Seasonal variation in total and soluble tissue nitrogen of Pleurophycus gardneri (Phaeophyceae: Laminariales) in relation to environmental nitrate

Seasonal variations in tissue nitrogen (ethanol soluble nitrate and ninhydrin positive substances, as well as total nitrogen) of different thallus parts of Pleurophycus gardneri Setchell and Saunders were monitored simultaneously with ambient seawater nitrate from 1982 until 1984 in Bamfield, Vancouver Island, British Columbia, Canada. A trend of low, nearly zero levels in ambient nitrate typical for the area in late spring and early summer normally contrasts with average nitrate concentrations of 10 mol NO₃ ^- l^-1 in late fall and winter. Total nitrogen content was greater in the perennial thallus parts, stipe and holdfast than in the annual blade and peaked in fall and early winter. The longitudinal thallus distribution of nitrate revealed a distinct and significant concentration of nitrate in the haptera reaching at maximum 8% nitrate-N of the internal total nitrogen. Internal nitrate concentration ranged from 20 to 5 000 times the ambient nitrate concentration in the midrib, and from 40 to 3 100 times in the wing, while the range was greatest with 400 to 14 000 times in the haptera. P. gardneri contained at most about 7 mol NO₃ ^- g fresh wt^-1 in the blade, which corresponds to about 6% of total tissue nitrogen. Ninhydrin positive substances comprised the major portion of the soluble N pool in P. gardneri and showed a pronounced seasonality. Concentrations of ninhydrin positive substances ranged from 20 to 800 g N g fresh wt^-1 in the midrib and in the wing. In the stipe, ninhydrin positive substances varied from 180 to 2 200 g N g fresh wt^-1, and from 250 to 1 200 g N g fresh wt^-1 in the haptera. Evidence is given that (1) the perennial parts, stipe and haptera of P. gardneri contain the majority of nitrogen products independent of season and ontogenetic stage; (2) ninhydrin positive substances are the most abundant internal nitrogen constituents; (3) the low N values in the blade in summer suggest a nitrogen limited growth; and (4) nitrate may not be the predominant external nitrogen source.     

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.     

Nitrogen fixation by blue-green algae associated with the siphonous green seaweed Codium decorticatum: effects on ammonium uptake

Nitrogen fixation (acetylene reduction) at rates of up to 1.2 g N₂ g dry wt^-1 h^-1 was measured for the siphonous green seaweed Codium decorticatum. No nitrogenase activity was detected in C. isthmocladum. The nitrogenase activity was light sensitive and was inhibited by the addition of DCMU and triphenyl tetrazolium chloride. Additions of glucose did not stimulate nitrogen fixation. Blue-green algae (Calothrix sp., Anabaena sp., and Phormidium sp.) were implicated as the organisms responsible for the nitrogenase activity. They occurred in a reduced microzone within the C. decorticatum thallus where nitrogen fixation was optimized. Nitrogen fixation did not affect the kinetic constants for ammonium uptake in C. decorticatum (K_s=12.0 M, V_max=13.4 mol NH₃ g dry wt^-1 h^-1) determined using the perturbation method. Nevertheless, C. decorticatum thalli which fixed nitrogen had internal dissolved nitrogen concentrations which were over 1.4 times higher than in non-fixing thalli. This suggests that if C. decorticatum does derive part of its nitrogen requirement from the blue-green algae which it harbors, the transfer does not involve competition between this process and the uptake of ambient ammonium.     

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.     

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.     

Fluorescence induction in the macroalgae Chondrus crispus (Rhodophyceae) and Ulva sp. (Chlorophyceae)

The induction of in vivo chlorophyll a (chl a) fluorescence (change in fluorescence intensity during a time-scale of ms to s) was measured to determine the potential of this technique for assessing the physiological condition of the macroalgae Chondrus crispus and Ulva sp. A gradient in variable fluorescence (P-F ₀ =peak minus initial fluorescence, a measure of Photosystem II activity) was found along the frond of C. crispus, the values increasing with distance from the thallus apex. No gradient was observed for Ulva sp. thalli. Nitrogen- or phosphorus-depleted Ulva sp. required a longer dark-conditioning period and had lower values of P-F ₀ than did controls. In contrast, no differences were found in P-F ₀ of N- or P-depleted C. crispus unless values were normalized to chl a. The irradiance history of C. crispus strongly influenced P-F ₀ , even after dark-conditioning: P-F ₀ declined by about 70% as the mean daily natural irradiance increased between 2 February and 14 March 1986; a negative correlation was observed between P-F ₀ and the photon flux density 1 d prior to the measurement; P-F ₀ remained elevated when C. crispus was grown under a low photon flux density; and P-F ₀ decreased in thalli within 5 d of transfer from growth under natural irradiance to an incubator with artificial irradiance. Changes in variable fluorescence at different growth irradiances of C. crispus may be due to adaptive changes in the relative absorption cross-section of this alga. The influence of irradiance history on Ulva sp. was minimal in comparison.Issued as NRCC No. 28730Part of this study was carried out while employed by Focal Marine Ltd., Bedford, Nova Scotia, Canada     

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.     

Dietary effects on oocyte yolk-composition inCapitella sp. I (Annelida: Polychaeta): Fatty acids and sterols

In 1988, immatureCapitella sp. I (initial biomass 80µg dry wt) were raised on four diets: Gerber cereal, TetraMin fish food,Ulva sp., and benthic diatoms. After 2 wk of culturing different populations on these diets, eggs were dissected from gravid females, frozen and analyzed for fatty acid and sterol composition. Eggs produced by worms on different food types were discriminated by fatty acid composition in a principal component analysis (PCA), with 18:1⁹, 18:2, 20:5 and 20:4 fatty acids showing the greatest differences between experimental groups. The sterol profiles of all eggs were dominated by cholesterol (cholest-5-en-3-ol) and cholest-5,24-dien-3-ol (>60%). A PCA of egg sterols discriminated between adult diets with cholesterol, 23,24-dimethyl-cholesta-5-en-3-ol, cholest-5,24-dien-3-ol and a C-29 stenol showing the greatest differences. In field populations ofCapitella sp. I, oocytes produced at different times of the year may have different levels of lipids depending on their dietary availability during vitellogenesis. Variations in oocyte composition may influence larval growth and development and thus have an impact on population dynamics. Alternatively, variations in fatty acid and sterol composition of oocytes may allow us to identify the food resources utilized by this species.     

The association of N2-fixing bacteria with sea urchins

Dinitrogen fixation associated with bacteria in the gastrointestinal tract of sea urchins appears to be a widespread phenomenon: sea urchins from the tropics (Diadema antillarum, Echinometra lacunter, Tripneustes ventricosus), the temperature zone (Strongylocentrotus droebachiensis) and the arctic (S. droebachiensis) exhibited nitrogenase activity (C₂H₂ reduction). Pronounced seasonal variation was found in nitrogenase activity of temperate sea urchins feeding on kelp (Laminaria spp.) and eelgrass (Zostera marina). The mean monthly nitrogenase activity was inversely correlated with the nitrogen content of the sea urchin's food, which varied up to fivefold over the course of a year. The highest rate of nitrogenase activity recorded for a temperate sea urchin during the 14 month sampling period was 11.6g N fixed g wet wt^-1 d^-1, with a yearly mean activity of 1.36 g N fixed g wet wt^-1 d^-1. Studies with ¹⁵N confirmed the C₂H₂ reduction results and showed incorporation of microbially-fixed nitrogen into S. droebachiensis demonstrating that N₂ fixation can be a source of N for the sea urchin. Laboratory experiments indicated that part of the sea urchin's (S. droebachiensis) normal gastrointestinal microflora is responsible for the observed nitrogenase activity.     

Nitrogen assimilation by phytoplankton and other microorganisms in the surface waters of the central North Pacific Ocean

Assimilation rates of ¹⁵N-labelled ammonium, urea, and nitrate by plankton in the upper euphotic zone were measured in 2 summer, 2 winter, and 1 spring cruise in the central North Pacific Ocean. Average rates of ammonium plus urea assimilation could not be determined precisely, but were estimated to be 7 to 25 g-at. N m^-3 day^-1. Indirect evidence suggested that non-photosynthetic microorganisms contributed to these rates. Nitrate assimilation was negligible in the upper waters considered in this report (above the chlorophyll maximum and the nutricline). Potential, nitrate-saturated rates were in the range 1 to 8 g-at. N m^-3 day^-1. Seasonal comparison showed lowest rates of both carbon and nitrogen assimilation rates per chlorophyll a in winter.     

Marine diatoms grown in chemostats under silicate or ammonium limitation. II. Transient response of Skeletonema costatum to a single addition of the limiting nutrient

Skeletonema costatum was grown at different steady-state growth rates in ammonium or silicate-limited chemostats. The culture was perturbed from its steady-state condition by a single addition of the limiting nutrients ammonium or silicate. The transient response was followed by measuring nutrient disappearance of the liliting perturbation experiment indicate that three distinct modes of uptake of the limiting nutrient can be distinguished; surge uptake (V _s ), internally controlled uptake (V _i ), and externally controlled uptake (V _e ). An interpretation of these three modes of uptake is given and their relation to control of uptake of the limiting nutrient is discussed. The uptake rates of the non-limiting nutrients were shown to be depressed during the surge of the uptake of the limiting nutrient. Kinetic uptake parameters, K _s and V _max, were obtained from data acquired during the externally controlled uptake segment, V _e . The same V _max value of 0. 12 h^-1, was obtained under either silicate or ammonium limitation. Estimates of K _s were 0.4 g-at NH₄-N l^-1 and 0.7 g-at Si l^-1. Short-term ¹⁵N uptake-rate measurements conducted on nitrogen-limited cultures appear to be a combination of V _s or V _i , or at lower substrate concentrations V _s and V _e . It is difficult to separate these different uptake modes in batch or tracer experiments, and ensuing problems in interpretation are discussed.Contribution No. 882 from the Department of Oceanography, University of Washington, Seattle, Washington 98195, USA. This work represents portion of three dissertations submitted to the Department of Oceanography, University of Washington, Seattle, in partial fulfillment of the requirements for the Ph.D. degree.     

Nitrogen fixation (acetylene reduction) associated with macroalgae in a coral-reef community in the Bahamas

N₂ fixation (C₂H₂ reduction) was associated with several species of macroalgae on a coral reef near Grand Bahama Island. The highest rates were associated with Microdictyon sp. (Chlorophyceae) and Dictyota sp. (Phaeophyceae). Extensive mats of filamentous blue-green algae, not heterotrophic bacteria, were the N₂ fixing agents: in experiments with samples of Microdictyon sp., the activity was lightdependent and not stimulated by organic compounds under either aerobic or anaerobic conditions. Assays in situ, at 20 m depth, and on shipboard, gave similar rates of N₂ fixation; the cyanophytes presumably have pigment adaptations to function in blue light. The maximum rate of N₂ fixation, associated with Microdictyon sp., was 3.8 g N fixed g dry weight^-1 h^-1. Coral-reef communities flourish in nutrientimpoverished waters, and therefore any input of nitrogen is probably important in stabilizing such ecosystems.     

Urea as a nitrogen source for the phytoplankton in the Oslofjord

Ambient concentrations of urea in the inner Oslofjord, Norway, showed a pronounced yearly cycle in 1980, with values in the range 0.1 to 10.0 μg-at N l^-1; this cycle resemble that of ammonia although urea concentrations were usually lower. The uptake of urea by phytoplankton was investigated using ¹⁵N. Urea was usually a less important N source than NH ₄ ⁺ , and accounted for 0 to 53% (mean 19%) of summed NH ₄ ⁺ +NO ₃ ^- + urea uptake rates from April to October. Absolute as well as relative (specific) uptake rates of urea were higher in the summer (June–August) than at other times. Uptake of urea was inhibited by NH ₄ ⁺ concentrations higher than 1 to 2 μg-at N l^-1. The summed NH ₄ ⁺ +NO ₃ ^- + urea uptake rate was exponentially related to temperature.     

Marine diatoms grown in chemostats under silicate or ammonium limitation. IV. Transient response of Chaetoceros debilis,Skeletonema costatum,and Thalassiosira gravida to a single addition of the limiting nutrient

The kinetic response of ammonium- or silicate-limited and ammonium- or silicatestarved populations of Chaetoceros debilis, Skeletonema costatum, and Thalassiosira gravida was determined by a single addition of the limiting nutrient to a steady-state culture and subsequent monitoring of the nutrient disappearance of the limiting and non-limiting nutrients at frequent time intervals. The kinetic response of nonlimited (nutrient) populations of these three species was also determined. Three distinct modes of the uptake of the limiting nutrient were observed for ammonium-or silicate-limited populations of these three species, surge uptake (V _s ), internally (cellular) controlled uptake (V _i ), and externally (ambient limiting nutrient concentration) controlled uptake (V _e ). Non-limited populations did not exhibit the three distinct segments of uptake, V _s , V _i and V _e . Estimates of the maximal uptake rate (V _max) and the Michaelis constant (K _s ) were obtained from nutrient-limited populations during the V _e segment of the uptake curve. Pooled values of V _e for the three ammonium-limited populations yielded V _max and K _s estimates of 0.16 h^-1 and 0.5 g-at NH₄–N l^-1. Kinetic data derived from the V _e segment of the uptake curve for silicate-limited populations yielded different values of V _max and K _s for each of the three species. In a number of parameters that were measured, T. gravida was clearly different from C. debilis and S. costatum and its recovery from nutrient starvation was the slowest. Recovery of all species from silicate limitation or starvation was slower than from ammonium limitation or starvation. Ammonium-starved populations maintained a maximal uptake rate at a substrate concentration an order of magnitude lower (0.1 g-at NH₄–N l^-1) than that observed for NH₄-limited populations (1.0 g-at NH₄–N l^-1). Adaptation to the severity of the nutrient limitation occurred as changes in the magnitude of cellular characteristics, such as short-term uptake potential (V _s ) and affinity for the substrate (K _s ). The consequence of these results are discussed in terms of another possible mechanism to explain changes in species composition and succession in nutrient-depleted environments.Contribution No. 944 from the Department of Oceanography, University of Washington, Seattle, Washington 98195, USA.     

Seasonal effects of light,temperature, nutrient concentration and salinity on the physiology and growth of Caulerpa paspaloides (Chlorophyceae)

Caulerpa paspaloides (Bory) Greville were collected during the winter and summer (1978 to 1979) from the Florida Keys, USA. Thalli collected during the winter photosynthesized more efficiently at low light intensities (I_c<1, I_k=38 Exm^-2xs^-1) than did thalli collected in the summer (I_c=13, I_k=111 Exm^-2xs^-1). Summer thalli exhibited higher P_max values (2.20 mgO₂xg^-1 dry wtxh^-1) than winter thalli (1.70 mg O₂xg^-1 dry wtxh^-1). Rates of rhizome elongation and frond initiation were strongly inhibited by winter temperatures. The maximum lethal temperature for summer thalli was 37° to 38°C as measured by both growth and photosynthesis. No evidence of nitrogen or phosphorus limitation was found. Relatively minor reductions in salinity (3S) resulted in significant increases in rhizome apex motality. Results indicate that low winter temperatures are responsible for reduced winter growth rates previously reported for the Key Largo population. Increased photosynthetic efficiency at low light intensities and preferential maintenance of rhizome elongation over frond initiation appear to allow this tropical macroalga to optimize growth and survival during the winter.     

Silicon and the ecology of marine plankton diatoms. II. Silicate-uptake kinetics in five diatom species

The variation of the rate of silicate uptake with varying silicate concentration in the medium was investigated in short-term experiments with the following marine diatom species:Skeletonema costatum, Thalassiosira pseudonana, T. decipiens, Ditylum brightwellii, andLicmophora sp. The uptake conformed to Michaelis-Menten kinetics only after a correction had been made for reactive silicate that apparently could not be utilized by the diatoms. The magnitude of this correction was in the range of 0.3 to 1.3 g-at Si/l. Mean values of the half-saturation constant of silicate uptake were calculated for the different species. The lowest value was found inS. costatum (0.80 g-at Si/l) and the highest inT. decipiens (3.37 g-at Si/l). Growth limitation by low silicate concentrations could be a cause of species succession in marine plankton-diatom blooms.     

Responses of Leptasterias hexactis (Echinodermata: Asteroidea) to low salinity

The six-rayed starfish Leptasterias hexactis (Stimpson, 1862) is seasonally exposed to low salinities in southeastern Alaska. Individuals that were gradually exposed to reduced salinities in the laboratory had a 28-d TLm of 12.9 S. The activity of L. hexactis, as measured by its activity coefficient, varied directly with salinity. Individual feeding rates of the starfish on similarly exposed Mytilus edulis, measured daily for 21 d at salinities of 30, 20 and 15 S, also varied directly with salinity. The dry weight of mussel tissue consumed was 8.84, 8.49 and 0.58 mg ·starfish^-1·d^-1 at 20, 20 and 15 S. Expressed as percent of dry starfish weight, the daily feeding rate was 1.35, 0.76 and 0.10% at 30, 20 and 15 S. Absorption efficiency decreased from 64% at 30 S to 49% at 20 S, further reducing the energy available for metabolism. Growth, measured in terms of changes in total dry weight or dry weight of soft tissues, also varied directly with salinity. Although exposure to hyposmotic conditions did induce stress responses, as indicated by reductions in activity, feeding, absorption efficiency and growth rates, L. hexactis maintained positive growth for at least a 3-wk period in the laboratory at 20 S and 13°CC. The population of L. hexactis investigated must be considered euryhaline and brief periods of exposure to hyposmotic conditions should not limit its distribution.     

Seasonal photosynthetic characteristics of Ascoseira mirabilis (Ascoseirales,Phaeophyceae) from King George Island,Antarctica

Monthly variation in photosynthesis, dark respiration, chlorophyll a content and carbon: nitrogen (C:N) ratios in different lamina sections of adult plants of Ascoseira mirabilis Skottsberg from King George Island, Antarctica, was investigated between September 1993 and February 1994. Light saturated net photosynthesis (P _max) showed maximum values in September (12 to 25 mol O₂ g^-1 fr wt h^-1), and decreased towards the summer to values ranging between 2.0 and 5.0 mol O₂ g^-1. In the distal section, however, a second optimum occurred in December (25 mol O₂ g^-1 fr wt h^-1). Dark respiration rates were also highest in October and November and decreased strongly in December to February (6.0 and 1.0 mol O₂ g^-1 fr wt h^-1, respectively). Gross photosynthesis exhibited high values between September and December. Concomitant with the seasonal decrease of photosynthetic efficiency () from mean values of 1.2 mol O₂ g^-1 fr wt h^-1 (mol photons cm^-2 s^-1)^-1 in September to 0.3 mol O₂ g^-1 fr wt h^-1 (mol photons cm^-2 s^-1)^-1 in January, the initial light saturating point (I _k) gradually increased from 19 to 60 mol photons m^-2 s^-1. Likewise C:N ratios were low in spring (12 to 13) and increased in summer (20). In general, the photosynthetic parameters P _max, gross photosynthesis, and Chl a concentrations were significantly higher in the distal section of the thallus. In contrast, C:N ratios were lower in the distal section of the lamina. The results show that photosynthesis obviously strongly supports growth of the alga in late winter to spring, as it does in some morphologically related brown algae from temperate and polar regions. The question whether growth is additionally powered     

Two heavy metal-binding proteins in the midgut gland of the crab Carcinus maenas

Two heavy metal-binding proteins occur naturally in the midgut glands of Carcinus maenas (L.) collected from the Firth of Clyde, Scotland. These proteins, of approximately 27,000 and 11,500 MW (molecular weight) have previously been described as Cd-binding proteins after their induction by high concentrations of cadmium in the laboratory. The 27,000 MW heavy metal-binding protein is bound to about 0.10 g-at of Cd, 0.70 g-at of Zn and 0.31 g-at of Cu per mole of protein; 7.7, 7.9 and 1.1%, respectively, of the soluble Cd, Zn and Cu in the midgut gland are associated with this 27,000 MW protein (6.7, 1.6 and 0.9% of the total midgut gland Cd, Zn and Cu). The 11,500 MW protein is bound to about 0.04 g-at of Cd, 0.37 g-at of Zn and 1.54 g-at of Cu per mole of protein; 29.3, 31.3 and 41.7%, respectively, of the soluble Cd, Zn and Cu in the midgut gland are associated with this protein (25.7, 6.3 and 34.4% of the total midgut gland Cd, Zn and Cu). Neither heavy metal-binding protein is bound to measurable amounts of lead.     

In-situ measurements of nitrogenous nutrient uptake by kelp (Ecklonia maxima) and phytoplankton in a nitrate-rich upwelling environment

Nitrogen uptake by the kelp Ecklonia maxima Osbeck and phytoplankton was examined under different conditions of nutrient availability in a kelp bed off the Cape of Good Hope by measuring nutrient depletion in large plastic bags by the kelp and ¹⁵N uptake by phytoplankton. E. maxima took up nitrate and ammonia, but not urea, and showed only a weak preference for reduced nitrogen. Phytoplankton absorbed all three forms of nitrogen available, with a preference for ammonia and urea. Ambient nitrate concentration exhibited a marked and rapid decrease with northerly winds and an increase in response to offshore southerly winds. Nitrogen uptake by E. maxima was linearly related to ambient concentration and did not saturate even at nitrate concentrations >20g-at N l^-1, resulting in a significantly higher tissue nitrogen content under upwelling conditions. Nitrate imported by upwelling was the chief source of nitrogen utilised within the kelp bed. Locally regenerated nitrogen (ammonia and urea) was calculated to contribute only ca 4% of total nitrogen uptake during upwelling and 30% during the relaxation or downwelling phase.