Evolutionary transition towards permanent chloroplasts? - Division of kleptochloroplasts in starved cells of two species of <i>Dinophysis</i> (Dinophyceae).

Species within the marine toxic dinoflagellate genus Dinophysis are phagotrophic organisms that exploit chloroplasts (kleptochloroplasts) from other protists to perform photosynthesis. Dinophysis spp. acquire the kleptochloroplasts from the ciliate Mesodinium rubrum, which in turn acquires the chloroplasts from a unique clade of cryptophytes. Dinophysis spp. digest the prey nuclei and all other cell organelles upon ingestion (except the kleptochloroplasts) and they are therefore believed to constantly acquire new chloroplasts as the populations grow. Previous studies have, however, indicated that Dinophysis can keep the kleptochloroplasts active during long term starvation and are able to produce photosynthetic pigments when exposed to prey starvation. This indicates a considerable control over the kleptochloroplasts and the ability of Dinophysis to replicate its kleptochloroplasts was therefore re-investigated in detail in this study. The kleptochloroplasts of Dinophysis acuta and Dinophysis acuminata were analyzed using confocal microscopy and 3D bioimaging software during long term starvation experiments. The cell concentrations were monitored to confirm cell divisions and samples were withdrawn each time a doubling had occurred. The results show direct evidence of kleptochloroplastidic division and that the decreases in total kleptochloroplast volume, number of kleptochloroplasts and number of kleptochloroplast centers were not caused by dilution due to cell divisions. This is the first report of division of kleptochloroplasts in any protist without the associated prey nuclei. This indicates that Dinophysis spp. may be in a transitional phase towards possessing permanent chloroplasts, which thereby potentially makes it a key organism to understand the evolution of phototrophic protists

Resilience to temperature and pH changes in a future climate change scenario in six strains of the polar diatom Fragilariopsis cylindrus

The effects of ocean acidification and increased temperature on physiology of six strains of the polar diatom Fragilariopsis cylindrus from Greenland were investigated. Experiments were performed under manipulated pH levels (8.0, 7.7, 7.4, and 7.1) and different temperatures (1, 5, and 8 °C) to simulate changes from present to plausible future levels. Each of the 12 scenarios was run for 7 days, and a significant interaction between temperature and pH on growth was detected. By combining increased temperature and acidification, the two factors counterbalanced each other, and therefore no effect on the growth rates was found. However, the growth rates increased with elevated temperatures by 20-50% depending on the strain. In addition, a general negative effect of increasing acidification on growth was observed. At pH 7.7 and 7.4, the growth response varied considerably among strains. However, a more uniform response was detected at pH 7.1 with most of the strains exhibiting reduced growth rates by 20-37% compared to pH 8.0. It should be emphasized that a significant interaction between temperature and pH was found, meaning that the combination of the two parameters affected growth differently than when considering one at a time. Based on these results, we anticipate that the polar diatom F. cylindrus will be unaffected by changes in temperature and pH within the range expected by the end of the century. In each simulated scenario, the variation in growth rates among the strains was larger than the variation observed due to the whole range of changes in either pH or temperature. Climate change may therefore not affect the species as such, but may lead to changes in the population structure of the species, with the strains exhibiting high phenotypic plasticity, in terms of temperature and pH tolerance towards future conditions, dominating the population

Accumulation, transformation and breakdown of DSP toxins from the toxic dinoflagellate Dinophysis acuta in blue mussels, Mytilus edulis

Okadaic acid (OA), dinophysistoxins (DTX) and pectenotoxins (PTX) produced by the dinoflagellates Dinophysis spp. can accumulate in shellfish and cause diarrhetic shellfish poisoning upon human consumption. Shellfish toxicity is a result of algal abundance and toxicity as well as accumulation and depuration kinetics in mussels. We mass-cultured Dinophysis acuta containing OA, DTX-1b and PTX-2 and fed it to the blue mussel, Mytilus edulis under controlled laboratory conditions for a week to study toxin accumulation and transformation. Contents of OA and DTX-1b in mussels increased linearly with incubation time, and the net toxin accumulation was 66% and 71% for OA and DTX-1b, respectively. Large proportions (≈50%) of both these toxins were transformed to fatty acid esters. Most PTX-2 was transformed to PTX-2 seco-acid and net accumulation was initially high, but decreased progressively throughout the experiment, likely due to esterification and loss of detectability. We also quantified depuration during the subsequent four days and found half-life times of 5–6 days for OA and DTX-1b. Measurements of dissolved toxins revealed that depuration was achieved through excreting rather than metabolizing toxins. This is the first study to construct a full mass balance of DSP toxins during both accumulation and depuration, and we demonstrate rapid toxin accumulation in mussels at realistic in situ levels of Dinophysis. Applying the observed accumulation and depuration kinetics, we model mussel toxicity, and demonstrate that a concentration of only 75 Dinophysis cells l−1 is enough to make 60 mm long mussels exceed the regulatory threshold for OA equivalents

Ecophysiological traits of mixotrophic Strombidium spp

This is a pre-copyedited, author-produced version of an article accepted for publication in Journal of Plankton Research following peer review. The version of record Maselli, M., Altenburger, A., Stoecker, D. K. & Hansen, P. J. (2020). Ecophysiological traits of mixotrophic Strombidium spp. Journal of Plankton Research, 42(5), 485-496 is available online at: https://doi.org/10.1093/plankt/fbaa041.Ciliates represent an important trophic link between nanoplankton and mesoplankton. Many species acquire functional chloroplasts from photosynthetic prey, being thus mixotrophs. Little is known about which algae they exploit, and of the relevance of inorganic carbon assimilation to their metabolism. To get insights into these aspects, laboratory cultures of three mixotrophic Strombidium spp. were established and 35 photosynthetic algal species were tested as prey. The relative contributions of ingestion and photosynthesis to total carbon uptake were determined, and responses to prey starvation were studied. Ciliate growth was supported by algal species in the 2–12 μm size range, with cryptophytes and chlorophytes being the best prey types. Inorganic carbon incorporation was only quantitatively important when prey concentration was low (3–100 μgCL−1), when it led to increased gross growth efficiencies. Chla specific inorganic carbon uptake rates were reduced by 60–90% compared to that of the photosynthetic prey. Inorganic carbon uptake alone could not sustain survival of cultures and ciliate populations declined by 25–30% during 5 days of starvation. The results suggest that mixotrophy in Strombidium spp. may substantially bolster the efficiency of trophic transfer when biomass of small primary producers is low

Modelling succession of key resource harvesting traits of mixotrophic plankton populations

Unicellular eukaryotes make up the base of the ocean food web and exist as a continuum in trophic strategy from pure heterotrophy (phagotrophic zooplankton) to pure photoautotrophy (‘phytoplankton'), with a dominance of mixotrophic organisms combining both strategies. Here we formulate a trait-based model for mixotrophy with three key resource-harvesting traits: photosynthesis, phagotrophy and inorganic nutrient uptake, which predicts the trophic strategy of species throughout the seasonal cycle. Assuming that simple carbohydrates from photosynthesis fuel respiration, and feeding primarily provides building blocks for growth, the model reproduces the observed light-dependent ingestion rates and species-specific growth rates with and without prey from the laboratory. The combination of traits yielding the highest growth rate suggests high investments in photosynthesis, and inorganic nutrient uptake in the spring and increased phagotrophy during the summer, reflecting general seasonal succession patterns of temperate waters. Our trait-based model presents a simple and general approach for the inclusion of mixotrophy, succession and evolution in ecosystem models

Functional ecology of aquatic phagotrophic protists - Concepts, limitations, and perspectives

Special issue Current trends in protistology – results from the VII ECOP - ISOP Joint Meeting 2015.-- 25 pages, 6 figures, 1 tableFunctional ecology is a subdiscipline that aims to enable a mechanistic understanding of patterns and processes from the organismic to the ecosystem level. This paper addresses some main aspects of the process-oriented current knowledge on phagotrophic, i.e. heterotrophic and mixotrophic, protists in aquatic food webs. This is not an exhaustive review; rather, we focus on conceptual issues, in particular on the numerical and functional response of these organisms. We discuss the evolution of concepts and define parameters to evaluate predator–prey dynamics ranging from Lotka–Volterra to the Independent Response Model. Since protists have extremely versatile feeding modes, we explore if there are systematic differences related to their taxonomic affiliation and life strategies. We differentiate between intrinsic factors (nutritional history, acclimatisation) and extrinsic factors (temperature, food, turbulence) affecting feeding, growth, and survival of protist populations. We briefly consider intraspecific variability of some key parameters and constraints inherent in laboratory microcosm experiments. We then upscale the significance of phagotrophic protists in food webs to the ocean level. Finally, we discuss limitations of the mechanistic understanding of protist functional ecology resulting from principal unpredictability of nonlinear dynamics. We conclude by defining open questions and identifying perspectives for future research on functional ecology of aquatic phagotrophic protistsHA was supported by grants from the German Research Foundation (DFG; AR 288/16) and from the Federal Ministry for Education and Research (BMBF: 03G0237B; 02WRM1364D). Project FERMI (CGL2014-59227-R) was awarded to AC from the Spanish Ministry of Economy and Competitiveness. RA was supported by the the European Union's Horizon 2020 research and innovation programme (Marie Sklodowska-Curie grant agreement No 658882). PJH was supported by the Danish Council for independent Reseach, project DDF-4181-00484. TW was financially supported by the Austrian Science Fund (FWF, projects P20118-B17 and P20360-B17). DJSM received no support for his efforts on this study, other than his salary provided by the University of LiverpoolPeer Reviewe

Respiratory Physiology of European Plaice (<i>Pleuronectes platessa</i>) Exposed to <i>Prymnesium parvum</i>

During the last century, the blooms of the toxic haptophyte Prymnesium parvum have been responsible for massive fish kills in both aquaculture and wild populations. Despite decades of research, the ichthyotoxic properties of P. parvum, and how this alga affects fish, is still debated. Using a novel device to measure the respirometry, ventilation volume, ventilation frequency, oxygen extraction, and oxygen consumption of undisturbed European plaice (Pleuronectes platessa) were acquired during exposure to two algal species as well as hypoxia. Fourteen fish (258 &#177; 44 g) were initially exposed to severe hypoxia and left to recover for at least 48 h. Half of these fish were then exposed to known harmful concentrations of P. parvum (median &#177; standard deviation (SD); 2.6 &#215; 105 &#177; 0.6 &#215; 105 cells mL&#8722;1), while the remaining half were exposed to the non-toxic alga Rhodomonas salina (median &#177; SD; 3.2 &#215; 105 &#177; 0.7 &#215; 105 cells mL&#8722;1). During exposure to severe hypoxia, all of the fish were able to maintain oxygen consumption by increasing the ventilation volume. The results from fish that were exposed to P. parvum showed a significant decrease in oxygen extraction (median &#177; SD; 52.6 &#177; 6.9 percentage points) from pre-exposure to the end of the experiment, as opposed to fish exposed to R. salina, which were unaffected. These results indicate that suffocation affects the European plaice when exposed to P. parvum. The observed severe decrease in oxygen extraction can be ascribed to either damage of the gill epithelia or increased mucus secretion on the gills, as both would limit the transfer of oxygen, and both have been observed

The Cost of Toxicity in Microalgae: Direct Evidence From the Dinoflagellate Alexandrium

Empirical evidence of the cost of producing toxic compounds in harmful microalgae is completely lacking. Yet costs are often assumed to be high, implying substantial ecological benefits with adaptive significance exist. To study potential fitness costs of toxin production, 16 strains including three species of the former Alexandrium tamarense species complex were grown under both carbon limitation and unlimited conditions. Growth rates, levels of intracellular paralytic shellfish toxins (PSTs), and effects of lytic compounds were measured to provide trade-off curves of toxicity for both PST and lytic toxicity under high light (300 μmol photons m−2 s−1) and under low light (i.e., carbon limited; 20 μmol photons m−2 s−1). Fitness costs in terms of reduced growth rates with increasing PST content were only evident under unlimited conditions, but not under carbon limitation, in which case PST production was positively correlated with growth. The cost of production of lytic compounds was detected both under carbon limitation and unlimited conditions, but only in strains producing PST. The results may direct future research in understanding the evolutionary role and ecological function of algal toxins. The intrinsic growth rate costs should be accounted for in relation to quantifying benefits such as grazer avoidance or toxin-mediated prey capture in natural food web settings