TWO MORPHOLOGICAL TYPES OF PARTICULATE INCLUSIONS IN MARINE DINOFLAGELLATES12

Two species of laboratory-reared nonbioluminescent dinoflagellates, Gymnodinium splendens (photo-synthetic) and Crypthecodinium cohnii (hetero-trophic), were observed to posses abundant quantities of randomly moving structures, which are contained in vacuoles. When viewed with the electron microscope, 2 shapes of regular, geometric particles, here termed particulate inclusions, were observed. The morphology of the particulate inclusions is described.     

THE OCCURRENCE AND SIGNIFICANCE OF ENDOSYMBIOTIC CHLOROPLASTS IN THE DIGESTIVE GLANDS OF HERBIVOROUS OPISTHOBRANCHS1

Intact algal chloroplasts have been found in the digestive glands of 5 species of Opisthobranchia belonging to the order Saccoglossa. Preliminary studies on 3 of these confirm their endosymbiotic nature. It is suggested that the occurrence of these endosymbiotic organelles may be widespread among related species of Saccoglossa. Their independent functional existence supports the view that chloroplasts possess a system of nonchromosomal inheritance.     

IDENTIFICATION OF MARINE DINOFLAGELLATES USING FLUORESCENT LECTINS1

Toxic and nontoxic species of marine dinoflagellates were characterized using fluorescent lectins. Lectin binding was detected by epifluorescence as well as spectrofluorometry. The binding assay of fluorescent lectins readily differentiated between morphologically similar species (i.e the toxic dinoflagellate Gymnodinium catenatum and the nontoxic Gymnodinium sp.). Lectins appear to be a useful tool to distinguish among different clones of the same species and, thus, possibly as a tool in dinoflagellate identification. Moreover, the lectins used show that thecate species have more binding sites and diversity in glycan moieties than athecate species.     

OCCURRENCE OF CHOLINE ESTERS IN THE MARINE DINOFLAGELLATE AMPHIDINIUM CARTERI12

Acrylylcholine, choline O-sulfate, and a yet unidentified choline ester have been isolated from cells of Amphidinium carteri. Natural and synthetic acrylyl choline had approximately 1/4,000 the activity of acetylcholine and choline O-sulfate about 1/20,000 the activity on isolated Mercenaria mercenaria heart.     

THE FATTY ACID AND STEROL COMPOSITION OF FIVE MARINE DINOFLAGELLATES

The fatty acid and sterol compositions of five species of marine dinoflagellates (Scrippsiella sp. Symbiodinium microadriaticum Freud, Gymnodinium sp., Gymnodinium sanguineum Hirasaki, and Fragilidium sp.) are reported. All contained the major fatty acids that are considered common in dinoflagellates, but the proportions were quite variable, and some species contained low contents of some polyunsaturated fatty acids. Concentration ranges for the major fatty acids were: 16:0 (9.0%–24.8%), 18:4(n-3) (2.5%–11.5%), 18:5(n-3) (7.0%–43.1%), 20:5(n-3) (EPA) (1.8%–20.9%), and 22:6(n-3) (DHA) (9.9%– 26.3%). Small amounts of novel very-long-chain highly unsaturated C₂₈ fatty acids occurred in all species. Each dinoflagellate contained a complex mixture of 4-methyl sterols and 4-desmethyl sterols. Four species contained cholesterol, although the amounts were highly variable (from 0.2% of total sterols in Scrippsiella sp. to 45.6% in Fragilidium sp.). All but G. sanguineum contained the 4-methyl sterol dinosterol, and all species contained sterols lacking a double bond in the ring system (i.e. stanols); in Scrippsiella sp. cholestanol composed 24.3% of the total sterols. Other common features of the 4-methylsterol profiles were the presence of 23,24-dimethyl alkylation and unsaturation at Δ²² in the side chain. In Scrippsiella sp., four steroidal ketones were identified: cholestanone, dinosterone, 4α,23,24-trimethyl-5α-cholest-8(14)-en-3-one, and dinostanone. The structures of these corresponded to the major sterols in this species, suggesting that the sterols and steroidal ketones are biosynthetically linked. Steroidal ketones were not detected in the other species. Although fatty acid profiles can be used to distinguish among algal classes, they were not useful for differentiating among dinoflagellate species. In contrast, whereas some taxonomic groupings of dinoflagellates display similar sterol patterns, others, such as the gymnodinoids studied here, clearly do not. The combination of fatty acid, sterol, and steroidal ketone profiles may be useful complementary chemotaxonomic tools for distinguishing morphologically similar species. The identification of steroidal ketones supports earlier suggestions that certain dinoflagellates might be a significant source of such components in marine environments.     

COMPARATIVE PHYSIOLOGICAL STUDY OF MARINE DIATOMS AND DINOFLAGELLATES IN RELATION TO IRRADIANCE AND CELL SIZE. I. GROWTH UNDER CONTINUOUS LIGHT1,2

Cell division rates and chlorophyll a and protein contents for ten diatom and dinoflagellate species were measured. Species were chosen to include a wide range of cell size in terms of both cell volume and cell protein: from 0.004 ng protein/cell for a small Chaetoceros sp. to 2.2 ng protein/cell for Prorocentrum micans Ehrenberg. Experiments were conducted in batch or semi-continuous cultures at 21 C under continuous illumination from 8–256 μEin ^.m^-2'^.s^-1. Light saturation of cell division occurred at 32–80 μEin m^-1 s^-1 for all species, with no observable difference between the two phylogenetic groups. When the light-saturated cell division rates were plotted against cell size as protein/cell, the diatoms and dinoflagellates fell on two separate lines with the diatoms having higher rates. Chl a /protein ratios (μg/μg) decreased with increasing irradiance. The diatoms had higher chl a per unit protein. The relationship between cell division rate and the chl a/protein ratio is discussed.     

OCCURRENCE AND ABUNDANCE OF GREEN-FLUORESCING DINOFLAGELLATES IN SURFACE WATERS OF THE NORTHWEST ATLANTIC AND NORTHEAST PACIFIC OCEANS1

We have cultured green fluorescing heterotrophic dinoflagellates whose continuous green fluorescence is due to an unidentified compound, probably a flavin, that excites with blue (～460 nm) light and emits green (～535 nm) light. No evidence of bioluminescence was found, but we note that compounds with similar fluorescence characteristics have been associated with bioluminescence in other taxa. These cells, all naked gymnodinoids, are widespread and abundant in the Northwest Atlantic and Northeast Pacific Oceans (10³–10⁵ L^?1). They comprise 4–100% of the total heterotrophic dinoflagellate component which, in turn, is usually equivalent magnitude to the phototrophic naked dinoflagellate component of the phytoplankton community.     

THE DETERMINATION OF THE FAR-RED EFFECT IN MARINE PHYTOPLANKTON1,2

A far-red effect exists in 4 marine phytoplankton species: the diatom Ditylum brightwelli, the coccolithophorid Coccolithus huxleyi, the green flagellate Dunaliella tertiolecta, and the dinoflagellate Pyrocystis lunula. The effect is reversible and is manifested through a change in cell division rate. Cultures of algae which received 30-min far-red (FR) light (750 nm) prior to the dark period were compared to controls which received, no FR. Reversal of the FR effect was studied by exposing experimental cultures to 30 min FR followed by 5-min red (R) light (650 nm) prior to the dark period. Controls received only FR. Cultures were exposed to light at 6 different enumerated wavelengths between 460 and 750 nm. A decrease in division rate runs evident only with light at 750 nm. These results give evidence for the presence of the phytochrome system in these phytoplankton species.     

MARINE BACTERIA OF CARDIGAN BAY. I. BACTERIA OF AN OFF-SHORE AREA

SUMMARY: A study of marine bacteria from a fixed off-shore area in Cardigan Bay showed there was little significance in the variation between numbers from different levels within 8 fathoms deep. There was a suggestion of seasonal variation in surface water in colony counts at 15–18°. In general, the bacterial content was highest over the summer months, but showed little direct correlation with temperature, pH or light.
Strains were mainly obligate halophytes and small Gram-negative rods predominated (96%). Nearly 50% of strains isolated liquefied gelatin; otherwise, biochemical reactions were weak. The proportion of pigmented strains was low.
Alien bacteria were present in irregularly varying numbers in all samples and showed no seasonal variation. Coli-aerogenes strains were recorded in small numbers, at all depths and in all seasons, but Bact. coli type I only rarely occurred.     

ENHANCEMENT OF PHYTOPLANKTON GROWTH BY MARINE BACTERIA12

Investigations were conducted into the effect of 3 marine bacteria, Vibrio anguillarum #19264, V. anguillarum #19109, and Escherichia coli, on the growth of 10 phytoplakters. A disc method on agar plates was used to evaluate growth responses. Growth enhancement of all algae in the presence of V. anguillarum #19264 occurred on an enriched agar medium; the other bacteria yielded variable responses. Evidence available is consistent with the hypothesis that growth enhancement of algae is related to the release of stimulating substances through bacterial hydrolysis of the agar.