Carsten J. Schubert

Linking crenarchaeal and bacterial nitrification to anammox in the Black Sea

Active expression of putative ammonia monooxygenase gene subunit A (amoA) of marine group I Crenarchaeota has been detected in the Black Sea water column. It reached its maximum, as quantified by reverse-transcription quantitative PCR, exactly at the nitrate maximum or the nitrification zone modeled in the lower oxic zone. Crenarchaeal amoA expression could explain 74.5% of the nitrite variations in the lower oxic zone. In comparison, amoA expression by γ-proteobacterial ammonia-oxidizing bacteria (AOB) showed two distinct maxima, one in the modeled nitrification zone and one in the suboxic zone. Neither the amoA expression by crenarchaea nor that by β-proteobacterial AOB was significantly elevated in this latter zone. Nitrification in the suboxic zone, most likely microaerobic in nature, was verified by 15NO2− and 15N15N production in 15NH4+ incubations with no measurable oxygen. It provided a direct local source of nitrite for anammox in the suboxic zone. Both ammonia-oxidizing crenarchaea and γ-proteobacterial AOB were important nitrifiers in the Black Sea and were likely coupled to anammox in indirect and direct manners respectively. Each process supplied about half of the nitrite required by anammox, based on 15N-incubation experiments and modeled calculations. Because anammox is a major nitrogen loss in marine suboxic waters, such nitrification–anammox coupling potentially occurring also in oceanic oxygen minimum zones would act as a short circuit connecting regenerated ammonium to direct nitrogen loss, thus reducing the presumed direct contribution from deep-sea nitrate.

Nitrogen and carbon isotopic composition of marine and terrestrial organic matter in Arctic Ocean sediments:: implications for nutrient utilization and organic matter composition

Abstract Relationships between organic carbon, total nitrogen and organic nitrogen concentrations and variations in δ 13 C org and δ 15 N org are examined in surface sediments from the eastern central Arctic Ocean and the Yermak Plateau. Removing the organic matter from samples with KOBr/KOH and determining residual as well as total N shows that there is a significant amount of bound inorganic N in the samples, which causes TOC/N total ratios to be low (4–10 depending on the organic content). TOC/N org ratios are significantly higher (8–16). This correction of organic TOC/N ratios for the presence of soil-derived bound ammonium is especially important in samples with high illite concentrations, the clay mineral mainly responsible for ammonium adsorption. The isotopic composition of the organic N fraction was estimated by determining the isotopic composition of the total and inorganic nitrogen fractions and assuming mass-balance. A strong correlation between δ 15 N org values of the sediments and the nitrate concentration of surface waters indicates different relative nitrate utilization rates of the phytoplankton in various regions of the Arctic Ocean. On the Yermak Plateau, low δ 15 N org values correspond to high nitrate concentrations, whereas in the central Arctic Ocean high δ 15 N org values are found beneath low nitrate waters. Sediment δ 13 C org values are close to −23.0‰ in the Yermak Plateau region and approximately −21.4‰ in the central Arctic Ocean. Particulate organic matter collected from meltwater ponds and ice-cores are relatively enriched in 13 C ( δ 13 C org =−15.3 to −20.6‰) most likely due to low CO 2 (aq) concentrations in these environments. A maximum terrestrial contribution of 30% of the organic matter to sediments in the central Arctic Ocean is derived, based on the carbon isotope data and various assumptions about the isotopic composition of the potential endmembers.

Aquatic Terrestrial Linkages Along a Braided-River: Riparian Arthropods Feeding on Aquatic Insects

Rivers can provide important sources of energy for riparian biota. Stable isotope analysis (δ13C, δ15N) together with linear mixing models, were used to quantify the importance of aquatic insects as a food source for a riparian arthropod assemblage inhabiting the shore of the braided Tagliamento River (NE Italy). Proportional aquatic prey contributions to riparian arthropod diets differed considerable among taxa. Carabid beetles of the genus Bembidion and Nebria picicornis fed entirely on aquatic insects. Aquatic insects made up 80% of the diet of the dominant staphylinid beetle Paederidus rubrothoracicus. The diets of the dominant lycosid spiders Arctosa cinerea and Pardosa wagleri consisted of 56 and 48% aquatic insects, respectively. In contrast, the ant Manica rubida fed mainly on terrestrial sources. The proportion of aquatic insects in the diet of lycosid spiders changed seasonally, being related to the seasonal abundance of lycosid spiders along the stream edge. The degree of spatial and seasonal aggregation of riparian arthropods at the river edge coincided with their proportional use of aquatic subsidies. The results suggest that predation by riparian arthropods is a quantitatively important process in the transfer of aquatic secondary production to the riparian food web.

Stable Carbon Isotopic Fractionations Associated with Inorganic Carbon Fixation by Anaerobic Ammonium-Oxidizing Bacteria

ABSTRACT Isotopic analyses of Candidatus “Brocadia anammoxidans,” a chemolithoautotrophic bacterium that anaerobically oxidizes ammonium (anammox), show that it strongly fractionates against 13C; i.e., lipids are depleted by up to 47‰ versus CO2. Similar results were obtained for the anammox bacterium Candidatus “Scalindua sorokinii,” which thrives in the anoxic water column of the Black Sea, suggesting that different anammox bacteria use identical carbon fixation pathways, which may be either the Calvin cycle or the acetyl coenzyme A pathway.

The Last Deglaciation Event in the Eastern Central Arctic Ocean

Oxygen isotope records of cores from the central Arctic Ocean yield evidence for a major influx of meltwater at the beginning of the last deglaciation 15.7 thousand years ago (16,650 calendar years B.C.). The almost parallel trends of the isotope records from the Arctic Ocean, the Fram Strait, and the east Greenland continental margin suggest contemporaneous variations of the Eurasian Arctic and Greenland (Laurentide) ice sheets or increased export of low-saline waters from the Arctic within the East Greenland Current during the last deglaciation. On the basis of isotope and carbon data, the modern surface- and deep-water characteristics and seasonally open-ice conditions with increased surface-water productivity were established in the central Arctic at the end of Termination lb about 7.2 thousand years ago or 6,000 calendar years B.C.).

Stable isotope stratigraphy, sedimentation rates, and salinity changes in the Latest Pleistocene to Holocene eastern central Arctic Ocean

A high-resolution study including oxygen and carbon stable isotopes as well as carbonate and total organic carbon contents, has been performed on undisturbed near-surface (0–40 cm) sediment sequences taken in the eastern Arctic Ocean during the international Arctic 91 Expedition. Based on the oxygen stable isotope records measured on Neogloboquadrina pachyderma (sin.) and AMS 14C dating, the upper 10 to 20 cm of the sediment sequences represent isotope stage 1, and the base of Termination I (15.7 ka) can be identified very well. Stage 1 sedimentation rates vary between 0.4 and >2.0 cm/kyr. In general, glacial stage 2 sedimentation rates are probably lower and vary between 0.4 and 0.7 cm/kyr. The glacial-interglacial shifts in δ18O values of N. pachyderma sin. may reach values of 1.3 to 2.5‰ indicating (1) that, in addition to the glacial-interglacial global ice-volume signal, changes in surface-water salinity have effected the isotope records and (2) that these salinity changes have varied laterally. Glacial-interglacial differences in salinity were small in the Lomonosov Ridge area (0–0.4‰) and relatively high in the Morris-Jesup-Rise area (up to 1.4‰). This implies that the supply of low-saline waters onto the Eurasian shelves and its further transport into the central Arctic Ocean via the Transpolar Drift should have continued during the last glacial and should have significantly influenced the surface water characteristics in parts of the central Arctic. On the Morris-Jesup-Rise, on the other hand, the glacial low-saline-water signal at that time was strongly reduced in comparison to the modern situation. At the glacial-interglacial stage 12 boundary, a strong meltwater signal is recorded in a sharp depletion in δ18O as well as δ13C. This central Arctic Ocean meltwater event can be correlated from the Makarov Basin through the Lomonosov Ridge and Amundsen Basin to the eastern Gakkel Ridge. The beginning of this event is AMS 14C dated at 15.7 ka, i.e., significantly older than the major decrease in the global ice-volume signal which occurs between 9 and 13.5 ka. Large amounts of freshwater/meltwater were probably supplied from the Eurasian continent due to the decay of the Barents-Sea-Ice-Sheet, causing this distinct early meltwater anomaly in the central Arctic Ocean. The extension of a well-oxygenated surface-near water mass in the Arctic Ocean and (at least seasonal) open-ice conditions and some increased bioproductivity were probably established at the end of Termination I, as indicated by the increase in δ13C to modern values as well as increased carbonate (i.e., foraminifers, coccoliths, ostracodes) and total organic carbon contents.

Nutrient regime shift in the western North Atlantic indicated by compound-specific δ15N of deep-sea gorgonian corals

Despite the importance of the nitrogen (N) cycle on marine productivity, little is known about variability in N sources and cycling in the ocean in relation to natural and anthropogenic climate change. Beyond the last few decades of scientific observation, knowledge depends largely on proxy records derived from nitrogen stable isotopes (δ15N) preserved in sediments and other bioarchives. Traditional bulk δ15N measurements, however, represent the combined influence of N source and subsequent trophic transfers, often confounding environmental interpretation. Recently, compound-specific analysis of individual amino acids (δ15N-AA) has been shown as a means to deconvolve trophic level versus N source effects on the δ15N variability of bulk organic matter. Here, we demonstrate the first use of δ15N-AA in a paleoceanographic study, through analysis of annually secreted growth rings preserved in the organic endoskeletons of deep-sea gorgonian corals. In the Northwest Atlantic off Nova Scotia, coral δ15N is correlated with increasing presence of subtropical versus subpolar slope waters over the twentieth century. By using the new δ15N-AA approach to control for variable trophic processing, we are able to interpret coral bulk δ15N values as a proxy for nitrate source and, hence, slope water source partitioning. We conclude that the persistence of the warm, nutrient-rich regime since the early 1970s is largely unique in the context of the last approximately 1,800 yr. This evidence suggests that nutrient variability in this region is coordinated with recent changes in global climate and underscores the broad potential of δ15N-AA for paleoceanographic studies of the marine N cycle.

Evidence for anaerobic oxidation of methane in sediments of a freshwater system (Lago di Cadagno).

Anaerobic oxidation of methane (AOM) has been investigated in sediments of a high alpine sulfate-rich lake. Hot spots of AOM could be identified based on geochemical and isotopic evidence. Very high fractionation of methane (α=1.031) during oxidation was observed in the uppermost sediment layers, where methane is oxidized most likely with sulfate-containing bottom waters. However, we could not exclude that other electron acceptors such as iron, or manganese might also be involved. Light carbon isotope values (δ¹³C = -10‰ vs. Vienna Pee Dee Belemnite [VPDB]) of sedimentary carbonates at 16-20 cm sediment depth are indicative of a zone where methane was oxidized and the resulting bicarbonate ions were used for carbonate precipitation. 16S rRNA gene analysis revealed the presence of sequences belonging to the marine benthic groups B, C, and D and to the recently described clade of AOM-associated archaea (AAA). Catalyzed reporter deposition-FISH analysis revealed a high abundance of Deltaproteobacteria, especially of free-living sulfate-reducing bacteria of the Desulfosarcina/Desulfococcus branch of Deltaproteobacteria in the AOM zone. Here, loose aggregations of AAA cells were found, suggesting that AAA might be responsible for oxidation of methane in Lake Cadagno sediments.

Methane sources and sinks in Lake Kivu

Unique worldwide, Lake Kivu stores enormous amounts of CH 4 and CO 2 . A recent study reported that CH 4 concentrations in the lake have increased by up to 15% in the last 30 years and that accumulation at this rate could lead to catastrophic outgassing by ∼2100. This study investigates the present-day CH 4 formation and oxidation in Lake Kivu. Analyses of 14C and 13C in CH 4 and potential carbon sources revealed that below 260 m, an unusually high ∼65% of the CH 4 originates either from reduction of geogenic CO 2 with mostly geogenic H 2 or from direct inflows of geogenic CH 4 . Aerobic CH 4 oxidation, performed by close relatives of type X CH 4 -oxidizing bacteria, is the main process preventing CH 4 from escaping to the atmosphere. Anaerobic CH 4 oxidation, carried out by CH 4 -oxidizing archaea in the SO 4 2--reducing zone, was also detected but is limited by the availability of sulfate. Changes in 14C CH4 and 13C CH4 since the 1970s suggest that the amount of CH 4 produced from degrading organic material has increased due to higher accumulation of organic matter. This, as well as the sudden onset of carbonates in the 1960s, has previously been explained by three environmental changes: (1) introduction of nonnative fish, (2) amplified subaquatic inflows following hydrological changes, and (3) increased external inputs due to the fast growing population. The resulting enhancement of primary production and organic matter sedimentation likely caused CH 4 to increase. However, given the large proportion of old CH 4 carbon, we cannot exclude an increased inflow of geogenic H 2 or CH 4 . Copyright 2011 by the American Geophysical Union.

Co‐occurrence of denitrification and nitrogen fixation in a meromictic lake, Lake Cadagno (Switzerland)

The nitrogen cycling of Lake Cadagno was investigated by using a combination of biogeochemical and molecular ecological techniques. In the upper oxic freshwater zone inorganic nitrogen concentrations were low (up to approximately 3.4 microM nitrate at the base of the oxic zone), while in the lower anoxic zone there were high concentrations of ammonium (up to 40 microM). Between these zones, a narrow zone was characterized by no measurable inorganic nitrogen, but high microbial biomass (up to 4 x 10(7) cells ml(-1)). Incubation experiments with (15)N-nitrite revealed nitrogen loss occurring in the chemocline through denitrification (approximately 3 nM N h(-1)). At the same depth, incubations experiments with (15)N(2)- and (13)C(DIC)-labelled bicarbonate, indicated substantial N(2) fixation (31.7-42.1 pM h(-1)) and inorganic carbon assimilation (40-85 nM h(-1)). Catalysed reporter deposition fluorescence in situ hybridization (CARD-FISH) and sequencing of 16S rRNA genes showed that the microbial community at the chemocline was dominated by the phototrophic green sulfur bacterium Chlorobium clathratiforme. Phylogenetic analyses of the nifH genes expressed as mRNA revealed a high diversity of N(2) fixers, with the highest expression levels right at the chemocline. The majority of N(2) fixers were related to Chlorobium tepidum/C. phaeobacteroides. By using Halogen In Situ Hybridization-Secondary Ion Mass Spectroscopy (HISH-SIMS), we could for the first time directly link Chlorobium to N(2) fixation in the environment. Moreover, our results show that N(2) fixation could partly compensate for the N loss and that both processes occur at the same locale at the same time as suggested for the ancient Ocean.