The Lability of Riverine Particulate Organic Carbon Delivered to the Ocean

The large annual flux of organic carbon from land to the ocean largely disappears in the sea. Reasons for this loss are not clear. Under this award, the PI will examine abiotic and biotic aspects of the lability to loss of riverine particulate organic matter. Work will focus on the Mississippi, as it demonstrates the clearest loss of particulate organic material of any North American river upon deposition in the ocean. Photochemical experiments will follow up on initial work showing significant dissolution of particulate organic matter subjected to solar levels of radiation. Interactions with metal cycling will be studied, as will a variety of other physicochemical variables. Biochemical lability will be explored via enzyme hydrolyses and bacterial incubations. Seasonal patterns will be examined, as will be the influence of deposition in freshwater environments

Organic Coatings on Sedimentary Mineral Grains

A pore protection hypothesis will be tested, provide a mechanistic basis for the observed patterns of organic carbon. Relationships between organic matter and mineral surfaces will be developed by: (1) Assessing the extent of organic matter coverage on mineral grains. Is organic carbon localized in small areas or is it widely dispersed? Wide dispersal implies that major fraction must reside in small pores. This question will be tested by completing development of a promising new method based on the of gas adsorption onto mineral surfaces. Completion work will focus on the nature of naked mineral surfaces, identifying variations that derive from mineralogy, microtopography, and source area; (2) Determining if natural organic matter resides preferentially inside small mesopores, by careful examination of the effects of removal of organic coatings on the size distribution of pores. This test will also involve the reverse process of adding artificial organic coatings to uncoated mineral grains, providing insight into the localization and mechanism of organic matter sorption. (3) Testing if these small mesopores on mineral grains can indeed exclude hydrolytic enzymes, by performing a series of size- exclusion and adsorption experiments. These approaches will be carried out on a wide variety of sediments from around the world. In addition to testing the pore hypothesis, results from this project will have important implications for carbon cycling, mineral-water interactions, and pollutant cycling

Photodissolution of Sedimentary Organic Matter

The field of marine photochemistry has previously focused on dissolved organic matter and trace metals. However, recent studies have shown that sediment suspensions in the ocean are also affected by sunlight and have the potential to dissolve most of their particulate organic carbon to the dissolved organic phase. A researcher from the University of Maine will determine the importance of photodissolution in the coastal Louisiana area, where riverine particulates are quickly deposited in shallow waters. Optical properties of the particulates will be examined to assess the photon fluxes and to determine the quantum yields of this photodissolution reaction. To determine the importance of this reaction, modeling of the turbid nearshore regimes, field-tests of the optical results, and developments of the markers from previous photodissolutions will be carried out. In addition to the optical properties of the particulates, the fate of the photodissolved organic matter will be assessed, addressing both photo-oxidation and biological mineralization reactions. In terms of the broader impacts, this research will provide new insights into the impact of photodegradation of particulate organic matter on the cycling of nutrients in the deltaic region of the Mississippi River. Outreach efforts based on the research activities at the K-12 and community level will be carried out. One graduate student and one undergraduate student will be supported and trained as part of the study

Food Substrates and Digestive Capabilitites of Marine Deposit Feeders

Deposit feeders play several important roles in determining whether organic material is demineralized or buried. These animals function to make surfaces available for microbial growth and move particles both horizontally and vertically within the seabed at a pace that far exceeds sedimentation. The central problem in understanding deposit feeders is to identify the materials that they utilize and to determine the sources of those materials. The interdisciplinary approach of this project is to combine a chemical reactor theory of digestion with measurements of the processing of enzymatically available amino acids, focusing on rates of hydrolysis in, and absorption from, the gut. In vitro and in vivo fluorometric methods will be developed to assay enzyme activity in the guts of small deposit feeders; these methods that will be applicable to planktonic animals as well. Chemical analyses and deposit-feeder bioassays will be conducted to test whether methionine or other essential amino acids become more limiting in laboratory simulations of diagenesis. In a shallow subtidal site the hypothesis that the holothuroid Parastichopus californicus is limited to feeding during times of relatively high methionine concentration will be tested. At a 200-m site the hypothesis that the burrowing urchin Brisaster latifrons will show greater hydrolytic and absorptive capacity for amino acids after the spring bloom than during winter will be tested

Sedimentary Metal Bioavailability Determined By the Digestive Constraints of Marine Deposit Feeders: Gut Retention Time and Dissolved Amino Acids

Contaminant metals bound to sediments are subject to considerable solubilization during passage of the sediments through the digestive systems of deposit feeders. We examined the kinetics of this process, using digestive fluids extracted from deposit feeders Arenicola marina and Parastichopus californicus and then incubated with contaminated sediments. Kinetics are complex, with solubilization followed occasionally by readsorption onto the sediment. In general, solubilization kinetics are biphasic, with an initial rapid step followed by a slower reaction. For many sediment-organism combinations, the reaction will not reach a steady state or equilibrium within the gut retention time (GRT) of the organisms, suggesting that metal bioavailability in sediments is a time-dependent parameter. Experiments with commercial protein solutions mimic the kinetic patterns observed with digestive fluids, which corroborates our previous study that complexation by dissolved amino acids (AA) in digestive fluids leads to metal solubilization (Chen & Mayer 1998b; Environ Sci Technol 32:770-778). The relative importance of the fast and slow reactions appears to depend on the ratio of ligands in gut fluids to the amount of bound metal in sediments. High ligand to solid metal ratios result in more metals released in fast reactions and thus higher lability of sedimentary metals. Multiple extractions of a sediment with digestive fluid of A. marina confirm the potential importance of incomplete reactions within a single deposit-feeding event, and make clear that bioavailability to a single animal is Likely different from that to a community of organisms. The complex kinetic patterns lead to the counterintuitive prediction that toxification of digestive enzymes by solubilized metals will occur more readily in species that dissolve less metals

Distribution and Efficiency of Bacteriolysis in the Gut of Arenicola Marina and Three Additional Deposit Feeders

A simple technique was developed to measure the bacteriolytic activities of the digestive fluids of the deposit-feeding polychaete Arenicola marina. Lysis of a cultured environmental isolate, incubated with extracts of gut luminal contents, was monitored spectrophotometrically. Concurrent direct counts were used to verify cell lysis. The ability of extracts from 8 longitudinal sections of the gut to lyse the bacterium was monitored. The digestive ceca, anterior stomach, and posterior stomach regions exhibited high lytic activities, whereas bacteriolytic activities in all other regions of the gut were negligible. Similarly, extracts of surface sediments and fecal castings showed negligible lytic capabilities. The sharply limited distribution of lytic activity implicates the ceca as the source of bacteriolytic agent and suggests a true plug-flow system, with little axial mixing. Questions regarding the fate of lytic agents, which disappear abruptly posterior to the stomach, remain unanswered. Localization of lysis in the gut coupled with estimates of gut residence time permit the calculation that ingested bacteria are exposed to strong lytic activity for approximately 20 min. Incubation of in situ sediment samples with gut fluids corroborates the distributional findings of the in vitro work although the efficiency of lysis is much reduced, possibly due to exopolymer capsules and slimes of natural sedimentary bacteria. Cross-phyletic comparisons of bacteriolytic activities reveal both qualitative and quantitative differences. Much less demarcation of lytic activity is observed in the guts of a holothuroid (Caudina arenata) and a hemichordate (Stereobalanus canadensis), with a pattern more similar to that of A. marina observed in another polychaete, Amphitrite johnstoni. Quantitatively, the polychaetes showed higher levels of activity with rates in A. marina exceeding those of the hemichordate and holothuroid by more than 10-fold

Nutrient Uptake by a Deposit-Feeding Enteropneust: Nitrogenous Sources

We measured carbon, nitrogen, protein, bacterial and microalgal abundance, and mineral-specific surface area in sediments from the feeding zone of undisturbed Saccoglossus kowalewskyi, as well as in their fresh egesta. Comparison of results using surficial material 1 mm) and the top 3 mm of sediments indicated ingestion of surficial material by the enteropneusts. Assuming the surficial sediment as a food source results in apparent absorption efficiencies of 15% for TOC, 35% for TON, 60% for protein and 86% for microalgae. The C:N ratio of the apparently absorbed material was 4.2, consistent with an amino acid-rich diet. Protein- nitrogen uptake, however, accounted for only about 28% of total nitrogen absorption, indicating a dominant use of non-protein nitrogen . Bacterial and microalgal contributions to dietary nitrogen uptake were no more than 3% and 4% respectively. Active worms maintain 2 foraging areas with an average total foraging volume of 0.9 cm3 and a volume ingestion rate of 0.06 to 0.12 cm3 ind.-1 h-1. If the preferred feeding zone of these enteropneusts is the nitrogen -enriched surficial layer, we estimate that their feeding activities will deplete the available food resources every 8 to 16 h and they may rely on biological and tidal redistribution of surface material

Seasonal Variability in the Bacteriolytic Capacity of the Deposit Feeder Arenicola Marina: Environmental Correlates

Although deposit-feeding macrofauna consume and digest sedimentary bacteria, it is unclear whether feeding rates and digestion efficiencies are high enough to significantly impact the composition and abundance of bacteria in marine sediments. It is likely that both feeding rates and efficiency of digestion vary markedly through space and time. We used a turbidimetric assay to compare the rate of bacteriolysis by digestive fluids collected seasonally from the deposit-feeding polychaete Arenicola marina. Under standardized, experimental conditions, bacteriolytic rates represent concentrations of lytic agents. This concentration was found to vary significantly throughout the year (p = 0.001), showing greater than a 2x range. Lytic agent concentration was positively correlated with bioavailable amino acid concentrations in the surface sediment (r = 0.85, p = 0.03) but showed no apparent relationship to other proxies for food resources (e.g, chl a), sediment temperature, or gut throughput time. In vitro, temperature has been shown to have a strong positive influence on bacteriolytic rate. Temperature has no influence, however, on the in situ concentration of lytic agent in gut fluids, thus it appears that compensation for this temperature dependence is unimportant. These findings, combined with previous kinetics studies with A. marina gut fluids, predict that the quantitative influence of deposit feeding on the microbial ecology of sediments will exhibit clear seasonal variation