Algal Prey Research Articles

Refurbishing the marine parasitoid order Pirsoniales with newly (re)described marine and freshwater free-living predators.

Pirsoniales is a stramenopile order composed of marine parasitoids of diatoms with unique life cycle. Until recently, a single genus, Pirsonia, uniting six species, was known. The recent identification of new free-living eukaryotrophic Pirsoniales Pirsonia chemainus, Feodosia pseudopoda, and Koktebelia satura changed our understanding of this group as exclusively parasitic. However, their cell ultrastructure and feeding preferences were not fully studied due to the death of the cultures. In this study, we re-isolated some of these Pirsoniales and established six new strains exhibiting predatory behavior, including a first freshwater representative. This allowed us to describe five new genera and species, as well as to emend the diagnosis of the order Pirsoniales. The 18S rRNA gene phylogenetic analysis revealed the position of new strains within Pirsoniales and their relationships with parasitoid relatives and environmental sequence lineages. Feeding experiments on novel Pirsoniales strains using diverse algal prey showed that they were not able to form trophosomes and auxosomes. The ability of cell aggregation in Pirsoniales was observed for the first time. One of the studied strains contained intracellular gammaproteobacteria distantly related to Coxiella. Ultrastructural analyses revealed a more complex cytoskeleton structure in Pirsoniales than previously thought and supported the monophyly of Bigyromonadea and Pseudofungi.

Prey species and abundance affect growth and photosynthetic performance of the polyphagous sea slug Elysia crispata.

Some sacoglossan sea slugs steal functional macroalgal chloroplasts (kleptoplasts). In this study, we investigated the effects of algal prey species and abundance on the growth and photosynthetic capacity of the tropical polyphagous sea slug Elysia crispata. Recently hatched sea slugs fed and acquired chloroplasts from the macroalga Bryopsis plumosa, but not from Acetabularia acetabulum. However, adult sea slugs were able to switch diet to A. acetabulum, rapidly replacing the great majority of the original kleptoplasts. When fed with B. plumosa, higher feeding frequency resulted in significantly higher growth and kleptoplast photosynthetic yield, as well as a slower relative decrease in these parameters upon starvation. Longevity of A. acetabulum-derived chloroplasts in E. crispata was over twofold that of B. plumosa. Furthermore, significantly lower relative weight loss under starvation was observed in sea slugs previously fed on A. acetabulum than on B. plumosa. This study shows that functionality and longevity of kleptoplasts in photosynthetic sea slugs depend on the origin of the plastids. Furthermore, we have identified A. acetabulum as a donor of photosynthetically efficient chloroplasts common to highly specialized monophagous and polyphagous sea slugs capable of long-term retention, which opens new experimental routes to unravel the unsolved mysteries of kleptoplasty.

Mixotrophic lifestyle of the ichthyotoxic haptophyte, Prymnesium parvum, offered different sources of phosphorus

Many harmful algae are mixoplanktonic, i.e. they combine phototrophy and phagotrophy, and this ability may explain their ecological success, especially when environmental conditions are not optimal for autotrophic growth. In this study, laboratory experiments were conducted with the mixotrophic and ichthyotoxic haptophyte Prymnesium parvum (strain CCAP 946/6) to test the effects of phosphorus (P) sufficiency and deficiency on its growth rate, phagotrophic and lytic activities. P-deficient P. parvum cultures were grown without or with addition of P in the form of inorganic phosphorus (nutrients) and/or living algal prey (i.e. the cryptophyte Teleaulax amphioxeia). The ingestion rate of P. parvum and prey mortality rate were calculated using flow cytometry measurements based on pigment-derived-fluorescence to distinguish between prey, predators and digesting predators. The first aim of the study was to develop a method taking into account the rate of digestion, allowing the calculation of ingestion rates over a two-week period. Growth rates of P. parvum were higher in the treatments with live prey, irrespective of the concentration of inorganic P, and maximum growth rates were found when both inorganic and organic P in form of prey were added (0.79 ± 0.07 d-1). In addition, the mortality rate of T. amphioxeia induced by lytic compounds was 0.2 ± 0.02 d-1 in the P-deficient treatment, while no mortality was observed under P-sufficiency in the present experiments. This study also revealed the mortality due to cell lysis exceeded that of prey ingestion. Therefore, additional experiments were conducted with lysed prey cells. When grown with debris from prey cells, the mean growth rate of P. parvum was similar to monocultures without additions of prey debris (0.30 ± 0.1 vs. 0.38 ± 0.03 d-1), suggesting that P. parvum is able to grow on prey debris, but not as fast as with live prey. These results provide the first quantitative evidence over two weeks of experiment that ingestion of organic P in the form of living prey and/or debris of prey plays an important role in P. parvum growth and may explain its ecological success in a nutrient-limited environments.

Euglenozoan kleptoplasty illuminates the early evolution of photoendosymbiosis

Kleptoplasts (kP) are distinct among photosynthetic organelles in eukaryotes (i.e., plastids) because they are routinely sequestered from prey algal cells and function only temporarily in the new host cell. Therefore, the hosts of kleptoplasts benefit from photosynthesis without constitutive photoendosymbiosis. Here, we report that the euglenozoan Rapaza viridis has only kleptoplasts derived from a specific strain of green alga, Tetraselmis sp., but no canonical plastids like those found in its sister group, the Euglenophyceae. R. viridis showed a dynamic change in the accumulation of cytosolic polysaccharides in response to light-dark cycles, and 13C isotopic labeling of ambient bicarbonate demonstrated that these polysaccharides originate insitu via photosynthesis; these data indicate that the kleptoplasts of R. viridis are functionally active. We also identified 276 sequences encoding putative plastid-targeting proteins and 35 sequences of presumed kleptoplast transporters in the transcriptome of R. viridis. These genes originated in a wide range of algae other than Tetraselmis sp., the source of the kleptoplasts, suggesting a long history of repeated horizontal gene transfer events from different algal prey cells. Many of the kleptoplast proteins, as well as the protein-targeting system, in R. viridis were shared with members of the Euglenophyceae, providing evidence that the early evolutionary stages in the green alga-derived secondary plastids of euglenophytes also involved kleptoplasty.

Life-history omnivory in the fairy shrimp Branchinecta orientalis (Branchiopoda: Anostraca)

Very little is known about the feeding of naupliar and juvenile life stages of omnivorous fairy shrimps (Crustacea: Anostraca). Here, we aim to reveal whether the fairy shrimp Branchinecta orientalis is an ontogenetic omnivore and at which age and ontogenetic stage they gain the ability to feed on zooplankton. We assess how food uptake rates change with age until reaching maturity by providing algae (pico- and nanoplanktonic unicellular algae) and zooplankton (rotifers and copepod nauplii) as food in individual experiments. We found that the fairy shrimp B. orientalis started to feed on both types of algal prey immediately after hatching. Nanoplanktonic algae likely represented the most important food source until reaching maturity. Moreover, fairy shrimps started to feed on zooplankton already when they were 7 days old. Slow-moving rotifers gradually gained importance in the fairy shrimp diet with time. Our results reveal an ontogenetic change in the prey spectrum of fairy shrimp. The systematic shift towards omnivory likely affects both phyto- and zooplankton community composition, possibly contributing to temporal changes in food web dynamics in fairy shrimp habitats, and temporary ponds, which may warrant more detailed investigations in future studies.

Impaired grazing of marine protozoa in sub-lethal exposure to the water accommodated fraction of crude oil and dispersant

Despite the advances in safety technology and the improved implementation of precautionary measures, crude oil pollution has been occurring in the oceans globally. The water accommodated fraction (WAF) of crude oil and chemical dispersant are hypothesized to cause sub-lethal adverse effects on marine protists that are pivotal consumers of primary production. Exposure experiments were conducted to investigate the effects of crude oil and dispersant pollutants on the growth and grazing, separately, of protozoa species in cultures. In exposure to 0–30 μL L−1 of chemically enhanced WAF (CEWAF), the heterotrophic dinoflagellate Protoperidinium sp. and the ciliate Metacylis sp. showed slower positive population growth or negative population growth even at low concentrations. The dose-response model showed that Protoperidinium sp. and Metacylis sp. were highly susceptible to the CEWAF toxicity (median inhibition concentrations (IC50) at 1.1 and 5.9 μL L−1, respectively) while one algal species Ditylum brightwellii was relatively tolerant to the toxicity (IC50 at 168.7 μL L−1). With suppressed growth and impaired grazing of the protozoan species at high CEWAF concentrations, accumulation of their algal prey in culture containers was observed, as reflected by higher final:initial prey ratios at high CEWAF concentrations. Additionally, exposure experiments to the treatments of WAF, dispersant alone (Disp), and CEWAF of the same concentration revealed that the heterotrophic dinoflagellate Oxyrrhis marina had reduced bulk grazing impact towards its algal prey population in all three treatments when compared to the control treatment (i.e., grazing mortality of prey at 1.05 d−1). Similarly, Protoperidinium sp. and Metacylis sp. had reduced per capita prey ingestion rates in exposure to WAF and CEWAF when compared to the control treatments. This study provides experimental evidence for the potential link between impaired grazing activities and the formation of algal blooms in sub-lethal exposure to crude oil pollutants.

Cascading effects of prey identity on gene expression in a kleptoplastidic ciliate.

Kleptoplastidic, or chloroplast stealing, lineages transiently retain functional photosynthetic machinery from algal prey. This machinery, and its photosynthetic outputs, must be integrated into the host's metabolism, but the details of this integration are poorly understood. Here, we study this metabolic integration in the ciliate Mesodinium chamaeleon, a coastal marine species capable of retaining chloroplasts from at least six distinct genera of cryptophyte algae. To assess the effects of feeding history on ciliate physiology and gene expression, we acclimated M. chamaeleon to four different types of prey and contrasted well-fed and starved treatments. Consistent with previous physiological work on the ciliate, we found that starved ciliates had lower chlorophyll content, photosynthetic rates, and growth rates than their well-fed counterparts. However, ciliate gene expression mirrored prey phylogenetic relationships rather than physiological status, suggesting that, even as M. chamaeleon cells were starved of prey, their overarching regulatory systems remained tuned to the prey type to which they had been acclimated. Collectively, our results indicate a surprising degree of prey-specific host transcriptional adjustments, implying varied integration of prey metabolic potential into many aspects of ciliate physiology.

Hawksbill presence and habitat suitability of a marine reserve in Honduras

Marine protected areas (MPAs) have been established to decrease the negative impacts of human activities on important marine habitats. Several sea turtle species reside within MPAs; however, analyses of habitat suitability for sea turtles within many MPAs is typically lacking. Habitat modeling and species mapping have become essential tools in determining areas suitable for species inhabitance. MaxEnt is a popular machine-learning program that uses presence-only occurrence data and is considered the best modeling method in discriminating between suitable and unsuitable habitats. Using MaxEnt, we created species distribution models (SDMs) from presence-only occurrence points for juvenile hawksbills and three environmental variables: boat traffic intensities, prey item distributions, and benthic characterizations, within the Sandy Bay West End Marine Reserve (SBWEMR) in Roatán, Honduras. Area under the receiving operator curve (AUC) values were greater than 0.75, indicating high model accuracy. Jackknife evaluations indicated algal prey item Dictyota spp., boat traffic intensity, and Geodia neptuni sponge abundance had the greatest influence on species distribution. Areas of suitability shifted based on time of day and ranged from a low probability of hawksbill presence (0.0) to a high probability of hawksbill presence (0.8). Regardless of time of day, the SBWEMR is classified as at least moderately suitable (>0.2–0.4), with least suitable habitat (0–0.2) prevalent in the most northeastern region of the reserve. The greatest suitability (>0.6–1) was evident within the mid-region and southern most regions of the reserve. Probability of hawksbill presence increased in accordance with increasing boat traffic, although there was no clear causative relationship between the two. Although the reserve overall is mostly moderately suitable, the SBWEMR remains an important local recruitment and foraging ground for juvenile hawksbills. Hawksbills have been considered resilient, continually functioning beneath their operational limits. However, with continued global depletion of essential habitats over time and lack of effective MPA management, areas of habitat suitable for hawksbill inhabitance may decrease below levels sufficient for hawksbill presence in protected areas of the Western Caribbean.

Ultrastructure of Myzocytosis and Cyst Formation, and the Role of Actin in Tubular Tether Formation in Colpodella sp. (ATCC 50594).

Free-living relatives of the Apicomplexa such as Colpodella species, Alphamonas species, and Voromonas pontica are predators that prey on ciliate, bodonid, and algal prey using the process of myzocytosis. During myzocytosis, the pseudoconoid is used to attach to the prey leading to aspiration of cytoplasmic contents of the prey into a posterior food vacuole formed in the predator, aided by secretions from the apical complex organelles. The conoid and associated proteins are conserved among the apicomplexa. However, the organization and function of the pseudoconoid during myzocytosis are not well understood. In this study, we investigated the morphology and ultrastructure of Colpodella sp. (ATCC 50594) during the stages of myzocytosis and cyst formation in the life cycle using light microscopy and transmission electron microscopy (TEM) in order to identify the organization of the tubular tether involved in nutrient aspiration by Colpodella sp. Tubular tethers of varying lengths were identified by light microscopy. We report that initial contact by Colpodella sp. trophozoites with Parabodo caudatus prey is by an area posterior to the apical tip of the rostrum that engulfs the membrane of the prey pulling it into the cytoplasm of the predator. The tubular tether that forms contains membranes of both predator and prey and is facilitated by microtubule organization and the cytoskeleton at the point of contact. Cytochalasin D treatment of diprotist cultures resulted in morphological distortions of trophozoites and the tubular tether suggesting a role of actin in the formation of the tubular tether. This mechanism of predation may provide insight into the mode of invasion observed in pathogenic apicomplexan zoites during host cell entry.

Dimethyl sulfide mediates microbial predator-prey interactions between zooplankton and algae in the ocean.

Phytoplankton are key components of the oceanic carbon and sulfur cycles1. During bloom events, some species can emit large amounts of the organosulfur volatile dimethyl sulfide (DMS) into the ocean and consequently the atmosphere, where it can modulate aerosol formation and affect climate2,3. In aquatic environments, DMS plays an important role as a chemical signal mediating diverse trophic interactions. Yet, its role in microbial predator-prey interactions remains elusive with contradicting evidence for its role in either algal chemical defence or in the chemo-attraction of grazers to prey cells4,5. Here we investigated the signalling role of DMS during zooplankton-algae interactions by genetic and biochemical manipulation of the algal DMS-generating enzyme dimethylsulfoniopropionate lyase (DL) in the bloom-forming alga Emiliania huxleyi6. We inhibited DL activity in E. huxleyi cells in vivo using the selective DL-inhibitor 2-bromo-3-(dimethylsulfonio)-propionate7 and overexpressed the DL-encoding gene in the model diatom Thalassiosira pseudonana. We showed that algal DL activity did not serve as an anti-grazing chemical defence but paradoxically enhanced predation by the grazer Oxyrrhis marina and other microzooplankton and mesozooplankton, including ciliates and copepods. Consumption of algal prey with induced DL activity also promoted O. marina growth. Overall, our results demonstrate that DMS-mediated grazing may be ecologically important and prevalent during prey-predator dynamics in aquatic ecosystems. The role of algal DMS revealed here, acting as an eat-me signal for grazers, raises fundamental questions regarding the retention of its biosynthetic enzyme through the evolution of dominant bloom-forming phytoplankton in the ocean.

Interactions Between the Kleptoplastidic Dinoflagellate Shimiella gracilenta and Several Common Heterotrophic Protists

The newly described dinoflagellate, Shimiella gracilenta, is known to survive for approximately 1 month on the plastids of ingested prey cells during starvation, indicating kleptoplastidy. To understand the population dynamics of this dinoflagellate in marine planktonic food webs, its growth and mortality rate due to predation should be assessed. Thus, we investigated the feeding occurrence of eight common heterotrophic protists on S. gracilenta. We also determined the growth and ingestion rates of Oxyrrhis marina and the naked ciliate, Rimostrombidium sp. on S. gracilenta as a function of the prey concentration. The common heterotrophic dinoflagellates (HTDs) Gyrodinium dominans, O. marina, and Pfiesteria piscicida and a naked ciliate Rimostrombidium sp. were able to feed on S. gracilenta; whereas the HTDs Aduncodinium glandula, Gyrodinium jinhaense, Oblea rotunda, and Polykrikos kofoidii were not. Shimiella gracilenta supported positive growth of O. marina and Rimostrombidium sp. but did not support that of G. dominans and P. piscicida. With increasing prey concentrations, the growth and ingestion rates of O. marina and Rimostrombidium sp. on S. gracilenta increased and became saturated. The maximum growth rates of O. marina and Rimostrombidium sp. on S. gracilenta were 0.645 and 0.903 day−1, respectively. Furthermore, the maximum ingestion rates of O. marina and Rimostrombidium sp. on S. gracilenta were 0.11 ng C predator day−1 (1.6 cells predator−1 day−1) and 35 ng C predator day−1 (500 cells predator−1 day−1), respectively. The maximum ingestion rate of O. marina on S. gracilenta was lower than that on any other algal prey reported to date, although its maximum growth rate was moderate. In conclusion, S. gracilenta had only a few common heterotrophic protist predators but could support moderate growth rates of the predators. Thus, S. gracilenta may not be a common prey species for diverse heterotrophic protists but may be a suitable prey for a few heterotrophic protists.

The amoebae of Idionectes vortex (Cutosea, Amoebozoa): Motility, cytoskeleton architecture and extracellular scales.

The Cutosea represent a deep-branching lineage within the phylum Amoebozoa that is still relatively poorly explored. Currently, there are four cutosean representatives known - the monotypic genera Armaparvus, Idionectes, Sapocribrum, and Squamamoeba - with marked genetic distances. Idionectes vortex is the deepest-branching species and differs markedly from the other Cutosea in ecology, life history, and most importantly, in its ability to form a flagellated swarmer with an exceptional swimming mechanism. As far as we know, the other Cutosea lack flagella and rather represent small, marine amoebae with a characteristic cell coat. The present paper focuses on the amoeboid life history stage of the algivorous amoeboflagellate Idionectes vortex to provide data for a first in-depth comparison with other Cutosea and to document structural specialties. The amoeboid stage of Idionectes is mainly associated with the specific feeding process, that is, the interaction with algal prey cells and phagocytosis of protoplast material. Yet, the present data from time-lapse microscopy, cytochemical stainings, and electron microscopy demonstrate clear similarities with the other cutosean species concerning amoeboid locomotion and cell coat ultrastructure. Furthermore, Idionectes amoebae exhibit a well-developed microtubular cytoskeleton, and an unusual basal apparatus that seems to undergo marked changes during the life history of this exceptional amoebozoan.

Heterotrophic Dinoflagellate Growth and Grazing Rates Reduced by Microplastic Ingestion

Microplastics are ubiquitous contaminants in marine ecosystems worldwide, threatening fisheries production, food safety, and human health. Ingestion of microplastics by fish and large zooplankton has been documented, but there are few studies focusing on single-celled marine predators, including heterotrophic dinoflagellates. In laboratory experiments, the heterotrophic dinoflagellate species Oxyrrhis marina and Gyrodinium sp. readily ingested both algal prey and polystyrene microplastic spheres (2.5–4.5 μm), while Protoperidinium sp. did not ingest microplastics. Compared to algae-only fed dinoflagellates, those that ingested microplastics had growth rates reduced by 25–35% over the course of 5 days. Reduced growth resulted in a 30–50% reduction of secondary production as measured as predator biomass. Ingestion rates of algal prey were also reduced in the microplastic treatments. When given a mixture of microplastics and algal prey, O. marina displayed a higher selectivity for algal prey than Gyrodinium sp. Observations in the coastal ocean showed that phylogenetically diverse taxa ingested microplastic beads, and thus heterotrophic dinoflagellates could contribute to trophic transfer of microplastics to higher trophic levels. The results of this study may suggest that continued increase in microplastic pollution in the ocean could lead to reduced secondary production of heterotrophic protists due to microplastic ingestion, altering the flow of energy and matter in marine microbial food webs.

Prey preferences of invasive (Hemigrapsus sanguineus, H. takanoi) and native (Carcinus maenas) intertidal crabs in the European Wadden Sea

AbstractInvasive predators can have wide-ranging effects on invaded ecosystems and identifying the prey spectra and preferences of invaders are important steps in assessing their potential impacts on native biota. In this study, we investigated prey preferences of two invasive crab species (Hemigrapsus sanguineus and Hemigrapsus takanoi) that recently invaded Europe's shores and compared them with preferences of native shore crabs (Carcinus maenas) of similar size. In laboratory experiments, all three crab species preferred animal over algal prey. In general, sessile mussels (Mytilus edulis) were preferred over motile amphipods (Gammarus locusta) by all three crab species but amphipod predation was lower in the invasive compared with the native crabs. For the two invasive crab species, this pattern was the same in treatments where prey was offered separately (no-choice treatments) or simultaneously (choice treatments), while for the native crabs, mussel preference disappeared in choice treatments. The general preference of mussels by all three crab species suggests that local invasions of crabs most likely lead to increased competition among crabs. In addition, given that local densities of invasive crabs are often much higher than those of native crabs, predation pressure on native mussels can be expected to strongly increase at invaded sites. In contrast, local predation pressure on amphipods may be less affected by the crab invasions. Further field studies are needed to establish the magnitude of competition and predation pressure exerted by the invaders under natural conditions.

Metabolic Reliance on Photosynthesis Depends on Both Irradiance and Prey Availability in the Mixotrophic Ciliate, Strombidium cf. basimorphum.

Many species of the ciliate genus Strombidium can acquire functional chloroplasts from a wide range of algal prey and are thus classified as generalist non-constitutive mixotrophs. Little, however, is known about the influence of irradiance and prey availability on their ability to exploit the photosynthetic potential of the chloroplasts, and how this may explain their spatial and temporal distribution in nature. In this study, inorganic carbon uptake, growth, and ingestion rates were measured for S. cf. basimorphum under three different irradiances (10, 40, and 120 μmol photons m–2 s–1) when acclimated to three different prey densities (5 × 103, 1 × 104, and 4 × 104 cells mL–1), as well as when allowed to deplete the prey. After prey depletion, cultures survived without prey longest (∼6 days) at the medium irradiance treatment (40 μmol photons m–2 s–1), while ciliate density, inorganic carbon uptake rates, and cellular chl-a content declined fastest at the highest irradiance treatment. This indicates that the ciliates may be unable to maintain the chloroplasts functionally without replacement at high irradiances. Ingestion rates were not shown to be significantly influenced by irradiance. The maximum gross growth efficiency (GGE) in this study (1.1) was measured in cultures exposed to the medium test irradiance and lowest prey density treatment (5 × 103 cells mL–1). The relative contribution of inorganic carbon uptake to the ciliate carbon budget was also highest in this treatment (42%). A secondary GGE peak (0.99) occurred when cultures were exposed to the highest test irradiance and the medium prey density. These and other results suggest that S. cf. basimorphum, and other generalist non-constitutive mixotrophs, can flexibly exploit many different environmental conditions across the globe.

Preferential grazing and repackaging of small polyethylene microplastic particles (≤ 5 μm) by the ciliate Sterkiella sp.

Microplastic (MP) particles are pollutants of global concern and are ubiquitously distributed in the ocean by physical and biological processes. It has been shown that zooplankton can ingest MP yet the interaction between ciliates and MP is still poorly understood. The discrimination and preferential uptake of MP rather than algal prey by ciliates was assessed in this study. The ciliate Sterkiella sp. was fed a diet that consisted of only Isochrysis galbana or a mixture of the same algae and similarly sized polyethylene beads in a 1:3 ratio. Significant, preferential MP grazing was observed in the Plastic-Algae treatment, which is the first reported evidence of proto-zooplankton preferentially ingesting MP over algal food. The mixed treatment contained fecal pellets with embedded MP. Preferential uptake of MP suggests that Sterkiella sp. is capable of ingesting and then “repackaging” MP that would otherwise be too small for larger taxa. This process would thus offer a mechanism for the reintroduction of MP into different compartments of the marine food web. As a consequence, it is necessary to account for small-sized MP (<5 μm) particles, that may have additional and yet unknown, impacts on marine food webs.

Limits to the cellular control of sequestered cryptophyte prey in the marine ciliate Mesodinium rubrum.

The marine ciliate Mesodinium rubrum is famous for its ability to acquire and exploit chloroplasts and other cell organelles from some cryptophyte algal species. We sequenced genomes and transcriptomes of free-swimming Teleaulax amphioxeia, as well as well-fed and starved M. rubrum in order to understand cellular processes upon sequestration under different prey and light conditions. From its prey, the ciliate acquires the ability to photosynthesize as well as the potential to metabolize several essential compounds including lysine, glycan, and vitamins that elucidate its specific prey dependency. M. rubrum does not express photosynthesis-related genes itself, but elicits considerable transcriptional control of the acquired cryptophyte organelles. This control is limited as light-dependent transcriptional changes found in free-swimming T. amphioxeia got lost after sequestration. We found strong transcriptional rewiring of the cryptophyte nucleus upon sequestration, where 35% of the T. amphioxeia genes were significantly differentially expressed within well-fed M. rubrum. Qualitatively, 68% of all genes expressed within well-fed M. rubrum originated from T. amphioxeia. Quantitatively, these genes contributed up to 48% to the global transcriptome in well-fed M. rubrum and down to 11% in starved M. rubrum. This tertiary endosymbiosis system functions for several weeks, when deprived of prey. After this point in time, the ciliate dies if not supplied with fresh prey cells. M. rubrum represents one evolutionary way of acquiring photosystems from its algal prey, and might represent a step on the evolutionary way towards a permanent tertiary endosymbiosis.

The loss of mixotrophy in Alexandrium pseudogonyaulax: Implications for trade‐offs between toxicity, mucus trap production, and phagotrophy

AbstractTwo important competitive traits in protists, particularly in harmful algal bloom forming species, are lytic toxicity and mixotrophy. This study focuses on Alexandrium pseudogonyaulax, a dinoflagellate species that is both toxic and mixotrophic, and in addition uses a mucus trap to immobilize and capture potential prey. A single strain of A. pseudogonyaulax was subcultured and grown under two different conditions; one subculture was grown autotrophically, while the other was periodically offered algal prey. After 3 yr, the substrain that was fed remained an avid mixotroph, while the autotrophically grown substrain almost completely lost its ability to feed. The emergence of these two “substrains,” with different nutritional strategies, allowed us to investigate possible trade‐offs between lytic toxicity, mixotrophy, and mucus trap production. Both substrains were still capable of producing mucus traps, and successfully captured other cells. Although the nonfeeding substrain lost its ability to feed, it was more lytic than the feeding substrain, which may suggest a trade‐off between lytic toxicity and feeding ability. However, this increased toxicity was not enough to outcompete other faster growing competitors such as Teleaulax acuta and Heterocapsa rotundata. In contrast, the feeding substrain was able to inhibit the growth of these same competing species. The results indicate that for A. pseudogonyaulax, the benefits of phagotrophy far outweigh those of lytic toxicity or the mucus trap alone. The nonfeeding strain used here provides evidence of the loss of mixotrophy under culture conditions, highlighting one of the obstacles in the study of mixotrophic protists.

Spatial-temporal distributions of the newly described mixotrophic dinoflagellate Gymnodinium smaydae in Korean coastal waters

Gymnodinium smaydae is a newly described mixotrophic dinoflagellate that feeds on only Heterocapsa spp. and Scrippsiella acuminata among 19 tested algal prey. It is one of the fastest growing dinoflagellates when feeding, but does not grow well without prey. To investigate its spatial-temporal distributions in Korean waters, we quantified its abundance in water samples that were seasonally collected from 28 stations along the Korean Peninsula from April 2015 to October 2018, using quantitative real-time polymerase chain reactions. This dinoflagellate had a wide distribution, as reflected by the detection of G. smaydae cells at 23 of the sampling stations. However, this distribution had a strong seasonality; it was detected at 21 stations in the summer and only one station in winter. The abundance of G. smaydae was significantly and positively correlated with chlorophyll a concentration as well as with water temperature. However, there were no significant correlations between the abundance of G. smaydae and salinity, concentrations of nutrients, or dissolved oxygen concentration. During the study period, G. smaydae was present when water temperatures were 7.6–28.0°C, salinities were 9.6–34.1, concentrations of NO3 were not detectable-106.0 μM, and concentrations of PO4 were not detectable-3.4 μM. The highest abundance of G. smaydae was 18.5 cells mL−1 in the coastal waters of Jinhae in July 2017 when the chlorophyll a concentration was 127 mg m−3 and water temperature was 23.8°C. Therefore, the spatial-temporal distribution of G. smaydae in Korean coastal waters may be affected by chlorophyll a concentration and water temperature. Key words: Gymnodiniaceae; harmful algal bloom; mixotrophy; protist; qPCR; red tide; specific primer; temperature

Ambient temperature and algal prey type affect essential fatty acid incorporation and trophic upgrading in a herbivorous marine copepod.

The essential fatty acids (EFA) eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) are critical nutrients for all organisms, and the temperature sensitivity of their trophic transfer in marine systems is of concern because of rising ocean temperatures. Laboratory-reared copepodites of the marine calanoid Calanus finmarchicus were used to test the effects of temperature (at 6°C, 12°C and increasing temperature stress) and prey type (the dinoflagellate Heterocapsa triquetra and the diatom Thalassiosira weissflogii) on the extent and efficiency of dietary EPA and DHA incorporation from phytoplankton to copepods in a set of feeding experiments using 13C labelling. Temperature was a significant determinant of C. finmarchicus copepodites' EFA incorporation and gross growth efficiency, defined as the fraction of ingested EFA retained in copepod tissue. Ingestion and incorporation of both EFA were higher at warmer temperature, except in the case of DHA in copepods feeding on diatoms. DHA-associated growth efficiency was higher at the higher temperature for copepodites consuming the dinoflagellate, but temperature-related variation in algal EFA content was also a predictive factor. Moreover, our results strongly suggest that copepodites are capable of synthesizing EPA when consuming an EPA-depleted diet. Our study implies that the copepod link of marine food webs is resilient in terms of EFA transfer when confronted with alterations of ambient temperature and prey type availability. Measurements presented here are critical for estimating how EFA transfer dynamics respond to intra- and interannual environmental variability. This article is part of the theme issue 'The next horizons for lipids as 'trophic biomarkers': evidence and significance of consumer modification of dietary fatty acids'.