Uta Passow

The abundance and significance of a class of large, transparent organic particles in the ocean

Polysaccharide-specific staining techniques reveal the existence and high abundance of a class of large, discrete, transparent particles in seawater and diatom cultures formed from dissolved exopolymers exuded by phytoplankton and bacteria. Transparent exopolymer particles (TEP), ranged from 28 to 5000 particles ml−1 and 3 to 100s μm in longest dimension at five coastal stations off California. A high percentage of seemingly free-living bacteria (28–68%) were attached to these transparent sheets and films, suggesting that they may alter the distributions and microenvironments of marine microbes in nature. Preliminary coagulation experiments demonstrated that TEP are major agents in the aggregation of diatoms and in the formation of marine snow. The existence of microbial exudates acting as large, discrete particles, rather than as dissolved molecules or as coating on other particles, suggests that the transformation of dissolved organic matter into particulate form in the sea can occur via a rapid abiotic pathway as well as through conventional microbial uptake. The existence of these particles has far reaching implications for food web structure, microbial processes, carbon cycling and particulate flux in the ocean.

Transparent exopolymer particles (TEP) in aquatic environments

Since the development of methods to quantify transparent exopolymer particles (TEP) 1993, it has been shown that these gel-particles are not only ubiquitous and abundant, but also play a significant role in the biogeochemical cycling of elements and the structuring of food webs. TEP may be quantified either microscopically or colorimetrically. Although data based on measurements using one or other of these methods are not directly comparable, the results are consistent. TEP abundances in fresh and marine waters are in the same range as those of phytoplankton, with peak values occurring during phytoplankton blooms. TEP are very sticky particles that exhibit the characteristics of gels, and consist predominantly of acidic polysaccharides. In marine systems the majority of TEP are formed abiotically from dissolved precursors, which are released by phytoplankton that are either actively growing or are senescent. TEP are also generated during the sloughing of cell surface mucus and the disintegration of colonial matrices. The impact of exopolymers in the creation of microhabitats and in the cycling of trace compounds varies with the state in which the polymers occur, either as particles or as solute slimes. As particles, TEP provide surfaces for the colonization by bacteria and transfer by adsorption, trace solute substances into the particulate pool. As dissolved polymers they are mixed with the water and can neither be filtered nor aggregated. Because of their high abundances, large size and high stickiness, TEP enhance or even facilitate the aggregation of solid, non-sticky particles. They have been found to form the matrices of all marine aggregates investigated to date. By aggregating solid particles, TEP promote the sedimentation of particles, and, because their carbon content is high, their direct contribution to fluxes of carbon into deep water is significant. The direct sedimentation of TEP may represent a mechanism for the selective sequestration of carbon in deep water, because the C:N ratios of TEP lie well above the Redfield ratio. The turnover time of TEP as a result of bacterial degradation appears to range from hours to months, depending on the chemical composition and age of TEP. TEP may also be utilized not only by filter feeders (some protozoans and appendicularian) but TEP-rich microaggregates, consisting of pico- and nano-plankton are also readily grazed by euphausiids, thus permitting the uptake of particles that would otherwise be too small to be grazed directly by euphausiids. This short-circuits food chains and links the microbial food-web to the classical food-web. It is suggested that this expansion of the concept of food webs, linking the microbial loop with an aggregation web will provide a more complete description of particle dynamics

The role of particulate carbohydrate exudates in the flocculation of diatom blooms

Diatom blooms are frequently terminated by mass aggregation of cells into large, rapidly sinking aggregates. It has been hypothesized that transparent exopolymer particles (TEP), abundant particles formed from the polysaccharides exuded by living cells, may be essential for this mass flocculation processes. We investigated the abundance of TEP and their role in the aggregation of diatoms in laboratory cultures and during a natural diatom bloom off California. TEP and dissolved carbohydrates accumulated appreciably over the growth cycle of Chaetoceros gracilis in the laboratory. The flocculation of C. gracilis in a laboratory flocculator was dominated by TEP, not cells, and large flocs, consisting predominantly of particulate polysaccharides, formed at a rate more than an order of magnitude higher than predicted by coagulation theory for cells alone. The frequency of interparticle attachment was three orders of magnitude higher for TEP than for cells. The pattern of flocculation of a natural diatom bloom was similar to that of laboratory cultures. Prior to bloom flocculation the abundance and total quantity of TEP and the concentration of particulate carbohydrates increased, while dissolved carbohydrate concentrations decreased. During the flocculation stage TEP aggregated into fewer, but much larger particles and concentrations of dissolved carbohydrates decreased further. The percentage of diatom cells which were attached to TEP increased during the flocculation period from 3 to 90% and TEP formed the matrix of all the natural diatom aggregates observed. During the late flocculation stage the quantity of TEP and TEP aggregates did not increase further and concentrations of diatoms decreased, presumably because large flocs sank out. Our findings indicate that TEP should be included in models of particle aggregation in the ocean. The abundance, large size and high sticking coefficient of TEP make them essential to the aggregation of diatom blooms. The extracellular release of polysaccharides by growing cells may be an adaptation for aggregation. The abiotic formation of particulate organic matter (TEP) from dissolved organic matter (DOC) may help to explain the extremely high turnover rates of DOC observed during blooms.

The origin of transparent exopolymer particles (TEP) and their role in the sedimentation of particulate matter

Seasonal changes in the concentration of suspended transparent exopolymer particles (TEP) and flux rates of TEP and other particles at 500m were measured at 2-week intervals at a station in the Santa Barbara Channel between May 1995 and June 1997 in order to investigate the hypothesis that the presence of TEP is necessary for the aggregation and subsequent sedimentation of particles from the pelagic zone. During the 2-year period phytoplankton appeared to be the most significant source of TEP. However, in association with phytoplankton, the concentration of TEP was also positively aected by bacteria abundance. Possibly bacteria enhance the production of TEP by phytoplankton. The presence of TEP was necessary for the sedimentation of diatoms. However, only 67% of the peaks in particulate organic carbon flux corresponded to peaks in TEP flux. Lithogenic silica sedimented only when scavenged by marine snow; either by TEP-rich diatom aggregates or by zooplankton-derived snow (larvacean houses). TEP were not involved in the sedimentation of foraminifera. Although sedimentation was the dominant loss processes of TEP out of the euphotic zone, other loss process must have been important at greater depth, as only a small fraction of the standing stock of TEP arrived at 500m. # 2001 Elsevier Science Ltd. All rights reserved.

The role of surface-active carbohydrates in the flocculation of a diatom bloom in a mesocosm

A study was undertaken to evaluate the role of exocellular polysaccharides in the flocculation of a marine diatom bloom in a large tank mesocosm. Surface-active organic matter was extracted from 1.0 μ-filtered tank water by bubble adsorption each day for 7 days of the experiment. In agreement with past studies, particles (3–51 urn equivalent spherical diameter) were readily formed by bubbling and became concentrated in the foam. At the beginning of the bubbling (0–0.5 h), both particles and surface-active carbohydrates were extracted at high rates; however, these rates dropped off steeply after about 0.5 h of bubbling. The rate of particle formation by bubbling could be modeled fairly well by second order kinetics. The extracted, surface-active material was enriched in deoxysugars and galactose, while the residual material was enriched in glucose. Extracted surface-active carbohydrates reached a maximum of 33% of the total dissolved sugars ( 0.99) with particle stickiness (alpha). In addition, the concentration of surface-active carbohydrates was well correlated (2 = 0.91) with the concentration of transparent exopolymer particles (TEP) in the tank, and it was demonstrated that TEP could be copiously formed by bubbling of 1.0 μm-filered seawater. The finding of a highly surface-active, deoxysugar-rich polysaccharide material that can be rapidly (<0.5 h) and selectively extracted by bubble adsorption is significant, as it is apparent that this material played important roles in particle stickiness and TEP formation in the tank, and thus it may, at times, play similar roles in particle aggregation in the sea.

Formation of transparent exopolymer particles, TEP, from dissolved precursor material

Transparent exopolymer particles (TEP) form from polysaccharides released by many phytoplankton species, but this process by which dissolved organic matter becomes particulate is poorly understood. Here, the abiotic formation of TEP from precursors <0.2 μm and the minimum molecular weight (MW) of TEP-precursors were studied. In most samples TEP formed from material <0.2 μm (polycarbonate membrane filters, Poretics) when exposed to laminar shear in Couette flocculators. This result was unexpected as no TEP formed from material <0.45 μm (polycap capsules, Whatman) due to surface coagulation onto bubbles (Zhou et al. 1998; Limnol Oceanogr 43:1860-1871). Some TEP-precursors were able to pass through dialysis bags with a nominal pore size of 8 kDa (natural cellulose, Spektrum), although their MW is presumably 2 orders of magnitude larger, suggesting that TEP-precursors can be fibrillar. It is suggested that freshly released precursors are fibrillar and that these fibrillar precursors form larger colloids and eventually TEP within hours to days after their release.

Marine phytoplankton temperature versus growth responses from polar to tropical waters – outcome of a scientific community-wide study

“It takes a village to finish (marine) science these days” Paraphrased from Curtis Huttenhower (the Human Microbiome project) The rapidity and complexity of climate change and its potential effects on ocean biota are challenging how ocean scientists conduct research. One way in which we can begin to better tackle these challenges is to conduct community-wide scientific studies. This study provides physiological datasets fundamental to understanding functional responses of phytoplankton growth rates to temperature. While physiological experiments are not new, our experiments were conducted in many laboratories using agreed upon protocols and 25 strains of eukaryotic and prokaryotic phytoplankton isolated across a wide range of marine environments from polar to tropical, and from nearshore waters to the open ocean. This community-wide approach provides both comprehensive and internally consistent datasets produced over considerably shorter time scales than conventional individual and often uncoordinated lab efforts. Such datasets can be used to parameterise global ocean model projections of environmental change and to provide initial insights into the magnitude of regional biogeographic change in ocean biota in the coming decades. Here, we compare our datasets with a compilation of literature data on phytoplankton growth responses to temperature. A comparison with prior published data suggests that the optimal temperatures of individual species and, to a lesser degree, thermal niches were similar across studies. However, a comparison of the maximum growth rate across studies revealed significant departures between this and previously collected datasets, which may be due to differences in the cultured isolates, temporal changes in the clonal isolates in cultures, and/or differences in culture conditions. Such methodological differences mean that using particular trait measurements from the prior literature might introduce unknown errors and bias into modelling projections. Using our community-wide approach we can reduce such protocol-driven variability in culture studies, and can begin to address more complex issues such as the effect of multiple environmental drivers on ocean biota.

Rapid formation and sedimentation of large aggregates is predictable from coagulation rates (half-lives) of transparent exopolymer particles (TEP)

Two hypotheses have been proposed to account for the precipitous formation of large, rapidly settling aggregates at the termination of phytoplankton blooms in nature; aggregation due primarily to cell-cell collisions, and aggregation resulting from the presence of abundant transparent exopolymer particles (TEP), a recently discovered class of particles formed from polysaccharides excreted by phytoplankton. The hypothesis of TEP-driven coagulation in three disparate systems, a freshwater lake, a coastal ocean, and a saltwater mesocosm was evaluated, by comparing TEP abundance to several related factors including phytoplankton concentrations, measured sediment fluxes, and abundances of large aggregates. The timing of large aggregate formation and sedimentation events was related to coagulation rates expressed in terms of particle half-lives, t12, calculated as the time for TEP or phytoplankton to decrease to half their concentration through shear coagulation. While TEP have been previously investigated only in marine systems, it is reported here that TEP also can be present in high concentrations (860 ml−1) in freshwater lakes (Lake Constance, Germany) and that high fluxes of particulate organic matter at depth coincide with the disappearance of abundant TEP from overlying waters. The half-lives of TEP in the three different systems indicate that large aggregate formation and massive sedimentation events following diatom blooms occur when the TEP half-life decreases to less than a few days. By comparing TEP and phytoplankton half-lives in these systems, it is concluded that the formation of rapidly sinking aggregates following blooms of mucous-producing diatoms is primarily controlled by concentrations of TEP, not phytoplankton.

Aggregation of a diatom bloom in a mesocosm: The role of transparent exopolymer particles (TEP)

The role of TEP (Transparent Exopolymer Particles) in the flocculation of a diatom bloom was studied under controlled conditions in a mesocosm. The concentration of TEP increased exponentially during growth, flocculation and senescence of the bloom. Aggregation began dominating the particle dynamics of TEP during the early growth phase of the bloom, several days prior to the appearance of large flocs and nutrient depletion. TEP aggregated with themselves and with phytoplankton due to the high stickiness of TEP, but phytoplankton was not observed to aggregrate with itself. The production of TEP, estimated from changes in concentration, did not increase after nutrients were depleted. The concentration of TEP was a linear function of chl a and particulate organic carbon (POC), indicating that production of TEP was linked to growth rather than standing stocks of phytoplankton. The ratio between TEP and phytoplankton appeared to be one of the factors determining the onset of the flocculation of the bloom. The concentration of TEP may have been decreased by bacterial degradation. Bacterial degradation of TEP may explain the low TEP to chl a values, the decrease in stickiness of particles as the bloom progressed, and the retarded onset of flocculation.

Marine snow formation in the aftermath of the Deepwater Horizon oil spill in the Gulf of Mexico

The large marine snow formation event observed in oil-contaminated surface waters of the Gulf of Mexico (GoM) after the Deepwater Horizon accident possibly played a key role in the fate of the surface oil. We characterized the unusually large and mucus-rich marine snow that formed and conducted roller table experiments to investigate their formation mechanisms. Once marine snow lost its buoyancy, its sinking velocity, porosity and excess density were then similar to those of diatom or miscellaneous aggregates. The hydrated density of the component particles of the marine snow from the GoM was remarkably variable, suggesting a wide variety of component types. Our experiments suggest that the marine snow appearing at the surface after the oil spill was formed through the interaction of three mechanisms: (1) production of mucous webs through the activities of bacterial oil-degraders associated with the floating oil layer; (2) production of oily particulate matter through interactions of oil components with suspended matter and their coagulation; and (3) coagulation of phytoplankton with oil droplets incorporated into aggregates. Marine snow formed in some, but not all, experiments with water from the subsurface plume of dissolved hydrocarbons, emphasizing the complexity of the conditions leading to the formation of marine snow in oil-contaminated seawater at depth.