Molecular characterization of mesophilic and thermophilic sulfate reducing microbial communities in expanded granular sludge bed (EGSB) reactors

The microbial communities established in mesophilic and thermophilic expanded granular sludge bed reactors operated with sulfate as the electron acceptor were analyzed using 16S rRNA targeted molecular methods, including denaturing gradient gel electrophoresis, cloning, and phylogenetic analysis. Bacterial and archaeal communities were examined over 450 days of operation treating ethanol (thermophilic reactor) or ethanol and later a simulated semiconductor manufacturing wastewater containing citrate, isopropanol, and polyethylene glycol 300 (mesophilic reactor), with and without the addition of copper(II). Analysis, of PCR-amplified 16S rRNA gene fragments using denaturing gradient gel electrophoresis revealed a defined shift in microbial diversity in both reactors following a change in substrate composition (mesophilic reactor) and in temperature of operation from 30°C to 55°C (thermophilic reactor). The addition of copper(II) to the influent of both reactors did not noticeably affect the composition of the bacterial or archaeal communities, which is in agreement with the very low soluble copper concentrations (3–310 μg l⁻¹) present in the reactor contents as a consequence of extensive precipitation of copper with biogenic sulfides. Furthermore, clone library analysis confirmed the phylogenetic diversity of sulfate-reducing consortia in mesophilic and thermophilic sulfidogenic reactors operated with simple substrates.     

Spatiotemporal variations in the abundances of the prokaryotic rRNA genes, <Emphasis Type="Italic">pmoA</Emphasis>, and <Emphasis Type="Italic">mcrA</Emphasis> in the deep layers of a peat bog in Sarobetsu-genya wetland,Japan

Spatiotemporal variations in microbial gene abundances were investigated to identify potential zones of methanotroph and methanogen biomass in a peat bog in Sarobetsu-genya wetland. The abundances of the bacterial and archaeal 16S rRNA genes, pmoA, and mcrA were 10⁷–10⁹, 10⁷–10⁸, 10⁴–10⁶, and 10⁴–10⁷ copies g⁻¹ dry peat, respectively. Correlation analysis based on microbial gene abundances and environmental factors showed that the spatiotemporal distributions of the abundances of the four microbial genes in peat layers were similar. The mcrA abundance showed a significant negative correlation with the dissolved organic carbon content and a significant positive correlation with the peat temperature. The pmoA abundance was not detectable during the spring thaw when the lowest peat temperature at a depth of 50 cm was recorded. At a depth of 200 cm, the peat temperature exceeded 6°C throughout the year, and the mcrA abundance exceeded 10⁴ copies g⁻¹ dry peat. These results indicate that the seasonal microbial activity related to methane should be evaluated in not only the shallow but also the deep peat layers in order to elucidate the methane dynamics in boreal wetlands.     

Microbial community structure in anaerobic co-digestion of grass silage and cow manure in a laboratory continuously stirred tank reactor

The impacts of feeding ratio and loading rate on the microbial community during co-digestion of grass silage with cow manure in an anaerobic laboratory continuously stirred tank reactor were investigated by 16S rRNA gene-based fingerprints. The microbial community remained stable when the reactor was fed with cow manure alone and with up to 20% of grass silage in feedstock at an organic loading rate (OLR) of 2 kg VS m⁻³ day⁻¹. Large changes in the bacterial community were observed when the loading ratio of grass was increased to 40%, while there was little change in the archaeal community. During the increase in OLR from 2 to 4 kg VS m⁻³ day⁻¹ the bacterial community structure showed few differences, whereas Archaea was undetectable. Sequencing of the major DGGE bands indicated that the phylum Bacteriodetes predominated in the bacterial community. Two unclassified bacteria with high abundance survived throughout the operation of the reactor.     

Copper enhances the activity and salt resistance of mixed methane-oxidizing communities

Effluents of anaerobic digesters are an underestimated source of greenhouse gases, as they are often saturated with methane. A post-treatment with methane-oxidizing bacterial consortia could mitigate diffuse emissions at such sites. Semi-continuously fed stirred reactors were used as model systems to characterize the influence of the key parameters on the activity of these mixed methanotrophic communities. The addition of 140 mg L⁻¹ NH₄⁺–N had no significant influence on the activity nor did a temperature increase from 28°C to 35°C. On the other hand, addition of 0.64 mg L⁻¹ of copper(II) increased the methane removal rate by a factor of 1.5 to 1.7 since the activity of particulate methane monooxygenase was enhanced. The influence of different concentrations of NaCl was also tested, as effluents of anaerobic digesters often contain salt levels up to 10 g NaCl L⁻¹. At a concentration of 11 g NaCl L⁻¹, almost no methane-oxidizing activity was observed in the reactors without copper addition. Yet, reactors with copper addition exhibited a sustained activity in the presence of NaCl. A colorimetric test based on naphthalene oxidation showed that soluble methane monooxygenase was inhibited by copper, suggesting that the particulate methane monooxygenase was the active enzyme and thus more salt resistant. The results obtained demonstrate that the treatment of methane-saturated effluents, even those with increased ammonium (up to 140 mg L⁻¹ NH₄⁺–N) and salt levels, can be mitigated by implementation of methane-oxidizing microbial consortia.     

Efficient degradation of rice straw in the reactors packed by carbon fiber textiles

We have reported for the first time that agricultural and cellulosic waste, i.e., rice straw was directly applied to methanogenic bioreactors containing carbon fiber textiles (CFT) as supporting material. Addition of CFT to the methanogenic bioreactors enhanced the conversion of dichromate chemical oxygen demand of the substrate to methane (41%) to a greater extent than bioreactors without CFT (9%). In addition, removal of rice straw as a suspended solid was increased from 31% (in bioreactors without CFT) to 57% (in those with CFT). Methanogenic 16S rRNA gene analysis showed that the abundance of acetoclastic methanogen, genus Methanosarcina, was about 11 times higher in bioreactors with CFT (suspended fraction plus retained fraction to CFT) than in bioreactors without CFT (suspended fraction), resulting in lower concentration of acetate in bioreactors with CFT (0.4 mM) than in those without CFT (29.7 mM). On the other hand, the abundance of hydrogenotrophic methanogen, genus Methanobacterium, in bioreactors with CFT was similar to those without CFT. Bacterial communities in bioreactors with CFT were different from those in bioreactors without CFT. Our results indicated that specific microbial community and cooperative relationships between microorganisms in reactors containing CFT facilitated efficient decomposition of rice straw and its conversion to methane.     

Long-term performance and microbial community analysis of a full-scale synthesis gas fed reactor treating sulfate- and zinc-rich wastewater

The performance of a full-scale (500 m³) sulfidogenic synthesis gas fed gas-lift reactor treating metal- and sulfate-rich wastewater was investigated over a period of 128 weeks. After startup, the reactor had a high methanogenic activity of 46 Nm³·h⁻¹. Lowering the carbon dioxide feed rate during the first 6 weeks gradually lowered the methane production rate. Between weeks 8 and 93, less than 1% of the hydrogen supplied was used for methanogenesis. Denaturing gradient gel electrophoresis analysis of polymerase chain reaction-amplified 16S rRNA gene fragments showed that the archaeal community decreased in diversity but did not disappear completely. After the carbon dioxide feed rate increased in week 88, the methane production rate also increased, confirming that methane production was carbon dioxide limited. Even though lowering the carbon dioxide feed appeared to affect part of the sulfate-reducing community, it did not prevent achieving the desired rates of sulfate reduction. The average sulfate conversion rate was 181 kg∙h⁻¹ for the first 92 weeks. After 92 weeks, the sulfate input rate was increased and from week 94 to 128, the average weekly sulfate conversion rate was 295 kg·h⁻¹ (SD ± 87). Even higher sulfate conversion rates of up to 400 kg·h⁻¹ could be sustained for weeks 120–128. The long-term performance and stability together with the ability to control methanogenesis demonstrates that synthesis gas fed reactor can be used successfully at full scale to treat metal and sulfate-rich wastewater.     

Decreasing ammonia inhibition in thermophilic methanogenic bioreactors using carbon fiber textiles

Ammonia accumulation is one of the main causes of the loss of methane production observed during fermentation. We investigated the effect of addition of carbon fiber textiles (CFT) to thermophilic methanogenic bioreactors with respect to ammonia tolerance during the process of degradation of artificial garbage slurry, by comparing the performance of the reactors containing CFT with the performance of reactors without CFT. Under total ammonia-N concentrations of 3,000 mg L⁻¹, the reactors containing CFT were found to mediate stable removal of organic compounds and methane production. Under these conditions, high levels of methanogenic archaea were retained at the CFT, as determined by 16S rRNA gene analysis for methanogenic archaea. In addition, Methanobacterium sp. was found to be dominant in the suspended fraction, and Methanosarcina sp. was dominant in the retained fraction of the reactors with CFT. However, the reactors without CFT had lower rates of removal of organic compounds and production of methane under total ammonia-N concentrations of 1,500 mg L⁻¹. Under this ammonia concentration, a significant accumulation of acetate was observed in the reactors without CFT (130.0 mM), relative to the reactors with CFT (4.2 mM). Only Methanobacterium sp. was identified in the reactors without CFT. These results suggest that CFT enables stable proliferation of aceticlastic methanogens by preventing ammonia inhibition. This improves the process of stable garbage degradation and production of methane in thermophilic bioreactors that include high levels of ammonia.     

A bioelectrochemical reactor containing carbon fiber textiles enables efficient methane fermentation from garbage slurry

A packed-bed system includes supporting materials to retain microorganisms and a bioelectrochemical system influences the microbial metabolism. In our study, carbon fiber textiles (CFT) as a supporting material was attached onto a carbon working electrode in a bioelectrochemical reactor (BER) that degrades garbage slurry to methane, in order to investigate the effect of combining electrochemical regulation and packing CFT. The potential on the working electrode in the BER containing CFT was set to −1.0 V or −0.8 V (vs. Ag/AgCl). BERs containing CFT exhibited higher methane production, elimination of dichromate chemical oxygen demand, and the ratio of methanogens in the suspended fraction than reactors containing CFT without electrochemical regulation at an organic loading rate (OLR) of 27.8 gCODcr/L/day. In addition, BERs containing CFT exhibited higher reactor performances than BERs without CFT at this OLR. Our results revealed that the new design that combined electrochemical regulation and packing CFT was effective.     

Effect of seed sludge microbial community and activity on the performance of anaerobic reactors during the start-up period

In this study, two laboratory-scale anaerobic batch reactors started up with different inoculum sludges and fed with the same synthetic wastewater were monitored in terms of performance and microbial community shift by denaturant gradient gel electrophoresis fingerprinting and subsequent cloning, sequencing analysis in order to reveal importance of initial quality of inoculum sludge for operation of anaerobic reactors. For this purpose, two different seed sludge were evaluated. In Reactor1 seeded with a sludge having less diverse microbial community (19 operational taxonomic unit (OTU’s) for Bacterial and 8 OTU’s for Archaeal community, respectively) and a methanogenic activity of 150 ml CH₄ g TVS⁻¹ day⁻¹, a chemical oxygen demand (COD) removal efficiency of 78.8 ± 4.17% was obtained at a substrate to microorganism (S/X) ratio of 0.38. On the other hand, Reactor2, seeded with a sludge having a much more diverse microbial community (24 OTU’s for Bacterial and 9 OTU’s for Archaeal communities, respectively) and a methanogenic activity, 450 ml CH₄ g TVS⁻¹ day⁻¹, operated in the same conditions showed a better start-up performance; a COD removal efficiency of over 98% at a S/X ratio of 0.53. Sequence analysis of Seed2 revealed the presence of diverse fermentative and syntrophic bacteria, whereas excised bands of Seed1 related to fermentative and sulfate/metal-reducing bacteria. This study revealed that a higher degree of bacterial diversity, especially the presence of syntrophic bacteria besides the abundance of key species such as methanogenic Archaea may play an important role in the performance of anaerobic reactors during the start-up period.     

Microbial diversity and activity in hypersaline high Arctic spring channels

Lost Hammer (LH) spring is a unique hypersaline, subzero, perennial high Arctic spring arising through thick permafrost. In the present study, the microbial and geochemical characteristics of the LH outflow channels, which remain unfrozen at ≥−18°C and are more aerobic/less reducing than the spring source were examined and compared to the previously characterized spring source environment. LH channel sediments contained greater microbial biomass (~100-fold) and greater microbial diversity reflected by the 16S rRNA clone libraries. Phylotypes related to methanogenesis, methanotrophy, sulfur reduction and oxidation were detected in the bacterial clone libraries while the archaeal community was dominated by phylotypes most closely related to THE ammonia-oxidizing Thaumarchaeota. The cumulative percent recovery of ¹⁴C-acetate mineralization in channel sediment microcosms exceeded ~30% and ~10% at 5 and −5°C, respectively, but sharply decreased at −10°C (≤1%). Most bacterial isolates (Marinobacter, Planococcus, and Nesterenkonia spp.) were psychrotrophic, halotolerant, and capable of growth at −5°C. Overall, the hypersaline, subzero LH spring channel has higher microbial diversity and activity than the source, and supports a variety of niches reflecting the more dynamic and heterogeneous channel environment.     

Change in ammonia-oxidizing microorganisms in enriched nitrifying activated sludge

In this study, sludge was taken from a municipal wastewater treatment plant that contained a nearly equal number of archaeal amoA genes (5.70 × 10⁶ ± 3.30 × 10⁵ copies mg sludge⁻¹) to bacterial amoA genes (8.60 × 10⁶ ± 7.64 × 10⁵ copies mg sludge⁻¹) and enriched in three continuous-flow reactors receiving an inorganic medium containing different ammonium concentrations: 2, 10, and 30 mM NH₄⁺–N (28, 140, and 420 mg N l⁻¹). The abundance and communities of ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB) in enriched nitrifying activated sludge (NAS) were monitored at days 60 and 360 of the operation. Early on, between day 0 and day 60 of reactor operation, comparative abundance of AOA amoA genes to AOB amoA genes varied among the reactors depending on the ammonium levels found in the reactors. As compared to the seed sludge, the number of AOA amoA genes was unchanged in the reactor with lower ammonium level (0.06 ± 0.04 mgN l⁻¹), while in the reactors with higher ammonium levels (0.51 ± 0.33 and 0.25 ± 0.10 mgN l⁻¹), the numbers of AOA amoA genes were deteriorated. By day 360, AOA disappeared from the ammonia-oxidizing consortiums in all reactors. The majority of the AOA sequences from all NASs at each sampling period fell into a single AOA cluster, however, suggesting that the ammonium did not affect the AOA communities under this operational condition. This result is contradictory to the case of AOB, where the communities varied significantly among the NASs. AOB with a high affinity for ammonia were present in the reactors with lower ammonium levels, whereas AOB with a low affinity to ammonia existed in the reactors with higher ammonium levels.     

Ovine Ruminal Microbes Are Capable of Biotransforming Hexahydro-1,3,5-Trinitro-1,3,5-Triazine (RDX)

Bioremediation is of great interest in the detoxification of soil contaminated with residues from explosives such as hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX). Although there are numerous forms of in situ and ex situ bioremediation, ruminants would provide the option of an in situ bioreactor that could be transported to the site of contamination. Bovine rumen fluid has been previously shown to transform 2,4,6-trinitrotoluene (TNT), a similar compound, in 4 h. In this study, RDX incubated in whole ovine rumen fluid was nearly eliminated within 4 h. Whole ovine rumen fluid was then inoculated into five different types of media to select for archaeal and bacterial organisms capable of RDX biotransformation. Cultures containing 30 μg mL⁻¹ RDX were transferred each time the RDX concentration decreased to 5 μg mL⁻¹ or less. Time point samples were analyzed for RDX biotransformation by HPLC. The two fastest transforming enrichments were in methanogenic and low nitrogen basal media. After 21 days, DNA was extracted from all enrichments able to partially or completely transform RDX in 7 days or less. To understand microbial diversity, 16S rRNA-gene-targeted denaturing gradient gel electrophoresis (DGGE) fingerprinting was conducted. Cloning and sequencing of partial 16S rRNA fragments were performed on both low nitrogen basal and methanogenic media enrichments. Phylogenetic analysis revealed similar homologies to eight different bacterial and one archaeal genera classified under the phyla Firmicutes, Actinobacteria, and Euryarchaeota. After continuing enrichment for RDX degraders for 1 year, two consortia remained: one that transformed RDX in 4 days and one which had slowed after 2 months of transfers without RDX. DGGE comparison of the slower transforming consortium to the faster one showed identical banding patterns except one band. Homology matches to clones from the two consortia identified the same uncultured Clostridia genus in both; Sporanaerobacter acetigenes was identified only in the consortia able to completely transform RDX. This is the first study to examine the rumen as a potential bioremediation tool for soils contaminated with RDX, as well as to discover S. acetigenes in the rumen and its potential ability to metabolize this energetic compound.     

Anaerobic co-digestion of dairy manure with mulched switchgrass for improvement of the methane yield

The owners of farm-scale anaerobic digesters are relying on off-farm wastes or energy crops as a co-digestion feedstock with animal manure in order to increase their production of methane and thus revenues. Switchgrass represents an interesting feedstock for Canadian digesters owners as it is a high-yielding low-maintenance perennial crop, well adapted to northern climate. Methane potential assays in batch tests showed methane production of 19.4 ± 3.6, 28.3 ± 1.7, 37.3 ± 7.1 and 45.7 ± 0.8 L kg⁻¹, for raw manure, blended manure, manure and mulched switchgrass, manure and pretreated switchgrass, respectively. Two 6-L lab-scale anaerobic digesters were operated for 130 days in order to assess the benefit of co-digesting switchgrass with bovine manure (digester #2), at a 20% wet mass fraction, compared with a manure-only operation (digester #1) The digesters were operated at an hydraulic retention time of 37 ± 6 days and at loads of 2.4 ± 0.6 and 2.6 ± 0.6 kg total volatile solids (TVS) L⁻¹ day⁻¹ for digesters #1 (D1) and #2 (D2), respectively. The TVS degradation reached 25 and 39%, which resulted in a methane production of 1.18 ± 0.18 and 2.19 ± 0.31 L day⁻¹ for D1 and D2, respectively. The addition of 20% on a wet mass ratio of switchgrass to a manure digester increased its methane production by 86%. The co-digestion of switchgrass in a 500 m³ manure digester could yield up to 10.2 GJ day⁻¹ of purified methane or 1.1 MWh day⁻¹ of electricity.     

Dry anaerobic ammonia–methane production from chicken manure

The effect of temperature on production of ammonia during dry anaerobic fermentation of chicken manure (CM), inoculated with thermophilic methanogenic sludge, was investigated in a batch condition for 8 days. Incubation temperature did not have a significant effect on the production of ammonia. Almost complete inhibition of production of methane occurred at 55 and 65°C while quite low yields of 8.45 and 6.34 ml g⁻¹ VS (volatile solids) were observed at 35 and 45°C due to a higher accumulation of ammonia. In order to improve the production of methane during dry anaerobic digestion of CM, stripping of ammonia was performed firstly on the CM previously fermented at 65°C for 8 days: the stripping for 1 day at 85°C and pH 10 removed 85.5% of ammonia. The first-batch fermentation of methane for 75 days was conducted next, using the ammonia-stripped CM inoculated with methanogenic sludge at different ratios, (CM: thermophilic sludge) of 1:2, 1:1, and 2:1 on volume per volume basis at both 35 and 55°C. Production of methane improved and was higher than that of the control (without stripping of ammonia) but the yield of 20.4 ml g⁻¹ VS was still low, so second stripping of ammonia was conducted, which resulted in 74.7% removal of ammonia. A great improvement in the production of methane of 103.5 ml g⁻¹ VS was achieved during the second batch for 55 days.     

Performance of a pilot-scale continuous flow microbial electrolysis cell fed winery wastewater

A pilot-scale (1,000 L) continuous flow microbial electrolysis cell was constructed and tested for current generation and COD removal with winery wastewater. The reactor contained 144 electrode pairs in 24 modules. Enrichment of an exoelectrogenic biofilm required ~60 days, which is longer than typically needed for laboratory reactors. Current generation was enhanced by ensuring adequate organic volatile fatty acid content (VFA/SCOD ≥ 0.5) and by raising the wastewater temperature (31 ± 1°C). Once enriched, SCOD removal (62 ± 20%) was consistent at a hydraulic retention time of 1 day (applied voltage of 0.9 V). Current generation reached a maximum of 7.4 A/m³ by the planned end of the test (after 100 days). Gas production reached a maximum of 0.19 ± 0.04 L/L/day, although most of the product gas was converted to methane (86 ± 6%). In order to increase hydrogen recovery in future tests, better methods will be needed to isolate hydrogen gas produced at the cathode. These results show that inoculation and enrichment procedures are critical to the initial success of larger-scale systems. Acetate amendments, warmer temperatures, and pH control during startup were found to be critical for proper enrichment of exoelectrogenic biofilms and improved reactor performance.     

Microbial and decomposition efficiencies of monoculture and polyculture vermireactors,based on epigeic and anecic earthworms

Efforts have been made to evaluate the microbial and decomposition efficiency of three different vermireactors: (i) polyculture (introducing equal numbers of anecic and epigeic earthworms), (ii) monoculture (anecic) and (iii) monoculture (epigeic), designed by using earthworms of two different ecological categories i.e. anecic (Lampito mauritii Kinberg) and epigeic (Eisenia fetida (Savigny)). The microbial load of vermireactors was measured through substrate-induced respiration rate (SIR), microbial biomass N content and rate of dehydrogenase activity, while mineralization rate was evaluated measuring some chemical parameters of the substrate. Earthworms caused a decrease (as compared to initial value) in pH (41.9–80.7%), organic C (10.3–14.2%) and C:N ratio (41.9–80.7%) and an increase in total N (29.1–58.8%), NH₄-N (876.1–1485.7%), NO₃-N (29081.8–56792.6%), available P (16–19.4%) and exchangeable K (9.8–13.5%) contents of the substrate. The mineralization efficiency of the reactors was in the order: polyculture (epigeic + anecic) > monoculture (anecic) > monoculture (epigeic). The polyculture reactor showed the maximum rate of SIR (2.91 ± 0.2 mg CO₂ g⁻¹ substrate), microbial biomass N (3108.1 ± 289.2 mg N g⁻¹ substrate), and dehydrogenase activity (2453.3 ± 379.8 μg g⁻¹ substrate 24 h), while in the monoculture (epigeic) the lowest values of the same parameters were observed. It is concluded that the observed differences among reactors were due to different feeding behaviour and niche structures of epigeic and anecic earthworms. Data suggests that burrowing earthworms in waste-decomposing-system not only enhance the microbial efficiencies, but at the same time also accelerate the organic matter mineralization in a vermireactor. However, most of the previous studies were based on monoculture reactors (using epigeic earthworms) which have been recommended for waste decomposition operations, but this study revealed that polyculture vermicomposting (adding of burrowing worms with epigeic earthworms in vermicomposting system) might be beneficial for rapid decomposition of organic wastes.     

Diverse communities of active Bacteria and Archaea along oxygen gradients in coral reef sediments

Microbial communities inhabiting highly permeable sediments of Checker Reef in Kaneohe Bay, Hawaii, were characterized in relation to porewater geochemistry (O₂, NO₃ ⁻, NO₂ ⁻, NH₄ ⁺, phosphate). The physiologically active part of the population, assessed by sequencing cDNA libraries of 16S rRNA amplicons, was very diverse, with an estimated ribotype richness ≥1,380 in anoxic sediment. Quantitative analysis of community structure by rRNA-targeted fluorescence in situ hybridization (FISH) indicated that the archaeal population (9–18%) was dominated by marine Crenarchaeota (5–9%). Planctomycetales were the most abundant group in the oxic and interfacial habitat (17–19%) but were a minority (<5%) in anoxic reef sediment, where γ-Proteobacteria were numerically dominant (18%). Another 9–14% of the microbial benthos belonged to β-Proteobacteria, predominantly within the order Nitrosomonadales, many cultured representatives of which are NH₄ ⁺ oxidizers. The results of this study contribute to the phylogenetic characterization of benthic microbial communities that are important in organic matter degradation and nutrient recycling in coral reef ecosystems.     

Response of benthic microbial communities to chitin enrichment: an in situ study in the deep Arctic Ocean

In situ enrichment experiments were carried out in the Arctic deep sea (Fram Strait region) to observe the response of benthic microbial communities to chitin supply. Chambers of a benthic lander were filled in July 2004 with deep-sea sediments enriched with 1.3–7.0 g m⁻² of chitin and the effects of chitin enrichment were assessed on the microbial hydrolytic activity potential, cell number and community structure after periods of 1 week and 1 year of in situ deployment. The input of chitin had no effect on microbial abundance and chitobiase activity after 7 days of incubation, whereas community structure in enriched sediments, determined by terminal restriction fragment length polymorphism analysis of 16S rRNA genes, was different from the controls. After 1 year, microbial numbers and activity significantly increased in sediments enriched with high chitin concentrations and bacterial community structure was different from that of the other treatments. The present study suggests that microbial community structure in Arctic deep-sea sediments can react quickly to sudden large chitin inputs into the sediments and that this appears to precondition subsequent enhanced growth and enzymatic activity changes. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Stratification of Archaeal communities in shallow sediments of the Pearl River Estuary,Southern China

Microorganisms are known to play fundamental roles in the biogeochemical cycling of carbon in the coastal environments. To get to know the composition and ecological roles of the archaeal communities within the sediments of the Pearl River Estuary, Southern China, the diversity and vertical distribution of archaea in a sediment core was reported based on the 16S rRNA and mcrA genes for the first time. Quantitative PCR analysis revealed that archaea were present at 10⁶–10⁷ 16S rRNA gene copies/g (wet weight) in the sediment core, and the proportion of mcrA versus 16S rRNA gene copies varied from 11 to 45%. 16S rRNA gene libraries were constructed and analyzed for the top layer (0–6 cm), middle layer (18–24 cm), sulfate-methane transition zone (SMTZ, 32–42 cm), and bottom layer (44–50 cm) sediments. The results indicated that Miscellaneous Crenarchaeotal Group (MCG) was the main component in the sediments. The MCG archaea could be further divided into six subgroups: MCG-A, B, C, D, E, and F. On the other hand, mcrA sequences from methanogens related to the order Methanomicrobiales and ANME-2 methanotrophs were detected in all sediment layers. Taken together, our data revealed a largely unknown archaeal community in which MCG dominated within the Pearl River estuarine sediments, while methanogens and methane-oxidizing archaea putatively involving in methane metabolism, were also found in the community. This is the first important step towards elucidating the biogeochemical roles of these archaea in the Pearl River Estuary.