New Tracer to Estimate Community Predation Rates of Phagotrophic Protists

Predation of eukaryotic microbes on prokaryotes is one of the most important trophic interactions on Earth, representing a major mortality term and shaping morphology and composition of prokaryotic communities. Here we introduce and validate a new tracer to determine predation rates on prokaryotes. Minicells of Escherichia coli marked with a bright green fluorescent protein (GFP) vector have many operational advantages over previously used prey analogs such as fluorescently labeled bacteria. GFP-minicells are similar in size to naturally occurring bacteria from a variety of environments including the oligotrophic open ocean and the deep sea. They are relatively stable against microbial and light degradation, are easy to grow and process, and can be produced inexpensively in large numbers. No chemical alteration of the particle surface due to heat killing and staining is involved. Grazing coefficients were compared between GFP-minicells and other GFP-modified bacteria, as well as 5-(4,6-dichlorotriazinyl) aminofluorescein (DTAF)stained cells. The grazing coefficients obtained from the removal of GFP-minicells compared favorably with estimates from tracer-independent estimates of grazing in the same experiments. Experiments with GFP-minicells resulted in community grazing coefficients similar to those reported for many different marine environments and those derived using various methods

Relevant Scales in Zooplankton Ecology: Distribution, Feeding, and Reproduction of the Copepod Acartia Hudsonica in Response to Thin Layers of the Diatom Skeletonema Costatum

We investigated the interaction of the copepod Acartia hudsonica in relation to thin layers of the diatom Skeletonema costatum. Thin layers have recently received much attention, since they are common and persistent features in the water column, often overlooked by traditional sampling methods. Their frequent abundance in coastal oceans and the high biomass associated with them has led to the assumption that they are important grazing sites of calanoid copepods. We employed 2-m tall tower tanks that allowed us to simulate thin layers. Three variables representative of three time scales were considered: the distribution of copepods in the tanks (time scale of minutes), fecal pellet production as a proxy for ingestion rate (time scale of hours), and egg production rate (time scale of \u3e12 h). A. hudsonica responded significantly but very little to the thin layers in terms of their distribution. Given a choice, there was a slightly higher tendency to swim through a patch of diatoms than to swim around it. Fecal pellet production was slightly lower in the thin-layer treatments than in the homogeneous controls. Egg production was not influenced by differential distribution of diatoms in the tanks, which indicated that the copepods dealt equally well with patchy food as when the same numbers of cells were available in a homogeneous distribution. Time series experiments showed that ingested carbon is integrated over time scales of \u3e12 h. Therefore, small-scale fluctuations of food in space and time do not necessarily translate into small-scale fluctuations in reproductive output

Absorption Efficiencies and Biochemical Fractionation of Assimilated Compounds in the Cold Water Appendicularian Oikopleura Vanhoeffeni

Using Ge-68:C-14 dual-labeling, we investigated the absorption efficiency of diatom carbon for the cold water appendicularian Oikopleura vanhoeffeni. The absorption efficiency of bulk carbon (mean = 67%) was not influenced by body size or ingestion rate. For the first time for a pelagic tunicate, food and feces were fractionated into their major biochemical constituents (i.e., low-molecular-weight compounds, lipid, protein, and polysaccharide), allowing calculation of absorption efficiencies for each fraction. Low-molecular-weight compounds and proteins were preferentially absorbed over lipids and polysaccharides. However, predicted C:N ratios of the fecal pellets of O. vanhoeffeni were in the lower range of C:N ratios reported for zooplankton feces. The results are relevant for modeling biogeochemical cycles because pelagic tunicates contribute greatly to vertical particulate organic carbon flux

Contribution of Zooplankton Lipids to the Flux of Organic Matter in the Northern Adriatic Sea

Analyses of particulate material collected by sediment traps moored at a location in the northern Adriatic Sea in 1991 revealed the presence of zooplankton fatty acids, even though zooplankton and other \u27swimmers\u27 killed by the trap\u27s preservative were carefully removed. Laboratory experiments were conducted to (1) prove the existence of zooplankton lipids within fecal pellets, (2) exclude the possibility of incomplete separation of swimmers and other material as eventual contamination with polyunsaturated fatty acids in fecal pellets, (3) evaluate the importance of zooplankton lipids to mass flux and (4) reveal the mechanisms which lead to excretion of undigested organic matter, in this case polyunsaturated fatty acids. Our results show that the main source of fatty acids found in mass flux were zooplankton lipid droplets inside fecal pellets. The predominant fatty acids of zooplankton fecal pellets were saturated acid 16:0, monounsaturated acid 18:1 and polyunsaturated acid 22:6. Lipid composition of fecal pellets was compared with those of zooplankton and phytoplankton. Aliquots of collected fecal pellets were stained with Nile Red in order to visualize lipid droplets within fecal pellets

The Aquatic Particle Number Quandry

Optical surveys of aquatic particles and their particle size spectra have become important tools in studies of light propagation in water, classification of water masses, and the dynamics of trophic interactions affecting particle aggregation and flux. Here, we demonstrate that typical settings used in image analysis vastly underestimate particle numbers due to the particle – gel continuum. Applying a wide range of threshold values to change the sensitivity of our detection system, we show that macrogels cannot be separated from more dense particles, and that a true particle number per volume cannot be ascertained; only relative numbers in relation to a defined threshold value can be reported. A quandary thus presents itself between choosing a detection threshold low enough to accurately record orders of magnitude more particles on one hand or selecting a higher threshold to yield better image quality of plankton on the other. By observing the dynamics of coagulation and dissolution steps unique to cation-bridged gels abundant in aquatic systems, we find naturally occurring gels, and microscopic particles attached to them, to cause the ill-defined particle numbers. In contrast, the slopes in particle number spectra remained largely unaffected by varying sensitivity settings of the image analysis. The inclusion of fainter particles that are not typically captured by imaging systems provides a window into the true microscale spatial heterogeneity at scales relevant to small plankton organisms and processes that are dependent on particle density such as surface-associated chemical reactions as well as particle coagulation and aggregation dynamics

Influence of Zooplankton Grazing on Free Dissolved Enzymes in the Sea

In the Northern Adriatic Sea, extracellular enzymatic activity was measured during a Lagrangian study following a drifting buoy for 40 h. Dissolved free enzymatic activity represented 20 to 70% of total activity depending on the type of enzyme. alpha- and beta-glucosidases exhibited a significantly higher free activity than proteolytic enzymes. In subsequent laboratory experiments we investigated the effect of zooplankton on the free enzyme pool. The 4-step approach included: (1) determination of the enzymatic activities in copepods (mainly Acartia clausi); (2) enzymatic activity in fecal pellets; (3) short- and long-term grazing experiments; and (4) degradability of free glucosidase in seawater. alpha- and beta-glucosidases, leu-aminopeptidase, lipase and chitinase were examined. Experiments in which zooplankton were selectively enriched revealed a significant increase in both particle-bound (due to the increase of bacterial density) and dissolved free enzymatic activity. Incubating water enriched in free enzymes released by zooplankton with natural bacterial consortia, we found that 70% of the original alpha- and beta-glucosidase activity remained after 22 h. The presence of microorganisms did not enhance the degradation of these enzymes as compared to autoclaved controls. We found that a considerable amount of free dissolved enzymes is lost by 0.2 mu m filtration using Nuclepore filters, thereby leading to an underestimation of dissolved enzymes by similar to 30% in our experiments. Based on our results we conclude that mesozooplankton contribute to the free enzymatic activity in natural waters especially during periods of high grazing activity

Development and Deployment of a Point-Source Digital Inline Holographic Microscope for the Study of Plankton and Particles to a Depth of 6000 m

Bochdansky, A. B., Jericho, M. H., & Herndl, G. J. (2013). Development and deployment of a point-source digital inline holographic microscope for the study of plankton and particles to a depth of 6000 m. Limnology and Oceanography: Methods, 11, 28-40. doi: 10.4319/lom.2013.11.2

Eukaryotic Microbes, Principally Fungi and Labyrinthulomycetes, Dominate Biomass on Bathypelagic Marine Snow

In the bathypelagic realm of the ocean, the role of marine snow as a carbon and energy source for the deep-sea biota and as a potential hotspot of microbial diversity and activity has not received adequate attention. Here, we collected bathypelagic marine snow by gentle gravity filtration of sea water onto μm filters from similar to 1000 to 3900 m to investigate the relative distribution of eukaryotic microbes. Compared with sediment traps that select for fast-sinking particles, this method collects particles unbiased by settling velocity. While prokaryotes numerically exceeded eukaryotes on marine snow, eukaryotic microbes belonging to two very distant branches of the eukaryote tree, the fungi and the labyrinthulomycetes, dominated overall biomass. Being tolerant to cold temperature and high hydrostatic pressure, these saprotrophic organisms have the potential to significantly contribute to the degradation of organic matter in the deep sea. Our results demonstrate that the community composition on bathypelagic marine snow differs greatly from that in the ambient water leading to wide ecological niche separation between the two environments

Estimating Carbon Flux From Optically Recording Total Particle Volume at Depths Below the Primary Pycnocline

Optical instruments can rapidly determine numbers and characteristics of water column particles with high sensitivity. Here we show the usefulness of optically assessed total particle volume below the main pycnocline to estimate carbon export in two systems: the open subarctic North Atlantic and the Ross Sea, Antarctica. Both regions exhibit seasonally high phytoplankton production and efficient export (i.e., a strong biological pump). Total particle volumes in the mesopelagic (200-300 m) were significantly correlated with those in the overlying surface mixed layer (50-60 m), indicating that most particles at depth reflect export from the surface. This connectivity, however, is modulated by the physical structure of the water column and by particle type (e.g., the presence of colonies of the haptophyte Phaeocystis antarctica versus diatoms). Evidence from both regions show that a strong pycnocline can delay or may even prevent particles from settling to deeper layers, which then succumb to disintegration, and microbial and zooplankton consumption. Strong katabatic winds in the Ross Sea may deepen the mixed layer, causing a rapid transfer of particles to mesopelagic depths through the mixed-layer pump. Independent estimates of seasonally integrated export production in the Ross Sea, based on upper water column carbon mass balance, were significantly correlated (in the order of shared variance) with (1) total particle volumes from images, (2) particulate organic carbon, and (3) chlorophyll fluorescence, all recorded at a depth range of 200-300 m. Carbon export was not significantly correlated with particle abundance measured by a Coulter counter at the same depth range. Measuring total particle volume below the primary pycnocline is therefore a useful approach to estimate carbon export at least in regions characterized by seasonally high particle export

Role of Macroscopic Particles in Deep-Sea Oxygen Consumption

Macroscopic particles (\u3e 500 µg), including marine snow, large migrating zooplankton, and their fast-sinking fecal pellets, represent primary vehicles of organic carbon flux from the surface to the deep sea. In contrast, freely suspended microscopic particles such as bacteria and protists do not sink, and they contribute the largest portion of metabolism in the upper ocean. In bathy- and abyssopelagic layers of the ocean (2,000-6,000 m), however, microscopic particles may not dominate oxygen consumption. In a section across the tropical Atlantic, we show that macroscopic particle peaks occurred frequently in the deep sea, whereas microscopic particles were barely detectable. In 10 of 17 deep-sea profiles (\u3e 2,000 m depth), macroscopic particle abundances were more strongly cross-correlated with oxygen deficits than microscopic particles, suggesting that biomass bound to large particles dominates overall deep-sea metabolism