MORPHOGENESIS OF MONOSTROMA OXYSPERMUM (KÜTZ.) DOTY (CHLOROPHYCEAE) IN AXENIC CULTURE,ESPECIALLY IN BIALGAL CULTURE1

The typical morphology of Monostroma oxyspermum (Kütz.) Doty is lost in axenic culture. In synthetic media of the ASP type, it grows as a colony-like mass composed of round cells with numerous rhizoids. Such a mass is a fragile structure which falls apart upon shaking, or slight touch, into small cell-groups and single cells or cells with a long rhizoid. Only temporary saccate or monostromatic fronds appear and reach 1–2 mm in length when grown in enriched seawater media, but disintegrate and become a colony-like mass. The typical morphology is easily restored by adding at specific intervals filtrates of bacterial cultures and supernatant medium from axenic brown and red algal cultures to the basal medium (ASP7), or by reinfecting the Monostroma with an appropriate bacterial flora. Furthermore, the typical morphology in also maintained by bialgal cultures between Monostroma and other axenic strains of various species of seaweeds except the species belonging to the Chlorophyceae. Monostroma thus appears to utilize some substances released by most species of brown and red algae for its typical growth. Active substances released by bacteria, brown and red algae have not yet been identified and purified. However, it is demonstrated that in axenic cultures many species of seaweeds produce active extracellular substances which play an important role in growth and Morphogenesis of other species of seaweeds.     

High rhenium enrichment in brown algae: a biological sink of rhenium in the sea?

Twenty-one species of seaweed from the California coast were analyzed for rhenium. For the first time, high enrichment (thousandfolds) of rhenium relative to seawater was found in brown algae, but not in green or red algae. Brown algae was suggested as a biological sink of rhenium in the sea and the analogous behavior of technetium to rhenium was found in marine algae. Preliminary incubation experiments with a common brown alga (Pelvetia fastigiata) showed that algal surface is not a major accumulating locus of rhenium.     

HABITAT MATTERS FOR INORGANIC CARBON ACQUISITION IN 38 SPECIES OF RED MACROALGAE (RHODOPHYTA) FROM PUGET SOUND,WASHINGTON, USA1

Measurements of pH drift were used to assess the ability of 38 red algal seaweeds to use bicarbonate and to deplete the dissolved inorganic carbon pool (DIC) from seawater medium. Subtidal algae were typically restricted to the use of DIC in the form of dissolved CO₂, reducing the initial DIC by only 9%. Intertidal species used both dissolved CO₂ and bicarbonate and reduced initial DIC by as much as 70%. DIC reductions and pH compensation points for the intertidal species tested were strongly correlated with their vertical zonation on the rocky shoreline (analysis of variance). DIC acquisition efficiency increased with tidal height, but species from the upper edge of the intertidal demonstrated a reversal of this trend. This general pattern associated with tidal height was observed not only among intertidal red algae in general, but also among four species of the genus Porphyra (P. torta V. Krishnamurthy, P. papenfussii Krishnamurthy, P. perforata J. Agardh, P. fucicola Krishnamurthy) and among four populations of the broadly distributed species Mastocarpus papillatus (C. Agardh). The Mastocarpus observations suggest either that individuals of this species may be able to express alternate strategies for carbon acquisition or that intertidal height may select for survivorship of genotypes with different carbon acquisition strategies. Taken together, these data suggest that the carbon acquisition strategy of intertidal red algae may be an important physiological set of adaptations that is under active evolutionary selection. These physiological differences were not related to phylogeny, tested as membership in red algal families and orders.     

Bioactive volatile compounds from marine algae: feeding attractants

Chemical communications play an important role in plants, fungi, and algae. Volatile organic compounds in marine algae are released into the seawater. These compounds play a role as either pheromones or allelochemicals. We observed that the turbinid gastropod Lunella coronata coreensis inhabits the intertidal zone and often grazes the green alga Ulva pertusa. Feeding tests and feeding preference studies were performed with green, brown and red algae or by using the powdered freeze-dried seaweed in agar. The snails fed on U. pertusa preferentially compared to the other marine algae, and recognized chemoreception compounds from the alga but not their structural or morphological differences. From feeding tests using artificial foods, it is suggested that the feeding attractants are in the essential oil of the alga U. pertusa.     

Recirculating Culture for Chondracanthus Exasperatus

The red alga Chondracanthus exasperatus is a source of the phycocolloid carrageenan as well as an ingredient referred to as ‘intralamellar gel’ in a recently developed cosmetic formula (US Patent 6136 329). The high value of the cosmetic product has sparked renewed interest in cultivation of this species. Previous cultivation methods for this species include open water culture on nets and immersed cultivation in tanks supplied with flow‐through pumped seawater. The installation of a high capacity seawater supply, pumping and drain system is a major cost for flow through systems. Recirculating or re‐use seawater systems that minimize seawater turnover may offer significant cost savings over single‐pass, flow through seawater systems. In this research several options for minimizing seawater use have been tested: recirculating batch culture in which nutrient replenished (carbon dioxide and mineral nutrients) natural or artificial seawater is used with minimal turnover and spray culture in which plants are suspended in air saturated with nutrient replenished natural or artificial seawater medium. Small volume (<2 L), single‐plant bioreactors and larger multiplant, 20, 80 and 320 L (sea water volume) immersion and spray systems have been developed and tested. Results from these systems will be presented. Research supported by Washington Sea Grant, Washington Biotechnology Center and Soliv International Corporation.     

Influence of Iodine and Bromine on Growth of Some Red Algae in Axenic Culture

Some red algae in axenic culture have been cultivated with different additions of iodine and bromine. Polysiphonia urceolata appeared to have an absolute demand for iodine. Additions could be made either as organically bound iodine or as inorganic iodine. A linear correlation between amount of added iodide and growth was found for iodide concentrations from 1 μmol up to at least 8 μmμmumol per 1. Nemalion proved to be indifferent to iodide additions, while Goniotrichum elegans was inhibited by concentrations higher than 0.4 μumol per 1, which corresponds to that of natural seawater. High additions of iodine generally inhibited growth of nonaxenic algae. Acrochaetium made an exception, being stimulated by 4 μumol per I. Bromine in the same concentration as that of seawater, viz. 814.3 μmol pa 1 inhibited growth of most species, but amounts smaller than 50 μumol had in some experiments a slightly increasing effect. Bromine seems, however, not to play an essential part in the metabolism of Polysiphonia urceolata.     

CHEMOSENSORY RESPONSES OF THE SYMBIOTIC DINOFLAGELLATE SYMBIODINIUM MICROADRIATICUM (DINOPHYCEAE)1

Motile Symbiodinium microadriaticum (Freudenthal 1962) were attracted to a variety of nitrogen-containing compounds, including ammonium, nitrate, urea and some amino acids. No chemosensory response to phosphate, sulphate, vitamins, trace metals or sugars was evident. Motile algae responded to concentrations of ammonium, nitrate, and urea at least as low as 10^?6 M. High concentrations (≥ 10^?2 M) of ammonium appeared to inhibit attraction of motile algae. Calculations using ammonium release rates from various aposymbiotic hosts suggest that motile S. microadriaticum can respond to released ammonium ca. 1 cm from the source. Cultured algae were not attracted to combined nitrogen cues for at least 2 days after inoculation into seawater with dissolved combined low nitrogen. Algae freshly isolated from starved animals were normally motile the day following isolation and attracted to ammonium and nitrate when maintained in seawater containing < 1 μM ammonium and nitrate. The algae lost their ability to orient to nitrogen attractants the day after incubation into culture medium containing high levels of ammonium and nitrate. These results suggest that chemosensory behavior is suppressed when nutrients are present in the ambient medium or are stored by the alga. There were few differences in chemosensory abilities in different strains of S. microadriaticum to the attractants assayed, suggesting that selection for a particular strain by a host species may not be due to differential chemosensory ability or cues. However, the absence of chemical attraction of motile S. microadriaticum to infected hosts may act to preserve strain selection occurring at other steps in the infection process of aposymbiotic hosts.     

Marine macroalgae as foods for fishes: an evaluation of potential food quality

Synopsis A revitalized view of feeding by herbivorous marine fishes is sought through two questions. First, What characteristics of major taxa of algae identify them as predictably high or low quality foods? Second, are marine algae valuable foods for fishes which do not mechanically disrupt cell walls and do not harbor specialized enzymes or microbes capable of lysing cell walls? Energy, ash and nutrient content of 16 species of marine algae were employed to assess food quality of fleshy red, green, brown and calcareous red algae. On the basis of ash, calories, total protein and total lipid content, fleshy algae should be superior to calcareous algae as foods for fishes; in addition, green algae should be superior to brown algae and brown algae superior to red algae. When the probable digestibility of storage and extracellular carbohydrates is considered, green and red algae are predicted superior to brown algae as food. Two species of damselfishes (Pomacentridae) from the Gulf of California,Eupomacentrus rectifraenum andMicrospathodon dorsalis, eat red and green algae and ignore brown and calcareous algae. They feed, therefore, in a fashion consistent with predictions based only on algal chemistry. These fishes absorb at least 20–24% of the biomass, 57–67% of the protein, 46–56% of the lipid and 37–44% of the carbohydrate contained in algae eaten in the wild. Since these damselfishes do not masticate their food, it appears that herbivorous fishes can digest major fractions of algal nutrients without mechanical destruction of algal cells.     

Nutrient-limited productivity of calcareous versus fleshy macroalgae in a eutrophic,carbonate-rich tropical marine environment

The results of a study of nutrient enrichment with nitrogen (N) and phosphorus (P) on productivity and calcification of fleshy and calcareous algae are reported in this study. Plants were collected from a nearshore eutrophic site in the Florida Keys (USA) and experimentally pulsed during the night with combinations of N and P. After several days of pulsing (7–10 days), net productivity, calcification, and alkaline phosphatase activity (APA), were measured. Productivity of fleshy algae were frequently enhanced by N, P, and N+P, during both summer and winter. Phosphorus limited the productivity of Hydroclathrus clathratus during winter and Ulva spp. during summer, whereas nitrogen limited the productivity of Laurencia intricata during both seasons. During summer, Dictyota cervicornis productivity was not enhanced by any nutrient enrichment. Nitrogen limited the productivity of the three calcareous species Penicillus capitatus, Penicillus dumetosus and Halimeda opuntia during winter and that of H. opuntia during summer. Neither N nor P enrichment increased calcification of calcareous species, and P enrichment greatly inhibited calcification of P. dumetosus during winter. Nutrient enrichment enhanced the productivity of the fleshy species to a greater extent than that of calcareous algae. The seawater DIN:SRP molar ratio was low at our eutrophic study site (molar ratio average of 3:1 during winter and 9:1 during summer) compared to more oligotrophic sites in the Florida Keys, suggesting that in carbonate-rich environments, eutrophication shifts nutrient regulation of productivity from P to N. APA activities of fleshy macroalage were higher than calcareous algae, and rates of all macro algae were 2- to 7-fold higher in summer compared to winter. Productivity was also about 3-fold higher in fleshy compared to calcareous species and about 2-fold higher in summer compared to winter. These results suggest that nutrient enrichment enhances productivity of fleshy algae to a greater extent than that of calcareous algae. Thus, overgrowth of calcareous algae by more opportunistic fleshy forms could reduce carbonate accretion in tropical coastlines experiencing increased eutrophication.     

SEASONAL VARIATION OF P CONTENT AND MAJOR N POOLS IN PALMARIA PALMATA (RHODOPHYTA)1

The annual variation of major nitrogen pools, phosphorus, carbon, ash, and thallus water content in relation to seasonal environmental changes was studied in two northern Spanish populations of the edible seaweed Palmaria palmata (Linnaeus) Kuntze. Observed patterns were investigated using Spearman rank order correlation coefficients. There were significant relationships between thallus nutrient content and nitrate and orthophosphate seawater concentration, irradiance, temperature, and wave force. The highest levels of total N and P and nitrogenous compounds were observed during autumn and winter because the thallus stored N‐ and P‐rich compounds in response to high nutrient seawater concentration when growth was limited by low light and temperature. Phycoerythrin and other proteins were the main N reserves. Thallus P content was higher in algae from the eutrophic site. During spring, reduced N and P thallus content and increased ash, water, and C content were observed in the growing fronds. N and P seawater concentrations were undetectable during summer when nutrient reserves were low and growth was reduced and eventually suppressed, suggesting nutrient limiting conditions. Palmaria palmata clearly could take advantage of elevated N and P concentrations to create storage reserves in winter to support early summer growth. This storage response reduced the dependence of algal nutrition on the external nutrient supply and supports the use of pulse fertilization to diminish summer nutrient limitation of cultured algae.     

Light control of the respiration of exogenous glycerol in the red macroalga Grateloupia doryphora

Heterotrophic activity in macroalgae has been little studied, but the red macroalga Grateloupia doryphora is known to grow in light at a higher rate in a glycerol-containing medium than in seawater. The effects of 0·1 M exogenous glycerol in seawater (SW90-gly) on the respiration rate of G. doryphora and the role played by light were investigated. The algae pretreated for 2 h in the light and in SW90-gly evolved oxygen and fixed carbon dioxide (H¹⁴CO₃ ^?), but also evolved radioactive ¹⁴CO₂ from [¹⁴C]glycerol. The rate of oxygen evolution was lower than that of samples in seawater, due to a high respiration rate and/or a partial inhibition of photosynthesis induced by glycerol. In contrast, the rate of inorganic carbon fixation was higher in SW90-gly than in control samples in seawater, suggesting that non-photosynthetic patterns were operating. In darkness, after pretreatment in the light in SW90-gly, samples showed a high oxygen uptake rate just after the light was turned off. Twenty minutes of darkness were enough to decrease this high respiration rate to that of samples in seawater. The oxygen uptake observed in all experiments with glycerol was mitochondrial as it was inhibited by potassium cyanide and salicylhydroxamic acid (SHAM). Pretreatment of samples in the light in SW90-gly with the photosynthetic inhibitor DCMU did not inhibit ensuing dark respiration, thus providing evidence for a non-photosynthetic effect of the light. The highest dark respiration rate was observed after the samples were pretreated in monochromatic blue light in glycerol-containing media.     

A NUTRITIONAL ANALYSIS OF A SUBLITTORAL DIATOM ASSEMBLAGE EPIPHYTIC ON ENTEROMORPHA FROM A LONG ISLAND SALT MARSH1,2

The diatom population structure of a salt marsh epiphytic community growing on Enteromorpha intestinalis was studied at one station throughout the summer. A total of 218 species or varieties were recognized. Six species— Fragilaria construens, Cocconeis scutellum, Cocconeis placentula, Achnanthes hauckiana varieties, Achnanthes pinnata, and Amphora coffeaeformis (var. acutiuscula)–dominated the Enteromorpha epiphytic community during the summer months and comprised ∼ 40% of the total populations. Melosira nummuloides, Opephora martyi, Synedra fasiculata var. tabulata, S. affinis, Navicula platyventris, and N. pavillardi were also very common species (13% of the total population). The distribution of many species in the community was seasonal. A series of differential media have been developed which are effective as tools for the isolation and nutritional characterization of the algae and bacteria from the community. Many diatom species can be recognized by their colony type or growth pattern on solidified media. A key and illustrated plates aid in identification. Changes in population structure of the community were reflected by changes in the nutritional patterns as judged by differences in the growth of diatoms and bacteria on the differential media tested. The nutritional requirements, selectivity, and rank order of media for individual species is given. Almost half of the colony types (32–33) grew on either unenriched seawater from the collection site or basal synthetic seawater. Erdschreiber was a poorer medium for the isolation of algae than seawater alone. Few algal colonies grew on either marine nutrient agar or trypticase soy agar. Nitrite inhibited algal growth. Media enriched with thiamine, biotin, or nitrate, phosphate, and B₁₂ were stimulatory. Soil extract or an acetone extract of Enteromorpha tripled growth; a complex vitamin mixture, or glycerol, or mannitol, or an autoclaved extract of Enteromorpha doubled colony numbers. The greatest numbers of diatom types (43) and total colonies (655) were recovered on media enriched with acetone extract of Enteromorpha, suggesting a possible nutritional relationship between Enteromorpha and its epiphytes. The representativeness of the isolations from the natural community as a function of time was also considered. Concentrations of selected dissolved potential growth-stimulating nutrients were measured within and external to Enteromorpha communities. We conclude that the algal assemblage growing epiphytically on Enteromorpha intestinalis has a diverse auxotrophic profile which contributes to the productivity and stability within this important component of the epiphytic community and that much of the organic substrates used by individual species originate within and are recycled among the community members.     

Culture of an autoflocculent microalga in a vertical tubular photobioreactor for phycoerythrin production

Summary A red microalgal, Rhodosorus marinus, was used to produce phycoerythrin. This autoflocculent species, grown in a vertical tubular photoreactor equipped with a gaz-lift system, achieved a growth rate of 0.029 h^-1 with a maximum biomass yield of 2 g.l^-1 dry weight. These values were close to those obtained in batch cultures of other red microalgae cultured as free cell suspensions. Results have shown that these algae could be grown efficiently in natural seawater enriched with nutrients and harvested by decantation followed by filtration.     

Photosynthetic inorganic carbon utilization and growth of Porphyra linearis (Rhodophyta)

Photosynthetic (oxygen evolution) and growth (biomass increase) responses to ambient pH and inorganic carbon (Ci) supply were determined for Porphyralinearis grown in 0.5 L glass cylinders in the laboratory, or in 40 L fibreglass outdoor tanks with running seawater. While net photosynthetic rates were uniform at pH 6.0–8.0, dropping only at pH 8.7, growth rates were significantly affected by pH levels other than that of seawater (c. pH 8.3). In glass cylinders, weekly growth rates averaged 76% at external pH 8.0, 13% at pH 8.7 and 26% at pH 7.0. Photosynthetic O₂ evolution on a daily basis(i.e. total O₂ evolved during day time less total O₂ consumed during night time) was similar to the growth responses at all experimental pH levels, apparently due to high dark respiration rates measured at acidic pH. Weekly growth rates averaged 53% in algae grown in fibreglass tanks aerated with regular air (360 mg L_-1 CO₂) and 28% in algae grown in tanks aerated with CO₂-enriched air (750 mg L^-1 CO₂). The pH of the seawater medium in which P. linear is was grown increased slightly during the day and only rarely reached 9.0. The pH at the boundary layer of algae submerged in seawater increased in response to light reaching, about pH 8.9 within minutes, or remained unchanged for algae submerged in a CO₂-free artificial sea water medium. Photosynthesis of P. linearissaturated at Ci concentrations of seawater (K_0.5560 μM at pH 8.2) and showed low photosynthetic affinity for CO₂(K_0.5 61 μM) at pH 6.0. It is therefore concluded that P. linearisuses primarily CO₂ with HCO₃ ^- being an alternative source of Ci for photosynthesis. Its fast growth could be related to the enzyme carbonic anhydrase whose activity was detected intra- and extracellularly. This revised version was published online in August 2006 with corrections to the Cover Date.     

Dopamine release by Ulvaria obscura (Chlorophyta): environmental triggers and impacts on photosynthesis,growth, and survival of the releaser

In the NE Pacific, Ulvaria obscura is a common component of “green tide” blooms. It is also the only alga known to produce dopamine, which is released into seawater on sunny days when Ulvaria is emersed and then rehydrated. To better understand the mechanisms associated with dopamine release, we experimentally determined whether light quantity and quality, desiccation, temperature, exudates from conspecifics, and dissolved dopamine caused dopamine release. We also examined the effects of desiccation on Ulvaria's ability to photosynthesize, grow, and survive. Desiccation was the only factor that caused significant amounts of dopamine to be lost from U. obscura tissues. The loss of water from Ulvaria tissues was strongly and positively correlated with the loss of dopamine after rehydration. Only 56% of desiccated algae survived for 1 week, compared to 100% of undesiccated control algae. Desiccated algae lost 77% of their pigmented surface area and grew only 15% as much as undesiccated algae, which remained fully pigmented. The oxygen saturation of water containing Ulvaria that was desiccated and then rehydrated was significantly lower than that of seawater containing undesiccated algae. Thus, desiccation, which is coupled with dopamine release, is associated with the deterioration and death of some, but not all, tissues in Ulvaria. Although dopamine released into seawater can reduce the survival or growth of potential competitors, its release is associated with significant physiological stress and tissue mortality. However, the survival and continued growth of some Ulvaria tissues indicates that a net fitness benefit to release dopamine following desiccation cannot be ruled out.     

A NEW,ARTIFICIAL SEA WATER THAT FACILITATES GROWTH OF LARGE NUMBERS OF CELLS OF ACETABULARIA ACETABULUM (CHLOROPHYTA) AND REDUCES THE LABOR INHERENT IN CELL CULTURE1

We designed a new, artificial seawater (Ace 25) in order to grow bulk cultures of Acetabularia acetabulum (L.) Silva with a minimum of labor. To this end, we modified the traditional recipe for cell growth (Müller's medium as modified by Schweiger et al.) We eliminated five of the inorganic chemicals and determined the optimum concentration for 16 of the remaining 18 inorganic chemicals from modified Müller's seawater. Ace 25 enables growth of A. acetabulum from the beginning of the juvenile phase through gametangial formation in 11 weeks at high cell densities without medium replenishment. This represents a 98% reduction in the seawater volume required to mature each cell, a 30–40% reduction of the duration of the life cycle, an estimated 80% reduction in labor, and a 50–95% reduction in the space required for culturing A. acetabulum as compared with traditional procedures. These improvements may facilitate studies that require large numbers of cells such as population studies, genetics, and biochemistry, contribute to understanding the nutritional requirements of marine algae, and extend the use of this cell to those who lack the space or manpower to grow the cells in the traditional manner.     

Interaction between the macrophyte <Emphasis Type="Italic">Stratiotes aloides</Emphasis> and filamentous algae: does it indicate allelopathy?

The aquatic macrophyte Stratiotes aloides Linnaeus, which has recently received attention in studies on allelopathy, has been shown to suppress phytoplankton growth. In the Netherlands, S. aloides often co-occurs with floating filamentous algae. However, filamentous algae are generally absent in close proximity to S. aloides, resulting in gaps in filamentous algae mats. We analyzed whether those gaps may be caused by allelopathic substances excreted by S. aloides or by nutrient depletion. We studied in a field survey the colonization of natural S. aloides by filamentous algae and determined in situ nutrient concentrations in natural S. aloides stands. To analyze the relative importance of allelopathy and nutrient competition in the interaction between S. aloides and filamentous algae, we carried out field experiments. Introduction of artificial (non-allelopathic) plants in natural S. aloides stands enabled us to compare the colonization by filamentous algae of both Stratiotes sp. and artificial plants. The filamentous algae were absent in close vicinity to S. aloides. Significantly lower concentrations of ortho-phosphate and potassium were observed close to S. aloides as compared with the filamentous algae. In the field experiments the artificial plants were rapidly colonized by filamentous algae, mainly Cladophera Kützing and Spirogyra Link, while all natural plants remained free of such algae. Additionally, most nutrient concentrations did not significantly differ in the proximity of artificial or natural stands of S. aloides. The concentrations of the major growth-limiting nutrients, phosphate and nitrate, were significantly higher and nonlimiting in natural Stratiotes stands. Our main conclusion is that, although allelopathic interactions between S. aloides and filamentous algae do occur under natural conditions, nutrient competition between the two can also be an important factor.     

The effect of pH upon the photosynthesis of littoral marine algae

Summary The photosynthetic rates of a number of littoral marine algae were determined over a pH range from 8.1 to 10.3, employing the Winkler technique to measure evolved oxygen. In a large number of red, brown and green algae, the rate fell to a low value, or to complete inhibition, at pH 9.5 and higher. This indicates, in conformity with general permeability theory, that the HCO₃ ^}-ion cannot readily penetrate the cells, and is hence not available for photosynthesis.However, in two genera of coralline red algae (Bossea andCorallina) photosynthesis persists up to pH 10 or higher (though at a considerably reduced rate). This can be interpreted as due to utilization of the HCO₃ ^}-ion (though probably not the CO₃=ion). But several other non-coralline red algae (Centroceras, Botryocladia andGastroclonium), as well as the brown algaPelvetia (and possiblyFucus), and the green algaeUlva andEnteromorpha display this same ability.Thus the utilization of HCO₃ ^}-ion cannot alone be ascribed as the cause of calcium carbonate deposition, since several other non-calcareous algae have this same power. The utilization of HCO₃ ^}-at least permits calcification, however. The question might be reversed, to ask whyUlva, Enteromorpha. Pelvetia and several red algae are not calcified. Some cell wall property may be involved.     

An inexpensive inorganic medium for the mass cultivation of freshwater microalgae

A new and inexpensive inorganic medium (‘DS medium’) for the mass cultivation of freshwater blue-green algae (Cyanobacteria) and green algae has been developed. It consists basically of distilled or demineralized water (90%) and seawater (10%) and requires only little addition of pure purchasable chemicals (phosphate, trace elements, if necessary nitrate). No addition of macronutrients (NaCl, MgCl₂ or MgSO₄, KCl or K₂SO₄, CaCl₂) and of boron is required because they are sufficiently provided by the seawater. Decalcified water may also be suitable instead of demineralized water. For the cultivation of green algae, a higher trace element concentration is recommended than for blue-green algae. Because of its low total salt concentration the DS medium is freshwater-like. It is easy to prepare and effects rapid algal growth. It may be of special value for algal mass culture in regions which are close to the sea.     

Inhibitory activity of brown algal phlorotannins against glycosidases from the viscera of the turban shell Turbo cornutus

The crude phlorotannins from the brown alga Eisenia bicyclis showed inhibitory activity against 10 of 13 kinds of glycosidases present in the viscera of the turban shell Turbo cornutus. Phloroglucinol and its oligomers – eckol (a trimer), phlorofucofuroeckol A (a pentamer), dieckol and 8,8′-bieckol (hexamers), and an unidentified tetramer – were isolated from the crude phlorotannins by column and thin-layer chromatography. Phlorofucofuroeckol A, dieckol and 8,8′-bieckol inhibited α-fucosidase, β-galactosidase and β-mannosidase partially purified from T. cornutus, while phloroglucinol, eckol and the unidentified tetramer were weakly active. Dieckol was a competitive inhibitor of α-fucosidase with an inhibition constant (K _i) of 0.12?mM. The amounts of phlorotannins released after the immersion of freshly collected E. bicyclis in seawater or deionized water were estimated by high-performance liquid chromatography. Nearly all the phlorotannins were exuded into the medium following the death of the algae, whereas no phlorotannins were detected in the medium of living algae. These findings indicate that the phlorotannins deter the feeding of marine herbivorous gastropods by inhibiting the glycosidases.