RELATIONSHIP BETWEEN PHOTOSYNTHESIS AND CELL SIZE OF MARINE DIATOMS12

Three photosynthetic parameters of 7 species of marine diatoms were studied using Na₂¹⁴CO₃ at 5–8 C using log phase axenic cultures. The cell volumes of the different species varied from 70 μm³ to 40 × 10⁵μm³. The present experiment is consistent with the interpretation that the initial slope α (mg C · [mg chl a]^?1· h^?1· w^?1· m²) of photosynthesis vs. light curves is controlled by self-shading of chlorophyll a in the cell. Pm, the rate of photosynthesis at light saturation (mg C · [mg cell, C]^?1· h^?1) and R, the intercept at zero light intensity (mg C · [mg cell C]^?1· H^?1) are both dependent on the ratio of surface area to volume of cell.     

A CARBON BUDGET FOR THE AUTOTROPHIC CILIATE MESODINIUM RUBRUM1

Extensive blooms of the autotrophic ciliate Mesodinium rubrum (Lohmann) occurred in the Peru coastal upwelling region at 15°S latitude in March through May 1977 and contributed significantly to the organic productivity of the region. From observations made during the JOINT-II oceanographic expedition, a budget of the carbon flux of these unusual photosynthetic organisms can be constructed. The light dependent C fixation was determined with short (1 h) incubations because of the organisms' sensitivity to confinement and rapid nutrient exhaustion. Maximum photosynthesis occurred at 50% of incident light with a maximum rate of particulate C synthesis of 2187 mg C · m^?3· h^?1. The specific carbon uptake rates were also high with a maximum light saturated value of 16.8 mg C ·(mg chl a)^?1· h^?1. The rate of excretion of dissolved organic C at the productivity maximum ranged from 16.1 to 181.1 mg C · m^?3· h^?1. The range of percent excretion was 1.8–12.5% the total C fixed, similar to the range found in both motile and nonmotile phytoplankton assemblages. Respiration, determined by the decrease in particulate C in the dark, averaged 4.6% of the previously fixed photosynthetic C · h^?1. M. rubrum actively took up amino acids and naturally occurring dissolved organic carbon. The C budget for this ciliate indicates that the daily contribution to the particulate food chain is large, although not as great as is indicated by short incubations. The contribution of M. rubrum to the productivity and elemental fluxes of upwelling and coastal ecosystems has been seriously underestimated.     

CELL AND GROWTH CHARACTERISTICS OF TYPES A AND B OF EMILIANIA HUXLEYI (PRYMNESIOPHYCEAE) AS DETERMINED BY FLOW CYTOMETRY AND CHEMICAL ANALYSES1

Two morphotypes of Emiliania huxleyi (Lohmann 1902) Hay et al. 1967, types A and B, known to be unequally distributed in the oceans, were grown in dilution cultures at a range of photon flux densities (PFDs) (1.5–155 μmol photons·m^?2·s^?1) and two temperatures (10° and 15° C). Calcite carbon and organic carbon content of the cells as well as instantaneous growth rate, cell size, chlorophyll fluorescence, and light-scatter properties clearly depended on growth conditions and differed considerably for the two morphotypes. The ratio between calcite carbon and organic carbon production showed an optimum of 0.65 in E. huxleyi type A cells at PFD = 17.5. The ratio increased slightly with a temperature increase from 10° to 15°C but remained < 1.0 at both temperatures in light-limited cells. In contrast, calcite carbon production exceeded organic carbon production (ratio: 1.4–2.2) in phosphate-deprived cultures. Emiliania huxleyi type B generally showed a higher calcite carbon/organic carbon ratio than E. huxleyi type A, but the relation with PFD was similar. The content of calcite carbon and organic carbon as well as the instantaneous growth rate, cell size, chlorophyll fluorescence, and light-scatter properties showed large diel variations that were closely related to the division cycle. Our results show the importance of mapping the structure of any sampled cell population with respect to the phase in the cell division cycle, as this largely determines the outcome of not only “per cell” measurements but also short time (less than 24 h) flux measurements. For instance, dark production of calcite by E. huxleyi was negatively affected by cell division. Slowly growing (phosphate-stressed) cultures produced calcite in the light and in the dark. In contrast, rapidly growing cultures at 10°C produced calcite only in the light, whereas in the dark there was a significant loss of calcite due to dissolution.     

EFFECTS OF VARIATIONS IN LIGHT INTENSITY ON THE EFFICIENCY OF GROWTH OF SKELETONEMA COSTATUM (BACILLARIOPHYCEAE) IN A CYCLOSTAT1

The relative importance of respiration and organic carbon release to the efficiency of carbon specific growth of Skeletonema costatum (Grev.) Clave was evaluated over a light range from 1500–15 μE · m^?2· s^?1. Net growth efficiency ranged from 0.45–0.69 with a maximum at 130 μE · m^?2· s^?1. Respiration was 93% or more of the variations in growth efficiency. Organic carbon release ranged from 0–7% of gross production and increased with light intensity. Carbon specific particulate production was a hyperbolic function of incident light intensity and was related exponentially to particulate carbon production per unit chlorophyll a. Full sunlight conditions, 1500 μE · m^?2· s^?1, did not induce photoinhibition of gross production. Variations in the efficiency of growth of S. costatum were minimized over a wide range of light intensities mainly because of variations in cellular pigments which permitted the efficient utilization of available light energy, and a reduction in the losses of carbon which increases the growth rate, possibly as a consequence of the recycling of respired carbon within the cell.     

CELL‐SPECIFIC EXTRACELLULAR PHOSPHATASE ACTIVITY OF DINOFLAGELLATE POPULATIONS IN ACIDIFIED MOUNTAIN LAKES1

High bulk extracellular phosphatase activity (PA) suggested severe phosphorus (P) deficiency in plankton of three acidified mountain lakes in the Bohemian Forest. Bioavailability of P substantially differed among the lakes due to differences in their P loading, as well as in concentrations of aluminum (Al) and its species, and was accompanied by species‐specific responses of phytoplankton. We combined the fluorescently labeled enzyme activity (FLEA) assay with image cytometry to measure cell‐specific PA in natural populations of three dinophyte species, occurring in all the lakes throughout May–September 2007. The mean cell‐specific PA varied among the lakes within one order of magnitude: 188–1,831 fmol · cell^?1 · h^?1 for Gymnodinium uberrimum (G. F. Allman) Kof. et Swezy, 21–150 fmol · cell^?1 · h^?1 for Gymnodinium sp., and 22–365 fmol · cell^?1 · h^?1 for Peridinium umbonatum F. Stein. To better compare cell‐specific PA among the species of different size, the values were normalized per unit of cell biovolume (amol · μm^?3 · h^?1) for further statistical analysis. A step‐forward selection identified concentrations of total and ionic Al together with pH as significant factors (P < 0.05, Monte Carlo permutation test), explaining cumulatively 57% of the total variability in cell‐specific PA. However, this cell‐specific PA showed an unexpected reverse trend compared to an overall gradient in P deficiency of the lake plankton. The autecological insight into dinophyte cell‐specific PA therefore suggested other factors, such as light availability, mixotrophy, and/or zooplankton grazing, causing further PA variations among the acidified lakes.     

Kocuria SM1 controls vibriosis in rainbow trout (Oncorhynchus mykiss,Walbaum)

Aims: To develop probiotics for the control of vibriosis caused by Vibrio anguillarum and Vibrio ordalii in finfish. Methods and Results: Kocuria SM1, isolated from the digestive tract of rainbow trout, was administered orally to rainbow trout (Oncorhynchus mykiss) for 2 weeks at a dose equivalent to c. 10⁸ cells per g of feed and then challenged intraperitoneally with V. anguillarum and V. ordalii. Use of SM1 led to a reduction in mortalities to 15–20% compared to 74–80% mortalities in the controls. SM1 stimulated both cellular and humoral immune responses in rainbow trout, by elevation of leucocytes (5·5 ± 0·8 × 10⁶ ml^?1 from 3·7 ± 0·8 × 10⁶ ml^?1), erythrocytes (1·2 ± 0·1 × 10⁸ ml^?1 from 0·8 ± 0·1 × 10⁸ ml^?1), protein (23 ± 4·4 mg ml^?1 from 16 ± 1·3 mg ml^?1), globulin (15·7 ± 0·2 mg ml^?1 from 9·9 ± 0·1 mg ml^?1) and albumin (7·3 ± 0·2 mg ml^?1 from 6·1 ± 0·1 mg ml^?1) levels, upregulation of respiratory burst (0·05 ± 0·01 from 0·02 ± 0·01), complement (56 ± 7·2 units ml^?1 from 40 ± 8·0 units ml^?1), lysozyme (920 ± 128·8 units ml^?1 from 760 ± 115·3 units ml^?1) and bacterial killing activities. Conclusions: Kocuria SM1 successfully controlled vibriosis in rainbow trout, and the mode of action reflected stimulation of the host innate immune system. Significance and Impact of the Study: Probiotics can contribute a significant role in fish disease control strategies, and their use may replace some of the inhibitory chemicals currently used in fish farms.     

PHYCOBILIN CONTENT OF THE CONCHOCELIS PHASE OF ALASKAN PORPHYRA (BANGIALES,RHODOPHYTA) SPECIES: RESPONSES TO ENVIRONMENTAL VARIABLES1

Variations of pigment content in the microscopic conchocelis stage of four Alaskan Porphyra species were investigated in response to environmental variables. Conchocelis filaments were cultured under varying conditions of irradiance and nutrient concentrations for up to 60 d at 11°C and 30 psu salinity. Results indicate that conchocelis filaments contain relatively high concentrations of phycobilins under optimal culture conditions. Phycobilin pigment production was significantly affected by irradiance, nutrient concentration, and culture duration. For Porphyra abbottiae V. Krishnam., Porphyra sp., and Porphyra torta V. Krishnam., maximal phycoerythrin (63.2–95.1 mg · g dwt^?1) and phycocyanin (28.8–64.8 mg · g dwt^?1) content generally occurred at 10 μmol photons · m^?2 · s^?1, f/4–f/2 nutrient concentration after 10–20 d of culture. Whereas for Porphyra hiberna S. C. Lindstrom et K. M. Cole, the highest phycoerythrin (73.3 mg · g dwt^?1) and phycocyanin (70.2 mg · g dwt^?1) content occurred at 10 μmol photons · m^?2 · s^?1, f nutrient concentration after 60 d in culture. Under similar conditions, the different species showed significant differences in pigment content. P. abbottiae had higher phycoerythrin content than the other three species, and P. hiberna had the highest phycocyanin content. P. torta had the lowest phycobilin content.     

Temperature × light interaction and tolerance of high water temperature in the planktonic freshwater flagellates Cryptomonas (Cryptophyceae) and Dinobryon (Chrysophyceae)

Using microcosm experiments, we investigated the interactive effects of temperature and light on specific growth rates of three species each of the phytoplanktonic genera Cryptomonas and Dinobryon. Several species of these genera play important roles in the food web of lakes and seem to be sensitive to high water temperature. We measured growth rates at three to four photon flux densities ranging from 10 to 240 μmol photon · m^?2 · s^?1 and at 4–5 temperatures ranging from 10°C to 28°C. The temperature × light interaction was generally strong, species specific, and also genus specific. Five of the six species studied tolerated 25°C when light availability was high; however, low light reduced tolerance of high temperatures. Growth rates of all six species were unaffected by temperature in the 10°C–15°C range at light levels ≤50 μmol photon · m^?2 · s^?1. At high light, growth rates of Cryptomonas spp. increased with temperature until the temperature optimum was reached and then declined. The Dinobryon species were less sensitive than Cryptomonas spp. to photon flux densities of 40 μmol photon · m^?2 · s^?1 and 200 μmol photon · m^?2 · s^?1 over the entire temperature range but did not grow under a combination of very low light (10 μmol photon · m^?2 · s^?1) and high temperature (≥20°C). Among the three Cryptomonas species, cell volume declined with temperature and the maximum temperature tolerated was negatively related to cell size. Since Cryptomonas is important food for microzooplankton, these trends may affect the pelagic carbon flow if lake warming continues.     

ROLE OF GLUTAMATE DEHYDROGENASE AND GLUTAMINE SYNTHETASE IN CHLORELLA VULGARIS DURING ASSIMILATION OF AMMONIUM WHEN JOINTLY IMMOBILIZED WITH THE MICROALGAE‐GROWTH‐PROMOTING BACTERIUM AZOSPIRILLUM BRASILENSE1

Enzymatic activities of glutamate dehydrogenase (GDH) and glutamine synthetase (GS) participating in the nitrogen metabolism and related ammonium absorption were assayed after the microalga Chlorella vulgaris Beij. was jointly immobilized with the microalgae‐growth‐promoting bacterium Azospirillum brasilense. At initial concentrations of 3, 6, and 10 mg · L^?1 NH₄⁺, joint immobilization enhances growth of C. vulgaris but does not affect ammonium absorption capacity of the microalga. However, at 8 mg · L^?1 NH₄⁺, joint immobilization enhanced ammonium absorption by the microalga without affecting the growth of the microalgal population. Correlations between absorption of ammonium per cell and per culture showed direct (negative and positive) linear correlations between these parameters and microalga populations at 3, 6, and 10 mg · L^?1 NH₄⁺, but not at 8 mg · L^?1 NH₄⁺, where the highest absorption of ammonium occurred. In all cultures, immobilized and jointly immobilized, having the four initial ammonium concentrations, enzymatic activities of Chlorella are affected by A. brasilense. Regardless of the initial concentration of ammonium, GS activity in C. vulgaris was always higher when jointly immobilized and determined on a per‐cell basis. When jointly immobilized, only at an initial concentration of 8 mg · L^?1 NH₄⁺ was GDH activity per cell higher.     

Releases of ingested phytoplankton carbon by Daphnia magna

1. The carbon budgets and assimilation efficiencies (AEs) of adults and juveniles of Daphnia magna were quantified using ¹⁴C as a tracer. Animals were fed pure Chlamydomonas reinhardtii or Scenedesmus obliquus at different food concentrations. Carbon AEs (46–70%) were comparable at food concentrations of 0.03–0.30 mg C L^?1 for both algal species, but decreased to 34–49% when the food concentration further increased by 10‐fold. The carbon AEs were significantly and negatively correlated with the food level. 2. During the postdigestive period, partitioning of ingested carbon into different compartments including excretion, respiration and egestion was not influenced by the food species and life stage. There was a negative correlation between respiration (as % of total loss) and food concentration and a positive correlation between egestion (as % of total loss) and food concentration. Dissolved organic carbon (DOC) and CO₂ accounted for 55–72% and 9–37%, respectively, of the total carbon loss from juveniles fed both algal diets. For adults, DOC and CO₂ contributed to 44–64% and 20–47% of the total carbon loss, respectively. Particulate organic carbon (POC) was a minor pathway for the overall carbon loss. 3. The turnover and release budget of structural carbon (as moults and neonate reproduction) were further evaluated in long‐term experiments at different algal concentrations. Food concentration did not affect the carbon efflux or the carbon allocation into different physiological compartments except for respiration. Juveniles had twofold lower carbon turnover rate (0.12–0.16 day^?1) than those of the adults (0.32–0.35 day^?1). In adults, comparable carbon was allocated into DOC (35–42%) and reproduction (27–35%), which were the dominant routes for carbon loss. For the juveniles, DOC accounted for 42–64% of the total carbon loss. 4. About 21–38% of the total DOC released by adults and juveniles was associated with the high molecular weight organic carbon fraction (>5 kDa). Our results show that carbon was mainly lost by D. magna in the form of DOC during assimilation process as well as from the structural materials. Reproduction or maternal transfer was another major drain of body carbon for adult D. magna.     

PHYSIOLOGICAL VARIATION AND ELECTROPHORETIC BANDING PATTERNS OF GENETICALLY DIFFERENT SEASONAL POPULATIONS OF SKELETONEMA COSTATUM (BACILLARIOPHYCEAE)1

Clones of Skeletonema costatum (Grev.) Cl. isolated from Narragansett Bay, R.I., during different seasons were grouped according to their electrophoretic banding patterns. The growth rates, pg chlorophyll · cell^?1, carbon uptake · cell^?1· h^?1, and carbon uptake · pg chl^?1· h^?1 were measured at 20°C, in a 14:10 h L:D cycle at 180 μE · m^?2· s^?1. Statistically significant sources of variation were found among groups of clones in growth rate, pg chl · cell^?1, and carbon uptake · pg chl^?1· h^?1. It was concluded that there is a significant relationship between the physiological characteristics of clones isolated from populations in different seasons and patterns of genetic variation inferred from the electrophoretic studies. However, genetic diversity detected by banding patterns tends to underestimate the total genetic diversity in natural populations. The groups of clones most common in summer bloom populations had significantly higher growth rates, lower values of pg chl · cell^?1, and higher rates of carbon uptake · pg chl^?1· h^?1 at 20°C than did the group of clones most common in winter bloom populations. However, differences among groups in these parameters at 20°C alone cannot account for the seasonal cycling of genetically variable populations of Skeletonema in Narragansett Bay. The range of growth rates among clones of this species is 0.1–5.0 divisions · d^?1 under a single set of temperature and light conditions. Chlorophyll concentrations range from 0.2–1.7 pg chl · cell^?1 and carbon uptake · pg chl^?1· h^?1 varies by a factor of 7 among clones. The range of physiological variation in this species means that it is difficult to use laboratory studies of single clones to analyze the responses of natural populations of Skeletonema.     

Increased synthesis of α‐tocopherol,paramylon and tyrosine by Euglena gracilis under conditions of high biomass production

Aims: To analyse the production of different metabolites by dark‐grown Euglena gracilis under conditions found to render high cell growth. Methods and Results: The combination of glutamate (5 g l^?1), malate (2 g l^?1) and ethanol (10 ml l^?1) (GM + EtOH); glutamate (7·15 g l^?1) and ethanol (10 ml l^?1); or malate (8·16 g l^?1), glucose (10·6 g l^?1) and NH₄Cl (1·8 g l^?1) as carbon and nitrogen sources, promoted an increase of 5·6, 3·7 and 2·6‐fold, respectively, in biomass concentration in comparison with glutamate and malate (GM). In turn, the production of α‐tocopherol after 120 h identified by LC‐MS was 3·7 ± 0·2, 2·4 ± 0·1 and 2 ± 0·1 mg [g dry weight (DW)]^?1, respectively, while in the control medium (GM) it was 0·72 ± 0·1 mg (g DW)^?1. For paramylon synthesis, the addition of EtOH or glucose induced a higher production. Amino acids were assayed by RP‐HPLC; Tyr a tocopherol precursor and Ala an amino acid with antioxidant activity were the amino acids synthesized at higher concentration. Conclusions: Dark‐grown E. gracilis Z is a suitable source for the generation of the biotechnologically relevant metabolites tyrosine, α‐tocopherol and paramylon. Significance and Impact of the Study: By combining different carbon and nitrogen sources and inducing a tolerable stress to the cell by adding ethanol, it was possible to increase the production of biomass, paramylon, α‐tocopherol and some amino acids. The concentrations of α‐tocopherol achieved in this study are higher than others reported previously for Euglena, plant and algal systems. This work helps to understand the effect of different carbon sources on the synthesis of bio‐molecules by E. gracilis and can be used as a basis for future works to improve the production of different metabolites of biotechnological importance by this organism.     

Kinetic Modeling for Simultaneous Organic Carbon Oxidation,Nitrification, and Denitrification of Abattoir Wastewater in Sequencing Batch Reactor

A laboratory-scale study was conducted in a 20.0-L sequencing batch reactor (SBR) to explore the feasibility of simultaneous removal of organic carbon and nitrogen from abattoir wastewater. The reactor was operated under three different combinations of aerobic-anoxic sequence, viz., (4+4), (5+3), and (5+4) h of total react period, with influent soluble chemical oxygen demand (SCOD) and ammonia nitrogen (NH₄⁺-N) level of 2200 ± 50 and 125 ± 5 mg L^?1, respectively. In (5+4) h cycle, a maximum 90.27% of ammonia reduction corresponding to initial NH₄⁺-N value of 122.25 mg L^?1 and 91.36% of organic carbon removal corresponding to initial SCOD value of 2215.25 mg L^?1 have been achieved, respectively. The biokinetic parameters such as yield coefficient (Y), endogenous decay constant (k_d), and half-velocity constant (K_s) were also determined to improve the design and operation of package effluent treatment plants comprising SBR units. The specific denitrification rate (q_DN) during anoxic condition was estimated as 6.135 mg N/g mixed liquor volatile suspended solid (MLVSS)·h on 4-h average contact period. The value of Y, k_d and K_s for carbon oxidation and nitrification were found to be in the range of 0.6225–0.6952 mg VSS/mg SCOD, 0.0481–0.0588 day^?1, and 306.56–320.51 mg L^?1, and 0.2461–0.2541 mg VSS/mg NH₄⁺-N, 0.0324–0.0565 day^?1, and 38.28–50.08 mg L^?1, respectively, for different combinations of react periods.     

STUDY ON BIOLOGICAL CHARACTERISTICS OF HETEROTROPHIC MARINE MICROALGA—SCHIZOCHYTRIUM MANGROVEI PQ6 ISOLATED FROM PHU QUOC ISLAND,KIEN GIANG PROVINCE,VIETNAM1

Schizochytrium sp. PQ6, a heterotrophic microalga isolated from Phu Quoc (PQ) Island in the Kien Giang province of Vietnam, contains a high amount of docosahexaenoic acid (DHA, C22:6n‐3). In this study, the culture conditions are developed to maximize biomass and DHA production. Nucleotide sequence analysis of partial 18S rRNA gene from genomic DNA showed that PQ6 has a phylogenetic relationship close to Schizochytrium mangrovei Raghu‐Kumar. The highest growth rate and DHA accumulation of this strain were obtained in 6.0% glucose, 1.0% yeast extract, 50% artificial seawater (ASW), and pH 7 at 28°C. In addition, carbon and nitrogen sources could be replaced by glycerol, ammonium acetate, sodium nitrate, or fertilizer N–P–K. Total lipid content reached 38.67% of dry cell weight (DCW), in which DHA and eicosapentaenoic acid (EPA, C20:5n‐3) contents accounted for 43.58% and 0.75% of the total fatty acid (TFA), respectively. In 5 and 10 L fermenters, the cell density, DCW, total lipid content, and maximum DHA yield were 46.50 × 10⁶ cells · mL^?1, 23.7 g · L^?1, 38.56% of DCW, and 8.71 g · L^?1 (in 5 L fermenter), respectively, and 49.71 × 10⁶ cells · mL^?1, 25.34 g · L^?1, 46.23% of DCW, and 11.55 g · L^?1 (in 10 L fermenter), respectively. Biomass of PQ6 strain possessed high contents of Na, I, and Fe (167.185, 278.3, and 43.69 mg · kg^?1 DCW, respectively). These results serve as a foundation for the efficient production of PQ6 biomass that can be used as a food supplement for humans and aquaculture in the future.     

Root respiration and associated costs in deciduous and evergreen species of Quercus

Root construction and maintenance costs were estimated in four evergreen and three deciduous Quercus species that are typical in the landscape of southern Spain. The cost quantification was based on analysis of the growth–­respiration ratio. Values observed for both construction cost (ranging from 1·17 to 1·29 g glucose g^?1 dry weight) and maintenance cost (ranging from 6·22 to 11·71 mg glucose g^?1 dry weight d^?1) were generally lower than those reported in other studies. The results showed non‐significant differences between deciduous and evergreen species. The lack of significant differences between species appeared to be due to the homogeneity of growth conditions. Hydroponic culture, with unrestricted nutrient and water supply, would lead to low tissue carbon content and low respiration rates, leading to the low costs observed. Furthermore, the fact that root organs are clearly importers of organic molecules inevitably entails some underestimation of the respiration associated with growth and, to a lesser extent, with maintenance respiration. This leads in turn to underestimation of the corresponding construction and maintenance costs. All this raises doubts as to the suitability of this method for studying root systems.     

DIFFERENCES IN THE PRODUCTION AND EXCRETION KINETICS OF OKADAIC ACID,DINOPHYSISTOXIN‐1, AND PECTENOTOXIN‐2 BETWEEN CULTURES OF DINOPHYSIS ACUMINATA AND DINOPHYSIS FORTII ISOLATED FROM WESTERN JAPAN1

We established clonal cultures of Dinophysis acuminata Clap. et Lachm. and D. fortii Pavill. isolated from western Japan and examined toxin production in them, focusing on intracellular production and extracellular excretion. At the end of incubations, the total amounts of pectenotoxin‐2 (PTX‐2), dinophysistoxin‐1 (DTX‐1), and okadaic acid (OA) in the D. acuminata cultures reached up to 672.7 ± 14.7 (mean ± SD), 88.1 ± 2.8, and 539.3 ± 39.7 ng · mL^?1, respectively, and the excreted extracellular amounts were equivalent to 5.1, 79.5, and 79.5% of the total amounts, respectively. Similarly, at the end of incubations, the total amounts of PTX‐2, DTX‐1, and OA in the D. fortii cultures reached up to 526.6 ± 52.6 (mean ±SD), 4.4 ± 0.4, and 135.9 ± 3.9 ng · mL^?1, respectively, and the excreted extracellular amounts were equivalent to 1.8, 80.1, and 86.6% of the total amounts, respectively. Further, we tested the availability of cell debris and dissolved organic substances that originated from the ciliate prey Myrionecta rubra for growth and toxin production in D. acuminata. Although no significant growth was observed in D. acuminata in the medium containing the cell debris and organic substances originated from M. rubra, the toxicity was significantly greater than that in the control (P < 0.05–0.001); this finding suggested the availability of organic substances for toxin production. However, toxin productivity was remarkably lower than that of Dinophysis species feeding on living M. rubra.     

COCCOLITH PRODUCTION AND DETACHMENT BY EMILIANIA HUXLEYI (PRYMNESIOPHYCEAE)1

Laboratory experiments were performed with the prymnesiophyte Emiliania huxleyi (Lohm.) Hay and Mohler, strain 88E, to quantify calcification per cell, coccolith detachment, and effects of coccolith production on optical scattering of individual cells. ¹⁴C incorporation into attached and detached coccoliths was measured using a bulk filtration technique. ¹⁴C-labeled cells also were sorted using a flow cytometer and analyzed for carbon incorporation into attached coccoliths. The difference between the bulk and flow cytometer analyses provided a ¹⁴C-based estimate of the rate of production of detached coccoliths. Coccolith production and detachment were separated in time in batch cultures, with most detachment happening well after calcification had stopped. Accumulation of coccoliths was maximum at the end of logarithmic growth with 50–80 coccoliths per cell (three to five complete layers of coccoliths around the cells). Net accretion rates of coccoliths were on the order of 7 coccoliths· cell^?1·d^?1 while net detachment rates were as high as 15 coccoliths· cell^?1·d^?1 for stationary phase cells. Equal numbers of coccoliths were attached and detached early in logarithmic growth, and as cells aged, the numbers of detached coccoliths exceeded the attached ones by a factor of 6. Our results demonstrate pronounced charges of forward angle light scatter and 90° light scatter of cells as they grow logarithmically and enter stationary phase. Counts of loose coccoliths in batch cultures are consistent with the detachment of coccoliths in layers rather than individual coccoliths.     

Bioenergetics of juvenile whitespotted bamboo shark Chiloscyllium plagiosum [Anonymous (Bennett)]

This study establishes the bioenergetics budget of juvenile whitespotted bamboo shark Chiloscyllium plagiosum by estimating the standard metabolic rate (R_S), measuring the effect of body size and temperature on the R_S, and identifying the specific dynamic action (R_SDA) magnitude and duration of that action in juvenile whitespotted bamboo sharks. The mean ±s .d . (R_S) of six fish (500–620 g) measured in a circular closed respirometry system was 30·21 ± 5·68 mg O₂ kg^?1 h^?1 at 18° C and 70·38 ± 14·81 mg O₂ kg^?1 h^?1 at 28° C, respectively. There were no significant differences in R_S between day and night at either 18 or 28° C (t‐test, P > 0·05). The mean ±s .d . Q₁₀ for 18–28° C was 2·32 ± 0·06 (n = 6). The amount of oxygen consumed per hour changed predictably with body mass (M; 295–750 g) following the relationship: (n = 40, r²= 0·92, P < 0·05). The mean magnitude of R_SDA was 95·28 ± 17·55 mg O₂ kg^?1 h^?1. The amount of gross ingested energy (E_I) expended as R_SDA ranged from 6·32 to 12·78% with a mean ±s .d . of 8·01 ± 0·03%. The duration of the R_SDA effect was 122 h. The energy content of juvenile whitespotted bamboo shark, squid and faeces determined by bomb calorimeter were 19·51, 20·3 and 18·62 kJ g dry mass^?1. A mean bioenergetic budget for juvenile whitespotted bamboo sharks fed with squid at 18° C was 100C = 29·5G + 31·9R_S+ 28·2R_SDA+ 6·7F + 2·1E + 1·6U, where C = consumption, G = growth, F = egestion, E = excretion and U = unaccounted energy.     

The use of copper and silver in carbon point-of-use filters for the suppression of Legionella throughput in domestic water systems

Aims: To evaluate throughput of seeded Legionella pneumophila bacteria in domestic point‐of‐use filters. Methods and Results: The filters were challenged with tap water seeded with Leg. pneumophila. After multiple challenge events (4·25 × 10¹¹ CFU per filter), the levels of Legionella were lower in the effluent from the filter containing both copper and silver (mean 4·48 × 10³ CFU ml^?1) than in the effluent from the filter containing copper only (1·26 × 10⁴ CFU ml^?1; P < 0·001). After a single challenge event of approx. 5 × 10⁹ CFU L. pneumophila per filter, there was no significant difference between the levels of Legionella in the effluents from a carbon filter containing copper and a carbon filter with no metals (mean 6·87 × 10² and 6·89 × 10² CFU ml^?1, respectively; P = 0·985). Conclusions: Legionella was detected in filter effluent up to 6 weeks after being challenged, indicating that while filters may reduce the levels during an initial contamination event, the exposure is extended as the accumulated bacteria slough off over time. Significance and Impact of the Study: This study has provided an understanding of the response of Legionella to the use of silver and copper in domestic point‐of‐use carbon filters.