Inhibitory effects of copper on marine dinoflagellates

The effect of copper on three species of marine dinoflagellates [Scrippsiella faeroense (Paulsen) Balech et Soares, Prorocentrum micans Ehrenberg, Gymnodinium splendens Lebour] was studied. It inhibited the growth of all species and was lethal to one species in batch cultures. The effect was more pronounced in semicontinuous culture than in batch cultures. Chlorophyll concentrations and rate of uptake of radioactive carbon by cells of S. faeroense were affected in a manner similar to cell numbers. Copper inhibited growth of cells, most probably either by arresting cell division or by penetrating inside the cell and affecting metabolism.     

Waste-water assay with continuous algal cultures: The effect of mercuric acetate on the growth of some marine dinoflagellates

The effect of mercuric acetate was studied in culture experiments with the dinoflagellates Scrippsiella faeroense (Paulsen) Balech et Soares, Prorocentrum micans Ehrenberg and Gymnodinium splendens Lebour. Impairment of growth rates, in vivo chlorophyll fluorescence, maximum cell densities and morphological changes served as criteria for assessing sublethal influences. Tests were made using the batch- and continuous-culture techniques. Addition of Hg at concentrations of 0.001 mg.1^-1 and higher resulted in reduction of relative growth rates. In a few cases populations recovered from the initial decline and showed new growth. Cell counts corresponded very closely to in vivo chlorophyll fluorescence measurements. Morphological variations were observed in S. faeroense, which responded (even in sublethal concentrations) by bursting it's thecae, releasing naked motile cells and forming vegetative resting stages. The problems of optimal algal-bioassay methods are discussed also, in the light of results obtained by other authors.     

Chemical composition of sergestid shrimps (Decapoda: Natantia) collected near Bermuda

Chemical composition was determined for Sergestes corniculum, S. grandis, S. japonicus, S. robustus and S. splendens collected in December, April and September. Protein content expressed as percent of dry weight varied from 65.7% in S. splendens to 46.8% in S. japonicus. Significant differences were noted among species in Apria and September, with highest protein values in April in S. corniculum, s. robustus and S. splendens. Lipid content expressed as percent dry weight ranged from 12.6% in S. robustus to 8.3% in s. splendens. Differences in percent lipid among species were significant in December and September with highest values in s. robustus and lowest values in S. splendens. Carbohydrate content varied from 1.0% in S. grandis to 2.5% in S. splendens. Ash and chitin were only determined for S. corniculum, S. grandis and S. splendens. Although protein and lipid content were correlated with size of shrimp analyzed, protein and lipid values in S. splendens, a species known to occur near the surface only at night, appeared to be related to seasonal zooplankton abundance in near-surface waters. Differences in lipid and protein content among species suggest that higher lipid and lower protein content occurs in species with shorter ranges of diel vertical migration.     

Laboratory experiments on the diurnal vertical migration of marine dinoflagellates through temperature gradients

The diurnal vertical migrations of 4 marine dinoflagellates (Cachonina niei, Ceratium furca, Gymnodinium splendens, and Prorocentrum micans) were studied in a thermally-stratified Plexiglas column (1.70 m deep, 0.61 m diameter). All species migrated through temperature gradients exceeding 7 C°. Some species exhibited altered migration patterns at different salinities either between experiments or compared to observations by other investigators. P. micans exhibited complex changes in swimming speed that depended both on temperature and on the light cycle. These changes provide insight into the possible time course of a field organism's diurnal exposure to physical variability in the water column. P. micans exhibited compositional changes in response to the light cycle, but not to the cross-thermocline temperature differential. A multi-parameter response matrix is required to model coupling between organisms and biologically-active physical mechanisms realistically.     

In vitro establishment of continuous cell cultures and cell lines from ten colonial cnidarians

Continuous cell cultures from the ten taxa of sedentary colonial marine cnidariansStylophora pistillata, Porites lutea, Favia favus (Anthozoa, Hexacorallia, Madeporaria),Parerythropodium fulvum fulvum, Dendronephthya hemprichi, Nephthya sp.Heteroxenia fuscescence (Anthozoa, Octocorallia, Alcyonacea),Clathraria rubrinoides, Plexaura A (Anthozoa, Octocorallia, Gorgonacea) andMillepora dichotoma (Hydrozoa) were established in vitro. Primary cultures of various cell types (5 to 20 m) were obtained from colony fragments and/or planula larvae (collected in 1993 from coral reefs at Eilat, Red Sea and from the San Blas islands, Panama) using three dissociation approaches: a mechanical approach, chemical approach and a novel approach, spontaneous dissociation. Cells were cultured in a modified Leibowitz L15 medium, with 5 to 10% heat-inactivated fetal bovine serum, diluted in seawater. Cell proliferation was observed in primary cultures within 7 to 20 d following dissociation. When developed to secondary cultures, spindle-shaped cells (5 to 10 m) gradually replaced all other cell types seen in primary cultures ofS. pistillata, C. rubrinoides, P.f. fulvum andM. dichotoma. The spindle-shaped cells differentiated to several cell types when the diluted medium was replaced with a concentrated one, or when microbial contamination occurred. Secondary cultures of spindle-shaped cells from three species (S. pistillata, C. rubrinoides andM. dichotoma) were cloned. They gave rise to continuously proliferating cell lines (NIO-SPP-1, NIO-CR-1 and NIO-MD-1, respectively). A clone fromPlexaura planula, originated from one rounded cell, started to differentiate and gave rise to a heterogeneous culture. Samples of the above four cultures were frozen for future work. Rounded cells of various sizes (5 to 30 m) dominated secondary cell cultures ofH. fuscescence, D. hemprichi, Nephthya sp.,P. f. fulvum andF. favus. Several cell cultures fromP. lutea developed flattened cell interconnected by ectoplasmic networks characteristic of the eukaryotic unicellular organisms of the phylum Labyrinthulomycota. We propose that this culture methodology may be used as a ubiquitous protocol for producing tissue cultures from other cnidarians. The established cell lines may be used in a variety of disciplines in cnidarian biology and ecology.     

Dispersal-generated sexual selection in a beetle-riding pseudoscorpion

Summary After several generations within a decaying tree (Ficus spp.), populations of the pseudoscorpion Cordylochernes scorpioides disperse by climbing under the elytra of harlequin beetles (Acrocinus longimanus) eclosing from the tree. Because the beetles then fly to newly-decaying Ficus for mating and oviposition, they act as effective dispersal agents. Field experiments and observations indicate that this dispersal mode has been exploited by males who compete to remain on a beetle as a strategic site for inseminating females dispersing on it and on other beetles. Whereas beetles just eclosed from old trees carried large, female-biased groups of dispersing pseudoscorpions, beetles captured after their maiden flight generally carried a single large male. Multivariate morphometric analyses indicated that these beetle-riding males were much larger than individuals randomly sampled from trees, with fighting traits exhibiting the greatest potential for selection (highest phoretic differentials). In females, sexual receptivity was significantly higher at the beginning of dispersal than at the end, suggesting that mating occurs on beetles. Field experiments confirmed that on-beetle insemination does take place and that small males are displaced from the subelytral space by larger rivals. By contrast, laboratory experiments suggest that large male size may not confer high mating success under the low-density conditions which characterize populations within trees. Thus, in addition to indicating a novel role for dispersal in the evolution of exaggerated male traits, this study suggests that oscillating sexual selection may be important in the maintenance of the extreme phenotypic variation exhibited by males of this species.     

Salinity dependence of sexual and asexual reproduction in the rotifer Brachionus plicatilis

A salinity dependent mictic response was observed in a clone of Brachionus plicatilis cultured in the 2 to 4 salinity range. This response was related to asexual exponential reproduction rates (G) and could be divided into three categories: (a) no mixis occurred at a salinity of 35 S and above, where G values were lower than 0.30 d^-1, (b) low mictic levels in rotifers cultured at 2 and 30 S, where G values ranged between 0.40 to 0.50 d^-1, and (c) high mictic levels in rotifers cultured at salinities ranging between 4 and 20 S, where G values ranged between 0.50 to 0.85 d^-1. Fluctuations in mictic levels varied with time during the course of the experiments. Results suggest that salinity conditions leading to optimal parthenogenic reproduction also support mixis.     

Vertical distribution of phytoplankton during investigations of a natural surface film

An interdisciplinary research program (Kombiniertes Oberflächenfilmprojekt KOFF), in the North Sea area about 75 km west of the Island of Sylt, is directed towards gaining further knowledge on the biological importance of the natural surface film. The investigations include the water body below the film as well as dynamic processes at the surface. Persistence of calm weather conditions (cloudless sky) before and during the investigations permitted observations of the diurnal species-specific vertical migration of Dinophyceae. Two groups could be distinguished: (1) Prorocentrum micans and Ceratium furca; (2) C. horridum, C. macroceros and C. fusus. The active accumulation of P. micans in the slick was verified by a significant correlation coefficient—linearly correlated with the time of day. The results presented reveal continuity between surface film and underlying water column.This work was supported by the Sonderforschungsbereich (SFB) 94, Hamburg, of the Deutsche For-schungsgemeinschaft.     

Osmotic and ionic regulation in the fiddler crab Uca rapax acclimated to dilute and hypersaline seawater

Adult male Uca rapax, collected from the central coast of Venezuela in early 1994 were gradually acclimated to salinities ranging from 1.7 to 139S. The hemolymph osmolality (791±15 mOsmol in crabs from 26S) changed less than three-fold over the entire range of concentrations tested. The urine was isosmotic with the hemolymph in crabs exposed to dilutions <26S, and significantly hyperosmotic in those exposed to media >34.8S. The hemolymph levels of Na⁺, Cl^–, K⁺, Ca²⁺ and Mg²⁺ (320±13, 405±17, 7.8±0.7, 7.2±0.1 and 31±2.2 mmol l^–1, respectively, in crabs acclimated to 26S) were maintained fairly constant over the range from 8.7 to 99S, decreasing by 15% in the more dilute media or increasing sharply to about twice those values in crabs from 139S. The excretory organs contributed to the osmoionic regulation of the hemolymph in crabs exposed to <3.5 or to >34.8S, by means of the partial reabsorption or excretion, respectively, of salts from or into the urine. The results described place U. rapax among the most powerful hypo/hyper-regulating crustaceans known.     

Feeding mechanism of Pelagia noctiluca (Scyphozoa: Semaeostomeae); laboratory and open sea observations

The behaviour of the oral arms of Pelagia noctiluca (Forsskål) has been investigated in open sea predation and in predation induced both in the laboratory and the natural environment. Specimens were first studied in the field and then collected from coastal aggregations in the Gulf of Trieste, North Adriatic Sea, in December 1985 (Lat. 13°40 E, Long. 45°42 N) and in June 1986 (Lat. 13°39 E, Long. 45°43 N). The results of laboratory experiments and in situ observations, recorded on videotapes and photographs, show that the marginal tentacles are utilized to: (1) paralyze the prey; (2) contract and bend inward towards the nearest oral arm. Occasionally the tentacle does not contract and the prey is released; thus prey selection may occur. The oral arms are therefore involved in the: (1) transport of prey from the tentacle to the gastric cavity; (2) catching of motionless prey; (3) anchoring the medusa to the substratum. A similar feeding pattern can explain the survival of several specimens of P. noctiluca near the bottom during the winter of 1985/1986 in the Gulf of Trieste (North Adriatic Sea) in spite of the severe climatic conditions.     

Carbon fixation in marine phytoplankton: carboxylase activities and stable carbon-isotope ratios; physiological and paleoclimatological aspects

We measured the activity of three carboxylases: RuBP carboxylase, PEP carboxylase and PEP carboxykinase of marine phytoplankton species in culture and in natural communities. Activities of the three carboxylases were measured simultaneously with stable carbon-isotope ratios. The enzymatic activities have been used to estimate the importance of carboxylation and its impact on the ¹³C:¹²C ratio (expressed as ¹³C). The marine phytoplankton species in culture were Fragilariopsis kerguelensis, Nitzschia turgiduloides, Skeletonema costatum, Phaeodactylum tricornutum, Isochrysis galbana, Dunaliella marina, and Prorocentrum micans, and the field samples were collected from different depths off the coast of Portugal (August/September, 1981). Our results indicate that, as in terrestrial plants, the ¹³C value is a good indicator of the extent of carboxylation. RuBP carboxylase activity was always predominant, whereas the ¹³C value never reached values typical of the C₄ pathway. The carboxylases could be PEP carboxylase (in dinoflagellates) or PEP carboxykinase (in diatoms). carboxylation increased at the end of the exponential growth phase in a diatom culture and with increased biomass in natural samples. We interpret these increases as an adaptative response mechanism to poor environmental conditions, especially to low light intensity.     

Effects of changes in water salinity upon exercise and cardiac performance in the European seabass (<Emphasis Type="Italic">Dicentrarchus labrax</Emphasis>)

The European seabass is an active euryhaline teleost that migrates and forages in waters of widely differing salinities. Oxygen uptake (M_O2) was measured in seabass (average mass and forklength 510 g and 34 cm, respectively) during exercise at incremental swimming speeds in a tunnel respirometer in seawater (SW) at a salinity of 30 and temperature of 14°C, and their maximal sustainable (critical) swimming speed (U_crit) determined. Cardiac output (Q) was measured via an ultrasound flow probe on their ventral aorta. The fish were then exposed to acute reductions in water salinity, to either SW (control), 10, 5, or freshwater (FW, 0), and their exercise and cardiac performance measured again, 18 h later. Seabass were also acclimated to FW for 3 weeks, and then their exercise performance measured before and at 18 h after acute exposure to SW at 30. In SW, seabass exhibited an exponential increase in M_O2 and Q with increasing swimming speed, to a maximum M_O2 of 339±17 mg kg^–1 h^–1 and maximum Q of 52.0±1.9 ml min^–1 kg^–1 (mean±1 SEM; n=19). Both M_O2 and Q exhibited signs of a plateau as the fish approached a U_crit of 2.25±0.08 bodylengths s^–1. Increases in Q during exercise were almost exclusively due to increased heart rate rather than ventricular stroke volume. There were no significant effects of the changes in salinity upon M_O2 during exercise, U_crit or cardiac performance. This was linked to an exceptional capacity to maintain plasma osmolality and tissue water content unchanged following all salinity challenges. This extraordinary adaptation would allow the seabass to maintain skeletal and cardiac muscle function while migrating through waters of widely differing salinities.Communicated by S.A. Poulet, Roscoff     

Growth and metamorphosis of Aplysia oculifera larvae in laboratory culture

Eggs of Aplysia oculifera (Adams and Reeve, 1850) were incubated in the laboratory. They hatched 8 to 9 d after spawning. Shell length (SL) of the hatched larvae was 102±2 m. Larvae were fed on the unicellular algae Isochrysis galbana in a concentration of 10⁴ cell ml^-1, and after 45 to 60 d grew to a maximum SL of 385±11 m. Larvae survived up to 330 d. A total of 12 species of algae from the natural habitat of A. oculifera were examined as metamorphosis inducers. Red algae Dasia sp., Jania sp., Hypnea sp. and Liagora sp. induced metamorphosis in 66.7±21.2, 28.3±17.7, 26.0±18.5 and 4.0±8.0% of the larvae, respectively. Green algae Enteromorpha intestinalis and Ulva sp. induced metamorphosis in 37.0±11.0 and 9.0±10.4% of the larvae, respectively. Cladophora sp. and Codium dichotomum, and the brown algae Padina pavonia, Colpomenia sinuosa, Hydroclathrus clathratus and Cystoseira sp. did not induce metamorphosis. There was no significant difference in the rate of metamorphosis between young (2 to 4 mo) and old (6 to 8 mo) larvae. Postmetamorphic juveniles grew and developed only when fed with E. intestinalis. They grew to a body length of>8 mm in 50 d. Postmetamorphic juveniles did not survive on other algae. The longevity of the planktonic A. oculifera larvae supports the hypothesis that the larvae can exist in the plankton and survive for several months until the next recruitment. The advantage of non-specificity in metamorphosis induction is discussed.     

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.     

Implications of salinity fluctuation for growth and nitrogen metabolism of the marine diatom Ditylum brightwellii in comparison with Skeletonema costatum

In 1987 effects of salinity fluctuations on growth of Ditylum brightwellii (West) Grunow, isolated from the Eastern Scheldt estuary (SW Netherlands) in 1981, were studied. D. brightwellii was grown in a 12 h light: dark cycle at constant salinity in brackish media. Ammonium-limited cultures were subjected to a salinity fluctuation. By decreasing the salinity to 4.8 photosynthesis and cell division were inhibited; cells were deformed. Protein and carbohydrate contents increased slightly, dark respiration was stimulated and cellular levels of glucose decreased at low salinity; this indicated a possible role of sugars in osmoregulation. Ammonium was accumulated in cultures, amino acids may have been stored; the role of the vacuole as a storage compartment was discussed. Both the ammonium uptake capacity and the affinity for ammonium decreased. Nitrogen limitation was relieved in the transient state. [With the activity of the nitrogen assimilation enzymes glutamine synthetase (GS) and glutamate synthase (GOGAT) being uninhibited by lower salinity.] Recovery from hypo-osmotic stress during a salinity increase was initiated by stimulated photosynthesis; chlorophyll a increased, but persistant contractions of cytoplasm (with chloroplasts) may have delayed cell growth. The glutamate dehydrogenase (GDH) activity decreased further whereas the cellular level of alanine increased in the presence of large ammonium pools; this may indicate a temporary activity of ADH (alanine dehydrogenase). Skeletonema costatum (Greville) Cleve, recovered faster from hypoosmotic stress than did D. brightwellii. Due to an osmotic shock from 13.6 to 7.1 S both species excreted amino acids and glucose; S. costatum accumulated more glucose, D. brightwellii accumulated more amino acids. S. costatum may with the competition for nitrogen in waters with an unstable salinity; it will replace D. brightwellii.Contribution no. 427 Delta Institute for Hydrobiological Research, Yerseke, The Netherlands     

Une approche vers l'estimation de la production potentielle du phytoplancton par analyse des cinétiques d'induction de fluorescence

In vivo chlorophyll fluorescence is particularly interesting ot ecologists because of various concepts (biomass, productivity, physiological state) associated with it. Using a modified spectrophotofluorometer, we have studied the kinetics of fluorescence in unialgal cultures and in a natural population of marine phytoplankton. Our apparatus did not achieve satisfactory results with cell suspensions having a chlorophyll concentration less than 10 g l^-1. We have also tested a method for estimating kinetics of diluted cultures and marine phytoplankton using cells collected on glass-fibre filters. For unialgal cultures in the exponential growth phase, the method proved satisfactory, and results obtained from both cell suspensions and filters were in good agreement. However, for aged cultures (principally diatoms) and natural marine phytoplankton the method proved unsuitable. The kinetics of fluorescence induction vary according to taxonomic position of the cells, light intensity of the measuring excitation beam and productiveness of the culture medium. The importance of the kinetics of fluorescence induction for characterization of phytoplankton activity is discussed.     

Lethal effects of Northwest Atlantic Ocean isolates of the dinoflagellate, Scrippsiella trochoidea, on Eastern oyster (Crassostrea virginica) and Northern quahog (Mercenaria mercenaria) larvae

The thecate dinoflagellate Scrippsiella trochoidea is a cosmopolitan, bloom-forming alga that has been generally considered non-toxic. Here, we report that environmentally relevant cell densities (10⁴ cells mL⁻¹) of Scrippsiella trochoidea strains isolated from the Northwest Atlantic Ocean caused 100% mortality in Eastern oyster (Crassostrea virginica) larvae during 3-day exposures while parallel control larvae exhibited 100% survival. S. trochoidea also exhibited lethal effects on Northern quahog (Mercenaria mercenaria) larvae (70% mortality during 3-day exposure) but were non-toxic to juvenile fish (Cyprinodon variegates). The cultures of S. trochoidea were more lethal to Northern quahog larvae than ten other species of harmful algae, including the highly toxic species Cochlodinium polykrikoides. Scrippsiella trochoidea cultures within later stages of growth were more toxic than exponential growth stages to bivalve larvae, and the toxicity was dose dependent. Furthermore, toxicity was maintained in the cultures that were sonicated, boiled, and frozen as well as in resuspended residues of the culture but was significantly lower in cell-free culture media. Collectively, these results suggest that S. trochoidea causes mortality in bivalve larvae through a physicochemical rather than strictly chemical mechanism, such as clogging of larval feeding apparatuses by materials produced by S. trochoidea (e.g., lipids, extracellular polysaccharides, and/or cell debris) which accumulate as cells in culture or blooms age. This is the first report of the lethal effects of Scrippsiella trochoidea on shellfish larvae.     

Interactions between two species of marine diatoms: Effects on their individual copper tolerance

The purpose of this study was to examine the effect of the interaction of two species of marine diatoms,Skeletonema costatum (Cleve) Greville andNitzschia thermalis (Ehrenberg) Auerswald, on their individual copper tolerances. The two species, obtained from stock cultures in 1989, were grown together at three copper concentrations (1, 4 and 5 × 10^–7 M added total copper). In the unialgal cultures that were used as controls, the two species grew as predicted from their tolerance tests. However, in mixed cultures,N. thermalis was the only species that exhibited growth, regardless of the copper concentration in the medium. Growth retardation ofS. costatum in the presence ofN. thermalis was attributed to an inhibitory exudate. The effect of the exudate appeared to have been temporary, as demonstrated by the extended lag phase and subsequent satisfactory exponential growth rate ofS. costatum. It is suggested that the exudate degraded within a period of 5 d (=lag phase) because exponential growth rate was resumed. It thus appears that the interaction between the two diatom species is more important in determining the survival ofS. costatum than its individual copper tolerance. This is not the case forN. thermalis. Such interactions would be unaccounted for in single-species toxicity tests. On the other hand, if they are known, prediction of how a community that includes these two species would respond to copper additions becomes possible.     

Orientation ofRhizophagus grandis (Coleoptera: Rhizophagidae) to oxygenated monoterpenes in a species-specific predator-prey relationship

Summary The predator,Rhizophagus grandis, is linked to its specific prey,Dendroctonus micans, by semiochemical signals regulating oviposition and long range orientation. A mixture of simple oxygenated monoterpenes [(–)—fenchone, (–)—pinocamphone, rac. camphor, terpinene-4-ol, borneol, fenchol and verbenone], identified from the frass ofD. micans has been found to be extremely active in a flight windtunnel as a long—range attractant for the predator. The mixture elicited 84 % of the response to larval frass of the prey. Excluding pinocamphone from the mixture did not influence its activity. Also, changing the absolute configurations of some of the components (fenchol, terpinene-4-ol and borneol) did not influence the predators' response. However, the addition of (–)—-terpineol increased the attractivity of the synthetic blend to almost that of larval frass of the prey. Ecological implications of the identified semiochemicals and their use in pest management are discussed.     

Effects of in situ tidal salinity fluctuations on osmotic and lonic composition of body fluid in Southeastern Alaska Rocky intertidal fauna

Tidal fluctuations in salinity and temperature were monitored at a location on Lynn Canal, north of Juneau, Alaska. Organisms were collected from 4 tidal levels during each slack water over a period of 24 h. Body-fluid milliosmolality as well as sodium, potassium, calcium, magnesium and chloride levels were determined. Mytilus edulis were collected at the +1.0 m tidal level, where ambient salinity ranged from 7.9 to 25.1% and body fluids from 356±62 to 730±17 mOsm/kg water. Cucumaria vegea and Katherina tunicata were collected at the +0.6 m level, where ambient salinity ranged from 13.5 to 24.9 and body fluids from 461±27 to 662±50 and 443±31 to 616±38 mOsm/kg water, respectively. Eupentacta quinquesemita and Evasterias troschelii were collected at the -0.9 m level, where ambient salinity ranged from 11.0 to 28.2 and body fluids from 504±32 to 632±51 and 316±31 to 664±37 mOsm/kg water, respectively. Strongylocentrotus droebachiensis were collected subtidally at the -3.7 m level, where ambient salinity ranged from 14.1 to 28.0 and perivisceral fluids from 448±35 to 661±51 mOsm/kg water. Ion levels of the perivisceral fluid of c. vegae were not determined. Potassium appeared to be regulated by all animals except Evasterias troschelii. Little regulation occurred for calcium, magnesium, sodium and chloride by the organisms. Eupentacta quinquesemita appeared to be regulating all ions determined, but this may have been due to its microhabitat. Water temperature cycled inversely to salinity at all tidal levels. Ambient-water sodium, chloride and calcium levels deviated from levels that would be expected upon dilution of seawater with deionized water. There was not indication that body-fluid osmoconcentration of the species collected intertidally increased on exposure to air.Communicated by J.S. Pearse, Santa Cruz