Microzooplankton grazing and algae growth responses to increasing pCO₂ levels (350, 700 and 1050 μatm) were investigated in nitrate and phosphate fertilized mesocosms during the PeECE III experiment 2005. Grazing and growth rates were estimated by the dilution technique combined with taxon specific HPLC pigment analysis. Microzooplankton composition was determined by light microscopy. Despite a range of up to 3 times the present CO₂ levels, there were no clear differences in any measured parameter between the different CO₂ treatments. During days 3–9 of the experiment the algae community standing stock, measured as chlorophyll a (Chl-a), showed the highest instantaneous grow rates (k=0.37–0.99 d⁻¹) and increased from ca. 2–3 to 6–12 μg l⁻¹, in all mesocosms. Afterwards the phytoplankton standing stock decreased in all mesocosms until the end of the experiment. The microzooplankton standing stock, that was mainly constituted by dinoflagellates and ciliates, varied between 23 and 130 μg C l⁻¹ (corresponding to 1.9 and 10.8 μmol C l⁻¹), peaking on day 13–15, apparently responding to the phytoplankton development. Instantaneous Chl-a growth rates were generally higher than the grazing rates, indicating only a limited overall effect of microzooplankton grazing on the most dominant phytoplankton. Diatoms and prymnesiophytes were significantly grazed (12–43% of the standing stock d⁻¹) only in the pre-bloom phase when they were in low numbers, and in the post-bloom phase when they were already affected by low nutrients and/or viral lysis. The cyanobacteria populations appeared more affected by microzooplankton grazing which generally removed 20–65% of the standing stock per day.

Diatoms are key phytoplankton organisms and one of the main primary producers in aquatic ecosystems. However, many diatom species produce a series of secondary metabolites, collectively termed oxylipins, that disrupt development in the offspring of grazers, such as copepods, that feed on these unicellular algae. We hypothesized that different populations of copepods may deal differently with the same oxylipin-producing diatom diet. Here we provide comparative studies of expression level analyses of selected genes of interest for three Calanus helgolandicus populations (North Sea, Atlantic Ocean and Mediterranean Sea) exposed to the same strain of the oxylipin-producing diatom Skeletonema marinoi using as control algae the flagellate Rhodomonas baltica. Expression levels of detoxification enzymes and stress proteins (e.g. glutathione S-transferase, glutathione synthase, superoxide dismutase, catalase, aldehyde dehydrogenases and heat shock proteins) and proteins involved in apoptosis regulation and cell cycle progression were analyzed in copepods after both 24 and 48 hours of feeding on the diatom or on a control diet. Strong differences occurred among copepod populations, with the Mediterranean population of C. helgolandicus being more susceptible to the toxic diet compared to the others. This study opens new perspectives for understanding copepod population-specific responses to diatom toxins and may help in underpinning the cellular mechanisms underlying copepod toxicity during diatom blooms.

13 pages, 1 tableFrom 3 to 6 November 2002, a colloquium was convened at the Benthos Laboratory of the Stazione Zoologica Anton Dohrn on Ischia, Italy, with the goal of evaluating the present status of the effects of diatoms on their main consumers, planktonic copepods, and to develop future research strategies to enhance our understanding of such interactions. These included toxic effects of diatom metabolites on copepods, particularly reproduction, and nutritional effects of diatoms on juvenile to adult copepods. Key issues involved in the impact of diatoms on the dynamics of natural plankton communities in situ were also addressed. During the plenary session, the most recent advances on this topic were presented. The plenary session was followed by 3 working groups on (1) production of aldehydes by phytoplankton, (2) toxic and nutritional effects of diatoms on zooplankton, and (3) the chemistry of diatom defense, as well as of their nutritional quality. These working groups focused on suggesting future research needs for the different topics. As a result, several recommendations were outlined, including experimental studies. It became evident that interdisciplinary efforts are needed, involving chemists, oceanographers and experimentalists, since many of the biological observations under controlled conditions and in situ require an integrated approach, including chemical causation. Extensive field observations based on common protocols are also recommended for investigation of the intrinsic variability of such effects and their environmental controls. Laboratory experiments are seen to be essential for the full understanding of environmentally occurring processes.Peer reviewe

Diatoms are dominant photosynthetic organisms in the world's oceans and are considered essential in the transfer of energy through marine food chains. However, these unicellular plants at times produce secondary metabolites such as polyunsaturated aldehydes and other products deriving from the oxidation of fatty acids that are collectively termed oxylipins. These cytotoxic compounds are responsible for growth inhibition and teratogenic activity, potentially sabotaging future generations of grazers by inducing poor recruitment in marine organisms such as crustacean copepods. Here we show that two days of feeding on a strong oxylipin-producing diatom (Skeletonema marinoi) is sufficient to inhibit a series of genes involved in aldehyde detoxification, apoptosis, cytoskeleton structure and stress response in the copepod Calanus helgolandicus. Of the 18 transcripts analyzed by RT-qPCR at least 50% were strongly down-regulated (aldehyde dehydrogenase 9, 8 and 6, cellular apoptosis susceptibility and inhibitor of apoptosis IAP proteins, heat shock protein 40, alpha- and beta-tubulins) compared to animals fed on a weak oxylipin-producing diet (Chaetoceros socialis) which showed no changes in gene expression profiles. Our results provide molecular evidence of the toxic effects of strong oxylipin-producing diatoms on grazers, showing that primary defense systems that should be activated to protect copepods against toxic algae can be inhibited. On the other hand other classical detoxification genes (glutathione S-transferase, superoxide dismutase, catalase, cytochrome P450) were not affected possibly due to short exposure times. Given the importance of diatom blooms in nutrient-rich aquatic environments these results offer a plausible explanation for the inefficient use of a potentially valuable food resource, the spring diatom bloom, by some copepod species.