Ammonium excretion and respiration rate of tropical copepods and euphausiids measured during METEOR cruise M93, M97 and Maria S. Merian cruise MSM22

Respiration and ammonium excretion rates at different oxygen partial pressure were measured for calanoid copepods and euphausiids from the Eastern Tropical South Pacific and the Eastern Tropical North Atlantic. All specimens used for experiments were caught in the upper 400 m of the water column and only animals appearing unharmed and fit were used for experiments. Specimens were sorted, identified and transferred into aquaria with filtered, well-oxygenated seawater immediately after the catch and maintained for 1 to 13 hours prior to physiological experiments at the respective experimental temperature. Maintenance and physiological experiments were conducted in darkness in temperature-controlled incubators at 11, 13 or 23 degree C (±1). Before and during experiments, animals were not fed. Respiration and ammonium excretion rate measurements (both in µmol h-1 gDW-1) at varying oxygen concentrations were conducted in 12 to 60 mL gas-tight glass bottles. These were equipped with oxygen microsensors (ø 3 mm, PreSens Precision Sensing GmbH, Regensburg, Germany) attached to the inner wall of the bottles to monitor oxygen concentrations non-invasively. Read-out of oxygen concentrations was conducted using multi-channel fiber optic oxygen transmitters (Oxy-4 and Oxy-10 mini, PreSens Precision Sensing GmbH, Regensburg, Germany) that were connected via optical fibers to the outside of the bottles directly above the oxygen microsensor spots. Measurements were started at pre-adjusted oxygen and carbon dioxide levels. For this, seawater stocks with adjusted pO2 and pCO2 were prepared by equilibrating 3 to 4 L of filtered (0.2 µm filter Whatman GFF filter) and UV - sterilized (Aqua Cristal UV C 5 Watt, JBL GmbH & Co. KG, Neuhofen, Germany) water with premixed gases (certified gas mixtures from Air Liquide) for 4 hours at the respective experimental temperature. pCO2 levels were chosen to mimic the environmental pCO2 in the ETSP OMZ or the ETNA OMZ. Experimental runs were conducted with 11 to 15 trial incubations (1 or 2 animals per incubation bottle and three different treatment levels) and three animal-free control incubations (one per experimental treatment). During each run, experimental treatments comprised 100% air saturation as well as one reduced air saturation level with and without CO2. Oxygen concentrations in the incubation bottles were recorded every 5 min using the fiber-optic microsensor system and data recording for respiration rate determination was started immediately after all animals were transferred. Respiration rates were calculated from the slope of oxygen decrease over selected time intervals. Chosen time intervals were 20 to 105 min long. No respiration rate was calculated for the first 20 to 60 min after animal transfer to avoid the impact of enhanced activity of the animal or changes in the bottle water temperature during initial handling on the respiration rates and oxygen readings. Respiration rates were obtained over a maximum of 16 hours incubation time and slopes were linear at normoxia to mild hypoxia. Respiration rates in animal-free control bottles were used to correct for microbial activity. These rates were < 2% of animal respiration rates at normoxia. Samples for the measurement of ammonium concentrations were taken after 2 to 10 hours incubation time. Ammonium concentration was determined fluorimetrically (Holmes et al., 1999). Ammonium excretion was calculated as the concentration difference between incubation and animal-free control bottles. Some specimens died during the respiration and excretion rate measurements, as indicated by a cessation of respiration. No excretion rate measurements were conducted in this case, but the oxygen level at which the animal died was noted.

Seawater carbonate chemistry and microbial polysaccharide degradation during experiments with phytoplankton Emiliania huxleyi (strain PML B92/11) and natural bacteria community, 2010

With the accumulation of anthropogenic carbon dioxide (CO2), a proceeding decline in seawater pH has been induced that is referred to as ocean acidification. The ocean's capacity for CO2 storage is strongly affected by biological processes, whose feedback potential is difficult to evaluate. The main source of CO2 in the ocean is the decomposition and subsequent respiration of organic molecules by heterotrophic bacteria. However, very little is known about potential effects of ocean acidification on bacterial degradation activity. This study reveals that the degradation of polysaccharides, a major component of marine organic matter, by bacterial extracellular enzymes was significantly accelerated during experimental simulation of ocean acidification. Results were obtained from pH perturbation experiments, where rates of extracellular alpha- and beta-glucosidase were measured and the loss of neutral and acidic sugars from phytoplankton-derived polysaccharides was determined. Our study suggests that a faster bacterial turnover of polysaccharides at lowered ocean pH has the potential to reduce carbon export and to enhance the respiratory CO2 production in the future ocean.

Chlorophyll content and temperature-dependent net photosynthesis and respiration rate in the lichen Cetraria aculeata

We studied polar and temperate samples of the lichen Cetraria aculeata to investigate whether genetical differences between photobionts are correlated with physiological properties of the lichen holobiont. Net photosynthesis and dark respiration (DR) at different temperatures (from 0 to 30 °C) and photon flux densities (from 0 to 1,200 ?mol/m**2/s) were studied for four populations of Cetraria aculeata. Samples were collected from maritime Antarctica, Svalbard, Germany and Spain, representing different climatic situations. Sequencing of the photobiont showed that the investigated samples fall in the polar and temperate clade described in Fernández-Mendoza et al. (2011, doi:10.1111/j.1365-294X.2010.04993.x). Lichens with photobionts from these clades differ in their temperature optimum for photosynthesis, maximal net photosynthesis, maximal DR and chlorophyll content. Maximal net photosynthesis was much lower in Antarctica and Svalbard than in Germany and Spain. The difference was smaller when rates were expressed by chlorophyll content. The same is true for the temperature optima of polar (11 °C) and temperate (15 and 17 °C) lichens. Our results indicate that lichen mycobionts may adapt or acclimate to local environmental conditions either by selecting algae from regional pools or by regulating algal cell numbers (chlorophyll content) within the thallus.

Fatty acids and intact polar lipids of laminated microbial sediments from St. Peter Ording, German Wadden Sea

Hidden for the untrained eye through a thin layer of sand, laminated microbial sediments occur in supratidal beaches along the North Sea coast. The inhabiting microbial communities organize themselves in response to vertical gradients of light, oxygen or sulfur compounds. We performed a fine-scale investigation on the vertical zonation of the microbial communities using a lipid biomarker approach, and assessed the biogeochemical processes using a combination of microsensor measurements and a 13C-labeling experiment. Lipid biomarker fingerprinting showed the overarching importance of cyanobacteria and diatoms in these systems, and heterocyst glycolipids revealed the presence of diazotrophic cyanobacteria even in 9 to 20 mm depth. High abundance of ornithine lipids (OL) throughout the system may derive from sulfate reducing bacteria, while a characteristic OL profile between 5 and 8 mm may indicate presence of purple non-sulfur bacteria. The fate of 13C-labeled bicarbonate was followed by experimentally investigating the uptake into microbial lipids, revealing an overarching importance of cyanobacteria for carbon fixation. However, in deeper layers, uptake into purple sulfur bacteria was evident, and a close microbial coupling could be shown by uptake of label into lipids of sulfate reducing bacteria in the deepest layer. Microsensor measurements in sediment cores collected at a later time point revealed the same general pattern as the biomarker analysis and the labeling experiments. Oxygen and pH-microsensor profiles showed active photosynthesis in the top layer. The sulfide that diffuses from deeper down and decreases just below the layer of active oxygenic photosynthesis indicates the presence of sulfur bacteria, like anoxygenic phototrophs that use sulfide instead of water for photosynthesis.

Studies of the microbial diversity on analyzed samples from from Peru Margin, Equatorial Pacific, Hydrate Ridge, and Juan de Fuca Ridge

Marine sediments harbor an enormous quantity of microorganisms, including a multitude of novel species. The habitable zone of the marine sediment column begins at the sediment-water interface and probably extends to depths of several thousands of meters. Studies of the microbial diversity in this ecosystem have mostly relied on molecular biological techniques. We used a complementary method - analysis of intact polar membrane lipids - to characterize the in-situ microbial community in sediments covering a wide range of environmental conditions from Peru Margin, Equatorial Pacific, Hydrate Ridge, and Juan de Fuca Ridge. Bacterial and eukaryotic phospholipids were only detected in surface sediments from the Peru Margin. In contrast, deeply buried sediments, independent of their geographic location, were dominated by archaeal diether and tetraether lipids with various polar head groups and core lipids. We compared ring distributions of archaeal tetraether lipids derived from polar glycosidic precursors with those that are present as core lipids. The distributions of these related compound pools were distinct, suggestive of different archaeal sources, i.e., the polar compounds derive from sedimentary communities and the core lipids are fossil remnants from planktonic communities with possible admixtures of decayed sedimentary archaea. This in-situ production of distinct archaeal lipid populations potentially affects applications of the TEX86 paleotemperature proxy as demonstrated by offsets in reconstructed temperatures between both pools. We evaluated how varying cell and lipid stabilities will influence the sedimentary pool by using a box-model. The results are consistent with (i) a requirement of continuous inputs of freshly synthesized lipids in subsurface sediments for explaining the observed distribution of intact polar lipids, and (ii) decreasing lipid inputs with increasing burial depth.

Dissolved lithium and silicon through time in microbial dissolution experiments on ODP Leg 209 peridotite

Using peridotite drilled during Ocean Drilling Program Leg 209, a series of enrichment cultures were initiated on board the ship to stimulate microbially enhanced dissolution of olivine. Dissolution was estimated by measured changes in dissolved Li and Si in the media through time (up to 709 days). The results suggest that there was no significant difference between the amounts of dissolved Li and Si in most of the inoculated microbial cultures compared to the control cultures. Alternative explanations for this are that 1. No microbes are living in the culture tubes that can affect the dissolution rates of olivine, 2. The control cultures have microbes effecting the dissolution of olivine as well as the inoculated cultures, 3. Not enough time has passed to build up a large enough microbial population to effect the dissolution of the olivine in the culture tubes, 4. Microbes act to suppress dissolution of olivine instead of enhancing dissolution, and 5. Abiotic dissolution overshadows microbially enhanced dissolution. Further work is required to test these alternatives.