Dissolved Organic Carbon: Patterns of Utilization and Turnover in Two Small Lakes

Planktonic bacterial utilization of ¹⁴C-labelled glucose and acetate was monitored by kinetic measurements throughout an annual period in a small lake. Resulting kinetic uptake data have shown that from 1–15% of the total dissolved organic carbon pool was removed chemo-organotrophically per day during the year by bacteria capable of metabolizing these substrates. The kinetic uptake of nine selected organic compounds was measured in a second small lake during summer thermal stratification. Metabolism of glucose, acetate, and glycollate was preferred. Respiration rates of the nine compounds varied generally between 20–60% of the total uptake. The uptake of these compounds accounted for removal of 3–8% of the total dissolved organic carbon pool per day.     

Production and Utilization of Dissolved Organic Carbon During in situ Phytoplankton Photosynthesis Measurements

Rates of phytoplankton photosynthesis, extracellular release of dissolved organic carbon, and production or utilization of dissolved organic carbon during in situ incubation were measured in a soft-water Vermont lake during summer thermal stratification. Phytoplankton photosynthesis rates were frequently in the range of 300–600 mg C m⁻² of lake surface day⁻¹; extracellular release of previously fixed organic carbon was generally in the range of 20–75% of the carbon incorporated into cell biomass, as determined by gas-phase radio-analysis. Rates of increase or decrease in total dissolved organic carbon occurring in light and dark incubated phytoplankton samples, during brief (4 hour) in situ measurements, indicate that a significant fraction of the total dissolved organic carbon „pool”︁ is probably labile and rapidly being cycled.     

Colloidal and Dissolved Organic Matter Excreted by a Mixotrophic Flagellate during Bacterivory and Autotrophy

Excretion of dissolved and colloidal organic carbon by a mixotrophic flagellate, the chrysophyte Poterioochromonas malhamensis, was studied. Flagellates were incubated either with ¹⁴C-labeled bacteria or with inorganic ¹⁴C, in order to compare organic exudates originating from primary production with exudates originating from ingested bacteria. Colloids of >0.02 μm constituted a larger fraction of the exudates originating from ingested bacteria, compared with exudates derived from primary production. Flagellate feeding on bacteria specifically labeled in different cell components was compared. Cell wall components gave rise to less colloidal organic carbon than did other cell constituents. To investigate the degradability of flagellate ¹⁴C-exudates, they were added to lake water and mineralization to ¹⁴CO₂ was monitored. Bacterially derived exudates were more recalcitrant than exudates originating from photosynthesis. The results support the hypothesis that bacterial utilization of labile organic compounds, followed by flagellate bacterivory and exudation, results in a transformation of labile organic matter into more recalcitrant forms.     

Evidence of in situ uptake and incorporation of bicarbonate and amino acids by a hydrothermal vent mussel

The hydrothermal vent mussel Bathymodiolus sp. is demonstrated to incorporate inorganic CO₂ from sea water. After ≈24 h incubation with H¹⁴CO⁻₂ the major part of the radioactivity is incorporated into macromolecules mostly in proteins but also in a notable lipidic fraction. 77 to 98% of this radioactivity is found in the gill and autoradiographs show that CO₂ fixation is only observed in cells containing high concentrations of bacteria. The results endorse the hypothesis that the associated bacteria might provide a nutritional source for the mussel.The mussel is also able to absorb and incorporate dissolved amino acids. Heterotrophic processes involving dissolved organic matter may interfere with the autotrophic pathways. Beside its capability of feeding on particulate material, the mussel may be thus able to live on reduced carbon and nitrogen compounds synthesized by its associated bacteria as well as on dissolved organic compounds present in sea water. The effective participation of the different processes is probably related to the ecological conditions experienced by the mussel in vent areas.     

Colloidal organic carbon and trace metal (Cd, Fe, and Zn) releases by diatom exudation and copepod grazing

Colloidal macromolecular organic compounds are important intermediaries between solution and particle phases and play a critical role in the biogeochemistry of trace metals and organic carbon. The releases of colloidal organic carbon and trace metals (Cd, Fe, and Zn) mediated by copepod grazing and decomposition, and direct diatom exudation, were examined using a radiotracer approach. The colloidal phase was operationally defined in this study as the size fraction between 5 kDa and 0.2 μm and the dissolved phase as the ≤0.2 μm filter passing phase. About 13-60% of dissolved carbon exuded by the diatom Thalassiosira pseudonana was partitioned into the colloidal phase, and this fraction increased considerably as the diatom cells grew older. A lower fraction of dissolved ¹⁴C (12-23%) excreted by the copepods Acartia erythraea was detected in the colloidal phase compared to carcass (13-35%) and feces decomposition (21-34%). In contrast to carbon, a lower fraction of regenerated dissolved Cd (1-11%) and Zn (0-20%) from copepods and diatoms was consistently detected in the colloidal phases. Copepod excretion and carcass decomposition resulted in more colloidal Fe (51-91%) than diatom exudation (46-62% for Thalassiosira weissflogii, and 3-33% for T. pseudonana) and copepod feces decomposition (16-30%). Copepod (Calanus sinicus) grazing reduced the colloidal fraction of dissolved ¹⁴C, although a higher concentration of the diatom's (T. weissflogii) carbon was regenerated into the dissolved phase. The grazing of these copepods did not have any influence on the colloidal metal partitioning. The release of trace metals and carbon was enhanced by a higher density of copepod's grazing. Thus, different biological processes (grazing, excretion, exudation, and decomposition) may contribute differently to the production and dynamics of colloidal carbon and metals in planktonic systems.     

Embryonic nutrition,growth and energetics inZoarces viviparus L. as indication of a maternal-fetal trophic relationship

Summary The embryos ofZoarces viviparus (L.) show linear growth during their intraovarian development. In early gestation before hatching, the embryos take up very small amounts of low molecular tracer compounds such as glucose, glycine or taurine. Later in gestation (two months after hatching), the embryos accumulate substantial amounts of the tracer compounds. The uptake rates of the tracer compounds in vitro are correlated with ambient concentrations of unlabelled compounds within the natural concentration range of the ovarian fluid. The highest uptake rates are found for glucose and the lowest for taurine. Release of¹⁴CO₂ and dissolved organic carbon (DO¹⁴C) from assimilated tracers in the embryos is low. Oxygen uptake and body weight of the embryos appear to be linearly correlated, and the average oxygen uptake is 4.20 (SD 0.73) mol O₂ g^–1 h^–1 WW at 11°C. The contribution of glucose respiration to total aerobic respiration is 13.9%. A growth to respiration ratio of 0.91 indicates a relatively high efficiency for converting food to growth.Symbols and abbreviations DW dry weight - WW wet weight - DO ¹⁴ C dissolved organic carbon (¹⁴C-labelled) - t _1/2 half life time - turnover time (replacement time)     

Physiology of diatom Skeletonema costatum (Grev.) Cleve photosynthetic extracellular release: evidence for a novel coupling between marine bacteria and phytoplankton

The photosynthesis of cellular materials by phytoplankton isaccompanied by release of organic molecules from the algal cellsinto the water. The patterns of carbon fixation in particulateand dissolved pools were investigated in Skeletonema costatumcultured under 12 h light/12 h dark cycles. The short-term production(1–15 min) of particulate organic carbon (POC) and extracellularorganic carbon (EOC) compounds was studied by measuring theuptake of ¹⁴C-labelled sodium bicarbonate and its subsequentincorporation and release into organic compounds. Slightly modifiedtraditional ¹⁴C radiotracer protocols were used, including separationby electrophoresis and thin-layer chromatography and detectionby autoradiography. Results indicated that there was a distinctdifference between radiolabelled compounds in the POC and EOCpools. Several metabolites found in the EOC pool were not presentin the POC pool, indicating the active release of these productsfrom the cells into the ambient water during short-term incubations,and indicating that inorganic carbon fixation pathways in marineautotrophs might be partly extracellular.     

Sorption of hydrophobic organic contaminants and trace metals on phytoplankton and implications for toxicity assessment

The partitioning of trace metals and hydrophobic organic contaminants to phytoplankton determines their toxicity as well as their fate and transport in aquatic ecosystems. Accurate impact assessments, therefore, depend on a good understanding of the factors regulating the sorption of these compounds to biotic particles. The accumulation of chlorinated organic compounds in phytoplankton is generally considered as being due solely to physical sorption, described by reversible equilibrium models based on Langmuir or Freundlich isotherms. On the other hand, the uptake of trace metals is a two phase process: a fast sorption component viewed as an ionexchange or a covalent bonding process with cell surface ligands, followed by an intracellular transport phase that is dependent on cellular metabolic activity. The uptake of inorganic and hydrophobic organic pollutants and their bioaccumulation are influenced in a complex manner by duration of exposure and cell density, by environmental factors such as pH, the concentration of cations and of dissolved and colloidal organic matter, as well as by phytoplankton physiological condition. High concentrations of H⁺, Ca²⁺, and Mg²⁺ ions will reduce trace metal sorption by directly competing for uptake sites on the cell's surface, whereas the presence of dissolved organic carbon such as natural and synthetic chelators and phytoplankton exudates will reduce the bioavailability of both trace metals and hydrophobic organic contaminants. Thus, the impact of toxic contaminants on phytoplankton may be determined as much by the factors influencing uptake and partitioning as by the potency of the toxicants and interspecies differences in sensitivity. Recommendations for improving toxicity assessments are presented.     

ORGANIC EXCRETION BY DUNALIELLA TERTIOLECTA1

Excretion of C¹⁴-labeled organic material by Dunaliella tertiolecta was observed under a variety of conditions. In healthy cultures, percent excretion tints related to cell density bill independent of light and preconditioning of the medium. Exaction was lower in nutrient-limited culture than in an enriched medium, and in both cases was minimal during logarithmic growth. Increases in dissolved carbohydrate during stationary phase represented an accumulation of high molecular weight compounds. The general composition of labeled excretory products was similar in both actively growing and stationary cultures. The data suggest that excretion may involve different processes at high and low cell densities.     

Substrate Use of Pseudovibrio sp. Growing in Ultra-Oligotrophic Seawater

Marine planktonic bacteria often live in habitats with extremely low concentrations of dissolved organic matter (DOM). To study the use of trace amounts of DOM by the facultatively oligotrophic Pseudovibrio sp. FO-BEG1, we investigated the composition of artificial and natural seawater before and after growth. We determined the concentrations of dissolved organic carbon (DOC), total dissolved nitrogen (TDN), free and hydrolysable amino acids, and the molecular composition of DOM by electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry (ESI FT-ICR-MS). The DOC concentration of the artificial seawater we used for cultivation was 4.4 μmol C L^-1, which was eight times lower compared to the natural oligotrophic seawater we used for parallel experiments (36 μmol C L ^-1). During the three-week duration of the experiment, cell numbers increased from 40 cells mL^-1 to 2x10⁴ cells mL ^-1 in artificial and to 3x10⁵ cells mL ^-1 in natural seawater. No nitrogen fixation and minor CO₂ fixation (< 1% of cellular carbon) was observed. Our data show that in both media, amino acids were not the main substrate for growth. Instead, FT-ICR-MS analysis revealed usage of a variety of different dissolved organic molecules, belonging to a wide range of chemical compound groups, also containing nitrogen. The present study shows that marine heterotrophic bacteria are able to proliferate with even lower DOC concentrations than available in natural ultra-oligotrophic seawater, using unexpected organic compounds to fuel their energy, carbon and nitrogen requirements.     

The impact of dissolved amino acids on protein and cellulose degradation in stream waters

Dissolved amino acids represent a significant carbon and nitrogen source for microorganisms in stream water environments, and may be utilized in preference to protein bound nitrogen in decomposing organic matter. Consequently, in streams with sufficiently high concentrations of dissolved amino acids, depolymerisation of nitrogen compounds may become delayed. This possibility was investigated in stream water samples incubated with ¹⁴C-labelled albumin and cellulose in presence of the indigenous microorganisms at different concentrations of dissolved amino acids.The experiments demonstrated at an average between 15 and 25% lower degradation of the compounds during a ten days incubation at an initial amino acid concentration of 1 mg l^–1. The reduction was most clearly expressed in a nutrient-poor stream water compared with a nutrient-rich. The kinetics of the degradation were most appropriately described by a first-order model, that is, the rate of transformation of the macromolecules was independent of the total number of bacteria in the water. The mechanism suggested for the retardation of macromolecules is a superproportional utilization of the dissolved amino acids at high concentrations, a phenomenon that can cause accumulation of slowly decomposing macromolecules in sediments affected by residual wastewater.     

ORGANIC COMPOUNDS RELEASED FROM A NATURAL POPULATION OF EPIBENTHIC BLUEGREEN ALGAE1

Extracellular release of dissolved organic compounds by the bluegreen algal community of a brackish marsh was studied using ¹⁴C techniques. Mannitol and trehalose were identified as the most commonly released compounds. The proportions of these two extracellular compounds varied in response to light intensity and the water potential of the environment. The presence of mannitol, in particular, suggests that excretion of organic compounds in natural situations is a function of osmotic adjustment.     

Synthesis of organic compounds from carbon monoxide and water by UV photolysis

The photolysis of water vapor with carbon monoxide at 1849 Å yields alcohols, aldehydes and organic acids, with an overall quantum yield of 3.3×10^–2. This rather high quantum yield could have led to a contribution of 10¹¹ organic molecules cm^–2 sec^–1 to the pool of organic material on the primitive Earth. The reactions are initiated by the photolysis of water molecules and the resulting hydrogen atoms reduce the carbon monoxide to a variety of one and two carbon compounds. The organic molecules are dissolved in water and thus escape destruction by photolysis. Photolysis of water vapor with carbon dioxide did not yield organic compounds under these conditions.     

Protein synthesis: a measure of growth for lake plankton

Problems in the interpretation of ¹⁴C-primary production estimates have resulted from our belief that all carbon compounds are created equal and few attempts have been made to include nighttime ¹⁴C-losses. The energetic costs of protein synthesis are high and once formed protein is not simply used as another form of energy but is conserved along food chains. A portion of the carbohydrate fraction is used to drive night protein synthesis and some, the so called colloidal dissolved organic carbon which represents a portion of the extracellular production, simply aggregates and settles out.Areal productivity measured over 24 hours was monitored in Lake Ontario and compared with losses through sedimentation and zooplankton grazing. The balance was related to NET changes in particulate organic carbon (POC) concentration. Prior to stratification, sedimentation accounted for most of the carbon losses from the trophogenic zone but during stratification, zooplankton grazing was coupled with plankton growth (determined as protein synthesis per unit particulate protein). This observation, supported by very low phosphorus sedimentation rates suggests that phosphorus is effectively recycled within the epilimnion.     

Incorporation of organic compounds into cell protein by lithotrophic,ammonia-oxidizing bacteria

Incorporation of organic compounds into cell protein by the obligate chemolithotrophs Nitrosomonas spec., Nitrosococcus oceanus, Nitrosococcus mobilis, Nitrosovibrio tenuis, Nitrosolobus spec., and Nitrosopira spec. was studied. In the presence of ammonia as energy source organic substrates were supplied. Distribution of ¹⁴C into cell amino acids arising from ¹⁴C-labelled glucose, Na-pyruvate, and Na-acetate was investigated. While carbon from glucose was distributed unrestricted, carbon from pyruvate preferably entered into the amino acids of the pyruvate and glutamate family and from acetate mainly into leucine and the glutamate family. Among the strains examined, slight differences were observed, but all should be included under group A of the scheme of Smith and Hoare (1977).     

Changes in dissolved lignin-derived phenols, neutral sugars, uronic acids, and amino sugars with depth in forested Haplic Arenosols and Rendzic Leptosols

A major part of the dissolved organic matter produced in the organic layers of forest ecosystems and leached into the mineral soil is retained by the upper subsoil horizons. The retention is selective and thus dissolved organic matter in the subsoils has different composition than dissolved organic matter leached from the forest floor. Here we report on changes in the composition of dissolved organic matter with soil depth based on C-to-N ratios, XAD-8 fractionation, wet-chemical analyses (lignin-derived CuO oxidation products, hydrolysable sugars and amino sugars) and liquid-state ¹³C nuclear magnetic resonance (NMR) spectroscopy. Dissolved organic matter was sampled directly beneath the forest floor using tension-free lysimeters and at 90cm depth by suction cups in Haplic Arenosols under Scots pine (Pinus sylvestris L.) and Rendzic Leptosols under European beech (Fagus sylvatica L.) forest. At both sites, the concentrations of dissolved organic carbon (DOC) decreased but not as strongly as reported for deeply weathered soils. The decrease in DOC was accompanied by strong changes in the composition of dissolved organic matter. The proportion of the XAD-8-adsorbable (hydrophobic) fraction, carboxyl and aromatic C, and the concentrations of lignin-derived phenols decreased whereas the concentrations of sugars, amino sugars, and nitrogen remained either constant or increased. A general feature of the compositional changes within the tested compound classes was that the ratios of neutral to acidic compounds increased with depth. These results indicate that during the transport of dissolved organic matter through the soils, oxidatively degraded lignin-derived compounds were preferentially retained while potentially labile material high in nitrogen and carbohydrates tended to remain dissolved. Despite the studied soils' small capacity to sorb organic matter, the preferential retention of potentially refractory and acidic compounds suggests sorption by the mineral soil matrix rather than biodegradation to govern the retention of dissolved organic matter even in soils with a low sorption capacity.     

Effects of Soil Water Content on Biodegradation of Phenanthrene in a Mixture of Organic Contaminants

Phenanthrene biodegradation was investigated at different soil water contents [0.11, 0.22, 0.33, 0.44 g H₂O (g soil)^?1] to determine the effects of water availability on biodegradation rate. A subsurface horizon of Kennebec silty loam soil was used in this study. [9-¹⁴C] phenanthrene was dissolved in a mixture of organic contaminants that consisted of 76% decane, 6% ρ-xylene, 6% phenanthrene, 6% pristane, and 6% naphthalene, and then added to the soil. The highest rate of mineralization, in which 0.23% of the [9-¹⁴C] phenanthrene degraded to ¹⁴CO₂ after 66 days of incubation, was observed at the soil water content of 0.44 g H₂O/g dry soil. Most of the ¹⁴C remained in the soil as the parent compound or as nonextractable compounds by acetonitrile at the highest water content. Concentrations of nonextractable compounds increased with water content, but residual extractable phenanthrene decreased significantly with increasing water content, which presumably indicates that bio-transformation occurred. The mineralization analysis of radiolabeled 9th carbon in phenanthrene underestimated phenanthrene biodegradation. The strong adsorption and low solubility of phenanthrene contributed to the low mineralization of phenanthrene 9th carbon. The other components were subject to higher biological and abiotic dissipation processes with increasing soil water content.     

Inorganic carbon fixation by chemosynthetic ectosymbionts and nutritional transfers to the hydrothermal vent host-shrimp Rimicaris exoculata

The shrimp Rimicaris exoculata dominates several hydrothermal vent ecosystems of the Mid-Atlantic Ridge and is thought to be a primary consumer harbouring a chemoautotrophic bacterial community in its gill chamber. The aim of the present study was to test current hypotheses concerning the epibiont''s chemoautotrophy, and the mutualistic character of this association. In-vivo experiments were carried out in a pressurised aquarium with isotope-labelled inorganic carbon (NaH¹³CO₃ and NaH¹⁴CO₃) in the presence of two different electron donors (Na₂S₂O₃ and Fe²⁺) and with radiolabelled organic compounds (¹⁴C-acetate and ³H-lysine) chosen as potential bacterial substrates and/or metabolic by-products in experiments mimicking transfer of small biomolecules from epibionts to host. The bacterial epibionts were found to assimilate inorganic carbon by chemoautotrophy, but many of them (thick filaments of epsilonproteobacteria) appeared versatile and able to switch between electron donors, including organic compounds (heterotrophic acetate and lysine uptake). At least some of them (thin filamentous gammaproteobacteria) also seem capable of internal energy storage that could supply chemosynthetic metabolism for hours under conditions of electron donor deprivation. As direct nutritional transfer from bacteria to host was detected, the association appears as true mutualism. Import of soluble bacterial products occurs by permeation across the gill chamber integument, rather than via the digestive tract. This first demonstration of such capabilities in a decapod crustacean supports the previously discarded hypothesis of transtegumental absorption of dissolved organic matter or carbon as a common nutritional pathway.     

Phytoplankton: a significant trophic source for soft corals?

Histological autoradiographs and biochemical analyses show that ¹⁴C-labelled microalgae (diatoms, chlorophytes and dinoflagellates) are used by the soft coral Dendronephthya sp. Digestion of the algae took place at the point of exit of the pharynx into the coelenteron. Ingestion and assimilation of the labelled algae depended on incubation time, cell density, and to a lesser extent on species-specificity. ¹⁴C incorporation into polysaccharides, proteins, lipids and compounds of low molecular weight was analysed. The ¹⁴C-labelling patterns of the four classes of substances varied depending on incubation time and cell density. ¹⁴C incorporation was highest into lipids and proteins. Dissolved labelled algal metabolites, released during incubation into the medium, contributed between 4% and 25% to the total ¹⁴C activity incorporated. The incorporated microalgae contributed a maximum of 26% (average of the four species studied) to the daily organic carbon demand, as calculated from assimilation rates at natural eucaryotic phytoplankton densities and a 1 h incubation period. The calculated contribution to the daily organic carbon demand decreased after prolonged incubation periods to about 5% after 3 h and to 1–3% after 9 h. Thus the main energetic demand of Dendronephthya sp. has to be complemented by other components of the seston. Received in revised form: 17 April 2001 Electronic Publication     

Organic carbon fluxes by precipitation,throughfall and stemflow in an olive orchard in Southern Spain

In forests and grasslands, canopy-derived carbon fluxes have been shown to be an important part of the carbon cycle, yet very few data are available for permanent agricultural crops. Concentration of total (TOC), dissolved (DOC) and particulate organic carbon (POC) was measured during an entire hydrological year in direct rainfall throughfall and stemflow in a mature olive orchard. Throughfall accounted for 68% of incoming rainfall, but TOC concentration was on average 14 times higher than in rainfall (7.63 vs. 106.12 mg/L). Stemflow on the other hand represented only a small fraction of gross precipitation, on average less than 1%, while its TOC concentration was on average 10-fold higher than in rainfall (74.13 mg/L). Dynamics of OC enrichment were mainly driven by precipitation patterns, as well as plant phenology, whereas a dramatic increase happened during flowering. Stemflow and throughfall were proven to be important sources of organic carbon supplying 13.5 g C/m²/year beneath the canopy of each tree, while 2.41 g C/m²/year reached the soil through gross precipitation. This large fraction of carbon is a mean of recirculating important carbon compounds that may help mobilize soil nutrients and maintain water holding capacity in the circumscribed area below olive canopies.