Food selectivity and grazing impact on toxic Dinophysis spp. by copepods feeding on natural plankton assemblages

Food selectivity and grazing impact by Acartia bifilosa, Temora longicornis and Centropages typicus on Dinophysis spp. plankton assemblages were experimentally investigated in the Baltic Sea. Toxin analyses were carried out on phyto- and zooplankton-dominated size fractions from field-collected samples to assess if toxins produced by Dinophysis spp. would end up in the zooplankton. All copepod species fed actively on toxic Dinophysis spp. (Dinophysis acuta and Dinophysis norvegica). Despite the non-selective feeding behaviour by T. longicornis and C. typicus, selectivity coefficients on D. acuta progressively decreased as food availability increased. Similar response was not observed for A. bifilosa, which displayed an even less selective behaviour. A. bifilosa had no significant negative effect on the net growth of D. norvegica at the lowest food concentration offered, whereas T. longicornis and C. typicus had significant negative effects on the net growth of D. acuta at low concentrations, similar to those observed in situ. Both species could potentially contribute as a substantial loss factor for Dinophysis spp. provided they are abundant at the onset of the blooms. The estimated grazing impact by the copepod populations was only considerable when C. typicus abundance was high and D. acuta population in sharp decline. Our results suggest that when high abundance of grazers and poor growth condition of prey populations prevail, grazing impact by copepods can contribute considerably to prevent Dinophysis spp. populations to grow or to cause the populations to decline. Okadaic acid was detected in the zooplankton size fraction at one occasion, but the concentration was far lower than the one expected from the ingested toxins. Thus, even if copepods may act as vectors of DSP-toxins to higher trophic levels, the amount of these toxins transported in the food web by copepods seems limited.     

Production and excretion of okadaic acid,pectenotoxin-2 and a novel dinophysistoxin from the DSP-causing marine dinoflagellate Dinophysis acuta – Effects of light,food availability and growth phase

Diarrhetic shellfish poisoning (DSP) toxins constitute a severe economic threat to shellfish industries and a major food safety issue for shellfish consumers. The prime producers of the DSP toxins that end up in filter feeding shellfish are species of the marine mixotrophic dinoflagellate genus Dinophysis. Intraspecific toxin contents of Dinophysis spp. vary a lot, but the regulating factors of toxin content are still poorly understood. Dinophysis spp. have been shown to sequester and use chloroplasts from their ciliate prey, and with this rare mode of nutrition, irradiance and food availability could play a key role in the regulation of toxins contents and production. We investigated toxin contents, production and excretion of a Dinophysis acuta culture under different irradiances, food availabilities and growth phases. The newly isolated strain of D. acuta contained okadaic acid (OA), pectenotoxins-2 (PTX-2) and a novel dinophysistoxin (DTX) that we tentatively describe as DTX-1b isomer. We found that all three toxins were excreted to the surrounding seawater, and for OA and DTX-1b as much as 90% could be found in extracellular toxin pools. For PTX-2 somewhat less was excreted, but often >50% was found extracellularly. This was the case both in steady-state exponential growth and in food limited, stationary growth, and we emphasize the need to include extracellular toxins in future studies of DSP toxins. Cellular toxin contents were largely unaffected by irradiance, but toxins accumulated both intra- and extracellularly when starvation reduced growth rates of D. acuta. Toxin production rates were highest during exponential growth, but continued at decreased rates when cell division ceased, indicating that toxin production is not directly associated with ingestion of prey. Finally, we explore the potential of these new discoveries to shed light on the ecological role of DSP toxins.     

Distinctly different behavioral responses of a copepod,Temora longicornis,to different strains of toxic dinoflagellates,Alexandrium spp.

Zooplankton responses to toxic algae are highly variable, even towards taxonomically closely related species or different strains of the same species. Here, the individual level feeding behavior of a copepod, Temora longicornis, was examined which offered 4 similarly sized strains of toxic dinoflagellate Alexandrium spp. and a non-toxic control strain of the dinoflagellate Protoceratium reticulatum. The strains varied in their cellular toxin concentration and composition and in lytic activity. High-speed video observations revealed four distinctly different strain-specific feeding responses of the copepod during 4 h incubations: (i) the ‘normal’ feeding behavior, in which the feeding appendages were beating almost constantly to produce a feeding current and most (90%) of the captured algae were ingested; (ii) the beating activity of the feeding appendages was reduced by ca. 80% during the initial 60 min of exposure, after which very few algae were captured and ingested; (iii) capture and ingestion rates remained high, but ingested cells were regurgitated; and (iv) the copepod continued beating its appendages and captured cells at a high rate, but after 60 min, most captured cells were rejected. The various prey aversion responses observed may have very different implications to the prey and their ability to form blooms: consumed but regurgitated cells are dead, captured but rejected cells survive and may give the prey a competitive advantage, while reduced feeding activity of the grazer may be equally beneficial to the prey and its competitors. These behaviors were not related to lytic activity or overall paralytic shellfish toxins (PSTs) content and composition and suggest that other cues are responsible for the responses.     

Copepod feeding behaviour and egg production during a dinoflagellate bloom in the North Sea

Feeding strategies of copepods were studied during a dinoflagellate-dominated bloom in the North Sea in August 2001. The aim of this study was to evaluate the importance of mesozooplankton grazing as a biological loss factor of harmful algal blooms under natural conditions. Therefore, ingestion, egestion and egg production experiments were performed with the most abundant copepod species Calanus helgolandicus, Temora longicornis and Acartia sp. feeding on the natural phytoplankton community. Dinophysis norvegica and Ceratium furca were the most abundant dinoflagellate species at the time of the experiments. Grazing experiments as well as examination of fecal pellet content revealed C. helgolandicus fed efficiently on D. norvegica. Ingestion rates up to 47 cells female⁻¹ h⁻¹ were measured and a large proportion of the C. helgolandicus fecal pellets contained intact D. norvegica cells. Dinophysis cells were rarely seen in fecal pellets produced by T. longicornis, and never observed in pellets produced by Acartia sp. The ingestion rate of C. furca, which was the dominating Ceratium species, mimicked that of D. norvegica. C. helgolandicus grazed significantly on C. furca (16 cells female⁻¹ h⁻¹), while the ingestion rate of T. longicornis was low and Acartia sp. was not able to graze on C. furca. Egg production experiments revealed that 92% of the C. helgolandicus females produced eggs. The specific egg production rate and the proportion of females producing eggs among T. longicornis were low. This field experiment clearly shows that some copepod species feed efficiently on D. norvegica and C. furca under natural conditions, which may affect the bloom development of these dinoflagellates.     

Seasonal distribution of species of the toxic dinoflagellate genus Dinophysis in Maizuru Bay (Japan), with comments on their autofluorescence and attachment of picophytoplankton

The seasonal distribution of the dinoflagellate genus, Dinophysis, in Maizuru Bay, Japan, was investigated from May 1997 to December 1999. Seven species of Dinophysis were detected, including the toxic species of Dinophysis acuminata and D. fortii. The most dominant species wasD. acuminata, detected year-around and more abundantly during periods when water temperatures were between 15 and 18 °C. No relationship was found between cell abundance of Dinophysis spp. and concentrations of dissolved inorganic nutrients. Phycoerythrin containing nano- and picophytoplankton (cryptophytes and cyanobacteria), suspected to be prey of mixotrophic Dinophysis, were enumerated simultaneously. A clear relationship was not found among the cell abundances of Dinophysis spp. and nano- and picophytoplankton. Autofluorescence of Dinophysis spp. (mainly D. acuminata and D. fortii) under blue-light excitation was usually of a yellow-orange color. Occasionally, Dinophysis spp. had red autofluorescencing and yellow-orange autofluorescencing particles. The proportion of cells possessing red autofluorescence tended to be higher in the warm season. Numerous coccoid cells of picophytoplankton (ca. 1–2 μm in diameter) were found attached to the cell surface of D. acuminata, D. fortii, etc. and food vacuole-like structures also observed. These observations suggest there is a close relationship between mixotrophic Dinophysis spp. and certain picophytoplankton. Based on our observations, the possibility that the picophytoplankton found to be attached onto Dinophysis cell surfaces are a food source for Dinophysis, and a source of DSP toxins, is discussed.     

Identification of unique microbiomes associated with harmful algal blooms caused by Alexandrium fundyense and Dinophysis acuminata

Biotic interactions dominate plankton communities, yet the microbial consortia associated with harmful algal blooms (HABs) have not been well-described. Here, high-throughput amplicon sequencing of ribosomal genes was used to quantify the dynamics of bacterial (16S) and phytoplankton assemblages (18S) associated with blooms and cultures of two harmful algae, Alexandrium fundyense and Dinophysis acuminata. Experiments were performed to assess changes in natural bacterial and phytoplankton communities in response to the filtrate from cultures of these two harmful algae. Analysis of prokaryotic sequences from ecosystems, experiments, and cultures revealed statistically unique bacterial associations with each HAB. The dinoflagellate, Alexandrium, was strongly associated with multiple genera of Flavobacteria including Owenweeksia spp., Maribacter spp., and individuals within the NS5 marine group. While Flavobacteria also dominated Dinophysis-associated communities, the relative abundance of Alteromonadales bacteria strongly co-varied with Dinophysis abundances during blooms and Ulvibacter spp. (Flavobacteriales) and Arenicella spp. (Gammaproteobacteria) were associated with cells in culture. Eukaryotic sequencing facilitated the discovery of the endosymbiotic, parasitic dinoflagellate, Amoebophrya spp., that had not been regionally described but represented up to 17% of sequences during Alexandrium blooms. The presence of Alexandrium in field samples and in experiments significantly altered the relative abundances of bacterial and phytoplankton by both suppressing and promoting different taxa, while this effect was weaker in Dinophysis. Experiments specifically revealed a negative feedback loop during blooms whereby Alexandrium filtrate promoted the abundance of the parasite, Amoebophrya spp. Collectively, this study demonstrates that HABs formed by Alexandrium and Dinophysis harbor unique prokaryotic and eukaryotic microbiomes that are likely to, in turn, influence the dynamics of these HABs.     

Selective grazing of Temora longicornis in different stages of a Phaeocystis globosa bloom—a mesocosm study

Selective grazing of a calanoid copepod Temora longicornis was measured during different stages of a Phaeocystis globosa bloom, in order to reveal (1) if T. longicornis feeds on single cells and/or colonies of P. globosa in the presence of alternative food sources, (2) if copepod food selection changes during the initiation, maintenance, collapse and decay of a P. globosa bloom and (3) if P. globosa dominated food assemblage provides a good diet for copepod egg production. Our results show low but constant feeding on small colonies of P. globosa, irrespective of the type or concentration of alternative food sources. In contrast, feeding on single cells was never significant, and the total contribution of P. globosa to carbon ingestion of T. longicornis was minor. T. longicornis fed most actively on the decaying colonies, whereas during the peak of the bloom copepods selected against P. globosa. Mostly, T. longicornis fed unselectively on different food particles: before the bloom, the major part of the diet consisted of diatoms, whereas during and after the bloom copepod diet was dominated by dinoflagellates and ciliates. Egg production was highest during the decay of the bloom, coinciding with highest proportional ingestion of heterotrophic organisms, but was not seriously reduced even during the peak of the bloom. We conclude that P. globosa blooms should not threaten survival of copepod populations, but the population recruitment may depend on the type (and concentration) of the dominant heterotrophs present during the blooms. Due to relatively unselective grazing, the impact of T. longicornis to the initiation of a Phaeocystis bloom is considered small, although grazing on decaying colonies may contribute to the faster termination of a bloom.     

Glutathione transferase activity and oocyte development in copepods exposed to toxic phytoplankton

Organisms present a series of cellular mechanisms to avoid the effects of toxic compounds. Such mechanisms include the increase in activity of detoxification enzymes [e.g., 7-ethoxyresorufin-O-deethylase (EROD) and glutathione S-transferase (GST)], which could explain the low retention of ingested toxins generally observed in copepods. In addition, decreasing gross growth efficiency (GGE) of copepods with increasing concentration of toxic diets could be caused either by a high expenditure coping with toxins (e.g., increase in the activity of detoxification enzymes) or by a deterioration of reproductive tissues. To assess the effect of toxic phytoplankton on the activity of detoxification enzymes and on oocyte maturation of Acartia tonsa and Temora longicornis, feeding and egg production experiments were carried out with a variety of toxic diets and an adequate non-toxic food control (Rhodomonas spp.) all provided as single species diets. Toxic diets included the nodularin-producing cyanobacterium Nodularia spumigena, the dinoflagellates Alexandrium minutum, and A. tamarense, which contained Paralytic Shellfish Poisoning (PSP) toxins, the dinoflagellate Prorocentrum lima with Diarrhetic Shellfish Poisoning (DSP) toxins and the haptophyte Prymnesium parvum, which produces ichtyotoxins with haemolytic activity. Feeding on toxic diets was lower than on Rhodomonas spp., except for A. minutum and A. tamarense. In addition, toxic diets negatively affected reproduction in both copepod species with the production of oocytes and oocyte development impaired with A. minutum and N. spumigena. While the negative effect of N. spumigena seemed to be connected to gonad atresia likely caused by severe food limitation (starvation), the negative effect of A. minutum could have been either caused by a direct effect of saxitoxins or nutritional inadequacy on oocyte production. We could not detect EROD activity in the copepods, while the activity of GST was generally higher with the non-toxic food control and positively related to the feeding and egestion rates, suggesting relation to feeding conditions rather than to exposure to toxic diets. No relationship was found between GGE and GST activity. Our results refute the hypothesis that toxic diets, provided at ecologically relevant levels, would induce cellular mechanisms in copepods regarding GST activity. GST activity thus seems to play no role in detoxification of copepods confronted with toxic phytoplankton. Toxin detoxification and its cost for copepods still remain an open question.     

Cell cycle regulation of the mixotrophic dinoflagellate Dinophysis acuminata: Growth,photosynthetic efficiency and toxin production

The mixotrophic dinoflagellate Dinophysis acuminata is a widely distributed diarrhetic shellfish poisoning (DSP) producer. Toxin variability of Dinophysis spp. has been well studied, but little is known of the manner in which toxin production is regulated throughout the cell cycle in these species, in part due to their mixotrophic characteristics. Therefore, an experiment was conducted to investigate cell cycle regulation of growth, photosynthetic efficiency, and toxin production in D. acuminata. First, a three-step synchronization approach, termed “starvation-feeding-dark”, was used to achieve a high degree of synchrony of Dinophysis cells by starving the cells for 2 weeks, feeding them once, and then placing them in darkness for 58 h. The synchronized cells started DNA synthesis (S phase) 10 h after being released into the light, initiated G2 growth stage eight hours later, and completed mitosis (M phase) 2 h before lights were turned on. The toxin content of three dominant toxins, okadaic acid (OA), dinophysistoxin-1 (DTX1) and pectenotoxin-2 (PTX2), followed a common pattern of increasing in G1 phase, decreasing on entry into the S phase, then increasing again in S phase and decreasing in M phase during the diel cell cycle. Specific toxin production rates were positive throughout the G1 and S phases, but negative during the transition from G1 to S phase and late in M phase, the latter reflecting cell division. All toxins were initially induced by the light and positively correlated with the percentage of cells in S phase, indicating that biosynthesis of Dinophysis toxins might be under circadian regulation and be most active during DNA synthesis.     

Epiphytic abundance and toxicity of Prorocentrum lima populations in the Fleet Lagoon, UK

Planktonic Dinophysis spp. and epiphytic Prorocentrum lima (Ehrenberg) Dodge are known dinoflagellate producers of okadaic acid (OA) and dinophysistoxins (DTX), causative phycotoxins of diarrhetic shellfish poisoning (DSP). Underestimation of toxic dinoflagellates associated with a toxic event may be due to the lack of sampling of species with epiphytic and epibenthic strategies, such as P. lima. As Dinophysis spp. is not found in the Fleet Lagoon, Dorset, but previous DSP events have closed the Crassostrea gigas oyster farm, P. lima is the most likely causative organism. A field assay for separating microalgal epiphytes and concentrating wild cells on to filters was successfully applied to sub-samples of a variety of macroalgae and macrophytes (seagrass) collected from the Fleet during summer 2002. P. lima was present in increasing cell densities on most substratum species, over the sampling period, from 10² to 10³ cells g⁻¹ fresh weight (FW) plant biomass. LC–MS analysis detected OA and DTX-1 in extracts of wild P. lima cells, in ratios characteristic of P. lima strains previously isolated from the Fleet. No toxins, however, were detected in oyster flesh.     

DSP toxin production de novo in cultures of Dinophysis acuminata (Dinophyceae) from North America

For decades, many aspects of Dinophysis biology have remained intractable due to our inability to maintain these organisms in laboratory cultures. Recent breakthroughs in culture methods have opened the door for detailed investigations of these important algae. Here, for the first time, we demonstrate toxin production in cultures of North American Dinophysis acuminata, isolated from Woods Hole, MA. These findings show that, despite the rarity of Dinophysis-related DSP events in North America, D. acuminata from this area has the ability to produce DSP toxins just as it does in other parts of the world where this species is a major cause of DSP toxicity. In our cultures, D. acuminata cells were observed feeding on Myrionecta rubra using a peduncle. Culture extracts were analyzed using LC–MS/MS, providing unequivocal evidence for the toxin DTX1 in the Dinophysis cultures. In addition, a significant amount of an okadaic acid diol ester, OA-D8, was detected. These results suggest that this Dinophysis isolate stores much of its OA as a diol ester. Also, toxin PTX-2 and a hydroxylated PTX-2 with identical fragmentation mass spectrum to that of PTX-11, but with a different retention time, were detected in this D. acuminata culture. This demonstration of toxin production in cultured North American Dinophysis sets the stage for more detailed studies investigating the causes of geographic differences in toxicity. It is now clear that North American Dinophysis have the ability to produce DSP toxins even though they only rarely cause toxic DSP events in nature. This may reflect environmental conditions that might induce or repress toxin production, genetic differences that cause modifications in toxin gene expression, or physiological and biochemical differences in prey species.     

Omnivory in the calanoid copepod Temora longicornis: feeding, egg production and egg hatching rates

We measured in laboratory experiments the ingestion, egg production and egg hatching rates of female Temora longicornis as a function of diet. The diets consisted of a diatom (Thalassiosira weissflogii), an autotrophic dinoflagellate (Heterocapsa triquetra), and a bacterivorous ciliate (Uronema sp.) given as sole foods, or combinations of these single-food items: diatom+dinoflagellate, diatom+ciliate, dinoflagellate+ciliate, and diatom+ciliate+dinoflagellate. For the three single-item diets, the functional response was similar; i.e., ingestion rate increased linearly with food concentration (food range: ∼25 to ∼600 μg C l⁻¹). When all diets were considered, maximum daily carbon ration (∼70% of body weight) was independent of food type. However, the maximum daily egg production rate (12% of body carbon) was obtained with the diatom diet. For all diets, both ingestion and egg production rates increased with food concentration. Ingestion and egg production rates were affected differently by the interaction of food concentration and food type: at low food concentrations, ingestion rates were highest on diets containing the diatom. At high food concentrations, egg production rates were highest on the two phytoplankter diets and their combination. The presence of the ciliate in the diet did not enhance ingestion rate or egg production. Mixed-food diets did not enhance egg production relative to single-food diets. Hence, dietary diversity did not appear to be particularly advantageous for reproduction. Carbon-specific egg production efficiency (EPE; egg production/ingestion) was independent of food concentration and type, and equaled 9%. Egg hatching success was low (mean<30%) and independent of food concentration and type, and egg production rates. Our results are consistent with previous observations that egg production in T. longicornis is enhanced during diatom blooms. However, the relatively low EPE and egg hatching success suggest that reproduction and recruitment in this study were severely constrained by the biochemical composition of the diet, or the physiological condition of the females towards the end of their season of growth in Long Island Sound.     

Latitudinal differentiation in the effects of the toxic dinoflagellate Alexandrium spp. on the feeding and reproduction of populations of the copepod Acartia hudsonica

Blooms of the dinoflagellate Alexandrium spp. increase in their frequency, toxicity and historical presence with increasing latitude from New Jersey (USA) to the Gaspé peninsula (Canada). Biogeographic variation in these blooms results in differential exposure of geographically separate copepod populations to toxic Alexandrium. We hypothesize that the ability of copepods to feed and reproduce on toxic Alexandrium should be higher in copepods from regions that are frequently exposed to toxic Alexandrium blooms. We tested this hypothesis with factorial common environment experiments in which female adults of the copepod Acartia hudsonica from five separate populations ranging from New Jersey to New Brunswick were fed toxic and non-toxic strains of Alexandrium, and the non-toxic flagellate Tetraselmis sp. Consistent with the hypothesis, when fed toxic Alexandrium we observed significantly higher ingestion and egg production rates in the copepods historically exposed to toxic Alexandrium blooms relative to copepods from regions in which Alexandrium is rare or absent. Such differences among copepod populations were not observed when copepods were fed non-toxic Alexandrium or Tetraselmis sp. These results were also supported by assays in which copepods from populations both historically exposed and naïve to toxic Alexandrium blooms were fed mixtures of toxic Alexandrium and Tetraselmis sp. Two-week long experiments demonstrated that when copepods from populations naïve to toxic Alexandrium were fed a toxic strain of Alexandrium they failed to acclimate, such that their ingestion rates remained low throughout the entire two-week period. The differences observed among populations suggest that local adaptation of populations of A. hudsonica from Massachusetts (USA) to New Brunswick (Canada) has occurred, such that some populations are resistant to toxic Alexandrium.     

Characterization of Dinophysis spp. (Dinophyceae,Dinophysiales) from the mid-Atlantic region of the United States1

Due to the increasing prevalence of Dinophysis spp. and their toxins on every US coast in recent years, the need to identify and monitor for problematic Dinophysis populations has become apparent. Here, we present morphological analyses, using light and scanning electron microscopy, and rDNA sequence analysis, using a ~2-kb sequence of ribosomal ITS1, 5.8S, ITS2, and LSU DNA, of Dinophysis collected in mid-Atlantic estuarine and coastal waters from Virginia to New Jersey to better characterize local populations. In addition, we analyzed for diarrhetic shellfish poisoning (DSP) toxins in water and shellfish samples collected during blooms using liquid-chromatography tandem mass spectrometry and an in vitro protein phosphatase inhibition assay and compared this data to a toxin profile generated from a mid-Atlantic Dinophysis culture. Three distinct morphospecies were documented in mid-Atlantic surface waters: D. acuminata, D. norvegica, and a “small Dinophysis sp.” that was morphologically distinct based on multivariate analysis of morphometric data but was genetically consistent with D. acuminata. While mid-Atlantic D. acuminata could not be distinguished from the other species in the D. acuminata-complex (D. ovum from the Gulf of Mexico and D. sacculus from the western Mediterranean Sea) using the molecular markers chosen, it could be distinguished based on morphometrics. Okadaic acid, dinophysistoxin 1, and pectenotoxin 2 were found in filtered water and shellfish samples during Dinophysis blooms in the mid-Atlantic region, as well as in a locally isolated D. acuminata culture. However, DSP toxins exceeded regulatory guidance concentrations only a few times during the study period and only in noncommercial shellfish samples.     

Acquired phototrophy in Mesodinium and Dinophysis – A review of cellular organization,prey selectivity,nutrient uptake and bioenergetics

Acquired phototrophy, i.e. the use of chloroplasts from ingested prey, can be found among some species of dinoflagellates and ciliates. The best studied examples of this phenomenon in these groups are within the ciliate genus Mesodinium and the dinoflagellate genus Dinophysis, both ecologically important genera with a worldwide distribution. Mesodinium species differ considerably in their carbon metabolism. Some species rely almost exclusively on food uptake, while other species rely mostly on photosynthesis. In Mesodinium with acquired phototrophy, a number of prey organelles in addition to chloroplasts may be retained, and the host ciliate has considerable control over the acquired chloroplasts; Mesodinium rubrum is capable of dividing its acquired chloroplasts and can also photoacclimate. In Dinophysis spp., the contents of ciliate prey are sucked out, but only the chloroplasts are retained from the ingested prey. Some chloroplast house-keeping genes have been found in the nucleus of Dinophysis and some preliminary evidence suggests that Dinophysis may be capable for photoacclimation. Both genera have been claimed to take up inorganic nutrients, including NO₃⁻, indicating that processes beyond photosynthesis have been acquired. M. rubrum seems to depend upon prey species within the Teleaulax/Plagioselmis/Geminigera clade of marine cryptophytes. Up until now, Dinophysis species have only been maintained cultured on M. rubrum as food, but other ciliates may also be ingested. Dinophysis spp. and M. rubrum are obligate mixotrophs, depending upon both prey and light for sustained growth. However, while M. rubrum only needs to ingest 1–2% of its carbon demand per day to attain maximum growth, Dinophysis spp. need to obtain about half of their carbon demand from ingestion for maximum growth. Both Mesodinium and Dinophysis spp. can survive for months in the light without food. The potential role for modeling in exploring the complex balance of phototrophy and phago-heterotrophy, and its ecological implications for the mixotroph and their prey, is discussed.     

Zooplankton community composition and copepod grazing on the West Florida Shelf in relation to blooms of Karenia brevis

Blooms of the toxin producing dinoflagellate Karenia brevis occur routinely on the West Florida Shelf of the Gulf of Mexico. Nutrient supplies are thought to play a large role in the formation and maintenance of these blooms. The role of top-down control has been less well studied, but grazing, or the lack thereof, on these toxic species may also enhance the formation of large biomass blooms in this region. Zooplankton community structure and copepod species composition were analyzed from samples collected on the West Florida Shelf (WFS) during a NOAA funded ECOHAB regional Karenia Nutrient Dynamics project during October 2007–2010. In 2008 there was no statistical difference in the abundance of zooplankton at bloom and non-bloom stations, however in 2009 there was a statistically significant difference (p < 0.05) between the abundance of zooplankton at stations with Karenia present. To investigate copepod ingestion rates in relation to K. brevis, shipboard and laboratory experiments of the single label method of ¹⁴C labeled phytoplankton culture, and time course ingestion experiments with isolated copepods were performed. Calculated ingestion rates suggest that the copepod species Centropages velificatus, and Acartia tonsa ingested K. brevis, however rates were variable among collection sites and K. brevis strains. Parvocalanus crassirostris did not ingest K. brevis in any of the experiments.     

Testing for resistance of pelagic marine copepods to a toxic dinoflagellate

With few exceptions, the evolutionary consequences of harmful algae to grazers in aquatic systems remain unexplored. To examine both the ecological and evolutionary consequences of harmful algae on marine zooplankton, we used a two-fold approach. In the first approach, we examined the life history responses of two geographically separate Acartia hudsonica (Copepoda Calanoida) populations reared on diets containing the toxic dinoflagellate Alexandrium fundyense . One copepod population was from a region, Casco Bay, Maine, USA, that has experienced recurrent blooms of highly toxic Alexandrium spp. for decades; whereas the other population from Great Bay, New Jersey, USA, has never been exposed to toxic Alexandrium blooms. The life history experiment demonstrated that when the copepod population from New Jersey was reared on a diet containing toxic A. fundyense it exhibited lower somatic growth, size at maturity, egg production and survival than the same population reared on a diet without toxic A. fundyense . In contrast, toxic A. fundyense did not affect the life-history traits of the Maine population. Fitness, finite population growth rate (), was significantly reduced in the New Jersey population, but not in the Maine population. These results are consistent with the hypothesis of local adaptation (resistance) of the historically exposed copepod population to the toxic dinoflagellate. In the second approach, we further tested the resistance hypothesis with a laboratory genetic selection experiment with the naïve New Jersey copepod population exposed to a diet containing toxic A. fundyense. This experiment demonstrated that the ingestion and egg production of adult females of naïve copepods fed A. fundyense improved after three generations of being reared on a diet containing the toxic dinoflagellate. The results of the present study have important implications for understanding how grazer populations may respond to the introduction of toxic algae to their environment, and suggest that grazer resistance may be a feedback mechanism that may lead to bloom control.Co-ordinating editor: Hurst     

Toxicity of the dinoflagellate Gambierdiscus sp. toward the marine copepod Tigriopus japonicus

The toxic dinoflagellate Gambierdiscus sp. has recently been observed in temperate areas in the Southern Sea of the Republic of Korea. The purpose of this study was to examine the toxicity of Gambierdiscus sp. toward the marine copepod Tigriopus japonicus. T. japonicus adult females and nauplii were exposed to various concentrations of algae or culture medium to assess toxicity and analyze gene expression patterns in the copepod. Based on the toxicity tests, Gambierdiscus sp. increased the mortality of nauplii and the immobility of adult females. The survival and mobility of T. japonicus were not affected by culture medium lacking Gambierdiscus sp. cells in the toxicity test. However, based on the analysis of gene expression in the copepod, exposure of the copepod to culture medium affected the expression of stress or detoxification-related genes. Further studies to identify toxins in Gambierdiscus sp. are required to increase our understanding of dinoflagellate toxicity.     

Seasonal variability of lipophilic toxins during a Dinophysis acuta bloom in Western Iberia: Differences between picked cells and plankton concentrates

This work describes and compares the seasonal variability of toxin profiles and content, estimated by LC–MS analyses, in picked cell of Dinophysis acuta Ehrenberg, in plankton concentrates rich in this species, and in extracellular lipophilic toxins collected by adsorbent resins during weekly sampling in a Galician ría (Western Iberia) from October 2005 to January 2006. Picked cells of D. acuta—which exhibited a fairly stable OA:DTX2 ratio, close to 3:2, but a variable okadaates:PTX2 ratio—showed a 9-fold variation in cell toxin quota, which was partly related to cellular volume, with maximum values (19 pg cell⁻¹) observed during the exponential decline of the population. Large differences in toxin profiles and content were observed between picked cells and plankton concentrates (up to 73 pg cell⁻¹ in the latter), that were most conspicuous after the bloom decline. The toxin profile of picked cells was more similar to that observed in the adsorbent resins than to the profiles of plankton concentrates. Their continued detection several weeks after the disappearance of Dinophysis spp. indicates that these toxins may take a long time to be degraded. It is concluded that analyses of picked-cells are essential to determine the contribution of each species of Dinophysis to a toxic outbreak. Estimates of cellular toxin content from plankton concentrates can lead to considerable overestimates after Dinophysis blooms decay due to extracellular toxins that persist in the water column, possibly bound to organic aggregates and detritus, and are retained (>0.22 μm) in the filters.     

Bioaccumulation of PSTs produced by Alexandrium ostenfeldii in the northern Baltic Sea

Bioaccumulation of paralytic shellfish toxins (PSTs) produced by the dinoflagellate Alexandrium ostenfeldii was investigated in the northern Baltic Sea. The study was based on the assumption that the toxins released during high magnitude blooms of A. ostenfeldii will accumulate in the biota at the bloom site, especially in bivalves. To test this, experiments with blue mussels (Mytilus trossulus) exposed to toxic A. ostenfeldii in field conditions were carried out together with a field survey aimed to quantify natural distribution of PSTs in the biota. As hypothesized, PSTs accumulated in the tissues of the blue mussels during the incubations. Toxins were also detected in natural bivalve communities at the bloom site, the highest toxin concentrations found in the small Cerastoderma glaucum individuals, exceeding the EC safety limit for shellfish consumption. Relatively high total toxin concentrations were also detected from fish (Perca fluviatilis). These are the first records of PST transfer in the food web of the northern Baltic Sea.