Protistan Grazing Research Articles

Phytoplankton Community Dynamic: A Driver for Ciliate Trophic Strategies

Phytoplankton plays a key role as primary producers and mediating biogeochemical cycles in the water column. The understanding of the temporal dynamic of primary grazers channelling energy and carbon from primary producers is important for evaluating aquatic ecosystems functioning. This study investigates the coupling between phytoplankton and ciliates from live samples collected with approximately daily frequency during an almost two-year cycle. The study site is a nutrient-rich temperate estuary, Roskilde Fjord (Denmark). Our aim is to evaluate the importance of protist grazers, especially ciliates, as predators on phytoplankton and to evaluate differences among multiple nutritional strategies through different seasons. The phytoplankton community, was mostly dominated by small organisms (< 20 µm) with few observations of diatoms. In most of observations, heterotrophic dinoflagellates biomass was smaller than biomass of ciliates (< 10%), indicating that ciliates are the main component of microzooplankton. Except for the spring 2016, the ciliate community closely followed the phytoplankton community, showing a tight coupling between the primary producers and grazers during all seasons. This somehow contradicts the general assumption that ciliate dominance is restricted to periods of nutrient limitations dominated by the microbial food web and suggests a year-round key role of ciliates as consumers of phytoplankton biomass. Biomasses of ciliates increased during spring and were highest during summer. Relative importance of mixotrophs were high due to occurrence of Mesodinium rubrum blooms as well as other mixotrophic ciliates in late spring/early summer. M. rubrum biomass had the opposite pattern of the cryptophyte prey Teleaulax spp., and the coupling between the two populations was very strong in late spring. Ciliates that grazed on selected phytoplankton, had a smaller potential grazing impact regarding their biomasses, likely due to food limitation; conversely ciliates that feed on diverse prey items were less constrained by food limitation, and their seasonality appear to be driven by other factors. These findings suggest that the ciliate community structure and dynamics is important in structuring the phytoplankton community on short and seasonal scale.

Herbivorous protist growth and grazing rates at in situ and artificially elevated temperatures during an Arctic phytoplankton spring bloom.

To assess protistan grazing impact and temperature sensitivity on plankton population dynamics, we measured bulk and species-specific phytoplankton growth and herbivorous protist grazing rates in Disko Bay, West Greenland in April-May 2011. Rate estimates were made at three different temperatures in situ (0 °C), +3 °C and +6 °C over ambient. In situ Chlorophyll a (Chl a) doubled during the observation period to ∼12 µg Chl a L−1, with 60–97% of Chl a in the >20 µm size-fraction dominated by the diatom genus Chaetoceros. Herbivorous dinoflagellates comprised 60–80% of microplankton grazer biomass. At in situ temperatures, phytoplankton growth or grazing by herbivorous predators <200 µm was not measurable until 11 days after observations commenced. Thereafter, phytoplankton growth was on average 0.25 d−1. Phytoplankton mortality due to herbivorous grazing was only measured on three occasions but the magnitude was substantial, up to 0.58 d−1. Grazing of this magnitude removed ∼100% of primary production. In short-term temperature-shift incubation experiments, phytoplankton growth rate increased significantly (20%) at elevated temperatures. In contrast, herbivorous protist grazing and species-specific growth rates decreased significantly (50%) at +6 °C. This differential response in phytoplankton and herbivores to temperature increases resulted in a decrease of primary production removed with increasing temperature. Phaeocystis spp. abundance was negatively correlated with bulk grazing rate. Growth and grazing rates were variable but showed no evidence of an inherent, low temperature limitation. Herbivorous protist growth rates in this study and in a literature review were comparable to rates from temperate waters. Thus, an inherent physiological inhibition of protistan growth or grazing rates in polar waters is not supported by the data. The large variability between lack of grazing and high rates of primary production removal observed here and confirmed in the literature for polar waters implies larger amplitude fluctuations in phytoplankton biomass than slower, steady grazing losses of primary production.

Rhizosphere Protists Change Metabolite Profiles in Zea mays.

Plant growth and productivity depend on the interactions of the plant with the associated rhizosphere microbes. Rhizosphere protists play a significant role in this respect: considerable efforts have been made in the past to reveal the impact of protist-bacteria interactions on the remobilization of essential nutrients for plant uptake, or the grazing induced changes on plant-growth promoting bacteria and the root-architecture. However, the metabolic responses of plants to the presence of protists or to protist-bacteria interactions in the rhizosphere have not yet been analyzed. Here we studied in controlled laboratory experiments the impact of bacterivorous protists in the rhizosphere on maize plant growth parameters and the bacterial community composition. Beyond that we investigated the induction of plant biochemical responses by separately analyzing above- and below-ground metabolite profiles of maize plants incubated either with a soil bacterial inoculum or with a mixture of soil bacteria and bacterivorous protists. Significantly distinct leaf and root metabolite profiles were obtained from plants which grew in the presence of protists. These profiles showed decreased levels of a considerable number of metabolites typical for the plant stress reaction, such as polyols, a number of carbohydrates and metabolites connected to phenolic metabolism. We assume that this decrease in plant stress is connected to the grazing induced shifts in rhizosphere bacterial communities as shown by distinct T-RFLP community profiles. Protist grazing had a clear effect on the overall bacterial community composition, richness and evenness in our microcosms. Given the competition of plant resource allocation to either defense or growth, we propose that a reduction in plant stress levels caused directly or indirectly by protists may be an additional reason for corresponding positive effects on plant growth.

In situ and experimental evidence for effects of elevated pH on protistan and metazoan grazers

AbstractPlankton succession was studied in a hyper-eutrophic stratified estuary, Mariager Fjord, Denmark. Above the pycnocline (15 m) pH increased from 8.5 to 9.2 and the oxygen increased to super saturation after 5 d of sunny weather due to high primary production. The protistan grazers were dominated by heterotrophic dinoflagellates and mixotrophic and heterotrophic ciliates. Metazooplankton was dominated by meroplankton, rotifers and the copepod, Acartia tonsa, all with a relatively low biomass. Cirriped nauplii occupied the upper strata while polychaete larvae populated the whole water column. Bivalve larvae occurred occasionally above the pycnocline even at very high pH. In pH challenge experiments, the mixotrophic ciliate Mesodinium rubrum was the least pH tolerant species, followed by Strombidium spp., which did not cope well with seawater pH &gt; 8.5. Some heterotrophic dinoflagellates were more tolerant with net growth at pH &gt; 9. The predominant rotifer Synchaeta sp. tolerated up to pH 9.5 and the copepod survived pH 10 but stopped producing eggs at pH 9.5 with unaffected egg hatching success. The polychaete and cirriped larvae tolerated pH 9.5, but bivalve larvae showed decreased survival already at pH 8.5. In situ distribution patterns and pH challenge experiments suggest that pH indeed contribute to structuring zooplankton distribution.

Interactions between the mixotrophic dinoflagellate Takayama helix and common heterotrophic protists

The phototrophic dinoflagellate Takayama helix that is known to be harmful to abalone larvae has recently been revealed to be mixotrophic. Although mixotrophy elevates the growth rate of T. helix by 79%–185%, its absolute growth rate is still as low as 0.3d−1. Thus, if the mortality rate of T. helix due to predation is high, this dinoflagellate may not easily prevail. To investigate potential effective protistan grazers on T. helix, feeding by diverse heterotrophic dinoflagellates such as engulfment-feeding Oxyrrhis marina, Gyrodinium dominans, Gyrodinium moestrupii, Polykrikos kofoidii, and Noctiluca scintillans, peduncle-feeding Aduncodinium glandula, Gyrodiniellum shiwhaense, Luciella masanensis, and Pfiesteria piscicida, pallium-feeding Oblea rotunda and Protoperidinium pellucidum, and the naked ciliates Pelagostrobilidium sp. (ca. 40μm in cell length) and Strombidinopsis sp. (ca. 150μm in cell length) on T. helix was explored. Among the tested heterotrophic protists, O. marina, G. dominans, G. moestrupii, A. glandula, L. masanensis, P. kofoidii, P. piscicida, and Strombidinopsis sp. were able to feed on T. helix. The growth rates of all these predators except Strombidinopsis sp. with T. helix prey were lower than those without the prey. The growth rate of Strombidinopsis sp. on T. helix was almost zero although the growth rate of Strombidinopsis sp. with T. helix prey was higher than those without the prey. Moreover, T. helix fed on O. marina and P. pellucidum and lysed the cells of P. kofoidii and G. shiwhaense. With increasing the concentrations of T. helix, the growth rates of O. marina and P. kofoidii decreased, but those of G. dominans and L. masanensis largely did not change. Therefore, reciprocal predation, lysis, no feeding, and the low ingestion rates of the common protists preying on T. helix may result in a low mortality rate due to predation, thereby compensating for this species’ low growth rate.

Saharan Dust Deposition Effects on the Microbial Food Web in the Eastern Mediterranean: A Study Based on a Mesocosm Experiment

The effect of episodicity of Saharan dust deposition on the pelagic microbial food web was studied in the oligotrophic Eastern Mediterranean by means of a mesocosm experiment in May 2014. Two different treatments in triplicates (addition of natural Saharan dust in a single-strong pulse or in three smaller consecutive doses of the same total quantity), and three unamended controls were employed; chemical and biological parameters were measured during a 10-day experiment. Temporal changes in primary (PP) and bacterial (BP) production, chlorophyll a (Chla) concentration and heterotrophic bacteria, Synechococcus and mesozooplankton abundance were studied. The results suggested that the auto- and hetero-trophic components of the food web (at least the prokaryotes) were enhanced by the dust addition (and by the nitrogen and phosphorus added through dust). Furthermore, a 1-day delay was observed for PP, BP and Chla increases when dust was added in three daily doses; however, the maximal values attained were similar in the two treatments. Although the effect was evident in the first osmotrophic level (phytoplankton and bacteria), it was lost further up the food web, masked under the impact of grazing exerted by predators such as heterotrophic flagellates, ciliates and dinoflagellates. This was partly proved by two dilution experiments. This study demonstrates the important role of atmospheric deposition and protist grazing when evaluating the effect on oligotrophic systems characterised by increased numbers of trophic levels.

A Review of Protist Grazing Below the Photic Zone Emphasizing Studies of Oxygen-Depleted Water Columns and Recent Applications of In situ Approaches

Little is still known of the impacts of protist grazing on bacterioplankton communities in the dark ocean. Furthermore, the accuracy of assessments of in situ microbial activities, including protist grazing, can be affected by sampling artifacts introduced during sample retrieval and downstream manipulations. Potential artifacts may be increased when working with deep-sea samples or samples from chemically unique water columns such as oxygen minimum zones (OMZs). OMZs are oxygen-depleted regions in the ocean, where oxygen concentrations can drop to <20 µM. These regions are typically located near eastern boundary upwelling systems and currently occur in waters occupying below about 8% of total ocean surface area, representing ~1% of the ocean’s volume. OMZs have a profound impact not only on the distribution of marine Metazoa, but also on the composition and activities of microbial communities at the base of marine food webs. Here we present an overview of current knowledge of protist phagotrophy below the photic zone, emphasizing studies of oxygen-depleted waters and presenting results of the first attempt to implement new technology for conducting these incubation studies completely in situ (the Microbial Sampling- Submersible Incubation Device, MS-SID). We performed 24-h incubation experiments in the Eastern Tropical South Pacific (ETSP) OMZ. This preliminary study shows that up to 28% of bacterial biomass may be consumed by protists in waters where oxygen concentrations were down to ~3 µM and up to 13% of saturation at a station with nitrate accumulation where oxygen concentrations were undetectable. Results also show that shipboard measurements of grazing rates were lower than rates measured from the same water using the MS-SID, suggesting that in situ experiments help to minimize artifacts that may be introduced when conducting incubation studies using waters collected from below the photic zone, particularly from oxygen-depleted regions of the water column.

Alanine δ15N trophic fractionation in heterotrophic protists

AbstractWe evaluated differences in the 15N isotopic enrichment factors of trophic amino acids (AA) for protistan (microzooplankton) and metazoan (mesozooplankton) consumers, testing the hypothesis that δ15N of alanine (ala) increases in both consumer types, while glutamic acid (glu) enriches mainly in mesozooplankton. AA δ15N values were measured for dinoflagellate and ciliate grazers and their respective algal prey (Oxyrrhis marina/Dunaliella tertiolecta and Favella sp./Heterocapsa triquetra) in four two‐stage chemostat experiments, including treatments with different nitrogen : phosphorous nutrient ratios and light/dark recycling conditions. Propagation of AA 15N enrichment to a metazoan consumer was also assessed in two‐ and three‐stage chemostat experiments simulating simple “classical” (Calanus pacificus and the diatom Thalassiosira weissflogii) and “multivorous” (C. pacificus, O. marina, and D. tertiolecta) food chains. We found small or negligible 15N‐enrichment of glu for both protistan grazers, while ala enrichment was consistently greater and similar to that in metazoan consumers. Ala and glu δ15N values were both highly elevated in C. pacificus relative to prey, and enrichment was higher with autotrophic diets. These laboratory results suggest that ala may be used as an alternate, accurate isotopic proxy for quantifying protistan contributions to trophic structure in aquatic systems.

Growth and mortality rates of prokaryotes in the hypolimnion of a deep freshwater lake (Lake Biwa, Japan)

The presence of pico-sized cyanobacteria (genus Synechococcus) in hypolimnetic waters has been reported, and investigators have suggested that Synechococcus contribute more to ecological processes in the hypolimnion than previously hypothesized; however, the ecological role of Synechococcus in food webs and/or matter cycling in the hypolimnion remains unknown. To address this issue, we assessed protistan grazing and the virus-mediated mortality of Synechococcus in the oxygenated hypolimnion of a large freshwater lake (Lake Biwa, Japan) during the stratification period. In addition, we compared the carbon flux through mortality of Synechococcus to that of heterotrophic bacteria to evaluate the role of Synechococcus in ecological processes within a hypolimnetic ecosystem. Our results suggest that the biomass of Synechococcus and heterotrophic bacteria in the hypolimnion was removed primarily by protistan grazing. The abundance of Synechococcus was highest in August, when the average Synechococcus:bacteria carbon biomass and daily grazing loss ratios were 10.8% and 11.0%, respectively. Thus, the Synechococcus biomass is likely an important seasonal component of the carbon flux in the hypolimnetic microbial loop. Our results provide the first data on carbon flux through the mortality of both Synechococcus and bacteria in a hypolimnetic ecosystem.

Changes in bacterial community composition and soil respiration indicate rapid successions of protist grazers during mineralization of maize crop residues

Decomposition of organic matter is crucial for ecosystem functioning. Microorganisms, which are responsible for the mineralization of organic matter, are usually treated as a homogeneous functional guild, despite mineralization capacity can differ profoundly between taxa. In addition, a significant part of the microbial community is top-down controlled by microbial grazers, such as protist. Since protist grazing is selective, and selectivity differs among species, we hypothesized that protist taxa complement each other in grazing intensity and thereby affect bacterial community structure and mineralization rate. In a laboratory experiment the species richness of protist communities was manipulated in an arable field soil and the mineralization rate of maize litter residues followed during the decomposition of the labile (4 days) and recalcitrant (3 weeks) carbon (C) fractions. Mineralization rate overall increased in the presence of protists. Changes in microbial function could be related to changes in microbial community composition (measured by phospholipid fatty acids pattern). During microbial decomposition, different protist grazers gained influence on mineralization rates over consecutive time intervals, indicating that a succession of protists caused an enhanced bacterial C-mineralization of plant detritus. Protist identity and species richness affected the microbial community composition, but not the magnitude of its mineralization function. In general, protist identity appeared to be more relevant for the composition of the microbial communities at the beginning of decomposition while the protist species richness appeared to be more critical in the later, slow phase of decomposition. This study provides an example that the overall outcome of ecosystem processes, such as mineralization rate is regulated by the sum of positive and negative effects of complex species interactions operating at a very fine spatial and temporal scales.

Differential interactions between the nematocyst-bearing mixotrophic dinoflagellate Paragymnodinium shiwhaense and common heterotrophic protists and copepods: Killer or prey

To investigate interactions between the nematocyst-bearing mixotrophic dinoflagellate Paragymnodinium shiwhaense and different heterotrophic protist and copepod species, feeding by common heterotrophic dinoflagellates (Oxyrrhis marina and Gyrodinium dominans), naked ciliates (Strobilidium sp. approximately 35μm in cell length and Strombidinopsis sp. approximately 100μm in cell length), and calanoid copepods Acartia spp. (A. hongi and A. omorii) on P. shiwhaense was explored. In addition, the feeding activities of P. shiwhaense on these heterotrophic protists were investigated. Furthermore, the growth and ingestion rates of O. marina, G. dominans, Strobilidium sp., Strombidinopsis sp., and Acartia spp. as a function of P. shiwhaense concentration were measured. O. marina, G. dominans, and Strombidinopsis sp. were able to feed on P. shiwhaense, but Strobilidium sp. was not. However, the growth rates of O. marina, G. dominans, Strobilidium sp., and Strombidinopsis sp. feeding on P. shiwhaense were very low or negative at almost all concentrations of P. shiwhaense. P. shiwhaense frequently fed on O. marina and Strobilidium sp., but did not feed on Strombidinopsis sp. and G. dominans. G. dominans cells swelled and became dead when incubated with filtrate from the experimental bottles (G. dominans+P. shiwhaense) that had been incubated for one day. The ingestion rates of O. marina, G. dominans, and Strobilidium sp. on P. shiwhaense were almost zero at all P. shiwhaense concentrations, while those of Strombidinopsis sp. increased with prey concentration. The maximum ingestion rate of Strombidinopsis sp. on P. shiwhaense was 5.3ngC predator−1d−1 (41 cells predator−1d−1), which was much lower than ingestion rates reported in the literature for other mixotrophic dinoflagellate prey species. With increasing prey concentrations, the ingestion rates of Acartia spp. on P. shiwhaense increased up to 930ngCml−1 (7180cellsml−1) at the highest prey concentration. The highest ingestion rate of Acartia spp. on P. shiwhaense was 4240ngC predator−1d−1 (32,610 cells predator−1d−1), which is comparable to ingestion rates from previous studies on other dinoflagellate prey species calculated at similar prey concentrations. Thus, P. shiwhaense might play diverse ecological roles in marine planktonic communities by having an advantage over competing phytoplankton in anti-predation against potential protistan grazers.

Grazing of particle-associated bacteria-an elimination of the non-viable fraction.

Quantification of bacteria being grazed by microzooplankton is gaining importance since they serve as energy subsidies for higher trophic levels which consequently influence fish production. Hence, grazing pressure on viable and non-viable fraction of free and particle-associated bacteria in a tropical estuary controlled mainly by protist grazers was estimated using the seawater dilution technique. In vitro incubations over a period of 42 h showed that at the end of 24 h, growth coefficient (k) of particle-associated bacteria was 9 times higher at 0.546 than that of free forms. Further, ‘k’ value of viable cells on particles was double that of free forms at 0.016 and 0.007, respectively. While bacteria associated with particles were grazed (coefficient of removal (g) = 0.564), the free forms were relatively less grazed indicating that particle-associated bacteria were exposed to grazers in these waters. Among the viable and non-viable forms, ‘g’ of non-viable fraction (particle-associated bacteria = 0.615, Free = 0.0086) was much greater than the viable fraction (particle-associated bacteria = 0.056, Free = 0.068). Thus, grazing on viable cells was relatively low in both the free and attached states. These observations suggest that non-viable forms of particle-associated bacteria were more prone to grazing and were weeded out leaving the viable cells to replenish the bacterial standing stock. Particle colonization could thus be a temporary refuge for the “persistent variants” where the viable fraction multiply and release their progeny.

Protistan predation interferes with bacterial long-term adaptation to substrate restriction by selecting for defence morphotypes.

Bacteria that are introduced into aquatic habitats face a low substrate environment interspersed with rare productive 'hotspots', as well as high protistan grazing. Whereas the former condition should select for growth performance, the latter should favour traits that reduce predation mortality, such as the formation of large cell aggregates. However, protected morphotypes often convey a growth disadvantage, and bacteria thus face a trade-off between investing in growth or defence traits. We set up an evolutionary experiment with the freshwater isolate Sphingobium sp. strain Z007 that conditionally increases aggregate formation in supernatants from a predator-prey coculture. We hypothesized that low substrate levels would favour growth performance and reduce the aggregated subpopulation, but that the concomitant presence of a flagellate predator might conserve the defence trait. After 26 (1-week) growth cycles either with (P+) or without (P-) predators, bacteria had evolved into strikingly different phenotypes. Strains from P- had low numbers of aggregates and increased growth yield, both at the original rich growth conditions and on various single carbon sources. By contrast, isolates from the P+ treatment formed elevated proportions of defence morphotypes, but exhibited lower growth yield and metabolic versatility. Moreover, the evolved strains from both treatments had lost phenotypic plasticity of aggregate formation. In summary, the (transient) residence of bacteria at oligotrophic conditions may promote a facultative oligotrophic life style, which is advantageous for survival in aquatic habitats. However, the investment in defence against predation mortality may constrain microbial adaptation to the abiotic environment.

Experimental Warming Decreases the Average Size and Nucleic Acid Content of Marine Bacterial Communities.

Organism size reduction with increasing temperature has been suggested as a universal response to global warming. Since genome size is usually correlated to cell size, reduction of genome size in unicells could be a parallel outcome of warming at ecological and evolutionary time scales. In this study, the short-term response of cell size and nucleic acid content of coastal marine prokaryotic communities to temperature was studied over a full annual cycle at a NE Atlantic temperate site. We used flow cytometry and experimental warming incubations, spanning a 6°C range, to analyze the hypothesized reduction with temperature in the size of the widespread flow cytometric bacterial groups of high and low nucleic acid content (HNA and LNA bacteria, respectively). Our results showed decreases in size in response to experimental warming, which were more marked in 0.8 μm pre-filtered treatment rather than in the whole community treatment, thus excluding the role of protistan grazers in our findings. Interestingly, a significant effect of temperature on reducing the average nucleic acid content (NAC) of prokaryotic cells in the communities was also observed. Cell size and nucleic acid decrease with temperature were correlated, showing a common mean decrease of 0.4% per °C. The usually larger HNA bacteria consistently showed a greater reduction in cell and NAC compared with their LNA counterparts, especially during the spring phytoplankton bloom period associated to maximum bacterial growth rates in response to nutrient availability. Our results show that the already smallest planktonic microbes, yet with key roles in global biogeochemical cycling, are likely undergoing important structural shrinkage in response to rising temperatures.

Decoding Microbial “Relationship Statuses” at OSM 2016

Some of the ocean's smallest organisms were the stars of the “Microbial Interactions in Ocean Ecosystems: Ecology to Biochemistry” session at OSM, the focus of which was on the relationships between marine microbes. Tying together far-ranging topics such as toxic algal blooms and novel methods for detecting bacterial chemotaxis was the idea that these microscale interactions are not only highly specific but also ecologically important. We heard from many speakers about how interactions between specific groups or even species of microbes can have large effects on their growth and behavior. One of the most striking findings was the specificity, both spatially and temporally, of the bacterial communities associated with eukaryotic phytoplankton. Thalassiosira rotula cells isolated from water samples from across the globe possessed distinct bacterial assemblages depending on location, found Olivia Marjorie Ahern. Another study by Hanna Farnelid, focusing on one location but taking samples across time, found evidence for species-specific functional associations between picoeukaryotes and bacteria. Laboratory coculturing experiments by Marilou Sison-Magnus showed that as the number of cocultured bacteria in a diatom culture increase, diatom growth rate also increases—but only when certain combinations of bacteria are present. In contrast, other combinations of bacteria enhanced production of domoic acid, a toxin associated with Harmful Algal Blooms (HABs). The distinct phycospheres created by these microbial relationships, the speakers pointed out, could impact processes from primary production up to ecosystem-level energy transfers. New ways to observe and quantify these microbial relationships were also a focus during this session. For example, a novel method of determining intracellular B vitamin concentrations has allowed Chris Suffridge to examine which members of the microbial community are consuming these essential vitamins. He found that bacteria, not phytoplankton, were the dominant consumers of B12, while the community was likely synthesizing B1. Furthermore, the addition of each vitamin had a distinct effect on the microbial community, indicating that shifting B-vitamin concentrations impact not only growth rates but also community composition. Another novel method presented during this session was a microfluidic technique developed by Bennett Lambert and his lab that will allow scientists to create in situ bacterial chemotaxis assays. Bacteria that respond to microplumes of chemoattractants are caught in the device's wells, and these samples are assessed using flow cytometry and molecular techniques. This technique can be used to investigate the chemical preferences of marine microbes at the microscale, rather than using bulk parameters that merely record the sum of microbial interactions. After hearing about microbial relationships for much of the session, one question remained: why are these associations so specific? Ecology-based studies delved into the basis behind these interactions, demonstrating again how varied they are. Lauren Marie Seyler demonstrated that the basis for the coexistence of bacteria and archaea in the deep sea may be related both to resource partitioning and selective predation, while a study of how grazers impact blooms of Synechococcus in the Gulf of Maine by Peter Dylan Countway found that certain types of protist grazers were likely to respond to Synechococcus blooms and that these grazers appear to exert a strong degree of control on the abundance and duration of the annual bloom. Focusing less on the microscopic, Nicole Lopanik examined variations in symbiosis between the marine bryozoan Bugula neritina and its bacterial symbiont, finding that high latitude, symbiont-less bryozoans likely do without their symbiont because the benefits of the relationship, namely protection from predation, aren't as necessary in high-latitude environments because of lower inherent predation pressure. While this session helped us all to better decode the ocean's microbial relationships, it also highlighted how much we still have to learn. With the potential for impacts on both micro- and macroscale ecological and biochemical processes, there's no doubt we'll be seeing more on this topic at future ASLO meetings.

Distribution and seasonal variation of picoplankton in Sanggou Bay, China

Picoplankton abundance and biomass in Sanggou Bay, China, were investigated in 4 successive seasons (April, August and October 2011, January 2012). Different distribution patterns of picoplankton abundance and biomass were observed according to season and culture areas (bivalves or macroalgae). Synechococcus, picoeukaryotes and heterotrophic prokaryotes exhibited higher abundance and biomass in warm seasons (summer and autumn) than in cold seasons (spring and winter). Over all 4 seasons, picoplankton abundance was higher in the bivalve culture area than in the macroalgae culture area. Among picoplankton, picoeukaryotes contributed most to the carbon standing stock in summer and autumn. In spring and winter, the heterotrophic component biomass exceeded that of the autotrophic picoplankton. Picoeukaryotes were an important contributor (21-27%) to total phytoplankton carbon biomass in spring to autumn. In spring, heterotrophic prokaryote biomass accounted for more than 56% of total phytoplankton biomass, and even exceeded phytoplankton biomass at some stations. As revealed by multiple stepwise regression analysis, physicochemical factors and protist grazing were the most important variables that controlled picoplankton distribution and variation. The reduction in grazing pressure, as well as phosphorus release by bivalves, is likely to explain the higher abundance of picoplankton in the bivalve culture area of Sanggou Bay.

Protist-Bacteria Associations: Gammaproteobacteria and Alphaproteobacteria Are Prevalent as Digestion-Resistant Bacteria in Ciliated Protozoa

Protistan bacterivory, a microbial process involving ingestion and digestion, is ecologically important in the microbial loop in aquatic and terrestrial ecosystems. While bacterial resistance to protistan ingestion has been relatively well understood, little is known about protistan digestion in which some ingested bacteria could not be digested in cells of major protistan grazers in the natural environment. Here we report the phylogenetic identities of digestion-resistant bacteria (DRB) that could survive starvation and form relatively stable associations with 11 marine and one freshwater ciliate species. Using clone library and sequencing of 16S rRNA genes, we found that the protistan predators could host a high diversity of DRB, most of which represented novel bacterial taxa that have not been cultivated. The localization inside host cells, quantity, and viability of these bacteria were checked using fluorescence in situ hybridization. The DRB were affiliated with Actinobacteria, Bacteroidetes, Firmicutes, Parcubacteria (OD1), Planctomycetes, and Proteobacteria, with Gammaproteobacteria and Alphaproteobacteria being the most frequently occurring classes. The dominance of Gamma- and Alphaproteobacteria corresponds well to a previous study of Global Ocean Sampling metagenomic data showing the widespread types of bacterial type VI and IV secretion systems (T6SS and T4SS) in these two taxa, suggesting a putatively significant role of secretion systems in promoting marine protist-bacteria associations. In the DRB assemblages, opportunistic bacteria such as Alteromonadaceae, Pseudoalteromonadaceae, and Vibrionaceae often presented with high proportions, indicating these bacteria could evade protistan grazing thus persist and accumulate in the community, which, however, contrasts with their well-known rarity in nature. This begs the question whether viral lysis is significant in killing these indigestible bacteria in microbial communities. Taken together, our study on the identity of DRB sheds new light on microbial interactions and generates further hypotheses including the potential importance of bacterial protein secretion systems in structuring bacterial community composition and functioning of “microbial black box” in aquatic environments.

Evidence for Strain-Specific Exometabolomic Responses of the Coccolithophore Emiliania huxleyi to Grazing by the Dinoflagellate Oxyrrhis marina

The coccolithophore Emiliania huxleyi forms massive blooms and plays a critical role in global elemental cycles, sequestering significant amounts of atmospheric carbon dioxide on geological time scales via production of calcium carbonate coccoliths and emitting dimethyl sulfoniopropionate (DMSP) which has the potential for increasing atmospheric albedo. Because grazing in pelagic systems is a major top-down force structuring microbial communities, the influence of grazers on E. huxleyi populations has been of interest to researchers. Roles of DMSP (and related metabolites) in interactions between E. huxleyi and protist grazers have been investigated, however, little is known about the release of other metabolites that may influence, or be influenced by, such grazing interactions. We used high-resolution mass spectrometry in an untargeted approach to survey the suite of low molecular weight compounds released by four different E. huxleyi strains in response to grazing by the dinoflagellate Oxyrrhis marina. Overall, a strikingly small number of metabolites were detected from E. huxleyi and O. marina cells, but these were distinctly informative to construct metabolic footprints. At most, E. huxleyi strains shared 25% of released metabolites. Furthermore, there appeared to be no unified metabolic response in E. huxleyi strains to grazing; rather these responses were strain specific. Concentrations of several metabolites also positively correlated with grazer activities, including grazing, ingestion, and growth rates; however, no single metabolite responded uniformly across all strains of E. huxleyi tested. Regardless, grazing clearly transformed the constituents of dissolved organic matter produced by these marine microbes. This study addresses several technical challenges, and presents a platform to further study the influence of chemical cues in aquatic systems and demonstrates the impact of strain diversity and grazing on the complexity of dissolved organic matter in marine systems.

Biomass and composition of protistan grazers and heterotrophic bacteria in the Costa Rica Dome during summer 2010.

We investigated biomass and composition of heterotrophic microbes in the Costa Rica Dome during June-July 2010 as part of a broader study of plankton trophic dynamics. Because picophytoplankton (<2 μm) are known to dominate in this unique upwelling region, we hypothesized tight biomass relationships between size-determined predator-prey pairs (i.e. picoplankton-nano-grazers, nanoplankton-micro-grazers) within the microbial community. Integrated biomass of heterotrophic bacteria ranged from 180 to 487 mg C m-2 and was significantly correlated with total autotrophic carbon. Heterotrophic protist (H-protist) biomass ranged more narrowly from 488 to 545 mg C m-2, and was comprised of 60% dinoflagellates, 30% other flagellates and 11% ciliates. Nano-sized (<20 μm) protists accounted for the majority (57%) of grazer biomass and were positively correlated with picoplankton, partially supporting our hypothesis, but nanoplankton and micro-grazers (>20 μm) were not significantly correlated. The relative constancy of H-protist biomass among locations despite clear changes in integrated autotrophic biomass, Chl a, and primary production suggests that mesozooplankton may exert a tight top-down control on micro-grazers. Biomass-specific consumption rates of phytoplankton by protistan grazers suggest an instantaneous growth rate of 0.52 day-1 for H-protists, similar to the growth rate of phytoplankton and consistent with a trophically balanced ecosystem dominated by pico-nanoplankton interactions.

Interspecific competition and protistan grazing affect the coexistence of freshwater betaproteobacterial strains.

The competitive strength of four cosmopolitan freshwater betaproteobacterial isolates was investigated in the presence or absence of bacterivorous flagellates during continuous cultivation in artificial minimal medium at two dilution rates. Bacteria reached similar abundance and growth rate in monocultures, but in co-cultures, two strains (Acidovorax sp. and Massilia sp.) displayed significantly higher numbers and growth rates. These potential cross-feeding benefits were also supported by a high nutritional versatility of the two strains. In contrast, Hydrogenophaga sp. was seemingly less competitive or even inhibited by co-cultivation, and Limnohabitans planktonicus displayed striking abundance fluctuations. The latter two strains were least versatile in the uptake of different carbon sources and thus suffered more from interspecific competition. Moreover, remarkable strain-specific responses appeared when bacteria experienced increasing loss rates due to grazing and/or raised dilution rates. Limnohabitans planktonicus developed no successful defence strategy and was close to extinction. Massilia sp. formed grazing-resistant filaments exclusively at low dilution, but was highly reduced at increased flow-through. Acidovorax sp. was selectively ingested, but compensated grazing losses with accelerated growth rates and formed (co-)aggregates together with Hydrogenophaga sp. to escape predation at high flow-through. These species-specific interactions, growth responses and defence strategies strongly modulate mixed microbial assemblages and the microbial food web.