Characteristics of feeding on dinoflagellates by newly hatched larval crabs

The zoeal larvae of brachyuran crabs must feed soon after hatching on a diet that includes large micro- and mesozooplankton in order to satisfy nutritional requirements. However, newly hatched larvae have been shown to ingest a variety of dinoflagellates, perhaps using microbial carbon sources to sustain them until they encounter more favored prey. Ingestion of dinoflagellates by larval crabs has been documented previously under conditions in which the larvae were exposed to algae provided in monoculture or in defined mixtures of cells. We report here on experiments conducted on the hatching stage of five crab species to determine if ingestion of dinoflagellates occurred when they were provided in combination with Artemia sp. nauplii or after a period of feeding on mesozooplankton. Quantitative measurements of chl a in the larval guts provided evidence of ingestion of algal cells. Active ingestion of the dinoflagellate Prorocentrum micans at specified intervals during an extended feeding period was determined on larvae of two crab species using fluorescently labeled cells provided for brief periods at prescribed time intervals. Stage 1 larvae of four of the five crab species ingested dinoflagellates when they were provided in combination with nauplii and larvae of all five species ingested cells after feeding solely on nauplii for 24 h. Ingestion of algal cells was first evident in the larval guts after 6 h of feeding at both low (200 cell ml⁻¹) and high (1,000 cells ml⁻¹) prey densities. Higher prey densities resulted in higher gut chl a. Larvae continuously exposed to dinoflagellates actively ingested cells at every 3 h interval tested over a 36 h period. Results confirm previous studies that larvae will ingest dinoflagellates even when they are encountered in a mixed prey field or when having previously fed. Ingestion of cells may occur on a continual basis over time.     

Importance of food quality in determining development and survival of Calanus pacificus (Copepoda: Calanoida)

Mixed zooplankton were collected in June and July of 1985 and 1986 from La Jolla Bay, California, USA, and experiments were conducted to determine how selected dinoflagellates affect development and survival of nauplius larvae of Calanus pacificus. We raised nauplii from eggs on nine species of dinoflagellates at concentrations generally >300 g C l^-1, and compared their development and survival to controls reared using the diatom Thalassiosira weissflogii or filtered seawater. Experiments were conducted for 6 d at 17°C. Development and survival rates of the nauplii fell clearly into one of two groups, depending upon the phytoplankton used as food. The first group was characterized by high development rate (0.46 to 0.84 stage d^-1), and by >27% of the original cohort surviving to at least Nauplius IV or V. The five species producing this result were Gymnodinium simplex, G. splendens, Exuviaella marie-lebourae, Gyrodinium dorsum, and T. weissflogii. The second group was characterized by a development rate similar to that in filtered seawater (0.21 to 0.34 stage d^-1), and by nauplii generally failing to molt past the first feeding stage (Nauplius III), often accompanied by high mortality. The five species producing this result were Gyrodinium resplendens, Ptychodiscus brevis, Glenodinium sp., Amphidinium carterae, and Gonyaulax grindleyi. Development rate and survival were not related to cell size or cell carbon, nor to shape or texture (thecate vs athecate dinoflagellates). Poor growth could be related to the absence of some important, but unidentified, nutritional factors. Alternatively, it could be caused by the presence of plant secondary metabolites which are deleterious to growth, a factor we suspect in P. brevis in particular. Prefeeding nauplii exposed to P. brevis lost neuromuscular control prior to becoming lethargic and dying; nutritional deficiencies may not explain these effects. Methods employed in this study provide useful bioassays for detecting chemical interactions between marine plants and animals. Lethal or sublethal effects of dinoflagellates on their most likely potential predators — copepods — may partially explain why they form significant blooms.     

Effects of the dinoflagellates Karlodinium veneficum and Prorocentrum minimum on early life history stages of the eastern oyster (Crassostrea virginica)

The bloom-forming dinoflagellates Prorocentrum minimum and Karlodinium veneficum can have detrimental effects on some marine life, including shellfish, but little is known about their effects on early life history stages of bivalves. In the Chesapeake Bay region, blooms of these dinoflagellates overlap with the spawning season of the eastern oyster, Crassostrea virginica. In laboratory experiments, we compared the effects of P. minimum and K. veneficum on the survival and development of embryos and larvae of the eastern oyster. At 10⁴ cells ml⁻¹, P. minimum did not have a negative effect on embryos and larvae in 2-day exposures. The yield of D-hinge larvae was equal to or greater than in control treatments. At 2 × 10⁴ cells ml⁻¹ (approximately equal biomass to the P. minimum treatment) K. veneficum caused significant mortality to oyster embryos within 1 day and almost no embryos developed into D-hinge larvae. This effect was not alleviated by the provision of an alternate food source (Isochrysis sp.). Significant mortality was observed when larvae were exposed to K. veneficum at concentrations of 10⁴ cells ml⁻¹ (approximately 5 ng ml⁻¹ of karlotoxin). The K. veneficum cultures used in these experiments were relatively low in toxin content, more toxic strains could be expected to cause mortality at lower cell concentrations. Survival and maturation of embryos and larvae may be reduced when spawns of the eastern oyster coincide with high bloom densities of K. veneficum.     

Growth and herbivory by heterotrophic dinoflagellates in the Southern Ocean,studied by microcosm experiments

Growth and herbivory of heterotrophic dinoflagellates (Gymnodinium sp.) from the Weddell Sea and the Weddell/Scotia Confluence were studied in 1988 in 100-liter microcosms. The microcosms were screened through 200-µm or 20-µm mesh nets and incubated for 12 d at 1 °C under artificial light. Mean cell volume of dinoflagellates was 1 000 to 1 500µm³, and that of their phytoplankton prey 360 to 430µm³. Dinoflagellate growth rate followed a Holling type II functional response, with a maximum growth rate of 0.3 d^–1 and half-saturation food concentrations of 1.0µg chlorophylla l^–1, 50µg C l^–1, or 1 500 cells ml^–1. Carbon budgets based on¹⁴CO₂ assimilation and biomasses of phytoplankton and heterotrophic dinoflagellates suggested a balance between phytoplankton grazing loss and dinoflagellate consumption, assuming a dinoflagellate carbon conversion efficiency of 40%. Applying this to the functional response yielded estimates of maximum ingestion rate (0.8µg Cµg^–1 C d^–1, or 6 pg C dinoflagellate^–1 h^–1) and maximum clearance (0.8 to 1.2 × 10⁵ body volumes h^–1, or 80 to 120 nl ind.^–1 h^–1). The microcosm experiments suggested that heterotrophic dinoflagellates may contribute significantly to maintenance of low phytoplankton biomass in the Southern Ocean.     

Toxin accumulation and feeding behaviour of the planktonic copepod Calanus finmarchicus exposed to the red-tide dinoflagellate Alexandrium excavatum

The planktonic copepod Calanus finmarchicus is a dominant member of the zooplankton community in the lower St. Lawrence Estuary in eastern Canada. Blooms of the toxic marine dinoflagellate Alexandrium excavatum which produces high cellular levels of paralytic shellfish poisoning (PSP) toxins, occur during the period of high C. finmarchicus production in summer in this region. To study the feeding behaviour of C. finmarchicus in the presence of Alexandrium spp., experiments were conducted in which female adult copepods collected from the St. Lawrence Estuary between May and September 1991 were exposed under controlled conditions to two toxic isolates of A. excavatum (Pr18b and Pr11f) from the estuary and to a non-toxic control (PLY 173) of a closely related species, A. tamarense isolated from the Tamar Estuary, Plymouth, U.K. Clearance rates on non-toxic A. tamarense cells averaged 5.5 ml ind^-1 h^-1 but were nearzero with either toxic isolate. When presented with a mixture of A. excavatum and the non-toxic diatom Thalassiosira weissflogii in varying proportions, C. finmarchicus fed upon the diatom but avoided the toxic dinoflagellate. Although feeding rates on A. excavatum were very low, toxin analysis by high-performance liquid chromatography with fluorescence detection (HPLC-FD) revealed that the PSP toxins were accumulated in copepods exposed to toxigenic dinoflagellates.The toxin composition in copepods was similar to that of the toxic dinoflagellate, but not necessarily identical, particularly after short-term (2-h) exposure, when relatively elevated levels of N-sulfocarbamoyl toxins were detected. The evidence suggests that C. finmarchicus ingests toxic dinoflagellate cells, either mistakenly or during exploratory bouts of feeding, and accumulates PSP toxins in its gut system and perhaps in other tissues.     

Accumulation of paralytic shellfish poisoning toxins in planktonic copepods during a bloom of the toxic dinoflagellate<Emphasis Type="Italic"> Alexandrium tamarense</Emphasis> in Hiroshima Bay,western Japan

Concentrations of paralytic shellfish poisoning (PSP) toxins in toxic dinoflagellate cells and in marine planktonic copepods were monitored during the bloom of Alexandrium tamarense in Hiroshima Bay, western Japan. Concentration of the toxins retained by copepods was a function of the ambient toxin concentration, i.e. the product of A. tamarense cell density and cellular toxicity. The toxin concentration in copepods increased with the increase of toxicants in the seawater then leveled off, but decreased significantly at higher concentrations. In the field, the maximum toxin concentration was 1.2 pmol ind^-1, whereas in the laboratory, the copepod Acartia omorii accumulated a much higher concentration of PSP toxins (24 pmol ind^-1). Feeding avoidance against Alexandrium tamarense and a shift to alternative food sources such as diatoms in the field might keep their toxin levels lower than their potentially maximum level. The copepod toxin levels in the field were not so high as to cause an instantaneous lethal effect on their predator fishes but may reach possibly lethal levels after a few days' continuous feeding. Overall toxin retention by copepods after 12 h feeding and 2 h starvation was only 2.5% of total ingested toxins, which suggested that a significant amount of toxins was released into the seawater. Measurements of toxin reduction and gut evacuation suggested that the toxins were removed through both fecal evacuation and metabolism (e.g. excretion, decomposition and transformation). The results, as a whole, imply that copepods can be a link for PSP toxin flux in both pelagic and benthic food webs and can also be a sink for toxins by metabolizing and removing them from the environment.Communicated by T. Ikeda, Hakodate     

Impact of copepod grazing on the red-tide flagellate Chattonella antiqua

Although planktonic copepods are major suspension feeders in the sea, the impact of their grazing pressure upon red-tide flagellates has not been fully investigated. In the present study, the grazing of adult females of several copepod species is examined using three food types: viz. natural suspended particles, natural suspended particles mixed with cultured Chattonella antiqua, and cultured C. antiqua. The functional response on C. antiqua was investigated for five species of copepods (Acartia erythraea, Calanus sinicus, Centropages yamadai, Paracalanus parvus and Pseudodiaptomus marinus). Ingestion rates increased linearly with increasing cell concentrations until a maximum level was reached, beyond which the rates were constant. This cell concentration was higher for larger copepods. The weight-specific maximum ingestion rates were higher in the small species. In general, copepods tended to feed selectively on larger particles when feeding on natural particles. This tendency was strongest in a simulated red-tide environment. Thus, it can be surmised that copepods may selectively graze on C. antiqua during the outbreak of a red tide. Grazing pressure by the natural copepod community in Harima Nada, the Inland Sea of Japan, was calculated by integration of the laboratory determined feeding rates and field measurements of zooplankton biomass. The daily removal rate was 3.4 to 30.8% (mean: 12.3%) of C. antiqua biomass at 20 cells ml^-1 and decreased to 0.6–4.3% (mean: 1.8%) at 500 cells ml^-1. Therefore, the grazing pressure by the copepod community is important at the initial stage of the red tide.     

Trophic role of small cyclopoid copepod nauplii in the microbial food web: a case study in the coastal upwelling system off central Chile

Copepod grazing impact on planktonic communities has commonly been underestimated due to the lack of information on naupliar feeding behaviour and ingestion rates. That is particularly true for small cyclopoid copepods, whose nauplii are mainly in the microzooplankton size range (<200 μm). The trophic role of Oithona spp. nauplii was investigated off Concepción (central Chile, ~36°S) during the highly productive upwelling season, when maximum abundances of these nauplii were expected. Diet composition, ingestion rates, and food-type preferences were assessed through grazing experiments with different size fractions of natural planktonic assemblages (<3, <20, <100, and <125 μm) and cultures of the nanoflagellate Isochrysis galbana. When the Oithona spp. nauplii were offered a wide range of size fractions as food (pico- to microplankton), they mostly ingested small (2–5 μm) nanoflagellates (5–63 × 10³ cells nauplius⁻¹ day⁻¹). No ingestion on microplankton was detected, and picoplankton was mainly ingested when it was the only food available. Daily carbon (C) uptake by the nauplii ranged between 28 and 775 ng C nauplius⁻¹, representing an overall mean of 378% of their body C. Our relatively high ingestion rate estimates can be explained by methodological constraints in previous studies on naupliar feeding, including those dealing with “over-crowding” and “edge” effects. Overall, the grazing impact of the Oithona spp. nauplii on the prey C standing stocks amounts up to 21% (average = 13%) for picoplankton and 54% (average = 28%) for nanoplankton. These estimates imply that the nauplii of the most dominant cyclopoid copepods exert a significant control on the abundances of nanoplankton assemblages and, thereby, represent an important trophic link between the classical and microbial food webs in this coastal upwelling system.     

Investigations on epibenthic meiofauna I. Abundances on and repopulation of the tube-caps of Diopatra cuprea (Polychaeta: Onuphidae) in a subtropical system

Cryptic meiofauna populated the imbricate, shell-sediment matrix of the tube-caps produced by the polychaete Diopatra cuprea (Bose) in Tampa Bay, Florida, USA. Nematodes, copepods (adults and nauplii), polychaetes (adults and juveniles) and amphipods (adults and juveniles) were the most abundant taxa found on tube-caps. Meiofaunal densities on tube-caps were 4 to 19 times higher than equivalent volumes of sediment in cores taken adjacent to D. cuprea tubes. Recruitment onto tube structure occurred within 1 to 2 d after defaunated tube-caps were replanted into sediments in the field (February–November 1980). Repopulation of copepods (adults and nauplii) attained levels equal to or exceeding natural abundances on tube caps within 1 to 5 d; nematode recovery rates were inconsistent. Short-term experiments using a variety of defaunated tube treatments indicated that immigration onto above-sediment tube-caps proceeds via both water column and sediment pathways. Based on data on tube-cap longevity and construction as well as meiofaunal recruitment rates, we conclude that the generation of new tube-cap structure is exploited rapidly by meiofauna.     

Electrophoretic survey of genetic variation in Pecten maximus,Chlamys opercularis,C. varia and C. distorta from the Irish Sea

Food selection by young larvae of the gulf menhaden (Brevoortia patronus) was studied in the laboratory at Beaufort, North Carolina (USA) in 1982 and 1983; this species is especially interesting, since the larvae began feeding on phytoplankton as well as microzooplankton. When dinoflagellates (Prorocentrum micans), tintinnids (Favella sp.), and N1 nauplii of a copepod (Acartia tonsa) were presented to laboratory-reared, larval menhaden (3.9 to 4.2 mm notochord length), the fish larvae ate dinoflagellates and tintinnids, but not copepod nauplii. Larvae showed significant (P<0.001) selection for the tintinnids. Given the same mixture of food items, larger larvae (6.4 mm notochord length) ate copepod nauplii as well as the other food organisms. These feeding responses are consistent with larval feeding in the northern Gulf of Mexico, where gulf menhaden larvae between 3 and 5 mm in notochord length frequently ate large numbers of dinoflagellates (mostly P. micans and P. compressum) and tintinnids (mostly Favella sp.), but did not eat copepod nauplii. As larvae grew, copepod nauplii and other food organisms became important, while dinoflagellates and tintinnids became relatively less important in the diet. Since the tintinnids and nauplii used in the laboratory feeding experiments were similar in size as well as carbon and nitrogen contents, the feeding selectivity and dietary ontogeny that we observed were likely due to a combination of prey capturability and larval fish maturation and learning.Contribution No. 5575 of the Woods Hole Oceanographic Institution     

Planktonic bioluminescence in the pack ice and the marginal ice zone of the Beaufort Sea

An icebreaker cruise into the Beaufort Sea in the fall of 1986 provided a unique opportunity for studying planktonic bioluminescence in ice fields and in the marginal ice zone. Bathyphotometer casts (bioluminescence intensity, seawater temperature, beam attenuation coefficient, and salinity) and biological collections were made to a depth of 100 m. A light budget, which describes the planktonic species responsible for the measured bioluminescence, and a dinoflagellate species budget were constructed from the mean light output from luminescent plankton and plankton counts. The vertical distribution of bioluminescence among the ice stations was similar. The maximum intensities were 2 to 8×10⁶ photons s^-1 cm^-3 in the upper 50 m of the sea-ice interface. The marginal ice zone station (MIZ) exhibited a maximum intensity of 2 to 3×10⁸ photons s^-1 cm^-3 between 5 and 30 m depth. At Ice Station 2, Metridia longa and their nauplii contributed approximately 80% of stimulable bioluminescence in the upper 10 m but, overall, Protoperidinium spp. dinoflagellates contributed most of the light to a depth of 100 m. In the MIZ, Protoperidinium spp. dinoflagellates contributed 90% of the light within the upper 10 m, decreasing to 43% of the contributed light at a depth of 40 m. Below 40 m, dinoflagellate bioluminescence decreased to a few percent of the total to a depth of 90 m. Metridia spp. copepods contributed more than 50% of the light at depths from 40 to 90 m. Ostracods, larvaceans, and euphausiid furcilia contributed <1% of all bioluminescence at all depths sampled. Correlation analyses between measured bioluminescence (photons s^-1 cm^-3), the number of bioluminescent dinoflagellates and the light budget for the MIZ indicated highly significant associations: r=0.919, p=0.001, and r=0.912, p<0.001, respectively (Student's two-tailed t-tests). Bioluminescence was negatively correlated with seawater salinity at all stations (p=0.001). Maximum bioluminescence was measured in the less saline surface waters at all stations.     

High mortality in a copepod population caused by a parasitic dinoflagellate

Infection of copepods by parasitic dinoflagellates has been known for many years, but the ecological consequences of this parasitism have been largely neglected. We estimated mortality rates in the copepodParacalanus indicus Wolfenden due to parasitism by the dinoflagellateAtelodinium sp. by applying laboratory mortality rates to a field population of infected copepods in Port Phillip Bay, Australia, sampled in 1982–1985. Adult female copepods were most often infected, with an incidence of 0 to 28.5% (median 6.2%). Stage V female copepodites were less often infected, and males were never infected. The median mortality rate in females was about 7% d^–1, or about one-third of total mortality, and the maximum was 41% d^–1. The frequent occurrence of dinoflagellate parasitoids in some species of copepod implies an important, species-specific mechanism for the regulation of populations.     

Effects of suspended sediments on egg production of the calanoid copepod Acartia tonsa

The estuarine copepod Acartia tonsa was collected on several occasions between 4 April and 14 August 1985 from Terrebonne Bay, Louisiana (29°08N; 90°36W) and the effects in its diet of suspended sediments, collected from the same area, were measured at five different concentrations of sediment (100 to 1 000 ppm) and six phytoplankton concentrations (500 to 13 000 cells ml^-1 Thalassiosira weissflogii). Egg production rate was used as an index of diet quality. At low phytoplankton concentrations (500 cells ml^-1), and at intermediate phytoplankton concentrations (2 000 cells ml^-1) for previously starved copepods, egg production was reduced by up to 40% at a sediment concentration of 250 ppm and further reduced at higher sediment concentrations. At higher food concentrations (4 000 to 13 000 cells ml^-1), suspended sediment had no effect on egg production rates at sediment concentrations up to 500 ppm. Rates were reduced only at the highest sediment concentration of 1 000 ppm. Under most natural conditions, suspended sediment would not significantly affect egg production rates in A. tonsa.     

Effects of prey escape ability,flow speed,and predator feeding mode on zooplankton capture by barnacles

Experimental studies of feeding on zooplankton often involve the use of non-evasive Artemia spp. to represent zooplanktonic prey. Some zooplankton, however, such as copepods, are potentially evasive due to possession of effective predator-avoidance mechanisms such as high-speed escape swimming. In the present study, we compared the efficiencies with which non-evasive (A. salina) and evasive (copepods) zooplankton were captured by a sessile, suspension feeder, the coral-inhabiting barnacle Nobia grandis (Crustacea, Cirripedia). N. grandis specimens and zooplankton used in the present study were collected near Eilat, Israel in 1993. The effect of different flow speeds (from 0 to 14 cm s^-1) on captures of the two preys was also investigated. Additionally, we examined the effect of a flow-induced barnacle behavioral switch from active to passive suspension feeding, on zooplankton capture. Two video cameras were used to make close-up, three dimensional recordings of predator-prey encounters in a computer-controlled flow tank. Frame-by-frame video analysis revealed a highly significant difference (P< 0.001) in the efficiency with which A. salina and copepods were caught (A. salina being much more readily captured than copepods). After an encounter with cirri of feeding barnacles, copepods were usually able to swim out of the barnacles capture zone within one video frame (40 ms), by accelerating from a slow swimming speed (approximately 1.85 cm s^-1) to a mean escape swimming speed of 18.11 cm s^-1 (ca. 360 body lengths s^-1). This was not the case for A. salina nauplii, which usually remained in contact with cirri before being transferred to the mouth and ingested. Thus, experimental studies addressing the methodology of organisms feeding on zooplankton should consider that slow-swimming prey like Artemia sp. nauplii may only represent the non-evasive fraction of natural mesozooplankton assemblages.     

Summer copepod production in subtropical waters adjacent to Australia's North West Cape

Growth and secondary production of pelagic copepods near Australia's North West Cape (21° 49 S, 114° 14 E) were measured during the austral summers of 1997/1998 and 1998/1999. Plankton communities were diverse, and dominated by copepods. To estimate copepod growth rates, we incubated artificial cohorts allocated to four morphotypes, comprising naupliar and copepodite stages of small calanoid and oithonid copepods. Growth rates ranging between 0.11 and 0.83 day^–1 were low, considering the high ambient temperatures (23–28°C). Calanoid nauplii had a mean growth rate of 0.43±0.17 day^-1 (SD) and calanoid copepodites of 0.38±0.13 day^-1. Growth rates of oithonid nauplii and copepodites were marginally less (0.38±0.19 day^–1 and 0.28±0.11 day^–1 respectively). The observed growth rates were suggestive of severe food limitation. Although nauplii vastly outnumbered copepodite and adult copepods, copepodites comprised the most biomass. Copepodites also contributed most to secondary production, although adult egg production was sporadically important. The highest copepod production was recorded on the shelf break (60 mg C m^-2 day^-1). Mean secondary production over both shelf and shelf break stations was 12.6 mg C m^-2 day^-1. Annual copepod secondary production, assuming little seasonality, was estimated as ~ 3.4 g C m^-2 year^-1 (182 kJ m^-2 year^-1).Communicated by G.F. Humphrey, Sydney     

Zooplankton grazing on the coccolithophore Emiliania huxleyi and its role in inorganic carbon flux

Grazing and faecal pellet production by the copepods Calanus helgolandicus and Pseudocalanus elongatus, feeding on the coccolithophore Emiliania huxleyi, were measured under defined laboratory conditions, together with the chemical characteristics and sinking rates of the faecal pellets produced. Ingestion rates of both copepods were equivalent at comparable cell concentrations, the relationship between ingestion rate (I, cells copepod^-1 h^-1) and food concentration (C, cells ml^-1), being I=0.558C for both species. P. elongatus produced a larger number of smaller faecal pellets than C. helgolandicus, but egested a larger volume of material per individual. Only between 27 and 50% of the ingested coccolith calcite was egested in the faecal pellets, and it is possible that acid digestion in the copepod gut is responsible for these considerable losses. Average sinking rates of faecal pellets containing E. huxleyi coccoliths, produced by both species, were >100 m d^-1. The implications of the quantitative laboratory estimates for the vertical flux of inorganic carbon are considered using recently studied shelf-break and oceanic E. huxleyi blooms in the N. E. Atlantic as examples.     

Ingestion,growth and development of Penaeus indicus larvae as a function of Thalassiosira weissflogii cell concentration

Penaeus indicus larvae have been successfully reared in the laboratory using Thalassiosira weissflogii, Brachionus plicatilis and Artemia salina nauplii as food, with an average survival of 95.8% from nauplius 6 to postlarva 1. The effect of T. weissflogii cell concentration on larval ingestion, development and growth (total length) was investigated. Cell ingestion rates showed a saturation response to concentration. Both maximum ingestion rates and incipient limiting levels (the lowest concentration before ingestion rates were limited) were established for the feeding larval stages. Both were found to increase with progressive increase in larval development. Maximum ingestion rates increased from 0.25×10⁴ cells. larva^-1.h^-1 during protozoea 1 to reach a peak of 1.2×10⁴ cells. larva^-1.h^-1 during mysis 3 and then declined to 0.6×10⁴ cells. larva^-1.h^-1 at postlarva 1. Incipient limiting levels (ILLs) increased from approximately 0.6×10⁴ cells.ml^-1 during protozoea 2, to 0.65×10⁴ cells.ml^-1 during mysis 1, to 1.3×10⁴ cells. ml^-1 during mysis 3 to 1.6×10⁴ cells.ml^-1 at post-larva 1. Filter feeding efficiency was found to reach a maximum during mysis 1. Filter mechanisms are discussed. Generally, the most advanced larval development per unit time occurred at concentrations at and above the ILLs, while retarded development occurred below these levels. Growth increased asymptotically with cell concentration. Incipient growth limiting levels (IGLLs; the lowest concentration before growth was significantly limited) also increased with larval development and with the exception of mysis 3 they coincided with the ILLs. IGLLs increased from 0.55×10⁴ cells.ml^-1 during protozoea 2, to 0.66×10⁴ cells.ml^-1 during mysis 1, to 0.99×10⁴ cells.ml^-1 during mysis 3, to 1.62×10⁴ cells.ml^-1 at postlarva 1. Below the ILLs where ingestion was limited, animals were significantly smaller, with larval development and growth positively correlated to ingestion rates. When culturing penaeoid larvae, ambient cell concentrations should be kept above these known limiting levels to yield consistently good larval survival and growth.     

Comparisons among species ofAlexandrium (Dinophyceae) grown in nitrogen- or phosphorus-limiting batch culture

Three species of the dinoflagellate genusAlexandrium (Halim)-two strains of toxic.A. minutum, one each of nontoxicA. tamarense andA. affini-were grown in batch culture in either a low-nitrogen or a low-phosphate medium. Maximum carbon-specific growth rates forA. tamarense were lower (at <0.25 d^-1) than for the other strains, which all exceeded 0.38 d^-1. C-quotas (C content per cell) during exponential growth were similar for all strains (2.5 ng C cell^-1), with cells becoming smaller during the N-limiting stationary phase, but enlarging during prolonged P-deprivation. Values of ¹³C during the exponential phase were low (-25to-30), with most cells during the light phase swimming at the surface when nutrient-replete and migrating to the bottom of the flasks when nutrient-deplete with ¹³C rising to around-15. Biomass could not be estimated reliably from pigmentation, but could be estimated from biovolume (r>0.95), although this was complicated in cultures ofA. minutum by the presence of particles comprized of thecal plates of a similar size to intact cells. Alkaline phosphatase activity was not a reliable indicator a P-status. The most toxic strain tested (A. minutum AL1V) contained the highest concentrations of free amino acids, of arginine (a precursor of paralytic shellfish toxins) and of proline, and also had the lowest C:N mass ratio (at 4.3).A. affini contained the lowest concentrations of arginine, andA. tamarense the highest exponential phase C:N (7.8). For all strains, the mole ratio of intracellular glutamine: glutamate (Gln: Glu, which was abnormally high compared to other algae) could only be used to indicate the presence or absence of N-stress rather than the degree of stress. Additions of ammonium and phosphate resulted in increases in Gln: Glu within 20 min in N-stressed cells and also enhanced toxin content inA. minutum (mainly gonyautoxin) 4 over a 24 h period.     

Allelopathic interactions between Skeletonema costatum and Alexandrium minutum

Potential allelopathic interactions between Skeletonema costatum and Alexandrium minutum were investigated using mixed cultures and culture filtrate in nutrient-replete medium. A. minutum growth was inhibited when grown in S. costatum filtrate, with the inhibitory effect directly proportional to the percentage of filtrate added. This demonstrates that the release of allelopathic compounds caused the growth inhibition. In contrast, the filtrate of A. minutum exerted no allelopathic activity on S. costatum. An autoinhibitory compound (15(S)-HEPE) extracted and purified from S. costatum culture was added to cultures of both S. costatum and A. mintum. This substance could depress S. costatum growth, but showed no significant inhibitory activity on A. minutum. This documented a second type of allelochemical interaction, termed auto-allelopathy, caused by a different compound from the one or ones that affected A. minutum in the co-cultures with added crude filtrate. Further studies are needed to explore the relative importance of these two types of allelopathy as factors influencing the competition between S. costatum and A. minutum in the field. Furthermore, given the observed decrease in diatom dominance relative to dinoflagellates with increasing eutrophication, one can predict that toxic species like A. minutum might become more prevalent in the future in the East China Sea if the trend of increasing pollution of coastal waters continues.     

Reproduction and growth of Euterpina acutifrons (Copepoda: Harpacticoida) under experimental conditions

Individual specimens of Euterpina acutifrons (Copepoda: Harpacticoida) taken from the mass cultures of the C.N.E.N.-EURATOM Laboratory at Fiascherino, Italy, were reared in new culture media prepared with suspensions of several species of algae in filtered and sterilized sea water. All the experiments were carried out at a temperature of 18°C±1 C°. The influence of food concentrations on adult life-span and reproductive activity of E. acutifrons was analyzed. A good correlation was found between concentration of algal suspension and egg production. Maximum life-span was observed at intermediate values of food concentration. Other experiments were carried out to determine egg fertility and duration of the various embryonic and postembryonic development stages. Embryonic development time was calculated as approximately 2 days; the adult females appeared 10 to 12 days after hatching of Nauplius I. Production of nauplii by 6 females reared under conditions of excess food supply was also studied. These conditions were achieved by supplying high concentrations of a mixture of 4 different algal species and by completely renewing the culture medium at frequent intervals. Under these conditions, each female laid an average of 12.5 sacs and produced an average of 294.3 nauplii. An average production of 355.5 eggs per female was estimated. An analysis was made of growth in size and weight of the females: the average daily egg production in terms of dry weight corresponded to about 32% of the biomass of the adult female. Birch's (1948) method was used to calculate net reproduction rate (R _o=70.89), intrinsic rate of increase (r _m=0.161) and mean generation time (T=26.5 days).This study was performed at: Laboratorio per lo Studio della Contaminazione Radioattiva del Mare, C.N.E.N.-EURATOM, Fiascherino, I-19030 La Spezia, Italy.