Antimicrobial activities of extracts from Indo-Pacific marine plants against marine pathogens and saprophytes

This study is the second of two surveys designed to systematically screen extracts from marine plants for antimicrobial effects against ecologically relevant marine microorganisms, and to compare results on a geographical basis. In the preceding survey, extracts from tropical Atlantic marine algae and seagrasses were screened in growth inhibition assays against the pathogenic fungus Lindra thalassiae, the saprophytic fungus Dendryphiella salina, the saprophytic stramenopiles, Halophytophthora spinosa and Schizochytrium aggregatum, and the pathogenic bacterium Pseudoaltermonas bacteriolytica. In this study, the same assay microorganisms were used to examine the antimicrobial effects of lipophilic and hydrophilic extracts from 54 species of marine algae and two species of seagrasses collected from Indo-Pacific reef habitats. Overall, 95% of all species surveyed in this study yielded extracts that were active against one or more, and 77% yielded extracts that were active against two or more assay microorganisms. Broad-spectrum activity against three or four assay microbes was observed in the extracts from 50 to 21% of all species, respectively. Extracts from the green alga Bryopsis pennata and the red alga Portieria hornemannii inhibited the growth of all assay microorganisms. Given that antimicrobial activity was prevalent among extracts of Indo-Pacific marine plants, it is interesting to note that the inhibitory effects of each extract varied considerably between the assay microorganisms. Overall, H. spinosa and D. salina were the most susceptible while L. thalassiae, S. aggregatum, and P. bacteriolytica were the most resistant to the extracts tested. These results provide good evidence that antimicrobial chemical defenses are widespread among Indo-Pacific marine plants. Further, the activity profiles of plant extracts suggest that antimicrobial secondary metabolites can have pathogen-selective or broad-spectrum effects. To confirm these results, chemical studies will be needed to isolate and characterize the compounds responsible for the observed antimicrobial activities.     

Impact of conditions of cultivation and adsorption on antimicrobial activity of marine bacteria

The potency of free-living and animal-associated marine bacteria to produce antimicrobial substances has been studied in 491 strains isolated from northern and southern parts of the Pacific Ocean. A total of 26% (126 out of 491) of the strains examined produced antimicrobial compounds against 11 test bacterial strains (TBS) including the fish pathogens Aeromonas hydrophila and Vibrio anquillarum. Antimicrobial substances (AS) produced by marine bacteria were especially active against Staphylococcus epidermidis, Proteus vulgaris, Enterococcus faecalis, and Candida albicans. Twelve strains, isolated from different sources, were chosen as promising candidates, producing a number of AS. Production of AS varied within 24 to 72 h, increasing in a culture medium based on natural sea water with Br-ions, and after attachment to polymeric surfaces. In order to study the influence of adsorption, selected strains with a high potential for antimicrobial production were cultivated on polymeric surfaces with different hydrophobicities and chemical functionalities. These parameters of the surface hydrophobicity (measured by means of water contact angles) and chemical functionality of the surfaces were manipulated using the photo- and thermochemistry of a polymeric system (diazo-naphto-quinone/novolak) commonly used as a photoresistant material in semiconducto-manufacturing. The highest antimicrobial activities occurred on hydrophilic surfaces (standard exposed photoresistant films), whereas the number of attached cells was greater on hydrophobic surfaces, characterized as unexposed resistant films. These results suggest that the chemical nature of induced hydrophilicity may also be a major factor in controlling antimicrobial activity of adsorbed bacteria. Received: 5 March 1997 / Accepted: 24 August 1997     

Antimicrobial activity of extracts of Caribbean gorgonian corals

Extracts of 39 species of Caribbean gorgonians were tested for antimicrobial activity against 15 strains of marine bacteria. The bacteria consisted of three opportunistic pathogens, Vibrio parahaemolyticus, Leucothrix mucor, and Aerococcus viridans, and 12 strains isolated from either healthy or decayed gorgonians. Overall, only 15% (79 out of 544) of the tests resulted in antibacterial activity with 33% (13 out of 39) of the gorgonians inhibiting only one bacterial strain and 23% (9 out of 39) showing no activity. The extracts of four Pseudopterogorgia species showed relatively high levels of activity, inhibiting 43 to 86% of the bacterial strains. The potency of the active Pseudopterogorgia species was variable, however, and three additional Pseudopterogorgia species were inactive against all bacterial strains. With the exception of one sensitive strain, Vibrio species were resistant to gorgonian metabolites. Our results indicate that organic extracts of most Caribbean gorgonians do not possess potent, broad-spectrum antibacterial activity inhibitory to the growth of opportunistic marine pathogens and bacteria associated with healthy and decayed gorgonian surfaces. These findings suggest that the inhibition of bacterial growth is not the primary ecological function of gorgonian secondary metabolites and that bacteria may not be important selective agents in the evolution of gorgonian secondary chemistry.     

Utilisation of mangroves and seagrasses by fishes in the Negombo Estuary,Sri Lanka

In two years of sampling, 10 287 fishes belonging to 62 species (39 families) were collected from mangroves and seagrasses of the Negombo Estuary. Of the species collected, 70% were marine, 27% estuarine and 3% freshwater. Aplocheilus melastigma, Ambassis dayi, Etroplus suratensis, E. maculatus, Bathygobius fuscus and Arothron immaculatus constituted 90% of the catch. Sixteen species were caught only from mangroves, 6 only from seagrasses, and 40 from both. The number of species, individuals, biomass and diversity were higher in seagrasses than in mangroves. Gobies and some siganids moved to the mangrove habitat as they grew, but cichlids, triacanthids, some eleotrids and tetraodontids preferred seagrasses. The number of species and individuals caught increased at the beginning of the north-east monsoon when the rainfall was low. The number of species and individual numbers were positively correlated with salinity and dissolved oxygen and negatively correlated with water temperature. Of the 33 species examined, seagrasses formed >25% of the gut content in juvenile Lates calcarifer, Apogon thermalis, E. suratensis, E. maculatus, Siganus javus, Butis butis and Bathygobius fuscus. Mangrove litter was the main gut content of the family Syngnathidae. E. suratensis, E. maculatus and S. javus from seagrasses contained more filamentous algae than seagrasses in their stomachs. S. javus collected from mangroves also contained more seagrasses in their stomachs than mangrove litter, indicating that the association of juvenile fishes with mangroves or seagrasses may not be obligate in estuaries subject to low tidal fluctuations.     

Saturated and unsaturated hydrocarbons in marine benthic algae

Saturated and olefinic hydrocarbons were determined in 24 species of green, brown and red benthic marine algae from the Cape Cod area (Massachusetts, USA). Among the saturated hydrocarbons, n-pentadecane predominates in the brown and n-heptadecane in the red algae. A C₁₇ alkyleyclopropane has been identified tentatively in Ulvalactuca and Enteromorpha compressa, two species of green algae. Mono-and diolefinic C₁₅ and C₁₇ hydrocarbons are common. The structures of several new C₁₇, C₁₉ and C₂₁ mono-to hexaolefins have been elucidated by gas chromatography, mass spectrometry and ozonolysis. In fruiting Ascophyllum nodosum, the polyunsaturated hydrocarbons carbons occur exclusively in the reproductive structures. The rest of the plant contains n-alkanes from C₁₅ to C₂₁. A link between the reproductive chemistry of benthic and planktonic algae and their olefin content is suggested. An intriguing speculation is based on Paffenhöfer's (1970) observation that the sex ratio of laboratory reared Calanus helgolandicus depends upon the species of algae fed to the nauplii. The percentage of males produced correlates with our analyses of heneicosahexaene in the algal food. Our analyses of the hydrocarbons in benthic marine algae from coastal environments should aid studies of the coastal food web and should enable us to distinguish between hydrocarbon pollutants and the natural hydrocarbon background in inshore waters.Contribution No. 2582 of the Woods Hole Oceanographic Institution; and Contribution No. 227, Systematics-Ecology Program.     

Antimicrobial activity of Red Sea corals

Scleractinian corals and alcyonacean soft corals are the two most dominant groups of benthic marine organisms inhabiting the coral reefs of the Gulf of Eilat, northern Red Sea. Antimicrobial assays performed with extracts of six dominant Red Sea stony corals and six dominant soft corals against marine bacteria isolated from the seawater surrounding the corals revealed considerable variability in antimicrobial activity. The results demonstrated that, while the majority (83%) of Red Sea alcyonacean soft corals exhibited appreciable antimicrobial activity against marine bacteria isolated from the seawater surrounding the corals, the stony corals had little or no antimicrobial activity. From the active soft coral species examined, Xenia macrospiculata exhibited the highest and most potent antimicrobial activity. Bioassay-directed fractionation indicated that the antimicrobial activity was due to the presence of a range of compounds of different polarities. One of these antibiotic compounds was isolated and identified as desoxyhavannahine, with a minimum inhibitory concentration (MIC) of 48 μg ml⁻¹ against a marine bacterium. The results of the current study suggest that soft and hard corals have developed different means to combat potential microbial infections. Alcyonacean soft corals use chemical defense through the production of antibiotic compounds to combat microbial attack, whereas stony corals seem to rely on other means.     

Release of dissolved organic matter by some marine macrophytes

The amount of photoassimilated carbon released as dissolved organic carbon was investigated for 5 species of benthic marine macrophytes (Thalassia testudinum, Diplanthera wrightii, Acanthophora spicifera, Chondria dasyphylla, and Dictyota dichotoma) and 1 species of pelagic marine macrophyte (Sargassum natans). Release rates ranged between 0.006 and 0.053 mg C g^-1 h^-1. Percent release values ranged from 1.1 to 3.8%. Spermatophytes had slightly lower percent release values than algae. Investigations performed on the ability of release products to be utilized by heterotrophic organisms showed that 20 to 30% of the released organic carbon was assimilated within a 2-h period.     

Spatial distribution of bacteria associated with the marine sponge Tethya californiana

Microbial diversity and spatial distribution of the diversity within tissue of the marine sponge Tethya californiana was analyzed based on 16S rRNA gene sequences. One candidate division and nine bacterial phyla were detected, including members of all five subdivisions of Proteobacteria. Moreover, chloroplast-derived Stramenopiles- and Rhodophyta-affiliated 16S rRNA gene sequences were found and Stramenopiles represented the most abundant clones (30%) in the clone library. On the phylum-level, the microbial fingerprint of T. californiana showed a similar pattern as its Mediterranean relative T. aurantium. An interesting difference was that Cyanobacteria that were abundantly present in T. aurantium were not found in T. californiana, but that the latter sponges harbored phototrophic Stramenopiles instead. Surprisingly, the phototrophic microorganisms were evenly distributed over the inner and outer parts of the sponge tissue, which implies that they also reside in regions without direct light exposure. The other phyla were also present in both the outer cortex and the mesohyl of the sponges. These results were confirmed by analysis on the operational taxonomic unit level. This leads to the conclusion that from a qualitative point of view, spatial distribution of microorganisms in T. californiana tissue is quite homogeneous. Thirty-two percent of the operational taxonomic units shared less than 95% similarity with any other known sequence. This indicates that marine sponges are a rich source of previously undetected microbial life.     

Chemical composition of dietary species of marine unicellular algae and rotifers with emphasis on fatty acids

The lipid profiles of a few species of marine unicellular algae and yeast were studied with emphasis on fatty acids as part of a search for the nutritional value of plankton to the diet of marine fish larvae commonly used in marine hatcheries. The general proximate chemistry of rotifers was closely related to the proximate chemistry of the diet organism, exhibiting a higher content of protein and carbohydrate and a lower content of lipid. Major lipids in all algae, yeast and rotifers comprised mono-, di- and tri-glycerides and polar lipids. The algae Chaetoceros gracilis Schutt, Isochrysis galbana Parke and their respective algaefed rotifers exhibited higher amounts of neutral lipids, consisting mainly of cyclic and branched polyunsaturated components. Fatty acid composition of the algae was species-specific, with the highest ratio of polyethylenic to saturated and monoethylenic acid in I. galbana and Phaeodactylum tricornutum Bohlin, and the highest content (15%) of n-3 highly unsaturated fatty acids in Nannochloropsis salina and P. tricornutum. A closely mirrored distribution of the fatty acids, but with a lower amount of n-3 highly unsaturated fatty acids, was present in the respective algae-fed rotifers. Comparison of the fatty acid spectrum of Artemia sp. and Euterpina acutifrons grown in the laboratory on I. galbana with zooplankton samples of E. acutifrons and Oitona nana collected from the sea showed a higher concentration of docosahexaenoic acid (22:6 n-3) in the naturally collected sample. The results indicate that the efficacy of the food algae C. gracilis and I. galbana in increasing the survival of fish larvae in marine hatcheries is not obvious on the sole basis of fatty acid composition.     

Zooplankton feeding ecology

Extracts of 19 of 24 species of sponge collected from Queensland. Australia, inhibited the growth of test microorganisms in in-vitro assays. A similar result has been obtained by earlier workers for extracts of species of sponge obtained from temperate waters. Some of the extracts, including extracts of Thorecta vasiforis, Arenochalina mirabilis and Acanthella kleutha, showed activity against bacterial strains that was comparable with that exhibited by penicillin G and streptomycin against these strains. Gram-positive bacteria were expecially sensitive to many extracts. Little activity was exhibited by any extract against four species of fungi tested. Some of the extracts were markedly toxic to one or more of the test organisms (a fish, a crustacean and a hydroid) used, but no clear pattern linking toxicity to these organisms with antimicrobial properties of the extracts emerged. In general, there bas a negative correlation between antimicrobial activity and surface-fouling, raising the possibility of using freedom from surface-fouling as an indicator of antimicrobial activity. Four of five encrusting species from the undersides of coral boulders showed antimicrobial activity. This study confirmed the value of using methanol-toluene extracts in field-based screening programmes, but emphasised the need to use additional extracting media such as methylene chloride in order to augment the number of antimicrobial compounds detected. There are indications that antimicrobial activity may be widespread in the order Dictyoceratida, in the order Verongida and in the order Axinellida, but sporadic in other orders of Porifera.     

Hydrocarbons of marine phytoplankton

The hydrocarbon contents of 23 species of algae (22 marine planktonic), belonging to 9 algal classes, were analyzed. The highly unsaturated 3,6,9,12,15,18-heneicosalhexaene predominates in the Bacillariophyceae, Dinophyceae, Cryptophyceae, Haptophyceae and Euglenophyceae. Rhizosolenia setigera contains n-heneicosane, presumably derived from the hexaolefin by hydrogenation. Two isomeric heptadecenes have been isolated: the double bond is located in 5-position in the bluegreen alga Synechococcus bacillaris and in 7-position in 2 green algae. Our complete analyses are discussed in the context of earlier data; some generalizations appear no longer valid. Hydrocarbon analysis of marine algae should provide a tool for the investigation of the dynamics of the marine food chain. Knowledge now available provides the background needed for distinguishing between hydrocarbons of recent biogenic origin and hydrocarbon pollutants from fossil fuels.Contribution No. 2576 of the Woods Hole Oceanographic Institution, USA.     

Depletion of heavy isotopes of oxygen and hydrogen in tissue water of intertidal plants: implications for water economy

Stable oxygen and hydrogen isotope ratios of leaf and thallus water of the intertidal seagrasses Phyllospadix scouleri and P. torreyi and the marine algae Egregia menziesii, Gelidium coulteri, and Corallina vancouverensis from three locations in California, USA, were determined in 1987. Compared with subtidal seawater, most plant-water samples were depleted in the heavy isotopes ¹⁸O and deuterium. Depletion of heavy isotopes was greatest in plants growing at the highest intertidal elevations. This was an unexpected result, because enrichment of heavy isotopes occurs during evapotranspiration in terrestrial plants. Two possible mechanisms for this isotopic depletion are proposed: direct uptake of heavy isotope-depleted water vapor and preferential diffusion of ¹⁶O and ¹H into littoral plants from water remaining in the intertidal zone at low tide.     

Antimicrobial activities of marine sponges from the Mediterranean Sea

Extracts from marine sponges collected along the French coast of Mediterranean in 1983 have been tested for antibacterial and antifungal activities. The distribution of antimicrobial activities against five classes of microorganisms (5 Gram-positive and 5 Gram-negative terrestrial bacteria, 10 marine bacteria, 15 human pathogenic fungi and 5 phytopathogenic fungi) among 28 species of marine demosponges is reported. Antibacterial activity against Gram-positive bacteria (77%) is much more frequent than against Gram-negative bacteria (53%). Activity against marine bacteria and yeast is less frequent (32%). Those sponges which contain a large quantily of symbiotic bacteria display a weak antifungal activity.     

Relative growth of different species of marine algae in wasterwater-seawater mixtures

In recent studies, we developed a combined nutrient removal-marine aquaculture process for the tertiary treatment of wastewater and the production of commercially important shellfish. Part of this process consists of an outdoor mass cultivation system for marine algae. During our previous experiments we observed that marine diatoms almost exclusively are the dominant algal species in our outdoor cultures. To better understand this phenomenon of diatom dominance we grew 16 species of marine algae in continuous monoculture under laboratory conditions simulating to some degree the conditions prevailing in our outdoor experiments. Species such as Skeletonema costatum, Monochrysis lutheri and Tetraselmis sp., which were never dominant in our outdoor cultures, grew as well in monoculture, as Phaeodactylum tricornutum, frequently, the prevalent species outdoors. However, when monocultures of Dunaliella tertiolecta and Thalassiosira pseudonana (3H) were purposely contaminated with P. tricornutum, the latter species quickly became dominant. It is suggested that a complex interaction of environmental factors is usually responsible for the dominance of a particular species; one such factor may be the nitrogen source in the growth media. Under conditions of virtually, complete nitrogen assimilation, the carbon: nitrogen ratio in the algae was high (7 to 8) when NO ₃ ^- –N was the source of nitrogen, and low (4 to 6) when NH ₄ ⁺ –N was the prime form of nitrogen. When algal growth was low, resulting in a large inorganic nitrogen residue, the carbon:nitrogen ratio was low regardless of whether NO ₃ ^- –N or NH ₄ ⁺ –N was the main nitrogen source.Contribution No. 3297 from the Woods Hole Oceanographic Institution.     

Screening and bioassays for biologically-active substances from forty marine sponge species from San Diego,California, USA

Biologically-active substances were investigated from 35 demosponge species and 5 calcareous sponge species collected from intertidal or shallow subtidal habitats near San Diego, California, USA, from 1978 to 1980. Crude methanolic extracts of each species were tested for suppression of growth by bacteria (7 species) and a yeast. Antimicrobial activity was found in 26 Demospongiae and 2 Calcarea. Strong activity was found in 11 demosponges and, subsequently, 38 natural products with antimicrobial activity were isolated from 8 of these species (Aplysina fistularis, Dysidea amblia, Leiosella idia, Euryspongia sp., Toxadocia zumi, Axinella sp., Haliclona?cinerea and ?Pachychalina lunisimilis). Twenty-eight of these natural products (usually as pure compounds) were assayed for: (1) suppression of growth of marine fungi (3 spp.) and a red alga; (2) behavior modifications of invertebrate adults (4spp.); (3) toxicity to a goldfish; (4) inhibition of sexual reproduction of a brown alga; (5) inhibition of settlement and/or metamorphosis of late larvae or invertebrate juveniles (4spp.). Many of the natural products were also incorporated into pelleted fish food and tested for feeding-behavior modifications of fishes (5 spp.). Three of the compounds from Dysidea amblia were inactive in all tests. All other natural products were active in at least one assay, although none was active in all assays. The discussion relates the possession of biologically-active substances to the ecology of each sponge species; for example, sponges with antimicrobial substances are rarely overgrown.     

Distribution of glycosylglycerols and oligosaccharides in the marine environment and their ecological significance in the deep sea

Determination of low molecular weight carbohydrates in marine environments indicated that 1-O--D-galactosylglycerol, 6-O--D-galactosyl-1-O--D-galacto-sylglycerol, sucrose, laminaribiose and laminaritriose are widely distributed in seawaters, suspended and sinking particles, and sediments in coastal as well as in deep-sea waters [e.g. Mikawa Bay and Sagami Bay, Kumano Nada (offshore Japan), and northwest North Pacific Ocean, Bering Sea and Antarctic Ocean: collections during 1978–1984]. Identification of these glycosylglycerols and oligosaccharides in algal cells such as a flagellate (Olisthodisus luteus), blue-green algae (e.g.Trichodesmium sp.) and a diatom (Reptocylindrus denicus) strongly suggests that these sugars are photosynthetically produced by algae in the euphotic zone and are then rapidly transported to the deep sea as sinking particles which can be collected by sediment-trap experiments. The rapid decay rate of low molecular weight carbohydrates by microorganisms suggests that the transported sugars provide energy substrates for microorganisms living in the deep sea.Please address all requests for reprints to Dr. Handa at Nagoya University     

Protective effect of Kappaphycus alvarezii (Rhodophyta) extract against DNA damage induced by mercury chloride in marine fish

Marine organisms are continuously exposed to agents, both exogenous and endogenous, that damage DNA. Consequently, it is important to determine the ability of compounds to provide protection against damaging chemicals. The aim of this study was to evaluate the anti-genotoxic activity of crude aqueous extracts of Kappaphycus alvarezii (Rhodophyceae), collected from the Southeast coast of India. This study focused on possible anti-genotoxic potential of aqueous extract of K. alverazii to interfere with clastogenicity induced by mercury chloride (HgCl₂) in marine fish, Therapon jarbua as measured by cytogenetic endpoints such as cell viability and comet assay. In the first set of experiments, fish were exposed to a single treatment of Hg at 0.125, 0.25, 0.5, 1, or 2?ppm along with controls. Mercury exposure produced significant DNA damage in all comet classes, maximum as >79% (Class 4) at 0.5, 1, and 2?ppm exposure in a time dependent manner. Algal extract did not induce genotoxicity when given alone and prevented Hg-induced genotoxicity. The algal extract reduction in genotoxicity was significant but not time- and concentration-dependent. Results suggested that under present experimental conditions, K. alvarezii extract exhibit potent anti-genotoxicity effects in this fish model; and thus these extracts may be recommended as a supplement in fish meal and may benefit humans ingesting Hg-contaminated fish.     

Antibacterial and antimutagenic activitiy of extracts and essential oil from (Tunisian) Pistacia lentiscus

Aqueous and flavonoid-enriched extract as well as essential oil (EO) obtained from leaves of Pistacia lentiscus were assessed for antibacterial and antimutagenic activities. Antibacterial activity of different extracts and EO were evaluated against six bacterial strains. A marked inhibitory effect was observed against Salmonella typhimurium, whereas lower activity was observed against Staphylococcus aureus, Pseudomonas aeruginosa, and Salmonella enteritidis. EO showed significant inhibitory effects against Salmonella typhimurium, Salmonella enteritidis and Staphylococcus aureus. The antimutagenic activity of the different extracts against Aflatoxin B₁ (AFB₁) and sodium azide was demonstrated with the Salmonella typhymurium assay. The number of revertants per plate decreased significantly when the plant extracts were added to the assay system using Salmonella typhimurium TA100, TA98 and TA1535.     

Questions on the mechanism of temperature adaptation in marine phytoplankton

The rate of light-saturated photosynthesis in 3 marine algae [Phaeodactylum tricornutum Bohlin, Nitzschia closterium (Ehrenberg) Smith and Dunaliella tertiolecta Butcher] varies during growth in batch culture. The photosynthetic rare declines most rapidly during growth at the higher temperatures. Because of these changes in photosynthesis rate, the previously reported enhanced photosynthetic abilities caused by growth at lower temperatures (generally interpreted as evidence for higher enzyme levels) can only be observed when measurements are made late in the exponential phase or after the onset of the stationary phase of growth. When allowance is made for the earlier peak of photosynthetic ability in cultures growing at higher temperatures, there is no evidence for adaptation to lower temperatures being caused by increased levels of the enzymes required for carbon-dioxide fixation. When the changes due to growth in batch culture are taken into account, certain effects of temperature can be recognized. the dry weight: chlorophyll ratio of all 3 algae increases with decreasing growth temperatures. For P. tricornutum and N. closterium, growth at lower temperatures reduces the cellular content of chlorophyll a, but has little effect on the chlorophyll content of D. tertiolecta. The dry weight: cell-number ratio of D. tertiolecta and P. tricornutum increases with lower growth temperatures, but growth temperature has little effect on the cell mass of N. closterium. Growth of the 3 algae at lower temperatures does not increase their ability to photosynthesize at these lower temperatures. Rather, it reduces their ability to assimilate carbon dioxide at the higher temperatures.     

Cycling of selenite and selenate in marine phytoplankton

The marine phytoplankton Dunaliella tertiolecta, Cachonina niei, Thalassiosira nordenskioldii, Phaeodactylum tricornutum, and Chaetoceros sp. were incubated with a range of molar concentrations of sodium-selenite (Na₂-SeIVO₃) and sodium-selenate (Na₂-SeVIO₄) to examine further their role in metabolic cycling of selenium in ocean waters. At low selenium concentrations, approaching those found naturally in seawater (10^-10 to 10^-9 M), all species distinguished between selenite and selenate, and actively concentrated selenite from the incubating medium while only marginally accumulating selenate. At much higher concentrations (10^-8 to 10^-6 M), selenate was also taken up. At the highest concentration tested, i.e., 10^-5 M with C. niei, after an immediate rapid uptake in the first 24 h, the intracellular selenite and selenate levels dropped to about 35 to 50% of the initial peak values. These observations suggest an uptake mechanism in these algae which, at normal ambient concentrations of selenium (10^-9 M), preferentially selects selenite and excludes selenate. At much higher concentrations (10^-8 M), the mechanism becomes overloaded and both selenium species enter the cells. Intracellularly, selenite became associated primarily with protein and amino acid fractions, in approximately equal proportions, while only ca. 4% of total intracellular selenium was found in the lipid fraction. Trace amounts of selenate that entered the cells, mainly during the first minutes of exposure, also entered the protein and amino acid components, but over time were increasingly associated with the protein fraction only. At the end of a 10-d incubation of algal cells in selenite-spiked medium, less than 25% of total Se in the medium could in fact be identified analytically as selenite. This suggests the presence of a non-selenite metabolite, possibly released back into the medium from the algae.