Growth response of Chaetoceros calcitrans (Bacillariophyceae) in batch culture to a range of initial silica concentrations

Batch cultures of the marine unicellular centric diatom Chaetoceros calcitrans (Paulsen) Takano were maintained by serial subculturing every 4 d into nutrient-enriched natural sea-water medium supplemented with 350, 950 and 1 400 g-at Si l^-1. The diatom cultures removed initial silica concentrations of 350 and 950 g-at l^-1 from the medium within 2 and 3 d, respectively. About 30 g-at l^-1 of the highest initial concentration remained in the medium after 4 d. The mean final cell density with an enrichment of 350 g-at Si l^-1 was 3.43±0.26×10⁴ cells l^-1 (median cell volume = 77.5±5.0 m³); with 950 g-at Si l^-1, 8.55±0.55×10⁴ cells l^-1 (50.0±4.5 m³); and with 1 400 g-at Si l^-1, 9.72±0.48×10⁴ cells l^-1 (37.3±5.0 m³). There was no significant difference in the final total organic weight of cells produced, which was in the range of 170 to 190 mg per 250 ml culture. This consisted of proportionately more lipid and carbohydrate and less protein from the treatment with 350 g-at Si l^-1 than from the 1 400 g-at Si l^-1 treatment.     

Effects of light intensity on nitrate and nitrite uptake and excretion by Chaetoceros curvisetus

Michaelis-Menten uptake kinetics were observed at all light intensities. With constant illumination, the V_max and K₁ in nitrate uptake over the natural light intensity range of 0 to 2000 E were 0.343 g-at NO₃–N(g)^-1 at protein-N h^-1 and 26 E, respectively. Nitrate uptake was inhibited at higher light intensities. The K_s for nitrate uptake did not vary as a function of light intensity remaining relatively constant at 0.62 g-at NO₃–N 1^-1. With intermittent illumination, the V_mzx for light intensity in nitrate uptake over a light intensity range of 0 to 5000 E was 0.341 g-at NO₃–N(g)^-1-at protein-N h^-1. No inhibition of nitrate uptake was observed at higher than natural light intensities. Chaetoceros curvisetus will probably never experience light inhibition of nitrate uptake under natural conditions.     

Interactions of inorganic nitrogen in the uptake and assimilation by marine phytoplankton

The effect of ambient ammonium concentration on the nitrate uptake rate of marine phytoplankton was investigated. These studies consisted of laboratory experiments using unialgal species and field experiments using natural phytoplankton communities. In laboratory experiments, ammonium suppressed the uptake rates of nitrate and nitrite. Approximately 30 min were required for ammonium to exhibit its fully inhibitory effect on nitrate uptake. At high ammonium concentration (>3 g-at/l), a residual nitrate uptake rate of approximately 0.006 h^-1 was observed. When the ambient ammonium concentration was reduced to a value less than 1 g-at/l, the suppressed nitrate uptake rate subsequently attained a value comparable to that observed before the addition of ammonium. A range of 25 to 60% reduction in the nitrate uptake rate of natural phytoplankton communities was observed at ambient ammonium concentrations of 1.0 g-at/l. A mechanism is proposed for the suppression of nitrate uptake rate by ammonium through feedback control of the nitrate permease system and/or the nitrate reductase enzyme system. The feedback control is postulated to be regulated by the level of total amino acids in the cell.Contribution No. 936 from the Department of Oceanography, University of Washington, Seattle, Washington 98195, USA. This paper represents a portion of a dissertation submitted to the Department of Oceanography, University of Washington, Seattle, in partial fulfillment of the requirements for the Ph.D. degree.     

Biological production of nitrite in seawater

The relative importance of 3 different sources for biological production of nitrite in seawater was studied. Decomposition of fecal pellets of the copepod Calanus helgolandicus (at a concentration of approximately 12 g-at N/l), in seawater medium, released small amounts of ammonia over a 6 week period. It nitrifying bacteria were added to the fecal pellets nitrite was barely detectable over the same period. Decomposition of phytoplankton (present at a concentration of about 8 g-at particulate plant N/l) with added heterotrophic bacteria, released moderate amounts of ammonia over a 12 week period. If the ammonia-oxidizing bacterium Nitrosocystis oceanus was added to the decomposing algae, nitrite was produced at a rate of 0.2 g-at N/l/week. Heterotrophic nitrification was not observed when 7 open-ocean bacteria were tested for their ability to oxidize ammonia. The diatom Skeletonema costatum, either non-starved or starved of nitrogen, produced nitrite when growing with 150 or 50 g-at NO ₂ ^- -N/l at a light intensity of about 0.01 ly/min. When nitrate in the medium was exhausted, S. costatum assimilated nitrite. If starved of vitamin B₁₂, both non-N-starved and N-starved cells of S. costatum produced nitrite in the medium with 150 g-at NO ₃ ^- -N/l. Nitrate was not exhausted and cell densities reached 2x10⁵/ml due to vitamin B₁₂ deficiency. If light intensity was reduced to 0.003 ly/min under otherwise similar conditions, cells did not grow due to insufficient light, and nitrite was not produced. In the sea, it appears that, in certain micro-environments, decomposition of particulate matter releases ammonia with its subsequent oxidation to nitrite. The amounts of these nutrients and the rate at which they are produced are dependent upon the nature of the materials undergoing decomposition and the associated bacteria. In certain other areas of the sea, where phytoplankton standing stock is high and nitrate is non-limiting, excretion by these organisms is a major source of nitrite.     

Growth and survival of Mytilus edulis larvae exposed to low levels of dibutyltin and tributyltin

Two studies were conducted to observe effects of dibutyltin (DBT) and tributyltin (TBT) on larvae of Mytilus edulis for an exposure period of 25 d. Endpoints for evaluation were shell growth and mortality measured at 33 d. Larvae were cultured in a new laboratory assay chamber in a recirculating static test. The control, 2, 20, and 200 g/l DBT-treated populations had mean shell lengths of 527, 523, 417, and 180 m, respectively. Survival was 1% for the 200 g/l DBT-treated population, but ranged from 73 to 83% for controls, 2, and 20 g/l treatments. The no-observed-effect concentration (NOEC) was 2 g/l for DBT, while the lowest-observed-effect concentration (LOEC) was 20 g/l. The chronic toxicity value was 6.3 g/l. In the TBT bioassay, mean shell lengths for the control, 0.006, 0.050, and 0.130 g/l-treated populations were 565, 437, 385, and 292 m, respectively. Control survival was 74%, whereas TBT-treated populations survival ranged from 52 to 58%. The NOEC for TBT was 0.006 g/l TBT and the LOEC was 0.050. A chronic toxicity value of 0.017 g/l was calculated. The results of this study indicated that the toxicity of DBT was less than that of TBT. It was concluded that shell length was inversely related to exposure level in both DBT and TBT bioassays. In this study, we have observed TBT effects at lower exposure levels in the laboratory than previously reported, and also report the first data for DBT effects on mussel larvae.     

Effects of nitrogen and phosphorus enrichement on in vivo symbiotic zooxanthellae of Pocillopora damicornis

The reef coral Pocillopora damicornis (Linnaeus) was grown for 8 wk in four nutrient treatments: control, consisting of ambient, unfiltered Kaneohe Bay seawater [dissolved inorganic nitrogen (DIN, 1.0 M) and dissolved inorganic phosphate (DIP, 0.3 M)]; nitrogen enrichment (15 M DIN as ammonium); phosphorus enrichment (1.2 M DIP as inorganic phosphate); and 15 M DIN+1.2 M DIP. Analyses of zooxanthellae for C, N, P and chlorophyll a after the 8 wk experiment indicated that DIN enrichment increased the cellular chlorophyll a and excess nitrogen fraction of the algae, but did not affect C cell^-1. DIP enrichment decreased both C and P cell^-1, but the decrease was proportionally less for C cell^-1. the response of cellular P to both DIN and DIP enrichment appeared to be in the same direction and could not be explained as a primary effect of external nutrient enrichment. The observed response of cellular P might be a consequence of in situ CO₂ limitation. DIN enrichment could increase the CO₂ (aq) demand by increasing the net production per unit area. DIP enrichment could slow down calcification, thus decreasing the availability of CO₂ (aq) in the coral tissue.Hawaii Institute of Marine Biology Contribution No. 920     

Silicon and the ecology of marine plankton diatoms. I. Thalassiosira pseudonana (Cyclotella nana) grown in a chemostat with silicate as limiting nutrient

The small marine plankton diatom Thalassiosira pseudonana Hasle and Heimdal (Guillard's clone 3H) was grown in chemostats with silicate as the limiting nutrient. The calculated maximum growth rates were comparable to those previously reported for this species. The silica content of the diatom shells varied with the growth rate. As the growth rate approached zero, there were still measurable quantities of residual reactive silicate in the medium. In one of the two chemostats used, silicate assimilation by the cells was inefficient due to some unknown internal or external factor. In the other chemostat, statistically calculated half-saturation constants of growth were in the range of 0.5 to 0.8 g—at Si/l, depending on which kind of correction was made for residual silicate. Half-saturation constants of steady-state mean silicate uptake per cell and hour, calculated in a similar fashion, were in the range of 1.4 to 2.6 g—at Si/l. These results indicate that the silicate concentrations causing a reduced silicate uptake by this species in nature do not necessarily result in a correspondingly reduced growth rate. Growth in coastal waters is likely to become seriously limited by a shortage of silicate only when most of the silicate originally present has been removed in the course of a diatom bloom.     

Elemental composition (C,N,H) in larval and crab-1 stages of Pagurus bernhardus (Decapoda,Paguridae) and Carcinus maenas (Decapoda,Portunidae)

Analyses of individual content of carbon (C), nitrogen (N), and hydrogen (H) were carried out for all larval stages of Pagurus bernhardus and Carcinus maenas, and for newly metamorphosed crabs. Maximum range in total larval development is 12.8 to 165.8 g C, 3.2 to 35.1 g N, and 1.9 to 24.9 g H in P. bernhardus and 3.1 to 43.2 g C, 0.7 to 10.1 g N, and 0.4 to 6.3 g H in C. maenas. From these data energy equivalents were calculated. Maximum range in total larval life is 0.43 to 6.38 J ind. ^-1 in P. bernhardus and 0.1 to 1.49 J ind. ^-1 in C. maenas. There is a 32.4% mean loss of energy in P. bernhardus megalopa development; this seems to describe the normal developmental pattern in this stage. Biomass was determined as fresh and dry weight respectively. Individual dry weight is about 3.6 to 5.6 times higher in P. bernhardus (44 to 340 g) than in C. maenas (12 to 93 g) larvae.Contribution to research project Experimentelle marine Ökosystemanalyse sponsored by Bundesministerium für Forschung und Technologie, Bonn (Grant No. MFU-0328/1)     

Growth of plankton algae as a function of N-concentration,measured by means of batch technique

Employing a special batch technique which involves extremely low concentrations of the alga Selenastrum capricornutum, it is shown that a growth rate of ca. 75% of optimum growth occurs at a concentration of 0.02 g-at N(NH₄ ⁺) l^-1. The low concentrations of this N-source in the oligotrophic parts of the oceans (ca. 0.06 g-at l^-1) thus give rise to a borderline deficiency. As the half-saturation constant (K _S) for growth was recorded at 0.05 g-at N(NO₃ ^-), this ion can hardly be of much importance at the low concentrations present in these parts of the oceans. It is wrong to assert that N is the primary limiting nutrient. Phosphate also is found at concentrations giving rise to a borderline deficiency.     

Bioavailable iron species in seawater measured by macroalga (Laminaria japonica) uptake

The bioavailability of iron in seawater filtered through a 0.025-m filter was investigated using ⁵⁹Fe-labeled iron uptake by the macroalga Laminaria japonica (Areschoug: Phaeophyta) (collected in the northern Japan Sea 1993) as an assay. About 80% of the iron in the 0.025-m filtered coastal seawater was soluble and/or small colloidal organically bound iron, associated with natural organic ligands forming complexes with ferric ion. After decomposition of the organic matter by ultraviolet (UV) irradiation, 55% of the iron addition [or 0.6 nM, nearly the concentration of Fe(OH) ₂ ⁺ in equilibrium with amorphous hydrous ferric oxide in seawater at pH 8.0] in the 0.025-m filtered coastal seawater was taken up by the macroalga. Since the iron concentrations in the 0.025-m filtered coastal seawater are 0.1 to 2.0 nM and only 0.6 nM of the iron is likely available to biota over 1 to 2 d, we suggest that only small amounts of bioavailable iron exist in coastal seawater not affected by inflow from land and that a significant fraction of dissolved (<0.025 ) iron occurs in forms, such as organic iron complexes, other than the simple hydroxo-complex species predicted by thermodynamic models.     

Interactions between light and temperature on the physiological ecology ofGracilaria tikvahiae (Gigartinales: Rhodophyta)

Main effects and interactions of light and temperature on rates of growth (), net photosynthesis (P_s), and dark respiration (R) of the red seaweedGracilaria tikvahiae were investigated in outdoor, nutrient-replete continuous-flow seawater culture chambers. Below 15°C,G. tikvahiae did not grow and between 15° and 30°C, both main effects and interactions of light and temperature on and P_s were significant, which explains the occurrence of this alga as a summer annual in its northern range. Temperature interacted with light (I) through its influence on the vs I and P_s vs I curves. The initial slope of the vs I curve, , the light saturation intensity, I_s, and maximum growth rate, _max, were all significantly lowerat 15°C compared to 20°, 25°, or 30°C. Maximum values of _max, the P_s:R ratio and the net photosynthesis:gross photosynthesis ratio (P_s:P_g) all occurred at 25°C, suggesting that this is the best temperature for growth ofG. tikvahiae. Values for P_max increased up to 30°C, indicating that the temperature for maximum growth and net photosynthesis are not the same forG. tikvahiae. Significant photoinhibition of growth and photosynthesis at full incident sunlight (I₀) occurred at 15°C but not at 20°, 25°, or 30°C. Steele's equation fit the 15°C vs I data best, whereas the hyperbolic tangent function fit the 20°, 25°, and 30°C data best. Main effects and interactionof light intensity and temperature on rates of R were also significant (P<0.001). R was highly intercorrelated with and P_s (0.86r0.94), indicating that R inG. tikvahiae is primarily regulated by growth rate and not temperatureper se. Environmental factors that regulate growth, such as light intensity, exert a great influence on R inG. tikvahiae.     

Distribution of labile and residual particulate carbon in the Baltic Sea

During the September 1971 cruise of F. K. Alkor in the central Baltic Sea, the surface or summer water layer down to a depth of about 30 m was found to contain 187.4 g/l of particulate organic carbon, with a C:N (atoms) ratio of 8.97. This carbon was 44% (89.9 g/l) labile to bacterial decomposition, as determined by burning an aliquot of each sample in a CHN-analyser before and after maintenance in nutrientenriched sea water at 20°C for 3 months. The particulate material from the intermediate or winter water layer, with a depth ranging from 30 to 70 m, contained only 48% (43.1 g/l) of the labile carbon found in the summer surface layer, and had a significantly higher C:N ratio (11.25). These two facts indicate that a considerable breakdown of the organic material had taken place. The material, removed from the particulate state during this process, was 48% (89.7 g/l) of the original total particulate carbon, and was relatively nitrogen-rich, with a C:N ratio of 6.49. In this material, 52% (46.8 g/l) of the organic carbon was labile. The particulate material in the rest of the water column showed no significant changes until it reached the sediment. The slurry immediately above the sediment had a C:N ratio of 9.15, indicating the introduction of nitrogen from either dissolved or colloidal material. Labile carbon (44% of the total) was also present in sufficient quantities to support life and to make this an important diagenetic site.     

Grazing and ingestion rates of nauplii,copepodids and adults of the marine planktonic copepod Calanus helgolandicus

The average grazing and ingestion rates of all stages of the marine planktonic copepod Calanus helgolandicus (Calanoida) from nauplius stage IV to adults were measured experimentally at 15°C in agitated cultures. The chain-forming diatom Lauderia borealis and the unarmoured dinoflagellate Gymnodinium splendens were offered as food. The food concentrations were close to natural conditions and ranged from 36 to 101 g of organic carbon per liter. The medium body weights expressed in g of organic carbon of almost all larval stages raised at 49 g C/1 were identical with the weight of the same stages caught in the Pacific Ocean off La Jolla, California, USA. In a log-log system, grazing and ingestion rates increased almost linearly with increasing body weight. Grazing rates ranged from 4 to 21 ml/day/nauplius stage IV to 286 ml to 773 ml/day/female. Ingestion rates increased from 0.2 g to 0.8 g C/day/nauplius stage IV to 18 g to 69 g C/day/female. Grazing and ingestion rates per unit body weight decreased gradually with increasing body weight. The daily ingested amount of food decreased from 292 to 481% of the body weight (g C) of nauplius stage V to 28–85% of the body weight of adult females. Grazing and ingestion performances of all stages increased with increasing particle size. Grazing rates decreased and ingestion rates increased with increasing food concentrations. The published data on food intake of the different age groups of C. helgolandicus show that the young stages of herbivorous planktonic copepods can play a major part in the consumption of phytoplankton in the sea due to their high grazing and ingestion rates.     

Influence of food concentration on the physiological energetics and growth ofOstrea edulis larvae

Feeding, respiration and growth rates of oyster (Ostrea edulis L.) larvae reared at five food levels were measured throughout the entire larval period. Energy budgets were derived as a function of alga (Isochrysis galbana Parke) food concentration. Ingestion rate (IR, cells h^-1) and oxygen consumption rate ( , nl h^-1) were almost isometric functions of larval size [ash-free dry weight, (AFDW, g)], characterized by the equations: IR=803.9 AFDW^1.13 and =4.85 AFDW^1.09. Ingested ration was directly correlated to cell concentration up to a maximum at 200 cells l^-1, with further increases failing to support higher ingestion rates. Likewise, growth rate linearly increased with food ration up to 100 cells l^-1 (max. growth efficiency,K ₁=25%) and reached a maximum at 200 cells l^-1 (growth rate=5.6 m d^-1), with further increases in food not supporting significantly faster growth. Maintenance ration was 2 to 3% daily dry weight (DW); optimum ration increased during larval development from 5 to 20% DW; maximum ration was 20% DW. During larval rearing, an increasing feeding schedule of 50, 100 and 200 cells l^-1 from Days 0, 5 and 10, respectively, is recommended.     

Glucose uptake in ocean profiles with special reference to temperature

Glucose uptake was measured at three different stations in the northwestern Pacific Ocean. The measurements were made at various temperatures (5° to 35°C) and glucose concentrations (3 to 100 g glucose/l). The uptake of almost all samples taken from the surface to 1,500 m was affected primarily by temperature. The temperature dependence of samples from near the transition region between the Kuroshio and the Oyashio current waters was twice as large as that of both current samples. Furthermore, the samples from the former, transition, region and the aphotic samples of the latter, current, regions showed an optimal temperature 15° to 30°C higher than the in situ temperature; the optimal temperature was only 5° to 10°C higher in the photic samples of the stable region. The average potential glucose uptake at/near the optimal temperature was 0.026 g glucose/l/h in the photic zone and 0.015 g glucose/l/h in the aphotic zone. However, after correction by the in situ temperature, the uptake was 0.024 and 0.003 g glucose/l/h, respectively.     

Development of the pericalymma larva of Solemya reidi (Bivalvia: Cryptodonta: Solemyidae) as revealed by light and electron microscopy

Solemya reidi Bernard 1980 is a gutless protobranch bivalve known to possess intracellular chemoautotrophic bacterial symbionts in its gill. A light and electron microscope study on the embryology and larval development of S. reidi provides data for the bivalve Subclass Cryptodonta. S. reidi spontaneously spawned large eggs (271 m in diameter), which developed within individual gelatious egg capsules. The first several cleavages were equal and a distinct molluscan cross was formed at the animal pole of the embryo, features previously unreported in bivalve development. Lecithotrophic pericalymma larvae (similar to the larvae of paleotaxodont protobranch bivalves and aplacophoran molluscs) hatched at 18 to 24 h and remained in the water column for a further 5 d at 10°C. At hatching, larvae measured from 360 to 440 m in length and from 225 to 265 m in cross-sectional diameter. Definitive adult structures developed within an epithelial locomotory test entirely covered with compound cilia. The test histolysed at metamorphosis and was ingested throught the mouth into the perivisceral cavity. Length and height of the shell following metamorphosis was 433 m (±42 m, n=16) and 282 m (± 29 m, n=13), respectively. Primary data and data from the literature show that the type of larval development in both paleotaxodont and cryptodont bivalves cannot be reliably estimated from egg or prodissoconch sizes.     

Identification of Amazon River water at Barbados,W. Indies,by salinity and silicate measurements

Salinity and silicate concentrations were studied at about fortnightly intervals for 21 months at a station near Barbados, W. Indies; latitude 13°15 N, longitude 59°42 W. A sensitive inverse correlation was found to exist at 5 and 25 m, but not at greater depths. Salinity near the surface varied between 33.5 and 36.0, and silicate between a little less than 1 and 4 g at/l. Low salinity water, rich in silicate, was found from February to July; salinity increased and silicate decreased from September to December. It is argued that the low salinity water at Barbados can be identified with the areas of reduced salinity found by Ryther et al. (1967) about latitude 8° to 10° N, longitude 50° to 55° W, and that this water originates from the Amazon River. Local precipitation does not seem to be a significant factor.     

Automated procedure for determination of dissolved organic nitrogen and phosphorus in aquatic environments

A method is described for estimating dissolved organic nitrogen (DON) and dissolved organic phosphorus (DOP) in aquatic environments. The method is based on ultraviolet oxidation under successive acid and alkaline conditions, and uses continuous-flow techniques. A number of pure organic nitrogen compounds were tested in the concentration range 2 to 40 g-at Nl^-1. At the 10 g-at Nl^-1 level, recovery ranged from 60 to 100% in deionized water, and from 40 to 80% in seawater (30 S). In general, recovery decreased with increasing nitrogen concentration. For pure organic phosphorus compounds, recovery ranged from 73 to 100% at the 5 g-at Pl^-1 level. An application of the method to the surface water of a clay pond along the Atlantic coast of France for a period of 72 h under natural irradiance revealed very strong diel variations of dissolved organic nitrogen, but no significant trends for dissolved organic phosphorus.     

Lead from dust and water as exposure sources for children

Data from the Edinburgh Lead Study are used to estimate the respective contributions of water and dust lead to blood lead in 6–9 year old children. Both sources are significantly related to blood lead. An exposure of 100 g/l in kitchen cold water is estimated to be equivalent to 2700 g/g of lead in dust. In this population water is a more important source of lead than dust for the bulk of the population.This paper is also being published in Proceedings of the Conference on Lead in the Home Environment and is one of several selected from SEGH sponsored conferences for simultaneous publication in this journal.     

Arsenic tissue concentration of immature mice one hour after oral exposure to gold mine tailings

A potentially high bioavailability of arsenic in gold mine tailings from a site in northern California has been suggested by solubility studies. To help address this issue, an in vivo dosing study was conducted using 12dayold Swiss Webster mouse pups (n=8/group). A sample of sizefractionated mine tailings from the site (<20m particle size, 691g g^–1 arsenic) was prepared as an aqueous suspension and administered by gavage in a volume that provided 4mg As/kg body weight. The control group received the same volume of a commercial soil (1g g^–1 As) of similar particle size (<60m). No mortality or toxic signs were noted in either group. Tissue samples were collected 1h after gavage, freezedried, microwavedigested and analysed for arsenic by ICP/MS (detection limit 2ng As g^–1 dry weight). Arsenic concentrations (ng As g^–1 dry weight) in tissues from the pups who received mine tailings were significantly higher than in control tissues. The mean elevation in arsenic concentration was highest in the liver (3364% of control, p<0.0001), followed by blood (818 of control, p<0.0001), skin (207% of control, p=0.07), and brain (143% of control, p<0.0001). The carcass arsenic concentration (excluding the GI tract, liver, brain and skin) was 138 of control (p=0.02). The data indicate uptake of arsenic from weathered mine tailings by the immature mouse pups after oral exposure.