Effects of calcium,strontium, and magnesium on the coccolithophorid Cricosphaera (Hymenomonas) carterae. I. Calcification

The present investigation concerns the effects of calcium, strontium, and magnesium on calcification and mineralogy of the calcified bodies (coccoliths) of the coccolithophorid Cricosphaera (Hymenomonas) carterae. The capacity of cells to calcify in various concentrations of these ions was examined following preliminary decalcification in CO₂. At a concentration of 10^-2 M Ca, 75% of the cells formed coccoliths within 24 h and almost all cells were recalcified after 2 days. At 10^-3 or 10^-4 M Ca no recalcification occurred. However, with the addition of Sr to the Ca-deficient media, calcification took place as shown by observations of coccoliths and by analysis of Ca. The percentage of calcified cells increased with increasing concentrations of Sr. Strontium added to a Ca-deficient media was much more effective than an equivalent concentration of Ca. No Sr was deposited in the coccoliths. X-ray analysis demonstrated that calcite was deposited by cells in all concentrations of Ca and Sr at which calcification took place. At concentrations of Mg in the media from O. O to 4.2×10^-2 M, the cells retained their ability to calcify, although calcification was markedly reduced in the absence of Mg. In low Mg concentrations (1.3×10^-4 and 4.2×10^-6 M), the coccoliths were 60% calcite and 40% aragonite, and in teh absence of Mg, only calcite was formed.     

Effects of cadmium and low magnesium on cell division and calcification in the marine coccolithophorid Cricosphaera (Hymenomonas) carterae

Cadmium (Cd) added to normal media and to magnesium (Mg)-deficient media produced inhibitory effects on cell division and cell calcification in the marine coccolithophorid alga Cricosphaera (Hymenomonas) carterae. Compared with controls in normal media (with 25 mM Mg), cell growth decreased progressively with Cd at concentrations of 5–20 M. In Mg-deficient media (with 6 mM and 0.08 mM Mg) the inhibitory effects were more pronounced with complete arrest of cell division at 20 M Cd. The greatest Cd inhibition occurred in media with the lowest Mg concentration (0.08 mM). Cadmium (5–40 M) also decreased recalcification (coccolith formation) in cells previoasly decalcified with CO₂ with complete inhibition at 40 M Cd. Inhibition of recalcification in various Cd concentrations (5–40 M) was more pronounced in low-Mg medium (with 6 mM and 0.08 mM Mg) compared with normal medium (25 mM Mg). Partial or complete reversal of the inhibitory effects of Cd and low Mg media on cell division and calcification occurred following a wash and resuspension of the cells in normal control medium (25 mM Mg).     

Influence of algal concentration and temperature on the filtration rate of Mytilus edulis

The filtration rates of Mytilus edilis (=galloprovincialis; 40 mm) were determined in relation to food concentration and temperature, using pure suspensions of the unicellular alga Platymonas suecica in concentrations ranging from 3x10⁵ cells/l to 1.5x10⁸ cells/l. The rate of filtration (ml/h/mussel) generally decreased as cell concentrations increased, and dropped to low values when concentrations above 5x10⁷ cells/l were supplied. The amount of water swept clear varied continuously, and noticeable differences in the filtration activity of M. edulis were observed over short time intervals (5 min). Fluctuations of filtered volumes per unit time were greater with lower than with higher concentrations of algae. The influence of temperature on filtration activity was highest between 5°–15°C and 25°–30°C. A temperature increase from 15° to 25°C resulted in only a slight increase in filtration rate. At 5° and 30°C, filtration dropped to very low values, namely 350 and 100 ml/h, respectively. The temperature coefficients for the filtration rates of M. edulis were determined as: Q₁₀ (5° to 15°C)=4.96; Q₁₀ (10° to 20°C)=1.22. The amount of algae cells ingested per mussel per hour is directly related to food concentration. The maximum number of cells filtered/mussel/h in an algal suspension of 70x10⁶ cells/l was 21.5x10⁵ cells/h. Cell concentrations of up to 40x10⁶ cells/l were swept clear without producing pseudofaeces. The critical cell density for M. edulis was reached at algal concentrations of 70 to 80x10⁶ cells/l. Above these concentrations no normal filtration activity was observed.     

On the potential use of magnesium and strontium concentrations as ecological indicators in the calcite skeleton of the red coral (Corallium rubrum)

 The variability of magnesium, strontium and calcium concentrations in the two skeleton types (sclerites and axis) of the red coral (Corallium rubrum) was assessed by using X-ray fluorescence spectroscopy and microprobe analysis as a prerequisite for their use as ecological indicators. Axis cross sections showed light and dark circular growth bands corresponding to fast and slow growth, respectively. Using microprobe analyses the Mg and Sr concentrations ranged from 2.8 to 3.0% and from 0.21 to 0.29%, respectively, in the sclerites and from 2.4 to 3.0% and from 0.1 to 0.28%, respectively, in the axis. Mg/Ca and Sr/Ca ratios varied within sclerites (average, 12% for both ratios) and within the axis of single colonies (average, 20% for Mg/Ca and 48% for Sr/Ca). Sr/Ca ratios in the axis were lower in fast-growing branch tips than in older, more basal parts of the colonies, whereas the Mg/Ca ratio did not differ significantly between colony regions. Mg/Ca and Sr/Ca ratios in the axis decreased significantly with depth, and we estimated an increase of the Mg/Ca ratio of 0.004–0.006 per degree Celsius. In all depth layers, Sr/Ca ratios showed a significant direct relationship with skeleton density in axis cross sections, whereas there was no significant relationship for Mg/Ca. Overall, our data indicate that temperature promotes the incorporation of Mg in C. rubrum as in other calcite skeletons, whereas Sr concentrations are inversely related to growth rate. This preliminary study suggests that Mg and Sr concentrations in the axis of the red coral have a strong potential as ecological indicator for temperature and growth rate. Received: 3 March 2000 / Accepted: 30 August 2000     

Bioavailability of dissolved copper to the American oyster Crassostrea virginica. I. Importance of chemical speciation

The effect of complexation on the accumulation of dissolved copper by the American oyster Crassostrea virginica was determined in chemically defined exposure media. The speciation of copper was varied by varying the concentrations of total copper and model chelator, nitrilotriacetic acid (NTA). Accumulation of copper in 14-d experiments was related to the cupric ion activity and not the concentration of chelated copper. Rapid accumulation of copper occurred at cupric ion activities above 10^-10 M and there was no measurable accumulation at 10^-11 M.Southeast Fisheries Center Contribution No. 81-45B. Reprint request should be addressed to this author     

Toxicity evaluation of a complex metal mixture to the softshell clam Mya arenaria

Adults of the softshell clam Mya arenaria were continuously subjected to a flowing raw seawater solution containing a mixture of salts of manganese, zinc, lead, nickel, copper, and cadmium. Final calculated concentrations, in g l^-1, of the toxicant solution were 7200 Mn, 2500 Zn, 70 Pb, 50 Ni, 50 Cu and 1 Cd; these concentrations approximated highest measured levels within surficial interstitial sediment waters from mid-Narragansett Bay, Rhode Island. M. arenaria were also subjected to a 20% solution, i.e., 1440 Mn, 500 Zn, 14 Pb, 10 Ni, 10 Cu, and 0.2 g l^-1 Cd. One study was conducted for 112 days in winter at 0° to 10°C and another for 16 days in summer at 16° to 22°C. In the winter study, all clams exposed to a 100% solution died between the 4th and 10th week; soft parts of survivors at 6 weeks contained about 19 times more Pb, 15 x more Zn, 12 x more Cu, 10 x more Mn, 3 x more Ni and 0.1 x more Cd than controls; relatively minor changes in whole body elemental content of Ca, Cr, Fe, K, Mg, Na, Sr, and V were observed. Clams exposed to a 20% solution during winter survived the 112 day study; at that time these contained about 5 x more Cu, 4 x more Mn, 3 x more Zn and about 2 x more Pb than controls; comparatively minor changes were observed in other elements examined. In the summer study, all M. arenaria subjected to the 100% solution died between 6 and 14 days; survivors from this group at 7 days contained about 25 x more Pb, 13 x more Cu, 11 x more Zn, 7 x more Mn, and 3 x more Ni than controls; other changes in elemental content were not as pronounced. Mortality in the 20% group during summer was slightly higher than controls during the 16 day study; at 14 days survivors from this group contained about 12 x more Mn, 7 x more Pb, 7 x more Zn, 4 x more Cu, and 3 x more Ni than controls. Survival and bioaccumulation patterns were not altered through feeding a supplemental diet of algae. The significance of these findings are discussed in terms of potential environmental perturbations, especially local dredging practices.     

Inorganic nitrogen metabolism in marine bacteria: Nitrate uptake and reduction in a marine pseudomonad

Growth experiments in batch cultures indicated that the uptake of nitrate by the marine pseudomonad PL1 was inhibited in the presence of ammonia provided that the ammonia concentration was higher than 1 mM. At ammonia concentrations of less than about 1 mM, however, both nitrate and ammonia were utilised simultaneously. The saturation constants for nitrate and ammonia uptake were both 2.6x10^-4 M, and similar to the Michaelis constants of nitrate reductase for nitrate (2.9x10^-4 M) and glutamine synthetase for ammonia (2x10^-4 M). Nitrate reductase activity linked to NADH was detected in chemostat-grown cultures with nitrate as nitrogen source, and in cultures containing limiting concentrations of nitrate and ammonia, ammonia or glutamate. Enzyme synthesis appeared to be repressed in cultures containing an excess of ammonia or glutamate. Chemostat cultures utilised ammonia or glutamate in preference to nitrate, while there was no marked preference between ammonia and glutamate.     

Feeding,growth and bioluminescence of the heterotrophic dinoflagellate Protoperidinium huberi

Feeding, growth and bioluminescence of the thecate heterotrophic dinoflagellate Protoperidinium huberi were measured as a function of food concentration for laboratory cultures grown on the diatom Ditylum brightwellii. Ingestion of food increased with food concentration. Maximum ingestion rates were measured at food concentrations of 600 g C l^-1 and were 0.7 g C individual^-1 h^-1 (1.8 D. brightwelli cells individual^-1 h^-1). Clearance rates decreased asymptotically with increasing food concentration. Maximum clearance rates at low food concentration were ca. 23 l ind^-1 h^-1, which corresponds to a volume-specific clearance rate of 5.9x10⁵ h^-1. Cell size of P huberi was highly variable, with a mean diameter of 42 m, but no clear relationship between cell size and food concentration was evident. Specific growth rates increased with food concentration until maximum growth rates of 0.7 d^-1 were reached at a food concentration of 400 g C l^-1 (1000 cells ml^-1). Food concentrations as low as 10 g C l^-1 of D. brightwellii (25 cells ml^-1) were able to support growth of P. huberi. The bioluminescence of P. huberi varied with its nutritional condition and growth rate. Cells held without food lost their bioluminescence capacity in a matter of days. P. huberi raised at different food concentrations showed increased bioluminescence capacity, up to food concentration that supported maximum growth rates. The bioluminescence of P. huberi varied over a diel cycle, and these rhythmic changes persisted during 48 h of continuous darkness, indicating that the rhythm was under endogenous control.     

Metals in Cancer irroratus (Crustacea: Decapoda): Concentrations,concentration factors,discrimination factors,correlations

Nine metals were analysed in whole specimens of the decapod crustancean Cancer irroratus, and three in the exoskeleton of Carcinus maenas. In terms of concentrations (expressed as parts per millions) and concentration factors, 2 groups of metals could be distinguished in Cancer irroratus: Zn, Cu, Fe and Mn in one group, Ca, Mg, Sr, Na and K in the other group. Na excepted, all the metals had concentration factors significantly higher (P<0.01) than 1. The concentration factor of Na was significantly lower (P<0.01) than 1, demonstrating the existence in C. irroratus of a barrier to the movement of Na into the crustacean from its environment. Studies of coefficients of discrimination (D), in C. irroratus, for metals considered in pairs, showed, in each case that D was significantly different (Plt;0.01) from 1. Only the discrimination factor between, Zn and Mn was less significantly different (0.05lt;Plt;0.1) from 1. Correlations were made between the concentrations of metals and other parameters: water content, size, and ash content. Significant correlations were found, especially in the cases of the pairs Ca?Sr, Na?K, Cu?Zn, and Mn-animal size.     

Environmental and structural control of trace elements in barnacle shells

The shell composition of 5 species of barnacles [Balanus balanoides (L.), B. crenatus Bruguière, B. hameri Ascanius, Elminius modestus Darwin and Chthamalus stellatus (Poli)] collected from four different sites on the coast of North Wales and one site in the Irish Sea were studied using chemical, X-ray diffraction and electron-microprobe analyses. All the shells consisted of calcite. The trace substances studied were Mg, Sr, Na, Mn, PO₄, and SO₄. Other trace elements detected included Si, Li, Ba, Zn, Cl, and Al. For each species, Mn was the only element in the shell which varied significantly from one locality to another. Variation of concentration of Mn in the shells may be related to the variations of the concentration of Mn in the water, since concentrations in the shells were highest in (estuarine) waters containing high concentrations of Mn. In Balanus balanoides L. and Elminius modestus Darwin, the Mn:Ca and Mg:Ca ratios decreased with decreasing shore level and Sr:Ca varied in the opposite direction. When B. crenatus Bruguière, a subtidal and lower littoral species, and B. hameri Ascanius, a Continental-Shelf species, were compared with the intertidal, species B. balanoides and E. modestus, the Mn:Ca and Mg:Ca ratios followed the same decreasing trend, and the Sr:Ca ratios the same increasing trend with increasing depth. These variations may be related either to increased growth rate of the shells of each species or maxium temperature to which the species is exposed, or both. The differences between species are more consistently reflected, in the concentration of Mn. However, since the concentration, of Mn is correlated with the amount of organic matter in the shell, the specific variations may reflect only the variations in the amount of organic material in the shell of each species.     

Phosphorus metabolism of oceanic dinoflagellates: phosphate uptake,chemical composition and growth of Pyrocystis noctiluca

The phosphorus metabolism of Pyrocystis noctiluca Murray (Schuett) 1886 has characteristics which may enhance its potential for success in orthophosphate impoverished waters. The steady-state phosphate uptake rates were equal in the light and dark, and were directly proportional to both the phosphorus cell quota and the cell division rate. In contrast, nutrient-saturated uptake rates were multiphasic, faster in the light than the dark, 2 to 4 orders of magnitude greater than steady-state rates, and were inversely proportional to both the phosphorus cell quota and the cell division rate. These uptake characteristics suggest that P. noctiluca may take up phosphate coincidently at their typically low ambient concentrations as well as to exploit episodic nutrient events in nature. Cell division rates were a hyperbolic function of the ambient orthophosphate concentration. The shortest doubling time was 8.7 d, the phosphate concentration at half the maximum division rate was 0.15 M and the threshold, concentration for cell division was ca 0.05 M PO ₄ ^3- . Division rates of P. noctiluca in the ocean are much faster than predicted from the measured ambient orthophosphate concentrations. Since this dinoflagellate has high naturally occurring alkaline phosphatase activities, and can utilize organic-P compounds, we suggest that organic-P can be as important as orthophosphate in supporting the observed division rates of P. noctiluca in the sea.     

水中主要阳离子对铜绿微囊藻生长及多糖的影响

通过室内培养试验,在65μmol·m-2·s-1光照度和12 h∶12 h光暗比下,模拟野外水体中铜绿微囊藻（Microcystis aeruginosa）的生长,研究了水中主要阳离子Ca、Mg、K、Na质量浓度的变化对铜绿微囊藻生长以及多糖的影响。依据自然水体中Ca、Mg、K、Na的实际浓度水平结合已有研究得到的抑制浓度,试验分别设定了5个质量浓度梯度的培养基,其中Ca质量浓度梯度为0、10、20、50、100 mg·L-1,Mg质量浓度梯度为0、2、5、10、20 mg·L-1,K质量浓度梯度为5、10、20、50、100 mg·L-1,Na质量浓度梯度为18、30、50、100、200 mg·L-1。试验测定的生理生化指标包括培养周期内每天铜绿微囊藻的藻细胞密度和对数生长期内铜绿微囊藻溶解性胞外多糖（sEPS）、固着性胞外多糖（bEPS）和胞内多糖（IPS）的含量。试验结果表明：低质量浓度的Ca对微囊藻的生长没有明显影响,高质量浓度的Ca（〉50 mg·L-1）会抑制铜绿微囊藻的生长但同时铜绿微囊藻合成多糖总量（TPS）会增加,Ca质量浓度的增大对铜绿微囊藻胞外多糖的分泌呈现先促进后抑制的趋势,并在刺激铜绿微囊藻细胞分泌多糖的同时会促进其溶解。Mg缺失时,铜绿微囊藻的生长会受到显著的抑制,较高质量浓度的Mg（〉5 mg·L-1）也会抑制铜绿微囊藻的生长但同时铜绿微囊藻合成多糖总量（TPS）会增加,Mg在适宜质量浓度（5 mg·L-1）会抑制多糖分泌、防止多糖溶解。K离子质量浓度的变化对微囊藻的生长无明显影响但铜绿微囊藻TPS的量呈现先增加后减少的趋势,K对多糖的分泌并没有显著影响,但对多糖的溶解呈现先促进后抑制的作用。Na离子质量浓度的变化对铜绿微囊藻的生长以及合成TPS的量均无明显影响,Na质量浓度增大对多糖的分泌的影响与Ca的一致,但影响的程度明显小于Ca的影响,其质量浓度的增加对多糖的溶解过程有轻微的促进作用。     

Effects of Cadmium on Nutrient Uptake and Translocation by Indian Mustard

Plants that hyperaccumulate metals are ideal subjects for studying the mechanisms of metal and mineral nutrient uptake in the plant kingdom. Indian Mustard (Brassica juncea) has been shown to accumulate moderate levels of Cd, Pb, Cr, Ni, Zn, and Cu. In this experiment, 10 levels of Cd concentration treatments were imposed by adding 10–190 mg Cd kg^–1 to the soils as cadmium nitrate [Cd(NO₃)₂]. The effect of Cd on phosphorus (P), potassium (K), calcium (Ca), magnesium (Mg), and the micronutrients iron (Fe), manganese (Mn), copper (Cu), and zinc (Zn) in B. juncea was studied. Plant growth was affected negatively by Cd, root biomass decreased significantly at 170 mg Cd kg^–1 dry weight soils treatment. Cadmium accumulation both in shoots and roots increased with increasing soil Cd treatments. The highest concentration of Cd was up to 300 mg kg^–1 d.w. in the roots and 160 mg kg^–1 d.w. in the shoots. The nutrients mainly affected by Cd were P, K, Ca, Fe, and Zn in the roots, and P, K, Ca, and Cu in the shoots. K and P concentrations in roots increased significantly when Cd was added at 170 mg kg^–1, and this was almost the same level at which root growth was inhibited. Zn concentrations in roots decreased significantly when added Cd concentration was increased from 50 to 110 mg kg^–1, then remained constant with Cd treatments from 110 to 190 mg kg^–1. However, Zn concentrations in the shoots seemed less affected by Cd. It is possible that Zn uptake was affected by the Cd but not the translocation of Zn within the plant. Ca and Mg accumulation in roots and shoots showed similar trends. This result indicates that Ca and Mg uptake is a non-specific process.     

Impact of calcium and magnesium on growth and morphological acclimations of Nitella: implications for calcification and nutrient dynamics

The impact of calcium (Ca) and magnesium (Mg) on the growth and morphology of a Charophyte, Nitella pseudoflabellata, and the influence of Mg on calcification and phosphorous (P) speciation were studied in laboratory experiments for variable concentrations (≤q120 mg · L^?1) of Ca and Mg. It was clearly identified that Mg aided shoot elongation. An increase in Ca concentrations produced intensified shoot elongation also, but at a lesser rate than the equivalent levels of Mg. Depending on the availability of Ca and Mg, the morphological appearance differed significantly, suggesting significant levels of ecoplasticity. Furthermore, Mg was observed to produce less calcite encrustation. Plant P-speciation suggested a higher Mg concentration corresponding to a more water-soluble and less carbonate-bound P fraction. This indicates that upon senescence and decomposition, a large fraction of P is supplied to the water column; ultimately behaving similar to a typical vascular plant.     

Elemental composition of a tusk of a dugong, Dugong dugon, from Exmouth, Western Australia

Element analysis of the annual increments in a longitudinal section from the tusk of a female dugong Dugong dugon (Müller) from Exmouth, Western Australia, was carried out by X-ray fluorescence-imaging, inductively coupled plasma-atomic emission spectrometry (ICP-AES) and inductively coupled plasma-mass spectrometry (ICP-MS). Nine elements (Ba, Ca, Fe, Li, Mg, Na, P, Sr and Zn) were present in concentrations amenable to determination by these techniques. Most elements revealed both long-term trends and shorter, year-to-year, variations. For example, sodium concentrations increased from 0.55 to 0.72% with the age of the dugong. Strontium concentrations (mean 0.17%) were closely correlated (r = 0.86) with those of barium (mean 4.5 mg kg⁻¹). Zinc concentrations increased from ∼70 to 170 mg kg⁻¹ with dugong age, but also showed shorter-term fluctuations of ∼30 mg kg⁻¹ that were correlated (r = 0.41) with mean annual Fremantle sea level (a measure of the Southern Oscillation Index and strength of the Leeuwin Current). The concentrations of the elements and correlations with year and between pairs of elements are discussed. Received: 10 January 1997 / Accepted: 21 April 1997     

Influence of relative water motion on the growth,ammonium uptake and carbon and nitrogen composition of Ulva lactuca (Chlorophyta)

In a series of multifactorial laboratory experiments, Ulva lactuca discs were grown in an apparatus in which they were exposed simultaneously to 3 simulated current speeds (7.5, 15, 22.5 cm s^-1) and a still control, and either 3 ammonium concentrations (0–10, 35–45 and 115–145 M) under ample uniform light (ca 200 E m^-2 s^-1) or 3 light intensities (approximately 35, 90 and 270 E m^-2 s^-1) with uniform surplus, ammonium. Disc growth rates were determined in each experiment as well as tissue nitrogen and carbon composition and fluxes of NH₄, NO₃/NO₂ and PO₄ in media. In a supplementary series of field experiments, U. lactuca discs were simultaneously exposed to 2 different water motion regimes in adjacent chambers at several sites characterized by widely different ammonium concentrations. In field experiments, growth rates were calculated and analyzed as a function of water motion at the various sites. The application of simulated current consistently enhanced disc growth rates in the laboratory, except at the lowest light intensity. In most cases this enhancement was fully realized at the lowest applied simulated current (7.5 cm s^-1). Simulated current slightly enhanced ammonium uptake rates by U. lactuca discs, relative to rates in still water, except at the highest ammonium concentration. C:N ratios of discs generally declined with increases in simulated current, except at the highest ammonium concentration. This decline was primarily attributable to increases in per cent N and was, again, mainly realized at 7.5 cm s^-1. The results suggested that simulated current compensated for N limitation, except when light was sufficiently low to become the overriding limiting factor, but that the enhancement of growth by simulated current could not be explained in terms of N metabolism alone. Field experiments showed that the higher level of water motion consistently enhanced growth at sites with comparatively low ammonium concentrations, but not at sites with moderate or high ammonium concentrations.     

Growth and competition of the marine diatoms Phaeodactylum tricornutum and Thalassiosira pseudonana. I. Nutrient effects

Two marine diatoms, Phaeodactylum tricornutum (Bohlin) and Thalassiosira pseudonana (Hasle and Heimdal), were grown both separately and together in batch cultures on a mixture of waste water and seawater enriched with different components of f medium. At 17°C, the maximum division rates of the two species were statistically indistinguishable. The waste water-seawater mixture used proved to have insufficient Si, relative to N and P, for the growth of T. pseudonana, which requires approximately 5x10^-14 g-at Si cell^-1 to divide at a maximum rate. P. tricornutum, on the other hand, although capable of taking up nearly 9x10^-15 g-at Si cell^-1, could sustain maximum rates of division with 4.3x10^-18 g-at Si cell^-1 or less. No allelopathic interaction between the two species could be detected. We conclude that P. tricornutum enjoys a considerable competitive advantage over T. pseudonana in a waste water-seawater-based mariculture system that is not supplemented with Si. Although Si proved necessary for T. pseudonana to complete more successfully with the other diatom, the presence of excess amounts of Si is not necessarily sufficient for the maintenance of T. pseudonana in mixed continuous culture with P. tricornutum: other factors, such as light-related or photoperiod-related growth response, are believed to determine the ultimate outcome of competition between these algae in light-limited continuous culture.Contribution No. 3999, from the Woods Hole Oceanographic Institution.Communicated by M.R. Tripp, Newark     

Ultrastructure of the gonad and gametogenesis in the eastern oyster,Crassostrea virginica. II. Testis and spermatogenesis

The pelagic harpacticoid copepod, Macrosetella gracilis (A. Scott), is found in association with colonies of the nitrogen-fixing (diazotrophic), bloomforming cyanobacterium Trichodesmium spp. in tropical and subtropical waters. M. gracilis is one of the few direct grazers of these often toxic cyanobacteria. Experiments investigating NH ₊ ⁴ regeneration by M. gracilis were conducted in the Caribbean in September 1992 and the Coral Sea, Australia in November 1994. Rates of M. gracilis ingestion of Trichodesmium thiebautii labelled with ¹⁵N₂ measured in the eastern Caribbean indicated that M. gracilis could consume 33 to 45% of total T. thiebautii colony N d^-1 and >100% of new N fixed d^-1. We also measured the release of NH ₊ ⁴ by M. gracilis feeding on T. thiebautii, as well as by non-feeding copepods, using ¹⁵N isotope dilution methods. In non-feeding copepods, rates of NH ₊ ⁴ release increased as numbers of copepods were increased as both copepod numbers and food availability increased. In the presence of T. thiebautii colonies, M. gracilis had an average rate of NH ₊ ⁴ regeneration of 7.7±1.5 nmol N copepod^-1 h^-1 (±SE), which was significantly higher than when food was absent (1.9±0.7 nmol N copepod^-1 h^-1). Rates of M. gracilis excretion were relatively high based on excretion: ingestion ratios, which could be due to having a high-N food source readily available, to sloppy-feeding effects, or as a response to toxins in the cyanobacterium. Incubations of M. gracilis with and without T. erythraeum resulted in significant increases in [NH ₊ ⁴ ] as a function of copepod density only. Ammonium leakage from the cyanobacterium and/or microheterotroph associates was relatively low. M. gracilis, through excretion and possible mechanical breakage of cells while grazing, appears to provide a direct link between atmospherically derived new nitrogen and regenerated NH ₊ ⁴ in the oligotrophic systems where Trichodesmium spp. are abundant.     

Ingestion,growth and development of Penaeus indicus larvae as a function of Thalassiosira weissflogii cell concentration

Penaeus indicus larvae have been successfully reared in the laboratory using Thalassiosira weissflogii, Brachionus plicatilis and Artemia salina nauplii as food, with an average survival of 95.8% from nauplius 6 to postlarva 1. The effect of T. weissflogii cell concentration on larval ingestion, development and growth (total length) was investigated. Cell ingestion rates showed a saturation response to concentration. Both maximum ingestion rates and incipient limiting levels (the lowest concentration before ingestion rates were limited) were established for the feeding larval stages. Both were found to increase with progressive increase in larval development. Maximum ingestion rates increased from 0.25×10⁴ cells. larva^-1.h^-1 during protozoea 1 to reach a peak of 1.2×10⁴ cells. larva^-1.h^-1 during mysis 3 and then declined to 0.6×10⁴ cells. larva^-1.h^-1 at postlarva 1. Incipient limiting levels (ILLs) increased from approximately 0.6×10⁴ cells.ml^-1 during protozoea 2, to 0.65×10⁴ cells.ml^-1 during mysis 1, to 1.3×10⁴ cells. ml^-1 during mysis 3 to 1.6×10⁴ cells.ml^-1 at post-larva 1. Filter feeding efficiency was found to reach a maximum during mysis 1. Filter mechanisms are discussed. Generally, the most advanced larval development per unit time occurred at concentrations at and above the ILLs, while retarded development occurred below these levels. Growth increased asymptotically with cell concentration. Incipient growth limiting levels (IGLLs; the lowest concentration before growth was significantly limited) also increased with larval development and with the exception of mysis 3 they coincided with the ILLs. IGLLs increased from 0.55×10⁴ cells.ml^-1 during protozoea 2, to 0.66×10⁴ cells.ml^-1 during mysis 1, to 0.99×10⁴ cells.ml^-1 during mysis 3, to 1.62×10⁴ cells.ml^-1 at postlarva 1. Below the ILLs where ingestion was limited, animals were significantly smaller, with larval development and growth positively correlated to ingestion rates. When culturing penaeoid larvae, ambient cell concentrations should be kept above these known limiting levels to yield consistently good larval survival and growth.     

An exploratory study with the proton microprobe of the ontogenetic distribution of 16 elements in the shell of living oysters (Crassostrea virginica)

Study of the chemical composition of shell of exoskeletonous organisms in the past has required the sacrifice of the organism. Because the beam of the proton microprobe is relatively nondestructive and analyzes the surface layer of the shell, organisms do not have to be killed. The present paper presents results of a preliminary experiment in which distribution of elements (Na to Sr) in shell of living juvenile oysters, Crassostrea virginica (Gmelin), was studied in situ with a proton microprobe at monthly intervals for four months. The relative concentration of 16 elements was measured in the newly deposited prismatic edge of the right valve of three oysters reared in controlled laboratory conditions. Na, Mg, Al, Si, S, Cl, K, Ca, Ti, Cr, Mn, Fe, Cu, Zn, Br, and Sr were detected in concentrations as low as a few parts per million relative to the concentration of standards added to pure CaCO₃. Concentration of elements varied nominally among shells of the three individual oysters and in their successive ontogenetic stages. Fluctuations in concentration of Na, Mg, S, Cl, Ca, Mn, Fe, Cu, and Zn were generally similar in the two normally growing oysters, but differed from those in the oyster that stopped growing. Trends in concentration of Al, Si, and Sr were similar in the three oysters: those of Br were variable. Relative concentrations of Na, Cl, S, Mn, Fe, and Zn increased slightly with age of oysters, that of the other elements stayed relatively constant. Concentration of most elements was higher in shell than in seawater. Variable concentrations, especially of Na, Cl, and Si in valve edges, tend to support the hypothesis of earlier workers that separate mineral phases are present as impurities entrapped within the shell during calcification.