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.     

A polyunsaturated hydrocarbon (3, 6, 9, 12, 15, 18-heneicosahexaene) in the marine food web

An olefinic hydrocarbon (all-cis-3,6,9,12,15,18-heneicosahexaene, HEH)_was islated from marine planktonic plants and animals. Its structure was established by ultraviolet, infrared and mass spectrometry in combination with chemical techniques. The olefin occurs in many species of marine planktonic algae and is probably derived from the corresponding docosahexaenoic acid. Rhincalanus nasutus accumulates HEH nonselectively from its algal food together with the triglyceride lipids. Other, related copepods contain little or no HEH even when grown in cultures of algae that provide R. nasutus with that olefin. The presence of HEH in marine vertebrates suggests that, within the animal lipids, hydrocarbons are remarkably stable. This work has practical implications for studies of the marine food web and of marine pollution with persistent chemicals.Contribution No. 2485 of the Woods Hole Oceanographic Institution, Woods Hole, Massachusetts 02543, USA.     

Alkanes and alkenes in marine benthic algae

Saturated and olefinic hydrocarbons were determined in additional species of benthic marine algae from the Cape Cod (Massachusetts, USA) area (see: Youngblood et al., 1971). The distribution of homologous and isomeric olefins was studied in plants of different age and in morphologically different parts of the same specimen. With two minor exceptions, only normal alkanes and alkenes are present. The methylene-interrupted C₁₉- and C₂₁-polyolefins are particularly abundant; ¹-heneicosahexaene and the corresponding pentaene are common to all brown algae, while the corresponding ³-isomers occur in green algae. The hydrocarbon concentration, the alkene-to-alkane ratio and the polyolefin content are highest in young plants or in rapidly growing tissues of older plants. This suggests a deeper involvement in cell biochemistry of straight-chain hydrocarbons than previously considered. The biosynthesis of the plant polyolefins remains to be explored; no immediately obvious precursors of the ¹-polyolefins were found among the algal fatty acids. The hydrocarbon composition of these benthic algae differs greatly from that of fossil fuels in its simplicity and predominately unsaturated nature. The separation of the isomers by gas chromatography and their structural elucidation by mass spectrometry, alone and in combination with hydrogenation and ozonolysis, are discussed.Contribution No. 3155 of the Woods Hole Oceanographic Institution.     

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.     

Antimicrobial activities of extracts from tropical Atlantic marine plants against marine pathogens and saprophytes

Studies investigating disease resistance in marine plants have indicated that secondary metabolites may have important defensive functions against harmful marine microorganisms. The goal of this study was to systematically screen extracts from marine plants for antimicrobial effects against marine pathogens and saprophytes. Lipophilic and hydrophilic extracts from species of 49 marine algae and 3 seagrasses collected in the tropical Atlantic were screened for antimicrobial activity against five ecologically relevant marine microorganisms from three separate kingdoms. These assay microbes consisted of the pathogenic fungus Lindra thalassiae, the saprophytic fungus Dendryphiella salina, the saprophytic stramenopiles, Halophytophthora spinosa and Schizochytrium aggregatum, and the pathogenic bacterium Pseudoaltermonas bacteriolytica. Overall, 90% of all species surveyed 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 microorganisms was observed in the extracts from 48 and 27% of all species, respectively. The green algae Halimeda copiosa and Penicillus capitatus (Chlorophyta) were the only species to yield extracts active against all assay microorganisms. Among all assay microorganisms, both fungi were the most resistant to the extracts tested, with less than 21% of all extracts inhibiting the growth of either L. thalassiae or D. salina. In contrast, over half of all lipophylic extracts were active against the stramenopiles H. spinosa and S. aggregatum, and the bacterium P. bacteriolytica. Growth sensitivity to hydrophilic extracts varied considerably between individual assay microorganisms. While 48% of all hydrophilic extracts were active against H. spinosa, 27% were active against P. bacteriolytica, and only 14% were active against S. aggregatum. Overall, more lipophilic extracts inhibited microbial growth than hydrophilic extracts. The variability observed in the antimicrobial effects of individual extracts against each assay microorganism reflects the importance of choosing appropriate test microbes in assays from which ecologically relevant information is sought. Results from this survey demonstrate that antimicrobial activities are prevalent among extracts from marine algae and seagrasses, suggesting that antimicrobial chemical defenses are widespread among marine plants.     

Callus formation in seven species of agarophyte marine algae

Callus formation of seven species of the agarophyte marine algae Phyllophora nervosa, Gracilaria verrucosa, Furcellaria fastigiata, ceramium kondoi, Gelidium vagum, Laurencia paniculata, Rhodymenia pertusa was obtained from 1984 to 1986. A new sterile agent, chlorhexidin bigluconate, was used to get axenic explants from the algae thalli. The formation of callus from axenic explants depended on temperature of cultivation. In the case of F. fastigiata and G. vagum it occurred at 12° to 20°C, and with P. nervosa at 18° to 26°C. The exchange of sucrose with mannitol in the culture medium and the addition of some growth regulators stimulated the formation of callus. Mass formation of lumpy and warty extensions was observed in the explants from leaf blades of P. nervosa. The results might be useful for improving the aquaculture of agarophyte marine algae.     

Bioavailability of a hydrocarbon from water and sediment to the marine worm Arenicola marina

The uptake, accumulation and depuration of a labelled hydrocarbon (¹⁴C-1-naphthalene) by the marine annelid Arenicola marina were studied. Naphthalene was chosen because it is an important component of oil and is known to be toxic to marine organisms. The bioavailability of the hydrocarbon to the worm from contaminated sediment is examined and the results discussed with reference to the rehabilitation of oiled environments, as A. marina is thought to be capable, in high numbers, of greatly reworking sediments.     

Photosynthesis,photorespiration, and dark respiration in eight species of algae

The rates of photosynthesis and dark respiration for 7 marine algae and 1 fresh-water alga were measured and compared. The dinoflagellates Glenodinium sp. and zooxanthellae have high dark respiration rates relative to photosynthetic rates, which may decrease their net growth rates. Photorespiration in the 8 algal species was studied by examining the effects of the concentration of oxygen on the rates of photosynthesis, on the incorporation of ¹⁴CO₂ into the photorespiratory pathway intermediates glycine and serine, and on the postillumination burst of carbon dioxide production and oxygen consumption. A combination of these results indicates that all the algae tested can photorespire, but that Glenodinium sp., Thalassiosira pseudonana, and zooxanthellae either have a photorespiratory pathway different from that proposed for freshwater algae (Tolbert, 1974), or an additional pathway for glycolate metabolism.     

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.     

Availability of copper from phytoplankton and water for the bivalveMacoma balthica. II. Uptake and elimination from64Cu-labelled diatoms and water

The amount of copper taken up via algae and water byMacoma balthica from the Oosterschelde sea arm, S.W. Netherlands, was established using the radioisotope⁶⁴Cu. As far as we know, this isotope has never been used before in marine flod chain studies. As a model food source the marine diatomPhaeodactylum tricornutum was allowed to accumulate⁶⁴Cu for 1 d. These labelled algae were fed to the clams in the presence of the complexing agent EDTA (0.27mM). EDTA was added to prevent uptake of dissolved⁶⁴Cu that could be leaking from the labelled diatoms. In control experiments, unlabelled diatoms were fed toM. balthica in the presence of dissolved⁶⁴Cu (with and without EDTA) in order to assure a similar filtration activity. In repeated experiments with varying particulate/dissolved copper ratios, uptake through food always turned out to be at least as efficient as uptake from the water. It was concluded that Cu, associated with food, is well available for uptake byM. balthica.     

Effects of 9–10 dihydroanthracene and its biodegradation products on the marine diatom Phaeodactylum tricornutum

Growth, photosynthetic capacity and chlorophyll a content of the marine diatom Phaeodactylum tricornutum Bohlin were observed after exposure to the aromatic hydrocarbon 9–10 dihydroanthracene and its biodegradation products. Growth was inhibited after exposure to the aromatic hydrocarbon, whereas no inhibition occurred in the presence of the biodegradation products alone. The degradation products were found to enhance the chlorophyll a cellular content. Synergistic effects between dihydroanthracene and its biodegradation products increased the toxicity of this aromatic hydrocarbon.     

Studies on the growth of some marine unicellular algae under different artificial light sources

Documented evidence of the suitability of various artificial light sources for the culture of unicellular algae is scarce. This paper reports the results of a comparison of the growth rates of 5 species of marine unicellular algae at various light intensities produced by an incandescent filament source or daylight fluorescent tubes. Under light saturating conditions the maximum growth rate achieved by each species was the same under each artificial source. Under sub-optimal intensities, no difference in the growth rates under the 2 sources was detected in the case of Phaeodactylum tricornutum, Chlorella ovalis or Monochrysis lutheri. However, under these conditions both Brachiomonas submarina and Dunaliella primolecta required higher intensities of incident illumination from the fluorescent source than from the incandescent source to maintain a particular relative growth rate.     

Occurrence and biological role of light-induced chromatophore displacements in seaweeds

The occurrence of light-induced chromatophore displacements and concomitant transmittance changes in marine algae was investigated by microscope and photometrically with an automated recording microphotometer system; 16 brown, 6 green and 20 red algae were studied. In most of the brown algae, both phaeoplast displacements and transmittance changes were found. In some red algae which are frequently exposed to direct sun light during emergence at low tide, light-induced transmittance changes were measured, but they could not unequivocally be correlated with changes in the position of rhodoplasts. Among green algae, only Ulva lactuca shows chloroplast displacements which, however, follow circadian rhythms and are consequently not light-induced in the strict sense. The dose-response curves of light-induced chromatophore displacements were measured in the following Fucus and Laminaria species: F. spiralis, F. vesiculosus, F. serratus, L. digitata, L. saccharina and L. hyperborea. While in Fucus species correlations between light-induced transmittance changes and zonation of the intertidal area seem to exist, no significant differences have been found in the Laminaria species. The physiological role and ecological importance of light-induced chromatophore displacements for seaweeds living in the intertidal belt are discussed.     

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.     

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.     

Effects of various diets on the nutritional value of Artemia sp. as food for the prawn Penaeus monodon

The nutritional value of Artemia sp. as food for marine fish and crustacean larvae has been linked to the level of its polyunsaturated fatty acid (PUFA) content. Experiments in August 1984 were conducted to determine the effects of various artificial diets and algae on fatty acid composition of PUFA-deficient Artemia sp. (Utah GSL strain) and their resulting value as food for postlarvae of the prawn Penaeus monodon (Fabricius). Nauplii of the brine shrimp were grown on extracts of corn, copra, soybean and rice bran containing precursors (C18) to long-chain PUFA and also on algal species containing different levels of long-chain PUFA (C20). The nauplii were then used as food for P. monodon postlarvae. The results revealed that absence of C20 polyunsaturates from the feeds and their presence in the algae were reflected in the polyunsaturated fatty acid content of the tissues of Artemia sp. When fed with brine shrimp fed on algae, P. monodon displayed better postlarval survival and significantly higher growth; related to the content of polyunsaturated fatty acids in Artemia sp. A practical feeding approach in prawn hatcheries would be to grow Artemia sp. on a cheap diet such as rice bran, and then to enhance its nutritional value with a diet high in PUFA prior to harvesting, in order to improve hatchery production.     

Daily photosynthesis,respiration, and carbon budgets in a tropical marine jellyfish (Mastigias sp.)

From measured diel photosynthesis and respiration rates, using oxygen electrodes, estimates of carbon flux between symbiotic algae (zooxanthellae) and host animal are presented for the marine scyphomedusan Mastigias sp. from a marine lake in Palau, Western Caroline Islands, during February and March 1982. The carbon budgets calculated for these lake medusae indicate that carbon fixed photosynthetically by zooxanthellae and made available to the host may satisfy up to 100% of the host's daily metabolic carbon demand (CZAR). The stable carbon isotope (¹³C) signature of the mesogleal carbon of lake Mastigias sp. was close to that of the zooxanthellae, supporting the interpretation that while these medusae may feed holozoically, some of their carbon comes from their symbionts. The diel photosynthesis, respiration, and preliminary estimates of carbon budgets of three individuals of another ecotype of Mastigias sp. collected from nearby oceanic lagoons are also given. Photosynthesis of lagoon medusae was generally greater than that for lake medusae of similar size, and lagoon medusae were phototrophic with respect to carbon, with commensurately greater CZAR values. Carbon translocated from the symbiotic algae also may contribute to the growth requirements of both lake and lagoon medusae. From carbon flux data, the lake jellyfish were estimated to contribute about 16% to the total primary productivity of their marine lake habitat.     

Are some bacteria toxic for marine algae?

It was observed in vitro that some bacteria from different origins, including the sea, are capable of inhibiting the growth of several marine algae. However, there is too much difference between in vitro experiments and in situ environmental conditions to assume that bacterial poisons may rule a great deal of algac-bacteria relationships in the sea as a whole. In the author's opinion the importance of bacterial poison is limited to special and rare instances.     

New methodological strategies for detecting bacterial indicators

The development of monitoring strategies for the early warning of seawater pollution, with particular reference to faecal and hydrocarbon contamination, has been the specific goal of research carried out within the Cluster 10-SAM (Advanced Systems for Coastal Marine Monitoring) Project, funded by the Italian Ministry for University and Scientific Research. Advanced analytical approaches have been designed and applied to detect bacterial species that have been selected as potential indicators of pollution in seawater samples. We report the results obtained using the fluorescent antibody and enzymatic assays for the detection of Escherichia coli, and a real-time PCR protocol for monitoring marine areas for hydrocarbon pollution. Immunofluorescence and enzymatic methods revealed the occurrence of different faecal pollution levels, reaching 10⁵ E. coli cells 100 ml^?1 in the Straits of Messina. Real-time PCR results corresponded to the different degree of oil pollution of the analysed samples. The specificity and speed make these methods promising for the detection and evaluation of marine pollution.     

Under-ice feeding and diel migration by the planktonic copepodsCalanus glacialis andPseudocalanus minutus in relation to the ice algal production cycle in southeastern Hudson Bay,Canada

The marine planktonic copepodsCalanus glacialis Jaschnov andPseudocalanus minutus (Kroyer) typically dominate the copepod biomass in spring under the ice in southeastern Hudson Bay, Canada. Females of both species exhibited significant diel feeding cycles, as measured by gut pigment content, throughout a bloom of ice algae at the ice-water interface in 1986. Periods of grazing correlated well with a nighttime vertical migration by females to within 0.2 m of the ice-water interface, suggesting that feeding took place at or just below the thermohaline boundary between seawater and the interfacial layer containing the ice algae. Seasonal melting of the ice bottom in mid-May resulted in freshening of the surface layer and release of the ice algae into the water column. FemaleC. glacialis andP. minutus responded by ceasing migration to the interface. Gut pigment content, and by reasonable assumption, feeding activity in the water column, increased substantially immediately after this event. In mid-May, the water column phytoplankton consisted of flagellates, sedimenting ice algal cells, and diatoms (Navicula pelagica andChaetoceros sp.) previously found at the interface and then growing in the water column. We conclude that algae growing at the ice-water interface, and sedimenting or actively growing algae derived from this interfacial layer, are a regular and principal source of nutrition for these pelagic copepods during and immediately after the ice algal bloom.