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Diatom oxylipins have been observed to deleteriously impact copepod reproductive success. However, field studies have revealed very variable and case-dependent results. Therefore, the plasticity of diatom oxylipin metabolism was studied among four clones of the marine diatom Skeletonema marinoi Sarno et Zingone. Diatom oxylipin metabolism was studied by two lipoxygenase (LOX) activity assays carried out at different pH values and by oxylipin quantification. The four clones showed no major metabolic differences in terms of protein content or growth rate. However, two of the clones produced significantly higher levels of oxylipins than the other two. LOX activity measurements also indicated clonal variability in fatty acid oxidative metabolism. The presence of clone-specific differences in oxylipin metabolism may play a role in shaping diatom population dynamics by conferring selective advantages to certain clones.

Egg production rates and/or hatching success in the copepods Acartia clausi, Calanus helgolandicus and Temora longicornis were negatively affected by a late spring (May-June 2003) phytoplankton bloom in the North Adriatic Sea, dominated mainly by the large diatom Cerataulina pelagica. Highest total concentrations of 3.3- 104 cells-ml(-1) were located in the vicinity of the Po River, which also corresponded to the area where the highest numbers of phaeophorbides were measured (0.779, 0.528 and 0.419 mu g.l(-1), respectively, compared to an average of the remaining stations of 0.183 +/- 0.049 SD), suggesting some grazing on the bloom. Phytoplankton biomass in terms of carbon was dominated by diatoms, representing on average 42% of total phytoplankton carbon and more than 80% at several stations. Cerataulina pelagica, Cyclotella spp., Chaetoceros spp. and small unidentified centric diatoms dominated the diatom community numerically but C. pelagica was by far the dominant diatom in terms of carbon due to its large cell size. This species represented more than 60% of the diatom biomass at nine of the 14 stations sampled, and was absent only at one station, which was the most offshore station sampled during the cruise. Although polyunsaturated aldehydes (PUAs) were not detected, other oxylipins which are hydroxy and keto derivatives of eicosapentaenoic and docosahexaenoic acids that affect copepod reproduction were found in these samples. Hence, we can attribute the negative impact of diatoms not only to PUAs, as previously believed, but also to these compounds. This is the first direct evidence of the presence of oxylipins other than PUAs in marine blooms dominated by diatoms.

The effects of bioactive aldehydes from diatoms, unicellular algae at the base of the marine food web, were studied on fertilization and early development processes of the ascidian Ciona intestinalis. Using whole-cell voltage clamp techniques, we show that 2-trans-4-trans-decadienal (DD) and 2-trans-4-cis-7-cis-decatrienal (DT) inhibited the fertilization current which is generated in oocytes upon interaction with the spermatozoon. This inhibition was dose-dependent and was accompanied by inhibition of the voltage-gated calcium current activity of the plasma membrane. DD and DT did not inhibit the subsequent contraction of the cortex. Moreover, DD specifically acted as a fertilization channel inhibitor since it did not affect the steady state conductance of the plasma membrane or gap junctional (GJ) communication within blastomeres of the embryo. On the other hand, DD did affect actin reorganization even though the mechanism of action on actin filaments differed from that of other actin blockers. Possibly this effect on actin reorganization was responsible for the subsequent teratogenic action on larval development. The effect of DD was reversible if oocytes were washed soon after fertilization indicating that DD may specifically target certain fertilization mechanisms. Thus, diatom reactive aldehydes such as DD may have a dual effect on reproductive processes, influencing primary fertilization events such as gating of fertilization channels and secondary processes such as actin reorganization which is responsible for the segregation of cell lineages. These findings add to a growing body of evidence on the antiproliferative effects of diatom-derived aldehydes. Our results also report, for the first time, on the action of a fertilization channel blocker in marine invertebrates.

Confocal Laser Scanning Microscope techniques have been applied to study the developmental biology of marine copepods and decapod larvae. The lipophylic probes Dil and DiOC(6) were used to study both the external and internal morphology of these crustaceans, whereas the same DiOC(6) and the specific nuclear probe Hoechst 33342 were used to study embryonic development of copepods in vivo. To distinguish viable from non-viable copepod embryos, the vital dye dichlorodihydrofluorescein diacetate (H(2)DCFDA) was used. Major advantages and difficulties in the use of these non-invasive techniques in studies of the reproductive biology of marine crustaceans are discussed. (C) 2003 Wiley-Liss, Inc.

Evidence is presented that hatching failure in Temora stylifera eggs can depend on poor sperm quality. Three dinoflagellate diets, Prorocentrum micans, Gymnodinium sanguinium,and Gonyaulax polyedra,significantly modified spermatophore production and reduced the fertilization capacity of male sperm after 6‐12 d of continuous feeding. Two other diets, the dinoflagellate P. minimum and the prymnesiophycean Isochrysis galbana,had no effect on hatching success, which remained high (.89%) and stable with time. A reduction in fertilization capacity was neither due to maternal effects nor to male age since hatching success returned to normal upon the introduction of freshly caught wild males or males conditioned with a good diet such as P. minimum for the same length of time as couples fed with the poor diets, P. micans, G. sanguinium, and G. polyedra. Confocal microscope images of unhatched eggs colored with a nucleus-specific fluorescent dye confirmed that these eggs had not been fertilized. Experiments with Calanus helgolandicus females, which did not require reinsemination and which were fed the same diets that induced hatching failure in T. stylifera, showed no change in hatching success with time. Copepod fecundity has been shown to be strongly related