Fine structure of some prosobranch ospharadia

The fine structure of the osphradia of Buccinum undatum 1. three Hawaiian Conus spp., Nassarius reticulatus (L), Nucella lapillus (L) and Littorina littorea (L) was examined. There is a remarkable uniformity in the arrangement of ultrastructural cell types in all the ospharadia investigated. The marginal glandular region is characterised by a single layer of cells, bearing a regular pallisade of microvilli and including two types of mucous cell. The epithelium of the sensory region is several cells deep, and bears a complex layer of microvilli in many orientations. Besides indifferent cells it contains two types of cell bodies of ciliated nerve processes. One type ends at the surface of the sensory region. The other, containing an elaborate array of smooth membranes in the perikaryon, has a process which enters the transitional region between sensory and glandular areas. In the transitional region, specialised cells with motile cilia adjoin the glandular region, and similar, but unciliated cells, containing large pigment granules scattered throughout the cytoplasm, adjoin the sensory region. The distal surface of both cell types is dissected by elefts extending 2 to 3 proximally, then widening into extensive spaces which contain cytoplasmic processes. The ciliated neurites which enter the transitional region end in the spaces at the base of the unciliated transitional cell elefts. It is suggested that the neuroepithelial cell processes, both those distributed over the sensory region and those concentrated in the transitional region, are receptors.     

Cytological study and immunohistochemical location of PSP toxins in foot skin of the ormer, Haliotis tuberculata, from the Galician coast (NW Spain)

The ormer, Haliotis tuberculata Linnaeus, 1758, is able to accumulate paralytic shellfish toxins (PST) in the foot side skin. Epidermal and secretory cells are described separately for the pedal side and sole epithelia, as notable differences have been observed. Melanin and photosynthetic pigment granules in the epidermal cells of the foot side epithelium give this skin its appearance and characteristic color. This epidermis also has abundant secretory cells containing at least three kinds of secretory granules. Conversely, the epidermal cells of foot sole epithelium do not have pigment granules, and the secretory cells are mostly subepithelial. Immunohistochemical reaction against PST (saxitoxin and its derivatives) showed that the toxins are located in specific cells of the foot side epithelium, which are distinct from the epidermal cells. Immunofluorescence also reveals toxins throughout many areas of the mucilage covering the epithelium, suggesting the secretory nature of the cells containing the toxins. These results do not show any apparent relationship between melanin or photosynthetic pigment granules and PST content. PST were not detected in the foot sole epithelium either immunohistochemically or by HPLC. Considering the toxic characteristics of ormers described in previous studies and the results presented here, the authors discuss the role of toxins retained or produced by the foot side epithelium Received: 25 July 2000 / Accepted: 7 November 2000     

A chromatophore system in the hermatypic,deep-water coral Leptoseris fragilis (Anthozoa: Hexacorallia)

Ultrastructural evidence is presented of a chromatophoresystem in the zooxanthellae containing hermatypic, deep-water coral Leptoseris fragilis (Milne Edwards and Haime). It consists of multilobed cells which mainly occupy the intercellular space of the oral gastrodermis. The cellular processes are filled with electron-dense granules up to 1-m-long and 0.5-m-wide. Within the cytoplasm an elaborate system of microtubules is established. The ramifications of the pigment cells, containing the pigment granules, form a dense and nearly continuous layer close to the overlying zooxanthellae. It is speculated that host pigments may transform the violet portion of the incident light into longer wavelengths, thus increasing the photosynthetic efficiency of the zooxanthellae.     

双酚A对青岛大扁藻的干扰效应

为了探讨双酚A(bisphenol A,BPA)对海洋微藻的生态毒性效应,实验选择了以青岛大扁藻(Platymonas helgolanidica)作为受试物种,设置6个实验浓度(即0、2、4、6、8、10 mg·L-1)对微藻进行了96 h暴露处理,测定了不同浓度暴露下对青岛大扁藻的生长以及抗氧化系统酶活性等指标。研究结果表明,BPA对青岛大扁藻的96h-EC50为9.32 mg·L-1,属高毒类污染物。青岛大扁藻经过BPA暴露处理后,细胞密度下降,细胞色素含量降低,并且呈现明显的剂量-效应关系;细胞抗氧化系统中超氧化物歧化酶(SOD)、抗坏血酸过氧化物酶(APX)、单脱氢抗坏血酸还原酶(MDHAR)、脱氢抗坏血酸还原酶(DHAR)、谷胱甘肽还原酶(GR)活性均受到干扰。     

Identification of haemocytes and their role in clotting in the blue crab,Callinectes sapidus

Three types of haemocyte — hyaline cells, small granule and large granule haemocytes — have been identified in the circulation of blue crabs, Callinectes sapidus Rathbun, that were collected and processed in North Carolina, USA in February 1991. Differentiation of haemocytes begins in the haematopoietic tissue and continues in the circulation. Hyaline cells are distinguished morphologically by their relatively small size, high nucleocytoplasmic ratio and sparse cytoplasmic granule content. Small and large granule haemocytes are distinguished, respectively, by a preponderance of small and large electron dense granules in their cytoplasm. Furthermore, a light and electron microscopic examination of the clotting process performed in July 1991 revealed that hyaline cells initiated coagulation when exposed to seawater, whereas granular haemocytes did not undergo marked changes when so treated. These distinct functional and morphologic features suggest that the hyaline cell and granular haemocytes represent distinct cell lines. It remains unclear whether small granule haemocytes differentiate into large granule haemocytes.     

Histochemical and ultrastructural characterisation of mantle storage cells in the hydrothermal-vent bivalve Bathymodiolus azoricus

This study reports on two types of storage cells that are present in the mantle connective tissue of the deep-sea mussel Bathymodiolus azoricus. One type corresponds to the adipogranular cells, a kind of storage cell previously described in other bivalves. In these cells extensive regions of the cytoplasm are filled with glycogen deposits and these zones became strongly stained after histochemical (PAS) or ultrastructural detection of polysaccharides. Several lipid droplets and membrane bound granules containing homogeneous electron-dense material are also present in adipogranular cells. A second type of cell contains large lysosomes in addition to numerous lipid droplets, but lacking cytoplasmic glycogen deposits. Due to these characteristics we named them adipolysosomal cells. They can be identified in semi-thin sections stained with PAS reaction because the lysosomes are the only positively stained structures. In the connective tissue of the mantle, some cells containing many lysosomes and a few lipid droplets were also observed. These cells differ from the adipolysosomal cells mainly because they have a reduced amount of lipid reserves, and could be an initial stage in the development of adipolysosomal cells. The vesicular connective tissue cells that in other Mytilidae are specialised in glycogen storage were not detected in B. azoricus. The reserves accumulated in the two types of storage cells described in B. azoricus may be important for the survival of these hydrothermal-vent bivalves if their nutrition is affected by a temporary loss or reduction of endosymbiotic bacteria due to sulphide and/or methane shortage caused by oscillations in vent activity.     

Resting spore formation and biochemical composition of the marine planktonic diatom Chaetoceros pseudocurvisetus in culture: ecological significance of decreased nucleotide content and activation of the xanthophyll cycle by resting spore formation

The biochemical composition of vegetative cells and resting spores of Chaetoceros pseudocurvisetus Mangin was compared in cultures under various nutrient and light conditions. The cellular content of major nucleotides such as AMP, ADP, ATP and UTP decreased in the order: vegetative cells, nutrient-starved (vegetative) cells and resting spores, indicating that the general metabolism of resting spores is relatively inactive. ADP-glucose was only abundant in nutrient-starved vegetative cells, suggesting metabolic imbalance in these cells. The chl a–specific fluorescence yield of vegetative cells grown under all culture conditions was low, but very high in resting spores. The ratios of the cellular contents of diadinoxanthin to chl a and of diatoxanthin to chl a were higher in resting spores and nutrient-starved vegetative cells than in nutrient-replete vegetative cells. The diadinoxanthin–diatoxanthin xanthophyll cycle was active in resting spores; the xanthophyll cycle was synchronized with a 14 h light:10 h dark photoperiod. Also, the ratios of cellular content of diadinoxanthin and diatoxanthin to cellular content of chl a in resting spores were relatively high in high irradiance, and decreased gradually in conditions of darkness over long culture periods. Under conditions of strong light and high temperature, most resting spores survived more than 40 d while nutrient-starved vegetative cells died within 33 d. These results suggest that resting spore formation is a strategy for enhancing protection and lowering metabolic rate for survival. These physiological changes accompanying spore formation enable resting spores not only to overwinter but also to “oversummer” in the coastal euphotic layer. Received: 23 March 1999 / Accepted: 11 August 1999     

The effect of intensity and quality of illumination on the photosynthesis of some tropical marine phytoplankton

Using solar energy as a source of illumination, photosynthesis in 11 species of marine plankton algae was studied as a function of light intensity. From the photosynthesis-light curve for each organism, the saturation points (I _k ) in different organisms were determined. Among the diatoms and flagellates, the highest I _k (saturation point) values were found in Rhizosolenia styliformis and Dinophysis miles respectively. When the organisms were exposed to a portion of the visible spectrum starting from 700 m, the photosynthesis was found to be related to the radiant energy. The missing portions of the spectrum produced no significant change in the rate of photosynthesis. The photosynthetic response shown by the different organisms was strikingly similar, which signifies that, despite the qualitative dissimilarities which the organisms may possess in their pigment composition, they are capable of much chromatic adaptation.     

Potential histopathological and molecular changes in rat vas deferens inhaled by Boswellia papyrifera and Boswellia carterii

Boswellia papyrifera and Boswellia carterii released from smoke contaminate indoor environment and consequently adversely affect humans as evidenced by respiratory disturbances. The aim of this study was to determine the effects of these plants on pathological and biochemical changes in vas deferens of albino rats. Animals were administered 4 g/kg body weight B. papyrifera and B. carterii daily for 120 days along with controls. Significant changes were observed in epithelial cell types and some cells showed signs of degeneration. The ultrastructural studies revealed marked changes in cytoplasmic organelles. Microvilli were missing and lysosomes were found in the cytoplasm. In addition, all treated groups plasma fructose and other biochemical parameters were decreased indicating reduced energy necessary for motility and contractility of spermatozoa. Many spermatozoa were disorganized and agglomerated. Data suggest that smoke from these plants adversely affects vas deferens.     

Diatom Thalassiosira weissflogii in oligotrophic versus eutrophic culture: models and ultrastructure

Populations of marine diatom Thalassiosira weissflogii were grown in continuous cultures enriched with f/2 medium. One of the two contrasting cultures (‘eutrophic’) received 5.6 times more nutrients than the other (‘oligotrophic’). Two mathematical models are analyzed to estimate eutrophication differences. The second model based on the Michaelis–Menten uptake and Droop growth shows that cells in the eutrophic culture should have about 56% higher content of silica which is the limiting nutrient. Diatom samples were prepared for the transmission electron microscopy after cells have been kept in chemostats for 37 days. The structure of diatom cells was investigated and a comparison is made between cells grown in oligotrophic and eutrophic conditions. In eutrophic culture, dividing cells were encountered more frequently while cell concentration was approximately equal in both chemostats. The central vacuole of cells in eutrophic culture accumulated dispersed and compact material from amorphous to spherical shape. In some cells the large central vacuole had fibrilar and peppered dense materials in addition to translucent granules, vesicules and multivesicular bodies. In the cytoplasm we found increased number of multivesicular bodies, dense and lucent granules some of which enclose membrane particles and lucent vesicules. Dense material depositions observed in the vacuole are also seen in the cytoplasm associated with organelles, mitochondria and plasmalemma. Cells have well-developed, active and slightly increased number of dictyosomes (5–6). Some dictyosomes with dense secretory material in the cistern are apparently engaged in a granule formation process. Functional significance of dense material in the central vacuole, which has not been observed in cells grown in oligotrophic condition, is discussed.     

In situ characterization of sulphur in gill-endosymbionts of the shallow water lucinid Codakia orbicularis (Linné, 1758) by high-pressure cryofixation and EFTEM microanalysis

In this study, Codakia orbicularis gill-tissues were cryo-fixed by using high-pressure freezing, a freeze substitution process and finally by cryo-embedding in Lowicryl. Ultrathin sections were then used for an EFTEM microanalysis. Results show that intracellular bacterial symbionts contain elemental sulphur in periplasmic vesicles as indicated by conventional TEM. When sulphur is temporarily depleted in the environment, such structures may act as energy sources for bacterial metabolism. Moreover, sulphate was detected in the cytoplasm of the bacterial symbionts, suggesting the oxidation of elemental sulphur, located in periplasmic granules, to sulphate (the final step in sulphur oxidation) by these chemoautotrophic bacteria. To assess the effects of host starvation on the bacterial sulphur content, adult individuals of C. orbicularis were maintained in starvation for 6 weeks in sterile artificial seawater depleted in sulphur. During starvation, both (1) the number of bacteria inside the bacteriocytes and (2) the number of periplasmic granules per prokaryotic cell decreased. The content of the remaining periplasmic granules had been modified to sulphate. This observation suggests that bacterial gill-endosymbionts used the elemental sulphur in their periplasmic granules as stored substrate for oxidation in order to produce energy in case of sulphur depletion.     

The osphradium in Placopecten magellanicus and Pecten maximus (Bivalvia,Pectinidae): histology,ultrastructure, and implications for spawning synchronisation

The bivalve osphradium is a band of putatively sensory tissue located in the gill axis, whose function is uncertain. In the present study, extending from 1987 to 1994, anatomical, histological, and electron microscopical techniques were used to elucidate the structure and ultrastructure of the osphradium in hatchery Pecten maximus L. and Placopecten magellanicus (Gmelin) (collected from Passamaquoddy Bay, New Brunswick, Canada). The osphradium consists of two distinct regions which run longitudinally on both sides of each gill axis: the osphradial ridge, and the dorsal tuft cilia region. The osphradial ridge was largely devoid of cilia other than those of the few free nerve fibres. The dorsal tuft cilia region contained free nerve fibres and ciliary tufts, separated by undifferentiated epithelial cells. No paddle cilia were observed under isosmotic fixation conditions, although under hypotonic conditions such cilia were quite common, suggesting an artefactual nature. Most of the cells of the osphradial ridge were highly secretory, the principal products being large pigment granules (in Pecten maximus) directly secreted by the Golgi bodies, and numerous small, electron-dense vesicles. These vesicles were arranged along extensive microtubule arrays in the basal region, indicative of axonal transport. These data support and extend Haszprunar's hypothesis of the role of the osphradium in the reception of chemical spawning cues and in the synchronization of gamete emission. Together with independent data on nerve pathways, osphradial sensory modalities, and monoamine localisation, an anatomical pathway and neurophysiological mediator are postulated.     

Tissue and sub-cellular distribution of Fe,Cu, Zn and 210Po in the abalone Haliotis rubra

The tissue and sub-cellular distribution of Fe, Cu, Zn and the naturally occurring radionuclide polonium-210 was determined in the gastropod mollusc Haliotis rubra collected from Western Port Bay, Australia, between March and July 1988. The highest concentrations of the metals, with the exception of Cu, were found in the digestive gland. Copper was more uniformly distributed, with tissues that are more vasculated having higher concentrations. Ultrastructural examination of the digestive gland, gill and kidney showed dense membrane-bound granules within the cytoplasm. Elemental analysis of the granules by electron probe x-ray microanalysis indicated that the granules in the digestive gland and gill contained high concentrations of iron, with small amounts of copper and zinc. In contrast, the metal-containing granules in the kidney were predominantly composed of iron, copper and phosphorus, with variable contributions of sodium potassium, and calcium. Homogenisation and fractionation of the digestive gland by differential centrifugation confirmed that approximately 80, 10, 90 and 50% of the total homogenate Fe, Cu, Zn and ²¹⁰Po, respectively, sedimented at 1200xg. In the haemolymph, all the elements studied were associated with the soluble high molecular weight component of the serum, not with the amoebocytes. ²¹⁰Po was present in the mucus-secreting hypobranchial glands at about half the concentration found in gill tissue.     

The histology,histochemistry and ultrastructure of the cement apparatus of three adult sessile barnacles,Elminius modestus,Balanus balanoides and Balanus hameri

The histology, histochemistry and ultrastructure of the cement gland cells of 3 species of sessile barnacles are described. Cells of both Elminius modestus Darwin and Balanus balanoides (L.) (membranous bases) are similar histologically and ultrastructurally, and within the cells there are 2 distinct regions. One is the site of secretion accumulation, the other the site of secretion synthesis. In Balanus hameri Ascanius (calcareous base) the cells are different in that the cytoplasm is more homogeneous. In all 3 species protein is the final secretion product (cement) of the cement cells. The mode of discharge of secretion from the cells differs between the membrane based and calcareous based barnacles. In the former, there is a series of collecting canals within the cytoplasm of the cement cells (intracellular canals) which join with the larger extracellular cement ducts. Secretion passes into the intracellular canals and is moved along to the larger cement ducts, which have an inner chitin lining. In Balanus hameri (calcareous base) there are no intracellular collecting canals. Secretions are thought to pass from the cement cells into the cement duct cells direct. The cement ducts in this species are not chitin lined.     

Effects of light quality on initiation and development of meroplanktonic diatom blooms in a eutrophic shallow sea

We investigated the effects of light quality on resting stage cell germination and vegetative cell growth of meroplanktonic diatoms in a small port in Hakata Bay, Japan and in the laboratory. During the investigation over the year of 2006, the meroplanktonic diatom bloom first occurred in the end of May and then repeated wane and wax until October in the small port. From late April to middle May, light penetrating the water column was often strong and attenuations of all spectral lights were low. During this period, Skeletonema costatum, Thalassiosira minima, and Chaetoceros sp. appeared frequently, followed by the blooms of S. costatum and Chaetoceros sp. in late May. Thereafter, S. costatum and Chaetoceros sp. bloomed in late June but not in middle June, when pigmented flagellates bloom appeared. The attenuation of short-wavelength light such as violet and blue lights was markedly high during these diatom and flagellate blooms; all blooms disappeared within several days. Vegetative cell strains of the three diatoms under light emitting diodes (LEDs) with six different spectra (violet, blue, green, orange, red, and near-infrared) grew at a higher rate under short-wavelength light, violet and blue. On the other hand, when suspensions of bottom sediments from Hakata Bay were cultured under the same LEDs and in the dark, vegetative cells of S. costatum appeared under all LEDs except for orange and near-infrared, vegetative cells of T. minima appeared under all LEDs but not in the dark, and vegetative cells of Chaetoceros sp. appeared under violet and blue LEDs. However, vegetative cell densities of the three diatoms increased much more under violet light than under other LEDs within a short period (6 days). Our study indicates that underwater penetration by short-wavelength light, such as violet and blue, may be an important factor in the initiation and development of meroplanktonic diatom blooms.     

Occurrence and distribution of antioxidant enzymes in the haemolymph of the shore crab Carcinus maenas

The haemocytes and cell-free haemolymph of the shore crab Carcinus maenas were examined for the presence of the antioxidant enzymes, catalase (EC 1.11.1.6), glutathione peroxidase (GPX) (EC 1.11.1.9) and superoxide dismutase (SOD) (EC 1.15.1.1) by biochemical assay and immunocytochemistry. The results show that all three enzymes are present in the haemocytes, but vary in distribution and amount. Catalase was detected principally in the phagocytic hyaline cells, whereas GPX and SOD were found in both the hyaline and granular cells. Catalase and glutathione peroxidase activities were absent from the semi-granular cells, and although SOD was detected in some samples, there was enormous variation in the level of this enzyme between individuals. Cell-free haemolymph, by contrast, was found to be positive for GPX and SOD but lacked catalase activity. Immunocytochemical analyses further showed that only 2 to 16% of the cells stain positively for these enzymes, and where present the enzymes tended to be distributed throughout the cytoplasm. With the granular cells, however, there was some localization of GPX and SOD around the granules. Since the phagocytes of C. maenas are known to produce superoxide anion radicals, hydrogen peroxide, and possible other reactive oxygen species upon stimulation with phorbol myristate acetate, lipopolysaccharide, or concanavalin A in vitro, it is likely that catalase, GPX and SOD are present in the haemolymph to assist in protection of the host tissues against oxyradical damage during host defence in vivo.     

Diel feeding behavior of neritic copepods during spring and fall blooms in Akkeshi Bay,eastern coast of Hokkaido,Japan

In situ diel feeding behavior of neritic copepods was investigated using the gut fluorescence method, during spring and fall bloom periods in Akkeshi Bay, on the eastern coast of Hokkaido, Japan. Acartia omorii and Paracalanus sp. were the dominant species during the fall, and Pseudocalanus spp. and A. longiremis during the spring. During both bloom periods, diel rhythms were always observed for the gut pigment contents of these dominant copepods, although there were interspecific differences in the pattern. The maximum gut pigment content was always observed during the night and the minimum during the day. For all species, except Paracalanus sp., the average gut pigment content during the night was significantly higher (p<0.05) than during daytime by factors of between 1.5 and 2.7. There were no significant differences between the gut evacuation rate constants determined during the day and the night, and initial gut pigment content had no effect on the value of gut evacuation rate constants. The instantaneous ingestion rates of individual copepods calculated from gut pigment and the mean value of gut evacuation rate constants followed the same diel rhythms as gut pigment contents. Copepod daily ingestion rates were higher than the daily requirements for respiration during both bloom periods. Estimated daily ration was 40 to 91% of body carbon during the fall bloom, and 17 to 28% during the spring bloom. The higher daily rations during fall were probably due to the difference in in situ temperature (ca. 14°C).     

Life-form population responses of a marine planktonic diatom,Chaetoceros pseudocurvisetus,to oligotrophication in regionally upwelled water

Life-form population responses of a centric planktonic diatom,Chaetoceros pseudocurvisetus Mangin, were investigated in summer 1986 and 1988 in the Izu Islands, Japan, in regionally upwelled water where nutrient concentration changed from favorable to unfavorable conditions for active growth and reproduction (oligotrophication). Two types of life form were observed: vegetative cells of healthy and unhealthy looking conditions and resting spores. The observed life-form responses were experimentally evaluated along with a depletion of limiting nutrients. The algal population ceased vegetative growth and initiated resting spore formation with a disappearance of limiting nitrate, and this life-form response seemed to be triggered by the decrease of cellular nitrogen content below a certain level. Since a large amount of silicon was required for the resting spore formation, a part of vegetative cells were unable to form resting spores and formed unhealthy looking vegetative cells under insufficient concentrations of silicic acid. Percentage shares of the resting spores in the population were linearly related to the amounts of available silicic acid. Vegetative cells which did not form resting spores showed greater mortality than resting spores by attack of bacteria and protozoa; however, vegetative cells could respond quickly to possible nutrient replenishment.     

Ecological and physiological differences between two colour morphs of the coral Pocillopora damicornis

Pocillopora damicornis (Linnaeus) is a ubiquitous branching coral found throughout the Indo-Pacific region. Like many other species of coral, P. damicornis displays a large range of morphologies. At One Tree Island, it occurs as two distinct morphs that are easily distinguished by the presence or absence of pink pigmentation. The two colour morphs of P. damicornis were found to differ in their distribution and abundance in the One Tree Island Lagoon. The brown morph was more abudant than the pink morph in the shallows (<1 m),whereas the pink morph was more abundant at deeper sites (>3 m). The two morphs also differed physiologically. The brown morph tended to have a greater calcification rate than the pink morph, regardless of environmental conditions. However, the difference in the calcification rate between the two morphs became non-significant under shaded conditions (5% full sunlight), indicating some degree of physiological plasticity of the morphs. The pink colour in P. damicornis was due to a hydrophilic pigment with a major peak absorbance at 560 nm. The expression of pink pigment had both genetic and phenotypic components. The brown morph has a reduced genetic capacity to express the pigment relative to the pink morph. On the other hand, pigment expression could be induced by light in the pink morph. Although genetic differences ultimately determine the differences between the two morphs of P. damicornis, the extent of pigment expression is under some degree of environmental influence.     

Effects of methionine sulfoximine on growth and nitrogen assimilation of the marine microalga Chlorella autotrophica

Chlorella autotrophica Shihira and Krauss (clone 580), a euryhaline microalga from the marine coastal environment is subject to large fluctuations in external salinity and nitrogen supply. The alga exhibits maximum growth at salinities lower than 100% ASW (artificial seawater). Cells divide faster and show higher cell yields when the supply of either NH ₄ ⁺ or NO ₃ ^- is increased above 0.2 mM. Cells growing on NH ₄ ⁺ show high levels of NADPH-glutamate dehydrogenase (GDH) activity, and the levels of glutamine synthetase (GS) are decreased to very low levels under these conditions. Methionine sulfoximine (MSX), an inhibitor of GS, has little effect on cell division and nitrogen assimilation of cells growing on NH ₄ ⁺ . Cells growing on NO ₃ ^- , however, show marked inhibition (65%) in nitrogen assimilation in the presence of 5 mM MSX. This MSX concentration also causes growth retardation and a progressive decrease in cell protein and nitrogen content. GS is almost completely inhibited by 5 mM MSX in both NH ₄ ⁺ and NO ₃ ^- -grown cells. Cells growing on NH ₄ ⁺ maintain high levels of NADPH-GDH activity in the presence of MSX. NADPH-GDH activity in MSX-treated NO ₃ ^- -grown cells increases, and, in the presence of 5 mM MSX, reaches 40% of the level found in NH ₄ ⁺ -grown cells. These results are consistent with NADPH-GDH providing an alternate pathway for NH ₄ ⁺ assimilation by this marine Chlorella species.