Intact polar lipids (IPLs) are considered biomarkers for living biomass. Their degradation in marine sediments, however, is poorly understood and complicates interpretation of their occurrence in geological samples. To investigate the turnover of IPLs, a degradation experiment with anoxic sandy sediments from the North Sea was conducted. Intact cells of two organisms that do not naturally occur in North Sea sediments were chosen as IPL sources: (i) <i>Saccharomyces cerevisiae</i>, representative for ester-bound acyl lipids that also occur in <i>Bacteria</i>, and (ii) the archaeon <i>Haloferax volcanii</i>, representative for ether-bound isoprenoid lipids. Surprisingly, IPLs with phosphoester-bound head groups showed approximately the same degradation rate as IPLs with glycosidic head groups. Furthermore, the results indicate a relatively fast degradation of <i>S. cerevisiae</i> IPLs with ester-bound moieties (analogs of bacterial membrane lipids) and no significant degradation of archaeal IPLs with ether-bound moieties. Pore water and 16S rRNA-based DGGE analysis showed only a minor influence of the IPL source on microbial metabolism and community profiles. Due to our results, the IPL-based quantification of <i>Archaea</i> and <i>Bacteria</i> should be interpreted with caution

To follow the anaerobic degradation of organic matter in tidal-flat sediments, a stimulation experiment with 13 C-labeled Spirulina biomass (130 mg per 21 g sediment slurry) was conducted over a period of 24 days. A combination of microcalorimetry to record process kinetics, chemical analyses of fermentation products and RNA-based stable-isotope probing (SIP) to follow community changes was applied. Different degradation phases could be identified by microcalorimetry: Within 2 days, heat output reached its maximum (55 lW), while primary fermentation products were formed (in lmol) as follows: acetate 440, ethanol 195, butyrate 128, propionate 112, H 2 127 and smaller amounts of valerate, propanol and butanol. Sulfate was depleted within 7 days. Thereafter, methanogenesis was observed and secondary fermentation proceeded. H 2 and alcohols disappeared completely, whereas fatty acids decreased in concentration. Three main degraders were identified by RNA-based SIP and denaturant gradient gel electrophoresis. After 12 h, two phylotypes clearly enriched in 13 C: (i) Psychrilyobacter atlanticus, a fermenter known to produce hydrogen and acetate and (ii) bacteria distantly related to Propionigenium. A Cytophaga-related bacterium was highly abundant after day 3. Sulfate reduction appeared to be performed by incompletely oxidizing species, as only sulfate-reducing bacteria related to Desulfovibrio were labeled as long as sulfate was available.

Intact polar lipids (IPLs) are considered biomarkers for living biomass. Their degradation in marine sediments, however, is poorly understood and complicates interpretation of their occurrence in geological samples. To investigate the turnover of IPLs, a degradation experiment with anoxic sandy sediments from the North Sea was conducted. Intact cells of two organisms that do not naturally occur in North Sea sediments were chosen as IPL sources: (i) <i>Saccharomyces cerevisiae</i>, representative for ester-bound acyl lipids that also occur in <i>Bacteria</i>, and (ii) the archaeon <i>Haloferax volcanii</i>, representative for ether-bound isoprenoid lipids. Surprisingly, IPLs with phosphoester-bound head groups showed approximately the same degradation rate as IPLs with glycosidic head groups. Furthermore, the results indicate a relatively fast degradation of <i>S. cerevisiae</i> IPLs with ester-bound moieties (analogs of bacterial membrane lipids) and no significant degradation of archaeal IPLs with ether-bound moieties. Pore water and 16S rRNA-based DGGE analysis showed only a minor influence of the IPL source on microbial metabolism and community profiles. Due to our results, the IPL-based quantification of <i>Archaea</i> and <i>Bacteria</i> should be interpreted with caution

Abstract. To date, North Sea tidal-flat sediments have been intensively studied down to a depth of 5 m below seafloor (mbsf). However, little is known about the biogeochemistry, microbial abundance, and activity of sulfate reducers as well as methanogens in deeper layers. In this study, two 20 m-long cores were retrieved from the tidal-flat area of Spiekeroog Island, NW Germany. The drill sites were selected with a close distance of 900 m allowing to compare two depositional settings: first, a paleo-channel filled with Holocene sediments and second, a mainly Pleistocene sedimentary succession. Analyzing these cores, we wanted to test to which degree the paleo-environmental imprint is superimposed by present processes.In general, the numbers of bacterial 16S rRNA genes are one to two orders of magnitude higher than those of Archaea. The abundances of key genes for sulfate reduction and methanogenesis (dsrA and mcrA) correspond to the sulfate and methane profiles. A co-variance of these key genes at sulfate-methane interfaces and enhanced ex situ AOM rates suggest that anaerobic oxidation of methane may occur in these layers. Microbial and biogeochemical profiles are vertically stretched relative to 5 m-deep cores from shallower sediments in the same study area, but still appear compressed compared to deep sea sediments. Our interdisciplinary analysis shows that the microbial abundances and metabolic rates are elevated in the Holocene compared to Pleistocene sediments. However, this is mainly due to present environmental Correspondence to: M. Beck (m.beck@icbm.de) conditions such as pore water flow and organic matter availability. The paleo-environmental imprint is still visible but superimposed by these processes.

A novel approach was developed to follow the successive utilization of organic carbon under anoxic conditions by microcalorimetry, chemical analyses of fermentation products and stable-isotope probing (SIP). The fermentation of (13) C-labeled glucose was monitored over 4 weeks by microcalorimetry in a stimulation experiment with tidal-flat sediments. Based on characteristic heat production phases, time points were selected for quantifying fermentation products and identifying substrate-assimilating bacteria by the isolation of intact ribosomes prior to rRNA-SIP. The preisolation of ribosomes resulted in rRNA with an excellent quality. Glucose was completely consumed within 2 days and was mainly fermented to acetate. Ethanol, formate, and hydrogen were detected intermittently. The amount of propionate that was built within the first 3 days stayed constant. Ribosome-based SIP of fully labeled and unlabeled rRNA was used for fingerprinting the glucose-degrading species and the inactive background community. The most abundant actively degrading bacterium was related to Psychromonas macrocephali (similarity 99%) as identified by DGGE and sequencing. The disappearance of Desulfovibrio-related bands in labeled rRNA after 3 days indicated that this group was active during the first degradation phase only. In summary, ribosome-based SIP in combination with microcalorimetry allows dissecting distinct phases in substrate turnover in a very sensitive manner.