bacteria in slurries of salt marsh sediment

Experiments with a Warburg respirometer showed that a sediment slurry consumed hydrogen from a hydrogen atmosphere, and this consumption was not due to the activity of methanogenic bacteria. The hydrogren uptake was inhibited by the addition of 20 mM molybdate. Further experiments with sediment slurry held in conical flasks under an atmosphere of nitrogen showed that hydrogen accumulated in the headspace when bacterial sulfate reduction was inhibited either by the addition of 20 mM molybdate or by low (<5 mM) sulfate concentrations in the slurry. Methanogenesis was stimulated in the presence of a hydrogen atmosphere or by the addition of 20 mM molybdate. The results confirmed that hydrogren was an important electron donor for sulfate-reducing bacteria present in the sediment. The stimulation of methanogenesis by molybdate could be explained in part by a competition for hydrogen between sulfate-reducing bacteria and hydrogen-metabolizing methanogenic bacteria, but competition for another common substrate, possibly acetate, could also be significant.     

The influence of light and nutrient addition upon the sediment chemistry of iron in an arctic lake

The geochemical response of sediments to increased nutrient input to an Alaskan, arctic lake was examined using direct measurements of sediment-water chemical fluxes. An unexpected increase in Fe flux occurred when sediments were exposed to high incident radiation and nutrient concentrations. Correlation between light and acid-soluble Fe concentrations suggests that photoreduction of Fe(III) oxides may occur, but nutrient addition enhanced the effect indicating that primary productivity was also important. The processes controlling the flux of Fe from sediments in this lake were complex and included the redox potential (dissolved oxygen concentration) of the water, quality of organic matter present in the sediment, light, and nutrients supplied from the sediments and/or water column. These four factors together with the possibility of direct uptake of Fe by phytoplankton and the possible release of algal reductants may contribute to Fe cycling in this lake.     

Effect of 2,4‐dinitrotoluene on the anaerobic bacterial community in marine sediment

Aims: To study the impact of added 2,4‐dinitrotoluene (DNT) on the anaerobic bacterial community in marine sediment collected from an unexploded ordnance dumping site in Halifax Harbour. Methods and Results: Marine sediment was spiked with 2,4‐DNT and incubated under anaerobic conditions in the presence and absence of lactate. Indigenous bacteria in the sediment removed 2,4‐DNT with subsequent formation of its mono‐ and diamino‐derivatives under both conditions. PCR–DGGE and nucleotide sequencing were used to monitor the change in the bacterial population in sediment caused by the presence of 2,4‐DNT. The results showed that denaturing gradient gel electrophoresis banding patterns of sediment microcosms treated with 2,4‐DNT were different from controls that did not receive 2,4‐DNT. Bacteroidetes, Firmicutes and δ‐Proteobacteria were present in sediment incubated in the absence of 2,4‐DNT. However, several γ‐Proteobacteria became dominant in sediment in the presence of 2,4‐DNT, two of which were 99% similar to Shewanella canadensis and Shewanella sediminis. In the presence of both 2,4‐DNT and lactate, two additional δ‐Proteobacteria were enriched, one closely related (98% similarity) to Desulfofrigus fragile and the other affiliated (96% similarity) to Desulfovibrio sp. In contrast, none of the above four Proteobacteria were enriched in sediment incubated with lactate alone. Conclusions: Presence of 2,4‐DNT led to a significant change in bacterial population of marine sediment with the enrichment of several γ‐ and δ‐Proteobacteria. Significance and Impact of the Study: Our results provided the first evidence on the impact of the pollutant 2,4‐DNT on the indigenous bacterial community in marine sediment, and provided an insight into the composition of bacterial community that degrade 2,4‐DNT.     

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.     

Chlorophenol dechlorination and subsequent degradation in denitrifying microcosms fed low concentrations of nitrate

The potential of using nitrate as a terminal electron acceptor to stimulate anaerobic degradation of mixtures of monochlorophenols (MCPs) or dichlorophenols (DCPs) was evaluated. Contaminated and non-contaminated soils were added to water saturated anaerobic microcosms supplemented with 1 mM or 5 mM nitrate. Denitrification and dechlorination activity were present in three diverse soil types and were maintained upon refeeding both nitrate and the appropriate chlorophenol. However, dechlorination activity could only be serially transferred in enrichments with an added electron donor such as acetate. Dehalogenation activity in enrichments from four of the primary microcosms showed at least five different dechlorination reactions, each mediated by different microbial communities. Three of these are distinct ortho-dechlorinating paths; two are meta-dechlorinating and one is the para-dechlorination of 3,4-DCP. Simultaneous dechlorination and denitrification was observed and both activities could be maintained in microcosms but only in the presence of low nitrate concentrations. Dechlorination and denitrification were mediated by two separate microbial communities; one that dechlorinates without use of nitrate and one that denitrifies while oxidizing the dechlorinated aromatic ring. There was no evidence that dechlorination is mediated by the denitrifying community, however the maintenance of a denitrification potential using low (< 1 mM) nitrate concentrations may be useful for completing the food chain by stimulating the mineralization of phenol and benzoate.     

The influence of catchment characteristics on suspended sediment properties

In order to investigate the influence of catchment characteristics on suspended sediment properties, suspended sediment samples were collected from the outlets of 60 small catchments located throughout Southwest England over a 14-month period. The catchments were selected to provide a representative range of topographic, land use, soil and geological characteristics. The suspended sediment samples were collected using time-integrating trap samplers, emptied at monthly intervals. Laboratory analysis focused on a selection of suspended sediment properties, including particle size composition, cation exchange capacity, and organic carbon, nitrogen, metal, radionuclide and phosphorus content. The results presented show that despite the substantial variability of catchment characteristics associated with the 60 sampling sites, the suspended sediment properties generally fell within a relatively limited range. However, statistical analysis identified significant spatial variability of suspended sediment within Southwest England, which in turn can be linked to catchment characteristics.     

Biodegradation of cyclic nitramines by tropical marine sediment bacteria

Undersea deposition of unexploded ordnance (UXO) constitutes a potential source of contamination of marine environments by hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) and octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine (HMX). The goal of the present study was to determine microbial degradation of RDX and HMX in a tropical marine sediment sampled from a coastal UXO field in the region of Oahu Island in Hawaii. Sediment mixed cultures growing in marine broth 2216 (21°C) anaerobically mineralized 69% or 57% (CO₂, 25 days) of the total carbon of [UL-¹⁴ C]-RDX (100 M) or [UL-¹⁴ C]-HMX (10 M), respectively. As detected by PCR-DGGE, members of -proteobacteria (Halomonas), sulfate-reducing -proteobacteria (Desulfovibrio), firmicutes (Clostridium), and fusobacterium appeared to be dominant in RDX-enrichment and/or HMX-enrichment cultures. Among 22 sediment bacterial isolates screened for RDX and HMX biodegradation activity under anaerobic conditions, 5 were positive for RDX and identified as Halomonas (HAW-OC4), Marinobacter (HAW-OC1), Pseudoalteromonas (HAW-OC2 and OC5) and Bacillus (HAW-OC6) by their 16S rRNA genes. Sediment bacteria degraded RDX to N₂O and HCHO via the intermediary formation of hexahydro-1-nitroso-3,5-dinitro-1,3,5-triazine (MNX) and methylenedinitramine. The present findings demonstrate that cyclic nitramine contaminants are likely to be degraded upon release from UXO into tropical marine sediment.     

The kinetics of the degradation of chloroform and benzene in anaerobic sediment from the River Rhine

In anaerobic methanogenic sediment microcosms¹⁴C labelled chloroform was degraded mainly to carbon dioxide. At a concentration of 4 g.l^–1 the mineralization followed first order kinetics with a half life of 12 days at 10°C and 2.6 days at 20°C. At a concentration of 400 g.l^–1 the mineralization rate increased with time and followed logarithmic kinetics with a _max of 0.02.d^–1 at 10°C. The logarithmic kinetics can be explained by growth of the bacteria on the higher concentration of chloroform with a generation time of 35 days. Shaking and oxygenation did not inhibit the mineralization of chloroform, probably because of bacterial consumption of the dissolved oxygen. ¹⁴C labelled benzene was mineralized only for a small percentage to¹⁴C labelled carbon dioxide while other, not acid extractable, degradation products were formed. Under anaerobic conditions after one day when 5% of the benzene was degraded to carbon dioxide, the mineralization ceased, while the disappearance of benzene proceeded. With air in the headspace of the incubation bottles 25% of the benzene was mineralized to carbon dioxide. The anaerobic degradation of benzene at a concentration of 100 .l^–1 showed similar kinetics as the degradation at 1 g.l^–1. Hence no adaptation of the microflora in the sediment occurred during the 63 days of the experiment at 100 g.l^–1.     

The denitrification potential and density of denitrifying bacteria in the bottom sediments of Matupi Harbor (Papua New Guinea)

The denitrification potential and density of denitrifying bacteria were determined in suboxidized sediments of Matupi Harbor (Papua New Guinea). The sediments were characterized by low denitrification potentials, not exceeding 0.8 nM/(g h). The maximum density of denitrifying bacteria was recorded in the upper sediment horizons and was 10⁴ cells/g of wet ground.     

Seasonal variation of fungal biomass in the sediment of a salt marsh in New Brunswick

In a marsh in New Brunswick, Canada, belowground biomass of Spartina alterniflora consistently exceeded aboveground biomass by a factor of approximately 9. Both values peaked in July. Redox potential of the sediment was negative at all levels tested (2, 6, and 11 cm below surface), and was negatively correlated with depth. Concentrations of ergosterol, a sterol typical of higher fungi, were negatively correlated with redox potential and were highest in roots and rhizomes in July and August, 1–3 cm below the surface. These maxima corresponded to a fungal content of approximately 0.6% per ash-free dry mass of Spartina material. Balsa wood panels buried in anaerobic salt marsh sediment were colonized by fungi within 12 weeks. Eight fungal species isolated from S. alterniflora roots did not grow in the absence of oxygen, but were able to grow downward into an anaerobic medium.Offprint requests to: F. Bärlocher.     

厌氧处理低浓度污水颗粒污泥的形成和生物特性研究

用UASB反应器在室温条件下处理模拟低浓度污水,研究了颗粒污泥的形成过程和特性。UASB反应器进水浓度为100～200mgCOD／L,水温为25℃～9℃,60天内形成了成熟的颗粒污泥。颗粒污泥中以丝状菌为主,存在少量球状菌。     

Effect of an aquatic herbicide on sediment nutrient flux in a freshwater marsh

Three concentrations of the herbicide simazine were added to in situ macrophyte-free enclosures with and without sediment contact. Changes in the concentrations of total ammonia, total reactive phosphorus, and silicon were monitored, and net sediment flux was calculated from the difference in nutrient concentration between bottomed and unbottomed enclosures. Rates of sediment release for all three nutrients were unaltered by 0.1 mg · l^–1 simazine in relation to a control, whereas rates were increased proportionally at 1.0 and 5.0 mg · l^–1. These results suggest that increases in dissolved nutrients commonly observed following herbicide treatment of shallow waters may not be attributable solely to macrophyte decay, byt may also involve a complex interaction of biotic and abiotic sediment nutrient exchange processes.Contribution Number 103 from the University of Manitoba Field Station, Delta Marsh, Canada     

The influence of anaerobic pretreatment on the nitrogen removal from biosynthetic pharmaceutical wastewaters

The C:N ratio of the pharmaceutical wastewaters is usually suitable for a combination of the anaerobic pretreatment with the high COD removal and aerobic posttreatment with the efficient biological N removal. This kind of anaerobic-aerobic process was tested in semipilot scale by using a UASB reactor and an activated sludge system with a predenitrification (total volume 100 1). It was found that at a total HRT of 2.3 days an average of 97.5% of COD and 73.5% of total N was removed. The UASB reactor was operated at 30°C with a volumetric loading rate of 8.7 kg.m^-3.d^-1, the efficiency of COD removal was 92.2%. The processes, which take part in the biological removal of nitrogen, especially the nitrification, were running with lower rates than usually observed in aerobic treatment systems.Abbreviations AAO anaerobic anoxic oxic configuration - AOO anaerobic oxic oxic configuration - B _V volumetric organic loading rate (kg COD.m^-3. d^-1) - dB _x specific COD removal rate (mg COD. g^-1 VSS. d^-1) - DNR denitrification rate (mg N–NO₃. g^-1 VSS. h^-1) - E_COD efficiency of COD removal (%) - HRT hydraulic retention time (d) - NR nitrification rate (mg N–NO₃. g^-1 VSS. h^-1) - R recirculation ratio (%) - SBP specific biogas production (m³.kg^-1 removed COD) - SRT solids retention time; sludge age (d) - SS suspended solids (g.1^-1) - UASB upflow anaerobic sludge blanket reactor - VSS volatile suspended solids (g.1^-1)     

Metabolism of low molecular weight organic compounds by sulfate-reducing bacteria in a Delaware salt marsh

Oxidation of acetate, lactate, pyruvate, and ethanol to CO₂ in anaerobic salt marsh sediments was rapid, with the oxidation rate being significantly inhibited (60–90% decrease) in the presence of 2 mM sodium molybdate, an inhibitor of sulfate-reducing bacteria (SRB). 2-Bromoethanesulfonic acid (BES), an inhibitor of methanogenic bacteria, generally had no effect on the oxidation rate. Acetate was the only intermediate product detected in the oxidation of lactate and ethanol. Competition studies with lactate, acetate, and ethanol indicated that the preferred order of substrate utilization was lactate, then acetate, then ethanol. The turnover times of these three compounds in salt marsh sediments via the combined CO₂ plus acetate pool was rapid (10–13 hours) with a two- to threefold increase in the turnover time in the presence of molybdate. These results strongly suggest that SRB play a major role in the terminal metabolism of low molecular weight organic compounds in anaerobic salt marsh sediment.     

Metabolism of alkylbenzenes, alkanes, and other hydrocarbons in anaerobic bacteria

Aromatic and aliphatic hydrocarbons are the main constituents of petroleum and its refined products. Whereas degradation of hydrocarbons by oxygen-respiring microorganisms has been known for about a century, utilization of hydrocarbons under anoxic conditions has been investigated only during the past decade. Diverse strains of anaerobic bacteria have been isolated that degrade toluene anaerobically, using nitrate, iron(III), or sulfate as electron acceptors. Also, other alkylbenzenes such as m-xylene or ethylbenzene are utilized by a number of strains. The capacity for anaerobic utilization of alkylbenzenes has been observed in members of the -, -, - and -subclasses of the Proteobacteria. Furthermore, denitrifying bacteria and sulfate-reducing bacteria with the capacity for anaerobic alkane degradation have been isolated, which are members of the - and -subclass, respectively. The mechanism of the activation of hydrocarbons as apolar molecules in the absence of oxygen is of particular interest.The biochemistry of anaerobic toluene degradation has been studied in detail. Toluene is activated by addition to fumarate to yield benzylsuccinate, which is then further metabolized via benzoyl-CoA. The toluene-activating enzyme presents a novel type of glycine radical protein. Another principle of anaerobic alkylbenzene activation has been observed in the anaerobic degradation of ethylbenzene. Ethylbenzene in denitrifying bacteria is dehydrogenated to 1-phenylethanol and further to acetophenone; the latter is also metabolized to benzoyl-CoA. Naphthalene is presumably activated under anoxic conditions by a carboxylation reaction. Investigations into the pathway of anaerobic alkane degradation are only at the beginning. The saturated hydrocarbons are mostlikely activated by addition of a carbon compound rather than by desaturation and hydration, as speculated about in some early studies. An anaerobic oxidation of methane with sulfate as electron acceptor has been documented in aquatic sediments. The process is assumed to involve a reversal of methanogenesis catalyzed by Archaea, and scavenge of an electron-carrying metabolite by sulfate-reducing bacteria. Among unsaturated non-aromatic hydrocarbons, anaerobic bacterial degradation has been demonstrated and investigated with n-alkenes, alkenoic terpenes and the alkyne, acetylene.