Heterotrophic Nanoflagellates Research Articles

Feeding and growth of the marine heterotrophic nanoflagellates, Procryptobia sorokini and Paraphysomonas imperforata on a bacterium, Pseudoalteromonas sp. with an inducible defence against grazing.

Heterotrophic marine nanoflagellates are important grazers on bacteria in the water column. Some marine bacteria appear more resistant to grazing than do others. Marine nanoflagellates can be grown in the laboratory in batch cultures fed specific bacterial isolates. In some cultures, the flagellates appear unable to completely deplete the bacterial prey even when the bacterial strain otherwise is an excellent prey. This may indicate that some marine bacteria are able to induce defence mechanisms if they are grazed by nanoflagellates. Four morphologically distinct marine heterotrophic nanoflagellates, of which 3 were still identified as Procryptobia sorokini (Kinetoplastea) and one as Paraphysomonas imperforata (Chrysophyceae) were isolated from a coastal location along with 3 isolates of the marine bacterium Pseudoalteromonas sp. Flagellate growth and grazing on bacterial prey were analysed in batch cultures. Pseudoalteromonas was a suitable prey for all 4 flagellate isolates. They grazed and grew on Pseudoalteromonas as sole prey with maximal cell-specific growth rates of 0.1–0.25 h-1 and gross growth efficiencies of 38–61%. Exposure to dense flagellate cultures or their supernatants did, however, cause a fraction of the Pseudoalteromonas cells to aggregate and the bacterium became apparently resistant to grazing. Concentrations of suspended Pseudoalteromonas cells were therefore not decreased below 1,700–7,500 cells μL-1 by any of the flagellate isolates. These results indicate that Pseudoalteromonas sp. can be an excellent prey to marine nanoflagellates but also that is in possession of inducible mechanisms that protect against flagellate grazing.

Feeding by the heterotrophic nanoflagellate Katablepharis remigera on algal prey and its nationwide distribution in Korea

Heterotrophic nanoflagellates are ubiquitous in natural waters, and most heterotrophic nanoflagellates are known to grow on bacteria. Recently, the heterotrophic nanoflagellate Katablepharis japonica has been reported to be an effective predator of diverse toxic or harmful algal prey. To date, 7 Katablepharis species have been identified, and therefore important questions arise as to whether other Katablepharis species can feed on algal prey, and further whether the types of prey of other Katablepharis species differ from those of K. japonica. To answer these important questions, feeding by Katablepharis remigera on diverse algal prey was examined. Specific growth and ingestion rates of K. remigera feeding on the raphidophytes Heterosigma akashiwo and Chattonella subsalsa were determined. Furthermore, the abundance of K. remigera at 28 stations along the coastline of Korea from January 2015 to October 2017 was quantified using qPCR method and newly designed specific primer-probe sets. Among 25 potential algal prey tested, K. remigera fed on only H. akashiwo and C. subsalsa; however, it did not feed on a diatom, a prymnesiophyte, a prasinophyte, cryptophytes, dinoflagellates, Mesodinium rubrum, a mixotrophic ciliate, and another raphidophyte Fibrocapsa japonica. The number of prey types on which K. remigera could feed (2 species) was considerably smaller than that of K. japonica (14 species). With the increase in the mean prey concentration, the specific growth rates of K. remigera on H. akashiwo and C. subsalsa increased as well before becoming saturated. The maximum specific growth rates of K. remigera on H. akashiwo and C. subsalsa were 0.717 and 0.129 d−1, respectively. In addition, the maximum ingestion rates of K. remigera on H. akashiwo and C. subsalsa were 0.333 and 0.661 ng C predator−1 d−1 (3.33 and 0.23 cells predator−1 d−1), respectively. The results of this study clearly indicate that K. remigera is an effective predator of 2 red tide-causing raphidophyte species, and additionally, the feeding activity of K. remigera differs greatly from that of K. japonica. The abundance of K. remigera was ≥0.1 cells mL−1 at 24 stations located in the East, West, and South Sea of Korea. Thus, K. remigera has a nationwide distribution in Korea. The highest abundance of K. remigera in Korean waters was 24.9 cells mL−1 in March 2017, when there was no red tide caused by H. akashiwo or Chattonella spp. In most regions where red tides caused by H. akashiwo or Chattonella spp. occurred in 2000–2017, K. remigera was detected. Thus, the abundance of K. remigera may increase during red tides caused by H. akashiwo and Chattonella spp.

Seasonal dynamics of transparent exopolymer particles (TEP) and their drivers in the coastal NW Mediterranean Sea

Transparent Exopolymer Particles (TEPs) are a subclass of organic particles with high impact in biogeochemical and ecological processes, such as the biological carbon pump, air-sea interactions, or the microbial loop. However, the complexity in production and consumption makes TEP dynamics hardly predictable, calling for the need of descriptive studies about the in situ dynamics of these particles. We followed monthly TEP dynamics and combined them with a dataset of environmental variables during three years in a coastal site of the oligotrophic North Western Mediterranean (Blanes Bay). TEP concentration, ranging from 11.3 to 289.1μgXGeqL−1 (average 81.7±11.7μgXGeqL−1), showed recurrent peaks in early summer (June–July). TEP were temporally disconnected from chlorophyll a maxima, that occurred in late winter and early spring (maxima 1.21μgL−1), but they were significantly related to the abundance of specific phytoplankton groups (diatoms and dinoflagellates) and also coincided with periods of low nutrient concentrations. The fraction of particulate organic carbon in the form of TEP (the TEP:POC and TEP:PM ratios) were also highest in early summer, indicating that TEP-enriched particles of low density accumulate in surface waters during stratified periods. We hypothesize that the accumulation of these particles affects the microbial food web by enhancing the activity of specific prokaryotic extracellular enzymes (esterase, β-glucosidase and alkaline phosphatase) and promoting the abundance of heterotrophic nanoflagellates.

Effects of warming and nutrients on the microbial food web in shallow lake mesocosms

We analysed changes in the abundance, biomass and cell size of the microbial food web community (bacteria, heterotrophic nanoflagellates, ciliates) at contrasting nutrient concentrations and temperatures during a simulated heat wave. We used 24 mesocosms mimicking shallow lakes in which two nutrient levels (unenriched and enriched by adding nitrogen and phosphorus) and three different temperature scenarios (ambient, IPCC A2 scenario and A2+%50) are simulated (4 replicates of each). Experiments using the mesocosms have been running un-interrupted since 2003. A 1-month heat wave was imitated by an extra 5 °C increase in the previously heated mesocosms (from 1st July to 1st August 2014). Changes in water temperature induced within a few days a strong effect on the microbial food web functioning, demonstrating a quick response of microbial communities to the changes in environment, due to their short generation times. Warming and nutrients showed synergistic effects. Microbial assemblages of heterotrophic nanoflagellates and ciliates responded positively to the heating, the increase being largest in the enriched mesocosms. The results indicate that warming and nutrients in combination can set off complex interactions in the microbial food web functioning.

Cryptophyta as major bacterivores in freshwater summer plankton

Small bacterivorous eukaryotes play a cardinal role in aquatic food webs and their taxonomic classification is currently a hot topic in aquatic microbial ecology. Despite increasing interest in their diversity, core questions regarding predator–prey specificity remain largely unanswered, e.g., which heterotrophic nanoflagellates (HNFs) are the main bacterivores in freshwaters and which prokaryotes support the growth of small HNFs. To answer these questions, we fed natural communities of HNFs from Římov reservoir (Czech Republic) with five different bacterial strains of the ubiquitous betaproteobacterial genera Polynucleobacter and Limnohabitans. We combined amplicon sequencing and catalyzed reporter deposition fluorescence in situ hybridization (CARD-FISH) targeting eukaryotic 18 S rRNA genes to track specific responses of the natural HNF community to prey amendments. While amplicon sequencing provided valuable qualitative data and a basis for designing specific probes, the number of reads was insufficient to accurately quantify certain eukaryotic groups. We also applied a double-hybridization technique that allows simultaneous phylogenetic identification of both predator and prey. Our results show that community composition of HNFs is strongly dependent upon prey type. Surprisingly, Cryptophyta were the most abundant bacterivores, although this phylum has been so far assumed to be mainly autotrophic. Moreover, the growth of a small lineage of Cryptophyta (CRY1 clade) was strongly stimulated by one Limnohabitans strain in our experiment. Thus, our study is the first report that colorless Cryptophyta are major bacterivores in summer plankton samples and can play a key role in the carbon transfer from prokaryotes to higher trophic levels.

Sensitivity of the simulated Oxygen Minimum Zone to biogeochemical processes at an oligotrophic site in the Arabian Sea

Oxygen minimum zones (OMZs) are large, low-oxygen areas in the global oceans. Although OMZs represent a serious threat to ecosystem functioning and services, our capability of modelling the main biogeochemical processes driving OMZ dynamic are still limited. Here we performed a full sensitivity analysis of a complex ecosystem model to rank the most important biogeochemical parameters influencing the simulation of the OMZ at an oligotrophic site in the open Arabian Sea. We applied a one-dimensional configuration of the European Regional Seas Ecosystem Model (ERSEM) - here advanced by including denitrification - coupled with the General Ocean Turbulence Model (GOTM). The coupled model was skilled in simulating the vertical gradients of climatological data of oxygen and nutrients. The sensitivity analysis of the model was carried out in two steps: i) a preliminary Morris screening analysis of 207 ERSEM parameters, which selected the three most influential groups of parameters; and ii) a subsequent Monte Carlo sampling-based analysis for ranking the importance of the 38 parameters within the three selected groups. Overall, the four most important parameters for the simulation of the minimum oxygen concentration were found to be: 1) the cubic half saturation constant for oxygenic control of denitrification; 2) the parameter regulating the fraction of ingested matter excreted by heterotrophic nanoflagellates; 3) the bacterial efficiency at low oxygen levels; and 4) the specific rate of bacterial release of capsular material. Based on these findings, and assuming that the ranking of the model parameters reflects the relevance of the process they characterize, we present a conceptual model describing the most important biogeochemical processes affecting the OMZ at the study site. Our results suggest that including bacteria explicitly in ecosystem models is useful to simulate and predict OMZs, provided that efforts are invested in estimating parameters characterizing the microbial loop in marine ecosystems.

Evidence for Coupling of the Carbon and Phosphorus Biogeochemical Cycles in Freshwater Microbial Communities

Considerable attention has been given to the roles of the carbon and phosphate cycles in aquatic environments, but less attention has been given to an experimental analysis of the coupling of the C and P cycles in freshwater and marine ecosystems. Using laboratory microcosm experiments, prepared with natural pond-water microbial communities, evidence is presented for the coupling of dissolved organic C with microbial production of alkaline phosphatase driving the phosphorus cycle in freshwater microbial communities. The effects of glucose C-supplementation in microcosm microbial communities (including bacteria and heterotrophic nanoflagellates) on gains in microbial C-content and alkaline phosphatase activity (APA) were estimated in relation to control microcosms without C-supplementation. The C-supplementation increased total microbial APA (pmol min-1 µg-1 bacterial C) in the C-supplemented treatment (6.5 ± 0.6) compared to the non-supplemented cultures (5.1 ± 1.7). Microbial-bound APA in the C-supplemented treatment was particularly enhanced (4.4 ± 0.9) compared to control cultures (1.3 ± 0.8), but the amount of free (soluble) APA in the aquatic phase was less compared to the controls (n = 5, p < 0.001). Alkaline phosphatase activity was highly correlated (r = 0.97) with bacterial densities in the C-supplemented cultures, further supporting the hypothesis that C-supplementation can increase phosphorus remineralization through elevated production of microbial alkaline phosphatase. This laboratory-based, experimental study suggests that additional research on the coupling of the C and P cycles in freshwater and marine environments may yield productive insights into the finer details of the roles of these two biogeochemical cycles in aquatic microbial community dynamics.

Spatio-temporal reproducibility of the microbial food web structure associated with the change in temperature: Long-term observations in the Adriatic Sea

Global and atmospheric climate change is altering the thermal conditions in the Adriatic Sea and, consequently, the marine ecosystem. Along the eastern Adriatic coast sea surface temperature (SST) increased by an average of 1.03 °C during the period from 1979 to 2015, while in the recent period, starting from 2008, a strong upward almost linear trend of 0.013 °C/month was noted. Being mainly oligotrophic, the middle Adriatic Sea is characterized by the important role played by the microbial food web in the production and transfer of biomass and energy towards higher trophic levels. It is very important to understand the effect of warming on microbial communities, since small temperature increases in surface seawater can greatly modify the microbial role in the global carbon cycle. In this study, the Self-Organizing Map (SOM) procedure was used to analyse the time series of a number of microbial parameters at two stations with different trophic status in the central Adriatic Sea. The results show that responses of the microbial food web (MFW) structure to temperature changes are reproducible in time. Furthermore, qualitatively similar changes in the structure of the MFW occurred regardless of the trophic status. The rise in temperature was associated with: (1) the increasing importance of microbial heterotrophic activities (increase bacterial growth and bacterial predator abundance, particularly heterotrophic nanoflagellates) and (2) the increasing importance of autotrophic picoplankton (APP) in the MFW.

Interactions between the voracious heterotrophic nanoflagellate Katablepharis japonica and common heterotrophic protists

Interactions between the voracious heterotrophic nanoflagellate Katablepharis japonica and common heterotrophic protists

Bacterial prey food characteristics modulate community growth response of freshwater bacterivorous flagellates

AbstractDifferent bacterioplankton species represent different food quality resources for heterotrophic nanoflagellate (HNF) communities, potentially affecting HNF growth, community dynamics and carbon flow to higher trophic levels. However, our knowledge of such dynamics is still very limited. Here, we describe the results of 11 experiments with natural HNF communities from distinct seasonal phases in two freshwater habitats. The HNF communities were released from predation pressure of zooplankton and incubated with 16 distinct ecologically relevant prey bacterial strains from important Betaproteobacteria genera (Limnohabitans, Polynucleobacter, and Methylopumilus) and one Actinobacteria strain from the Luna 2 cluster. We observed remarkable prey‐ and season‐specific variability in community HNF growth parameters, i.e., doubling time, volumetric gross growth efficiency (GGE), and length of lag phase. All strains, except for the actinobacterium, supported rapid HNF population growth with an average doubling time of 10 h and GGE of 29%. Our analysis revealed that 59% of the variability in flagellate GGE data was explained by the length of lag phase after prey amendments. This indicates a considerable “adaptation time,” during which the predator communities undergo compositional shifts toward flagellate bacterivores best adapted to grow on the offered prey. Importantly, the rapid HNF growth detected on various bacteria tightly corresponds to doubling times reported for fast growing bacterioplankton groups. We propose a conceptual model explaining the tight linkages between rapid bacterial community shifts and succeeding HNF community shifts, which optimize prey utilization rates and carbon flow from various bacteria to the microbial food chain.

Ocean acidification impacts primary and bacterial production in Antarctic coastal waters during austral summer

Polar waters may be highly impacted by ocean acidification (OA) due to increased solubility of CO2 at colder water temperatures. Three experiments examining the influence of OA on primary and bacterial production were conducted during austral summer at Davis Station, East Antarctica (68°35′ S, 77°58′ E). For each experiment, six minicosm tanks (650L) were filled with 200μm filtered coastal seawater containing natural communities of Antarctic marine microbes. Assemblages were incubated for 10 to 12days at CO2 concentrations ranging from pre-industrial to post-2300. Primary and bacterial production rates were determined using NaH14CO3 and 14C-leucine, respectively. Net community production (NCP) was also determined using dissolved oxygen. In all experiments, maximum photosynthetic rates (Pmax, mgCmgchl a−1h−1) decreased with elevated CO2, clearly reducing rates of total gross primary production (mgCL−1h−1). Rates of cell-specific bacterial productivity (μgCcell−1h−1) also decreased under elevated CO2, yet total bacterial production (μgCL−1h−1) and cell abundances increased with CO2 over Days 0–4. Initial increases in bacterial production and abundance were associated with fewer heterotrophic nanoflagellates and therefore less grazing pressure. The main changes in primary and bacterial productivity generally occurred at CO2 concentrations >2× present day (>780ppm), with the same responses occurring regardless of seasonally changing environmental conditions and microbial assemblages. However, NCP varied both within and among experiments, largely due to changing nitrate+nitrite (NOx) availability. At NOx concentrations <1.5μM photosynthesis to respiration ratios showed that populations switched from net autotrophy to heterotrophy and CO2 responses were suppressed. Overall, OA may reduce production in Antarctic coastal waters, thereby reducing food availability to higher trophic levels and reducing draw-down of atmospheric CO2, thus forming a positive feedback to climate change. NOX limitation may suppress this OA response but cause a similar decline.

Microbial plankton communities in the coastal southeastern Black Sea: biomass, composition and trophic interactions

Summary We investigated biomass and composition of the pico-, nano- and microplankton communities in a coastal station of the southeastern Black Sea during 2011. We also examined trophic interactions within these communities from size-fractionated dilution experiments in February, June and December. Autotrophic and heterotrophic biomasses showed similar seasonal trends, with a peak in June, but heterotrophs dominated throughout the year. Autotrophic biomass was mainly comprised by nanoflagellates and diatoms in the first half of the year, and by dinoflagellates and Synechococcus spp. in the second half. Heterotrophic biomass was mostly dominated by heterotrophic bacteria, followed by nanoflagellates and microzooplankton. Dilution experiments suggest that nano- and microzooplankton were significant consumers of autotrophs and heterotrophic bacteria. More than 100% of bacterial production was consumed by grazers in all experiments, while 46%, 21% and 30% of daily primary production were consumed in February, June and December, respectively. In February, autotrophs were the main carbon source, but in December, it was heterotrophic bacteria. An intermediate situation was observed in June, with similar carbon flows from autotrophs and heterotrophic bacteria. Size-fraction dilution experiments suggested that heterotrophic nanoflagellates are an important link between the high heterotrophic bacterial biomass and microzooplankton. In summary, these results indicate that nano- and microzooplankton were responsible for comprising a significant fraction of total microbial plankton biomass, standing stocks, growth and grazing processes. This suggests that in 2011, the microbial food web was an important compartment of the planktonic food web in the coastal southeastern Black Sea.

Newly discovered role of the heterotrophic nanoflagellate Katablepharis japonica, a predator of toxic or harmful dinoflagellates and raphidophytes

Heterotrophic nanoflagellates are ubiquitous and known to be major predators of bacteria. The feeding of free-living heterotrophic nanoflagellates on phytoplankton is poorly understood, although these two components usually co-exist. To investigate the feeding and ecological roles of major heterotrophic nanoflagellates Katablepharis spp., the feeding ability of Katablepharis japonica on bacteria and phytoplankton species and the type of the prey that K. japonica can feed on were explored. Furthermore, the growth and ingestion rates of K. japonica on the dinoflagellate Akashiwo sanguinea—a suitable algal prey item—heterotrophic bacteria, and the cyanobacteria Synechococcus sp., as a function of prey concentration were determined. Among the prey tested, K. japonica ingested heterotrophic bacteria, Synechococcus sp., the prasinophyte Pyramimonas sp., the cryptophytes Rhodomonas salina and Teleaulax sp., the raphidophytes Heterosigma akashiwo and Chattonella ovata, the dinoflagellates Heterocapsa rotundata, Amphidinium carterae, Prorocentrum donghaiense, Alexandrium minutum, Cochlodinium polykrikoides, Gymnodinium catenatum, A. sanguinea, Coolia malayensis, and the ciliate Mesodinium rubrum, however, it did not feed on the dinoflagellates Alexandrium catenella, Gambierdiscus caribaeus, Heterocapsa triquetra, Lingulodinium polyedra, Prorocentrum cordatum, P. micans, and Scrippsiella acuminata and the diatom Skeletonema costatum. Many K. japonica cells attacked and ingested a prey cell together after pecking and rupturing the surface of the prey cell and then uptaking the materials that emerged from the ruptured cell surface. Cells of A. sanguinea supported positive growth of K. japonica, but neither heterotrophic bacteria nor Synechococcus sp. supported growth. The maximum specific growth rate of K. japonica on A. sanguinea was 1.01 d−1. In addition, the maximum ingestion rate of K. japonica for A. sanguinea was 0.13ngC predator−1d−1 (0.06 cells predator−1d−1). The maximum ingestion rate of K. japonica for heterotrophic bacteria was 0.019ngC predator−1d−1 (266 bacteria predator−1d−1), and the highest ingestion rate of K. japonica for Synechococcus sp. at the given prey concentrations of up to ca. 107 cells ml−1 was 0.01ngC predator−1d−1 (48 Synechococcus predator−1d−1). The maximum daily carbon acquisition from A. sanguinea, heterotrophic bacteria, and Synechococcus sp. were 307, 43, and 22%, respectively, of the body carbon of the predator. Thus, low ingestion rates of K. japonica on heterotrophic bacteria and Synechococcus sp. may be responsible for the lack of growth. The results of the present study clearly show that K. japonica is a predator of diverse phytoplankton, including toxic or harmful algae, and may also affect the dynamics of red tides caused by these prey species.

The Response of Heterotrophic Prokaryote and Viral Communities to Labile Organic Carbon Inputs Is Controlled by the Predator Food Chain Structure.

Factors controlling the community composition of marine heterotrophic prokaryotes include organic-C, mineral nutrients, predation, and viral lysis. Two mesocosm experiments, performed at an Arctic location and bottom-up manipulated with organic-C, had very different results in community composition for both prokaryotes and viruses. Previously, we showed how a simple mathematical model could reproduce food web level dynamics observed in these mesocosms, demonstrating strong top-down control through the predator chain from copepods via ciliates and heterotrophic nanoflagellates. Here, we use a steady-state analysis to connect ciliate biomass to bacterial carbon demand. This gives a coupling of top-down and bottom-up factors whereby low initial densities of ciliates are associated with mineral nutrient-limited heterotrophic prokaryotes that do not respond to external supply of labile organic-C. In contrast, high initial densities of ciliates give carbon-limited growth and high responsiveness to organic-C. The differences observed in ciliate abundance, and in prokaryote abundance and community composition in the two experiments were in accordance with these predictions. Responsiveness in the viral community followed a pattern similar to that of prokaryotes. Our study provides a unique link between the structure of the predator chain in the microbial food web and viral abundance and diversity.

Effects of changing nutrient inputs on the ratio of small pelagic fish stock and phytoplankton biomass in the Black Sea

Significant increases in nitrogen and phosphorus inputs to the Black Sea in the second half of the 20th century caused eutrophication and drastically decreasing Si:N and Si:P ratios. Combined with climate change, overfishing of top predators and a huge outbreak of the non-indigenous ctenophore Mnemiopsis, the pelagic food web was strongly modified and its efficiency for channeling primary production to higher trophic levels substantially reduced. We used the ratio between small pelagic fish stock and phytoplankton biomass on the Danube shelf and in the open Black Sea to investigate long-term changes in food web functioning. The ratio had 1) highest values for the pre-eutrophication period when diatoms and copepods dominated the pelagic food web (‘muscle food chain’), 2) decreased during the eutrophication period with stronger prevalence of autotrophic pico- and nanophytoplankton, bacteria, heterotrophic nanoflagellates, microzooplankton, Noctiluca and jellyfish (‘jelly food chain’ with increased importance of the microbial loop), 3) lowest values during the ecological crisis (1989–1992), when small pelagic fish stocks collapsed, and 4) increased after 1993, indicating that the ecosystem went out of the crisis and exhibited a trend of recovery. However, in the last period (1993–2008) the ratio remained close to values observed in the middle eutrophication phase, suggesting that the ecosystem was far from fully recovered. Since early 2000s, fluctuating pelagic fish stocks, with a tendency to decreasing fish landing again, have been observed in the Black Sea. Additionally, the quality of food for the small pelagic fish has deteriorated due to warming trends and the legacy of eutrophication, giving support for the ‘jelly food chain’, exhibiting low energy transfer and prevalence of organisms with high respiration rate and low nutritional value.

Patterns of microbial food webs in Mediterranean shallow lakes with contrasting nutrient levels and predation pressures

To elucidate the specific and combined effects of bottom-up and top-down control on the microbial community in warm lakes, we sampled microbial community along with physical–chemical and biological variables and performed in situ food web experiments, in 14 Turkish shallow lakes with contrasting nutrient levels and predation pressures. Our field results revealed that differences in microbial communities correlated with differences in zooplankton community structure, temperature (increasing nutrient concentrations, change in zooplankton composition), nutrient concentrations (increasing bacteria and heterotrophic nanoflagellate abundances with increasing nitrogen concentrations and temperature) and macrophyte coverage (ciliates as potential consumers of bacteria and HNF was strongest in macrophyte-dominated lakes). Our in situ experimental study revealed that the zooplankton not only affect the biomass and composition of microbial communities but also alter the microbial structure and trophic relationships. Our results therefore indicate that both bottom-up factors and top-down effects were important for the efficiency of the carbon transfer from bacteria to higher trophic levels in the study lakes. Due to an anticipated increase in eutrophication, temperature and alteration of the classical food web with climate warming, major changes in the microbial community of lakes are, therefore, expected in a warmer future in semi-arid Mediterranean climatic regions.

Mixotrophic Phytoflagellate Bacterivory Field Measurements Strongly Biased by Standard Approaches: A Case Study.

Bacterivory among small (≤20 μm) phytoflagellates (SP) is increasingly recognized as a globally relevant phenomenon, impacting a wide range of aspects from primary production levels to marine fisheries. However, to correctly parametrize mixotrophic SP in biogeochemical and food web models, a better understanding of the magnitude and regulation of in situ SP feeding is urgently needed. Current methods to determine SP bacterivory in the field may introduce biases by treating these organisms as equivalent to heterotrophic nanoflagellates (HNF). In the present case study we experimentally tested two generally employed assumptions of such studies: (A) bacterivory rates of the whole SP community and of distinct SP groups remain constant over ‘short’ time scales (hours to a day) and (B) SP community ingestion rates approximate the average ingestion rate of all feeding individuals. Food vacuole markers (acidotropic probes), were applied along the diel cycle at three stations in December 2015, and May and June 2016. In December and June, surrogate prey (fluorescently labeled bacteria) were used in parallel at one sampling station. Sampling at different times of day produced an up to fourfold difference in estimates of SP daily bacterivorous impact. In contrast, daily bacterivory estimates for HNF remained constant in almost all cases. The perceived principal SP bacterivorous groups also shifted strongly. As an example, picoeukaryotes dominated total SP bacterivory in daylight hours but completely ceased to feed at night. Finally, a large fraction of the SP community was not feeding at all time points tested. This lead to significant errors in estimated ingestion rates determined using the whole SP community, being up to 16 times lower than those determined solely for actively feeding mixotrophic SP. Overall, this case study indicates that applying the two commonly used premises outlined above can introduce significant biases and considerably alter our perception of mixotrophy in a given system.

Effects of coral-derived organic matter on the growth of bacterioplankton and heterotrophic nanoflagellates

Exudates derived from hermatypic corals were incubated with <2 µm filtered seawater containing heterotrophic bacteria and <10 µm filtered seawater containing bacteria and nanoflagellates (HNF) under dark conditions for 96 h to quantify the growth of both bacteria and HNF in response to coral-derived dissolved organic matter (DOM). The addition of coral-derived DOM caused significantly higher growth rates and production of bacteria and HNF compared to those in control seawater without coral exudates. During the incubation, HNF exhibited their peak in abundance 24–48 h after the peak abundance of bacteria. The growth efficiencies of both bacteria and HNF were significantly higher with coral-derived DOM, suggesting higher transfer efficiency from bacteria that is fueled by coral organic matter to HNF. Therefore, trophic transfer of coral-derived DOM from bacteria to HNF can contribute to efficient carbon flow through the microbial food web.

Microbial - Planktonic foodweb dynamics of a eutrophic Area of Concern: Hamilton Harbour

Hamilton Harbour, located on the western end of Lake Ontario, has a long history of cultural eutrophication as well as industrial contamination. We explored the structure and function of the microbial – planktonic foodweb during the growing seasons (May–October) of 2004 and 2006 in order to consider the flow of autochthonous production from lower to higher trophic levels. Our analyses included microscope based assessments of bacteria, heterotrophic nanoflagellates, ciliates, phytoplankton and zooplankton as well as radioisotope based measurements of primary productivity and bacterial growth. While routine measures of total phosphorus (avg: 25–33 µg l−1) and chlorophyll a (avg: 12–15 µg l−1) were indicative of eutrophy, mean phytoplankton biomass in 2004 (2.0 g m−3) and 2006 (2.2 g m−3) suggested mesotrophic conditions. However, the appearance of algal blooms in the summer of 2006 was an obvious indicator of cultural eutrophication. With respect to the microbial – planktonic foodweb, the organic carbon pool increased from a mean of 757.5 mg C m−3 in 2004 to 1160.3 mg C m−3 in 2006 and this increase was almost evenly split between autotrophs (198.7 mg C m−3) and heterotrophs (204.1 mg C m−3). The increased autotrophic carbon is readily attributable to the observed algal blooms driven by warmer temperatures and higher concentrations of soluble reactive phosphorus. However, the increase in heterotrophic carbon, primarily heterotrophic nanoflagellates, was apparent from the earliest observations in 2006 and remained consistently high throughout the year. We hypothesize that the increased heterotrophic carbon was the consequence of increased allochthonous carbon being shunted through the microbial foodweb; the energy generated was not likely transferred to zooplankton and passed on to higher trophic levels. More research into the dynamics of allochthonous and autochthonous carbon in eutrophic environments is called for.

Acanthoecid choanoflagellates from the Atlantic Arctic Region - a baseline study.

The examination and statistical analysis of loricate choanoflagellate material collected from Greenland waters during the period 1988–1998 represents a de facto baseline study of heterotrophic nanoflagellates from the Atlantic Arctic Region. The geographic sites sampled are Disko Bay (West Greenland) and the high-arctic North-East Water (NEW) and North Water (NOW) polynya. The analyses encompass close to 50 taxa. Some of these are described as new species, i.e. Acanthocorbis glacialis, A. reticulata and Diaphanoeca dilatanda. Two distinct clusters of species that are separated in time and space occur at all three sampling sites. A PCA analysis of NEW and NOW data points to that one community is linked to e.g. an early season high nutrient and low phytoplankton biomass scenario, whereas the other is predominant when nutrient levels are exhausted and the phytoplankton biomass high or declining. The material additionally allows for a comprehensive examination of e.g. the Cosmoeca ventricosa morphological variability encountered, as well as puts on record bimodal size variability within a number of species.