Spirolide composition of micro-extracted pooled cells isolated from natural plankton assemblages and from cultures of the dinoflagellate Alexandrium ostenfeldii

A novel micro-extraction technique was applied to the extraction of biologically active macrocyclic imines known as spirolides from pooled individual cells isolated from spirolide-rich plankton material. For comparison, this method was also applied to pooled individual cells isolated from a unialgal culture of the marine dinoflagellate Alexandrium ostenfeldii (Paulsen) Balech & Tangen, a species known to produce spirolides. Both athecate cells and motile forms of gonyaulacoid dinoflagellates derived from size-fractionated plankton material from Nova Scotia, Canada were sorted and pooled by the glass micropipette isolation technique and by flow cytometry. The development of a highly sensitive analytical method for spirolides (detection limit 2 ng ml(-1) for spirolide B) using liquid chromatography-mass spectrometry (LC-MS) and application to micro-extracted samples allowed the accurate determination of spirolide composition in as few as 50 cells. Total spirolide concentrations (fmol cell(-1)) calculated from pooled micropipette isolated cells were very consistent with those based upon bulk- or micro-extractions of A. ostenfeldii cells from unialgal batch cultures in exponential growth phase. The results of the pooled cell selection from field material from two sites in Nova Scotia confirmed the association of spirolides with vegetative cells of A. ostenfeldii and related athecate forms. Combining these techniques represents a highly sensitive method for the analysis of marine toxins within complex plankton matrices, even when the toxigenic species is in low abundance, by enrichment of the target organism.     

On the allelochemical potency of the marine dinoflagellate Alexandrium ostenfeldii against heterotrophic and autotrophic protists

Three strains of the marine dinoflagellate Alexandrium ostenfeldiiof different geographic origin were tested for their short-termdeleterious effects on a diversity of marine protists. All A.ostenfeldii strains were capable of eliciting an apparent allelochemicalresponse, but the various protistan target species were differentiallyaffected. Protists that were negatively affected by exposureto cells of A. ostenfeldii and associated extracellular metabolitescomprised both autotrophs (Rhodomonas sp., Dunaliella salina,Thalassiosira weissflogii) and heterotrophs (Oxyrrhis marina,Amphidinium crassum, Rimostrombidium caudatum). Observed effectsincluded immobilisation (e.g. of O. marina), morphological changes(e.g. in D. salina) and/or aberrant behaviour (e.g. of R. caudatum),mainly as preliminary stages of cell lysis. Immobilization andlytic effects against O. marina were strongly dependent on A.ostenfeldii cell concentrations. Effects also differed substantiallyamong strains and different batch cultures of the same strain.Values of EC₅₀, defined as the A. ostenfeldii cell concentrationcausing lysis of 50% of O. marina cells, ranged from 0.3 to1.9 x 10³ mL^–1, depending on the A. ostenfeldii strain.The autotrophic dinoflagellate Scrippsiella trochoidea reactedto exposure to A. ostenfeldii cells by formation of temporary(ecdysal) cysts, whereas, in contrast, the flagellates Emilianiahuxleyi and Prymnesium parvum and the ciliate Strombidium sp.were relatively refractory or even unaffected. As long as cellsdid not lyse, the fluorescence yield of target autotrophs, estimatedby pulse-amplitude modulation fluorometry, did not significantlychange during the first 3 h of incubation, suggesting that allelochemicalsproduced by A. ostenfeldii caused no short-term negative effectson the photosynthetic apparatus. Overall, the allelochemicalresponses of target species showed no obvious relationship tocell quota or extracellular concentrations of either toxic macrocyclicimines (spirolides) or tetrahydropurine neurotoxins (saxitoxinand analogues) produced by various strains of A. ostenfeldii.Instead, the potency of A. ostenfeldii, eliciting immobilizationand lytic species-specific responses in potential predatorsand competitors, is consistent with the existence of an allelochemicalmechanism unrelated to the bioactivity of known phycotoxinsof the genus Alexandrium.     

Paralytic shellfish toxins or spirolides? The role of environmental and genetic factors in toxin production of the Alexandrium ostenfeldii complex

Dinoflagellates of the Alexandrium ostenfeldii complex (A. ostenfeldii, A. peruvianum) are capable of producing different types of neurotoxins: paralytic shellfish toxins (PSTs), spirolides and gymnodimines, depending on the strain and its geographic origin. While Atlantic and Mediterranean strains have been reported to produce spirolides, strains originating from the brackish Baltic Sea produce PSTs. Some North Sea, USA and New Zealand strains contain both toxins. Causes for such intraspecific variability in toxin production are unknown. We investigated whether salinity affects toxin production and growth rate of 5 A. ostenfeldii/peruvianum strains with brackish water (Baltic Sea) or oceanic (NE Atlantic) origin. The strains were grown until stationary phase at 7 salinities (6–35), and their growth and toxin production was monitored. Presence of saxitoxin (STX) genes (sxtA1 and sxtA4 motifs) in each strain was also analyzed. Salinity significantly affected both growth rate and toxicity of the individual strains but did not change their major toxin profile. The two Baltic Sea strains exhibited growth at salinities 6–25 and consistently produced gonyautoxin (GTX) 2, GTX3 and STX. The two North Sea strains grew at salinities 20–35 and produced mainly 20-methyl spirolide G (20mG), whereas the strain originating from the northern coast of Ireland was able to grow at salinities 15–35, only producing 13-desmethyl spirolide C (13dmC). The effects of salinity on total cellular toxin concentration and distribution of toxin analogs were strain-specific. Both saxitoxin gene motifs were present in the Baltic Sea strains, whereas the 2 North Sea strains lacked sxtA4, and the Irish strain lacked both motifs. Thus sxtA4 only seems to be specific for PST producing strains. The results show that toxin profiles of A. ostenfeldii/peruvianum strains are predetermined and the production of either spirolides or PSTs cannot be induced by salinity changes. However, changes in salinity may lead to changed growth rates, total cellular toxin concentrations as well as relative distribution of the different PST and spirolide analogs, thus affecting the actual toxicity of A. ostenfeldii/peruvianum populations.     

COMPARISON OF EUTHECOSOMATOUS PTEROPODS IN THE PLANKTON AND SEDIMENTS OFF BARBADOS, WEST INDIES

The euthecosomatous pteropod fauna of Barbados plankton wascompared with euthecosome shells occurring in recent sediments.Twenty species of euthecosomes were collected in the same relativeabundance in both provinces. Differences in the average sizesof Spiratella inflata and Creseis virgula conica, the most commonspecies were not statistically significant although differencesin the size-frequency histograms were noted. It is concludedthat euthecosomes deposited in the upper sediment layers offBarbados accurately reflect the species composition and relativeabundance of euthecosome species in the plankton. ¹Supported by a Postgraduate Scholarship from the National ResearchCouncil of Canada and NRC Grant No. A5248 to Dr. C.M. Lalli. ²Present address: Department of Oceanography, Dalhousie University,Halifax, Nova Scotia, Canada.     

Characterization of spirolide producing Alexandrium ostenfeldii (Dinophyceae) from the western Arctic

Toxin producing dinoflagellates of the genus Alexandrium Halim represent a risk to Arctic environments and economies. This study provides the first record and a characterization of Alexandrium ostenfeldii in the western Arctic. During a cruise along the coasts of western and southern Greenland 36 isolates of the species were established in August 2012. Plankton samples taken at three different stations from the upper water layer at water temperatures of approx. 4–7 °C, contained low amounts of A. ostenfeldii. Sequencing of SSU and ITS-LSU rDNA and subsequent phylogenetic analyses identified all Greenland strains as members of a NW Atlantic spirolide producing phylogenetic clade. Molecular results were confirmed by morphological features typical for this group (=Group 5 of a recent ITS-LSU phylogeny of A. ostenfeldii). The Greenland isolates did not contain either Paralytic Shellfish Poisoning toxins or gymnodimines, but produced several spirolides. Altogether 12 different analogs were detected, of which only SPX-1, C, 20-meG and H have been described earlier. The remaining 8 spirolides have not been identified so far. Some of them were found to dominate the toxin profiles of a number of isolates. Among the 36 investigated strains spirolide composition varied considerably, particularly isolates from western Greenland (Station 516) exhibited a high diversity of analogs, with different profiles in nearly all 22 isolates. All of the 34 tested Greenland strains showed considerable lytic capacity when exposed to Rhodomonas salina.     

Morphology, phylogenetic position, and ecophysiology of Alexandrium ostenfeldii (Dinophyceae) from the Bohai Sea, China

Alexandrium ostenfeldii is a potentially toxic dinoflagellate that often occurs in coastal areas at high latitudes.Here we report the presence of A.ostenfeldii in the Bohai Sea,China,for the first time.The vegetative cells ofA.ostenfeldii are characterized by a narrow first apical plate and a large ventral pore located on the anterior right side.Partial large subunit sequence comparison revealed that the Chinese strain differs from the Finnish strains at only three positions,and from A.peruvianum of Spain at five positions.Maximum parsimony analysis revealed that A.ostenfeldii from China and Finland and A.peruvianum from Spain grouped together.They were the nearest sister group to a clade with A.ostenfeldii from New Zealand,Europe,and North America.In culture,growth did not occur at temperatures below 9 ℃ and occurred at salinities between 7 and 27 psu.It took 10-20 days for newly formed cysts to mature at 20 ℃.Lower temperature delayed germination,but the germination rate exceeded 90％ at temperatures from 12 to 24 ℃.No germination occurred below 9 ℃ after 1 month of incubation.The Chinese strain ofA.ostenfeldii produced neither spirolides nor paralytic shellfish poisoning toxins.     

Morphology,phylogenetic position and ecophysiology of Alexandrium ostenfeldii (Dinophyceae) from the Bohai Sea,China

Alexandrium ostenfeldii is a potentially toxic dinoflagellate that often occurs in coastal areas at high latitudes. Here we report the presence of A. ostenfeldii in the Bohai Sea, China, for the first time. The vegetative cells of A. ostenfeldii are characterized by a narrow first apical plate and a large ventral pore located on the anterior right side. Partial large subunit sequence comparison revealed that the Chinese strain differs from the Finnish strains at only three positions, and from A. peruvianum of Spain at five positions. Maximum parsimony analysis revealed that A. ostenfeldii from China and Finland and A. peruvianum from Spain grouped together. They were the nearest sister group to a clade with A. ostenfeldii from New Zealand, Europe, and North America. In culture, growth did not occur at temperatures below 9 °C and occurred at salinities between 7 and 27 psu. It took 10–20 days for newly formed cysts to mature at 20 °C. Lower temperature delayed germination, but the germination rate exceeded 90% at temperatures from 12 to 24 °C. No germination occurred below 9 °C after 1 month of incubation. The Chinese strain of A. ostenfeldii produced neither spirolides nor paralytic shellfish poisoning toxins.     

The toxigenic marine dinoflagellate Alexandrium tamarense as the probable cause of mortality of caged salmon in Nova Scotia

The toxins associated with paralytic shellfish poisoning (PSP) are potent neurotoxins produced by natural populations of the marine dinoflagellate Alexandrium tamarense. In early June 2000, a massive bloom (>7×10⁵ cells l⁻¹) of this dinoflagellate coincided with an unusually high mortality of farmed salmon in sea cages in southeastern Nova Scotia. Conditions in the water column in the harbour were characterised by the establishment of a sharp pycnocline after salinity stratification due to abundant freshwater runoff. In situ fluorescence revealed a high sub-surface (2–4 m depth) chlorophyll peak related to the plankton bloom. The intense bloom was virtually monospecific and toxicity was clearly related to the concentration of Alexandrium cells in plankton size fractions. Cultured clonal isolates of A. tamarense from the aquaculture sites were very toxic on a per cell basis and yielded a diversity of PSP toxin profiles, some of which were similar to those from plankton concentrates from the natural bloom population. The toxin profile of plankton concentrates from the 21–56 μm size fraction was complex, dominated by the N-sulfocarbamoyl derivative C2, with levels of other PSP toxins GTX4, NEO, GTX5 (=B1), GTX3, GTX1, STX, C1, and GTX2, in decreasing order of relative abundance. Although no PSP toxin was found systemically in the fish tissues (liver, digestive tract) from this salmon kill event, the detection of Alexandrium cells and low levels of PSP toxins in salmon gills provide evidence that the enhanced mortalities were caused by direct exposure to toxic Alexandrium cells and/or to soluble toxins released during the bloom.     

Toxic strains of the Alexandrium ostenfeldii complex in southern South America (Beagle Channel,Argentina)

During phytoplankton monitoring in the Beagle Channel (≈54°52′ S, 67°32′ W) a previously undetected Alexandrium species was observed in coincidence with mouse bioassay toxicity. Detailed thecal plates analysis using epifluorescence and scanning electron microscopy revealed the presence of the Alexandrium ostenfeldii species complex, showing a mixture of the diagnostic features usually used to discriminate between the morphospecies A. ostenfeldii and A. peruvianum. Cells of the A. ostenfeldii complex were commonly observed during spring after the main annual diatom bloom, when temperatures and salinities were respectively around 7.5–10 °C and 30–30.5 psu, and nutrients showed a seasonal decrease. Toxin analysis by liquid chromatography–mass spectrometry revealed the production of 13-desmethyl spirolide C and 20-methyl spirolide G in cell cultures. The cellular contain of spirolides during exponential phase growth was 0.5906 ± 0.0032 and 0.1577 ± 0.0023 pg cell⁻¹ for 13-desMe-C and 20-Me-G, respectively. A third unknown compound, with a structure resembling that of spirolides was also detected in culture. Moreover, an additional compound with a similar m/z (692) than that of 13-desMe-C but presenting a higher retention time (Rt = 40.5 min) was found in high proportions in mussel samples. PSP toxins were present at low concentration in mussels but were not detected in cultures. These results extend the world-wide distribution of toxic strains of the A. ostenfeldii complex to the Beagle Channel (southern South America), where toxic events have been traditionally linked to the presence of Alexandrium catenella. This is the first confirmed occurrence of spirolides in mussels and plankton from Argentina, which highlights the importance of monitoring these toxins and their producing organisms to protect public health and improve the management of shellfish resources.     

Contribution of heterotrophic plankton to nitrogen regeneration in the upwelling ecosystem of A Coruna (NW Spain)

The contribution of heterotrophic plankton to nitrogen (N) regenerationin the water column, and its significance for the requirementsof phytoplankton, were studied at the seasonal scale in thecoastal upwelling ecosystem of A Coruña (Galicia, NWSpain). During 1995–1997, monthly measurements were takenof hydrographic conditions, dissolved nutrients, and abundanceand biomass of microplanktonic heterotrophs (bacteria, flagellatesand ciliates), phytoplankton and mesozooplankton (>200 µm).Additionally, series of experiments were conducted to quantifyN fluxes, including primary production (¹⁴C method), phytoplanktonuptake of nitrate, ammonium and urea (¹⁵N-labelling techniques),microheterotrophic regeneration of ammonium, mesozooplanktongrazing (chlorophyll gut-content method) and excretion of ammoniumby mesozooplankton. Two N budgets were built for the averagesituations of high (>100 mg C m^-2 h^-1) and low (<100 mgC m^-2 h^-1) primary production. The results revealed that phytoplanktonrelied strongly on regenerated ammonium all year round (33 and43% of total N uptake in high and low production situations,respectively). This demand for ammonium was closely matchedby regeneration rates of microplankton (0.14–0.25 mmolN m^-2 h^-1), whereas zooplankton contributed on average <10%to N regeneration. Likewise, zooplankton grazing had littledirect control on phytoplanktonic biomass. The results obtainedindicate that in the A Coruña upwelling system, N biomassof heterotrophic plankton is generally higher than phytoplanktonN biomass. The high rates of N regeneration measured also suggestthat a large proportion of the organic matter produced afteran upwelling pulse is recycled in the water column through themicrobial food web.     

Top-down control on plankton components in an Antarctic pond: experimental approach to the study of low-complexity food webs

In order to address the top-down effect on the different phytoplankton size-fractions and ciliates, a survey at microcosm scale was conducted in a hypertrophic Antarctic pond, testing the hypotheses that (1) the picophytoplankton is regulated by a top-down control exerted by organisms of the bigger size-fractions, and (2) the nanoplankton fraction (algae and ciliates) is not regulated by a top-down control exerted by the microplankton. The treatments enclosed pond water that was filtered to obtain the different plankton sizes: (a) through 55 μm, (b) 20 μm, and (c) 3 μm pore size filters. The variation in the net growth rate (k′) of the phytoplankton size-fractions and ciliates was analysed after 4 days. The results determined a significant difference (P<0.011) in the k′ value of the picophytoplankton when nano and micro-sized fractions where removed. Conversely, nanophytoplankton and nanociliates were not affected by the removal of bigger size-fractions. We suggest that in this pond the top-down control of the picophytoplankton is relevant, and that the grazing impact is not a key factor in the regulation of the nano-sized (algae and ciliates) plankton components.     

Dinoflagellate cyst production at a coastal Mediterranean site

To assess the diversity and seasonality of dinoflagellate cystproduction, surface sediment and trap samples were studied inthe Gulf of Naples (Mediterranean Sea). A total of 59 differentcyst morphotypes were recorded. At the stations within the 70m isobath, sediment assemblages were dominated by calcareousPeridiniales (66–79%), while at the deepest stations non-calcareousPeri-diniales attained the highest percentages (40–49%).The sediment trap sampling, carried out fortnightly over twoannual cycles, revealed high production rates (up to 1.7 x 10⁶cysts m^–2 day^–1) from spring to late autumn of bothyears, with a distinct seasonal production pattern. Althoughrather similar in species composition, the total cyst flux differedmarkedly between the 2 years (1.26 and 0.55 x 108 cysts m^–2year^–1, respectively). Species-specific production patternswere observed: some species formed cysts over several months,others in restricted periods of the year. Cyst-forming speciesconstituted a small part of the planktonic dinoflagellate populationsrecorded in the area. A coupling between the trap material andsurface water plankton was observed for calcareous Peridiniales.This sampling approach allowed the detection of some speciesnever recorded before in the gulf, including two potentiallytoxic species: Alexandrium andersoni and Gymnodinium catenatum-likespecies.     

THE RELATIONSHIP BETWEEN ENVIRONMENTAL VARIABLES AND THE DENSITY OF THE MUD SNAIL HYDROBIA TOTTENI IN A NOVA SCOTIA SALT MARSH

The relationship between environmental variables and the densityof Hydrobia totteni was examined in a restricted area of LawrencetownMarsh, Nova Scotia. Densities averaged 50/m² in Spartina alterniflora,9800/m² in Zostera marina, and 19,000/m² in sand. Of 8 environmentalfactors considered, depth and sediment grain size were the mostimportant in explaining variations in snail density. Since thesefactors together explained only 42.0% of the total variationno single physical factor controls the density of Hydrobia ina restricted area, though salinity does control densities overa wide area. Salinity and temperature tolerances and the abilityof the snails to select appropriate sediment grain sizes weredetermined experimentally and related to the distribution ofthe snails in the marsh. ^*Present address: Western Australian Museum, Francis Street,Perth, Western Australia, 6000 (Received 20 June 1976;     

Morphogenetic diversity and biotoxin composition of Alexandrium (Dinophyceae) in Irish coastal waters

The diversity of Alexandrium spp. in Irish coastal waters was investigated through the morphological examination of resting cysts and vegetative cells, the determination of PSP toxin and spirolide profiles and the sequence analysis of rDNA genes. Six morphospecies were characterised: A. tamarense, A. minutum, A. ostenfeldii, A. peruvianum, A. tamutum and A. andersoni. Both PSP toxin producing and non-toxic strains of A. tamarense and A. minutum were observed. The average toxicities of toxic strains for both cultured species were respectively 11.3 (8.6 S.D.) and 2.3 (0.5 S.D.) pg STX equiv. cell⁻¹. Alexandrium ostenfeldii and A. peruvianum did not synthesise PSP toxins but HPLC–MS analysis of two strains showed distinct spirolide profiles. A cyst-derived culture of A. peruvianum from Lough Swilly mainly produced spirolides 13 desmethyl-C and 13 desmethyl-D whereas one of A. ostenfeldii, from Bantry Bay, produced spirolides C and D. Species identification was confirmed through the analyses of SSU, ITS1-5.8S-ITS2 and LSU rDNA genes. Some nucleotide variability was observed among clones of toxic strains of A. tamarense, which all clustered within the North American clade. However, rDNA sequencing did not allow discrimination between the toxic and non-toxic forms of A. minutum. Phylogenetic analysis also permitted the differentiation of A. ostenfeldii from A. peruvianum. Resting cysts of PSP toxin producing Alexandrium species were found in Cork Harbour and Belfast Lough, locations where shellfish contamination events have occurred in the past, highlighting the potential for the initiation of harmful blooms from cyst beds. The finding of supposedly non-toxic and biotoxin-producing Alexandrium species near aquaculture production sites will necessitate the use of reliable discriminative methods in phytoplankton monitoring.     

The seasonal productivity of phytoplankton in a hypertrophic, gravel-pit lake

The seasonal time course of phytoplankton primary productivitywas studied weekly in a hypertrophic, gravel-pit lake closeto Madrid, Spain. Chlorophyll a ranged 22–445 mg m^–2.Gross primary productivity attained 0.28±0.14 g C m^–2h^–1 (range: 0.06–0.60), its yearly value being 900g C m^–2, but the shallow euphotic depths and the highplankton respiration ensured that net productivity was generallylow. Respiration losses amounted to 0.31±0.24 g O₂ m^–2h^–1, with phytoplankton respiration roughly attainingone-half of overall plankton respiration. Areal phytoplanktonproductivity and plankton respiration followed a seasonal trendbut this was not the case for photosynthetic capacity. Surfacephotoinhibition was evenly distributed throughout the study.Quantum yields showed an increasing depth trend, but no seasonaltrend. Both P_max and I_k were both temperature- and irradiance-dependent.As compared with lakes of lesser trophic degree, phytoplanktonprimary production in hypertrophic lakes might be increasednot only by higher nutrient contents but also by low chlorophyll-specificattenuation coefficients and low background, non-algal attenuation,thereby allowing for higher areal chlorophyll contents and hencehigher areal productivity. Our study suggests that physical(irradiance and water column stability) as well as chemicalfeatures (dissolved inorganic carbon and soluble reactive phosphorus)may control seasonality of phytoplankton primary productionin this lake despite recent claims that only physical factorsare of significance in hypertrophic lakes. However, this doesnot explain all the variability observed and so a food web controlis also likely to be operating.     

Mycosporine-like amino acids in planktonic organisms living under different UV exposure conditions in Patagonian lakes

Mycosporine-like amino acids (MAAs) were studied in zooplanktonfrom 13 Argentinian lakes covering a broad range in altitude,maximum depth and physico-chemical properties of the water.Four to nine different MAAs (predominantly porphyra-334 andshinorine) were found in the copepods Boeckella gibbosa, B.gracilipes, B. meteoris and Parabroteas sarsi, and in the ciliateStentor amethystinus, while MAAs were undetectable in the cladoceranDaphnia middendorffiana. Among the different copepods, maximumMAA concentrations accounted for 0.25–1.31% of the dryweight, and contents were generally about three to seven times(up to 43 times) higher in the animals living in the clearestlakes compared to those occurring in low-UV systems. This variabilityin the content of MAAs was related to the lake altitude (r²= 0.71), and the fraction of the water column to which 1% ofthe surface UV radiation at 320 nm penetrated (r² = 0.57). Ourdata therefore underscore the role of MAAs as sunscreens todecrease the potential negative effects of solar radiation,but they also indicate that other environmental factors besidesUV transparency play a role in determining MAA concentrations.One lake was selected to obtain additional information on thequalitative composition of MAAs in seston of <100 µmbetween two sampling sites and over a 2 month study period (australsummer). Six different MAAs were detected in the samples, withporphyra-334 and palythine being predominant. In the copepodscollected simultaneously, there was low variation in MAA concentrationsbetween the two sites and over time. Thus, our results suggestthat under similar UV exposure conditions MAA contents of planktonicorganisms show low temporal variation.     

The distribution of microcrustacea in the littoral zone of a freshwater lake

The distribution of microcrustacea in the water column, sediments and on different macrophyte species was examined in the littoral zone of Jack Lake, Nova Scotia, Canada. Large numbers of microcrustacea occurred in association with macrophytes, suggesting that this habitat should receive greater attention in future studies of microcrustacean numbers, biomass, and production. The relative abundance of different microcrustacea varied considerably among sediments, macrophytes and water column samples. Although microcrustacean species composition differed among macrophyte groups, consistent differences in absolute numbers per gram could not be demonstrated. Epiphytic microcrustacean community structure also varied among depth strata in Jack Lake. Few epiphytic and benthic microcrustacea migrated into the water column on a diurnal basis.     

Simulated annual plankton production in the northeastern Pacific Coastal Upwelling Domain

A microcomputer simulation model is presented that describesthe generalized plankton production dynamics, in the surfacemixed layer, of the Juan de Fuca Eddy located on the southwesternBritish Columbia continental shelf. The Juan de Fuca Eddy simulationmodel evaluates how the annual biomass production of diatoms,copepods and euphausiids is forced by plankton feeding interactions,seasonal variability in upwelling, water temperature and solarradiation, and generalized fish predation. The model estimatesannual primary production of 345 g C m^–2 year^–1and secondary production of 19.4 g C m^–2 year^–1for copepods and 6 g C m^–2 year^–1 for euphausiids,during 1985–89; -90% of the annual plankton productionwas generated during the April-October upwelling season. Perturbationsof 22 abiotic and biotic parameters, one at a time by ±10%of nominal values, indicated that oceanic variability (e.g.upwelling rate) most strongly affected primary production. Conversely,zooplankton production was most sensitive to variability inbiological parameters describing zooplankton grazing potentialand growth (e.g. gross growth efficiency). Simulated seasonalbiomass patterns of diatoms, copepods and euphausiids were foundto closely match empirical data. However, euphausiid biomassproduction in the Juan de Fuca Eddy alone was unable to meetthe demands of estimated pelagic fish consumption. Local Eddyeuphausiid populations had to be supplemented, from regionaleuphausiids. by a mechanism that is proposed to be linked tothe seasonal pattern and intensity of positive Ekman transport(upwelling).