Rhythmic release of dissolved free amino acids from partly synchronized Thalassiosira rotula under nearly natural conditions

In outdoor tanks enclosed monocultures of Thalassiosira rotula showed partly synchronized cell divisions during exponential growth. The changes in division activity showed a characteristic frequency, which also could be found for cell length (pervalvar axis) and assimilation rate as well as for concentration changes of dissolved free amino acids (DFAA) and ammonia. Excretion of DFAA occurred preferentially during periods of reduced cell division activity, when assimilation was at a higher level. The concentration of ammonia decreased during these periods. The rates per hour of these parameters showed significant multiple correlations. The release of six individual amino acids (ala, -ala, gly, lys, orn and ser) seemed to be controlled by the physiological status of the diatoms. These amino acids amounted to a portion of about 70–80% of the total DFAA. Uptake and release rates of DFAA nitrogen and ammonia were in the same range with maxima of about 350 ng at nitrogen dm^-3 h^-1 and litre. These dynamics pointed to strong fluxes of organic bound nitrogen in marine micro-heterotrophs.     

Amino acid uptake by the toxic dinoflagellate Alexandrium fundyense

A toxic axenic strain of Alexandrium fundyense is shown to be capable of removing dissolved free amino acids (DFAAs) until concentrations are similar (low nM) to those found in natural waters. Uptake is greatest during exponential growth, rather than during C and/or N-stress as is usual in diatoms and other flagellates. A wide range of amino acids can be taken up, their concentration being decreased within a few hours to the levels observed prior to DFAA addition. The maximum rate of DFAA-N uptake, during early exponential phase, was 0.8 pmol-N cell⁻¹ h⁻¹, equivalent to ≃20% of the total N requirement. More typically, the contribution of DFAA-N was only ≃5%. However, these uptake rates are not sustainable. It is apparent that this organism cannot use amino-N to support significant growth, even though it can take up DFAAs. This, and the fact that the composition of the internal amino acid pool differed from that externally, is further evidence that the N-physiology of this genus is abnormal (differences to other dinoflagellates include an abnormally high concentration of glutamine and arginine, an effective absence of amine X, and release of nitrite during the␣concurrent assimilation of nitrate and ammonium in␣darkness). There is no evidence that the use of DFAAs enhance toxin content, except when cells are supplied with very high (unnatural) concentrations of arginine. Received: 8 May 1989 / Accepted: 14 September 1998     

Comparisons among species ofAlexandrium (Dinophyceae) grown in nitrogen- or phosphorus-limiting batch culture

Three species of the dinoflagellate genusAlexandrium (Halim)-two strains of toxic.A. minutum, one each of nontoxicA. tamarense andA. affini-were grown in batch culture in either a low-nitrogen or a low-phosphate medium. Maximum carbon-specific growth rates forA. tamarense were lower (at <0.25 d^-1) than for the other strains, which all exceeded 0.38 d^-1. C-quotas (C content per cell) during exponential growth were similar for all strains (2.5 ng C cell^-1), with cells becoming smaller during the N-limiting stationary phase, but enlarging during prolonged P-deprivation. Values of ¹³C during the exponential phase were low (-25to-30), with most cells during the light phase swimming at the surface when nutrient-replete and migrating to the bottom of the flasks when nutrient-deplete with ¹³C rising to around-15. Biomass could not be estimated reliably from pigmentation, but could be estimated from biovolume (r>0.95), although this was complicated in cultures ofA. minutum by the presence of particles comprized of thecal plates of a similar size to intact cells. Alkaline phosphatase activity was not a reliable indicator a P-status. The most toxic strain tested (A. minutum AL1V) contained the highest concentrations of free amino acids, of arginine (a precursor of paralytic shellfish toxins) and of proline, and also had the lowest C:N mass ratio (at 4.3).A. affini contained the lowest concentrations of arginine, andA. tamarense the highest exponential phase C:N (7.8). For all strains, the mole ratio of intracellular glutamine: glutamate (Gln: Glu, which was abnormally high compared to other algae) could only be used to indicate the presence or absence of N-stress rather than the degree of stress. Additions of ammonium and phosphate resulted in increases in Gln: Glu within 20 min in N-stressed cells and also enhanced toxin content inA. minutum (mainly gonyautoxin) 4 over a 24 h period.     

Variations in the biochemical composition of the diatom Coscinodiscus eccentricus with culture age and salinity

Cultures of the marine diatom Coscinodiscus eccentricus Ehrenberg were grown at four salinities, namely 20, 25, 30 and 35%. The changes in the carbohydrate, protein, silicon and pigment concentrations of the cells were studied throughout the growth cycles in these salinities. The carbohydrate content, expressed as a percentage of the cell dry weight, increased in all cultures during the lag and early log phases. It later fell, sharply at first, as it was used up faster than it was produced. It was not stored during the stationary phase. The protein percentage composition built up to a peak at different stages during the exponential or stationary growth stages, depending on the salinity, but was later respired. The silicon percentage of the dry weight usually decreased during the log phase, as the cells were dividing before becoming fully silicified, and remained low in the stationary phase due to an increase in the volume of the cells. The changes in the frustular thickness were also calculated. The values ranged from 0.24 to 0.35 μm (at the beginning and end of the experiments) to 0.15 μm during the log growth phase. The pigment content was always low. The lipid composition of the cells was calculated by difference. It was usually low during the exponential growth phase, but was accumulated during the stationary phase. The point at which this accumulation began was associated with the time when protein respiration started.     

Acides aminés libres chez les larves de Crangon septemspinosa (Caridea). Variation de leur taux de l'éclosion à la métamorphose. Leur rôle au cours du développement et leur importance dans la nutrition

Studies have been carried out on the variations in the pool of free amino-acids throughout the larval development of artificially reared Crangon septemspinosa, and their relationship with the nutrition of this species. Although quantitative variations are less apparent for essential than for non-essential amino acids, a certain uniformity appears. During larval development, the concentration of each amino acid reveals two optimal values: the first at the third larval stage; the second at the final larval stage, just before metamorphosis. Each of these peak values is followed by a more or less rapid drop in amino-acid concentration. An attempt is made to explain these variations in relation to their specific function in general metabolism and growth. By the addition of certain amino acids during phases of high metabolic activity, it may be possible to overcome difficulties encountered during critical stages of rearing experiments.     

Ammonium regeneration and carbon utilization by marine bacteria grown on mixed substrates

We examined the impact of exposing natural populations of marine bacteria (from seawater collected near Woods Hole, Massachusetts, USA) to multiple nitrogen and carbon sources in a series of batch growth experiments conducted from 1989 through 1990. The substrate C:N ratio (C:N_s) was varied from 1.5:1 to 10:1 either with equal amounts of NH ₄ ⁺ and different amino acids or an amino acid mixture, all supplemented with glucose to maintain the C:N_s ratio equal to that of the respective amino acid, or with combinations of glucose and NH ₄ ⁺ alone. A common feature of the experiments involving amino acids was the concurrent uptake of NH ₄ ⁺ and amino acids that persisted as long as a readily assimilable carbon source (glucose in our case) was taken up. There was no net regeneration of NH ₄ ⁺ , even though catabolism of amino acids occurred. Regeneration of NH ₄ ⁺ was evident only after glucose was completely utilized, which usually occurred at the end of exponential growth. The contribution of¹⁵NH ₄ ⁺ to total nitrogen uptake by the end of exponential growth varied from ~60 to 80% when individual amino acids were present and down to ~24% when the amino acid mixture was added. These estimates are conservative because we did not account for possible isotope dilution effects resulting from amino acid catabolism. When NH ₄ ⁺ and glucose were the sole nitrogen and carbon sources, there was a stoichiometric balance between glucose and NH ₄ ⁺ uptake over a wide range of C:N_s ratios, leading to a constant bacterial biomass C:N ratio (C:N_B) of ~4.5:1. As a result NH ₄ ⁺ usage varied from 50% when the C:N_s ratio was 3.6:1, to 100% when the C:N_s ratio was 10:1. Gross growth efficiency varied from ~60% when NH ₄ ⁺ plus glucose were added alone or with the amino acid mixture, to 47% when the individual amino acids were used in place of the mixture. It is thus evident that actively growing bacteria will act as sinks for nitrogen when a carbon source that can be assimilated easily is available to balance NH ₄ ⁺ uptake, even when amino acids are available and are being co-metabolized.     

Aerobic nitrogenase activity measured as acetylene reduction in the marine non-heterocystous cyanobacterium Trichodesmium spp. grown under artificial conditions

Aerobic nitrogenase activity in the marine non-heterocystous cyanobacterium Trichodesmium spp. NIBB 1067, isolated off the Izu Peninsula, Japan in 1983 and grown under artificial conditions, was assayed by the acetylene reduction method. This strain exhibited acetylene reduction activity under aerobic conditions when cells had been grown in the medium free of combined nitrogen. Activity was markedly enhanced by light, and dependent on the growth phase being higher during the exponential growth phase and lower during the late linear and stationary growth phases. Since typical colony formation occurred during the last growth phase, the present results contradict the idea that N₂-fixation depends on colony formation. The photosynthesis inhibitor DCMU at 10^-6 M inhibited light-dependent acetylene reduction completely. Acetylene reduction by Trichodesmium spp. was tolerant of O₂ as strongly as that in the heterocystous cyanobacteria. Even at a partial pressure of oxygen (p_{O 2}) of 3 atm, the activity still remained as high as half of the maximum. It was almost under anaerobic conditions. Maximum activity was obtained at pO₂ of ca. 0.1 atm.     

Use of high-performance liquid chromatography to investigate the production of paralytic shellfish toxins by Protogonyaulax spp. in culture

Procedures have been developed for the extraction and high-performance liquid chromatography (HPLC) analysis of paralytic shellfish poisoning (PSP) toxins from Protogonyaulax spp. grown in batch culture. Using these procedures, the toxin content of two isolates of P. tamarensis (NEPCC 183 and 255) and one isolate of P. catenella (NEPCC 355) were examined. Total toxin and individual toxin concentrations were measured for each isolate during the exponential and stationary phases of growth in batch culture. The total toxicity of each isolate as measured by HPLC analysis was found to agree with toxicity as determined by the standard mouse bioassay. Two of the isolates (255 and 355) were found to be toxic and the third (183) was non-toxic. The toxic isolates (255 and 355) both showed higher average total PSP toxin content during the exponential phase (35 and 23 fmol toxin cell^-1, respectively) than during the stationary phase (21 and 8 fmol toxin cell^-1, respectively). These cultures differed dramatically in their toxin composition. P. tamarensis (255) contained a large proportion of the N(21) sulfo toxins (B₁, B₂, C₁, C₂) while P. catenella (355) contained primarily Gonyautoxins 1 through 4. The percent composition of individual toxins was found to be constant throughout the growth cycle for both toxic isolates, even though the total toxin concentration varied. Our results suggest that PSP toxin profiles might be useful as chemotaxonomic indicators.     

Osmotic properties of eggs of the herring Clupea harengus

Yolk osmolality of developing eggs of the herring Clupea harengus L. is strongly hypoosmotic to seawater: about 440 mOsm during the first week of development decreasing to 360 mOsm before hatching. The perivitelline fluid (PVF) of the eggs is isoosmotic to the ambient medium. The PVF equilibrates within 10 min to changes in the ambient seawater. The content of Na⁺, K⁺, Cl^- and free amino acids amount to 26, 52, 48, and 54 n mol egg^-1, respectively, on Days 1 to 3 after fertilization, increasing to 63, 69, 80, and 79 n mol egg^-1, respectively, prior to hatching (Days 18–20). The apparently conflicting findings of a decreasing yolk osmolality and a simultaneous increase in the amount of egg solutes are at present unexplained.     

Croissance de la larve de l'oursin Paracentrotus lividus

Paracentrotus lividus (Lamarck) larvae were reared to metamorphosis. The larvae were fed on the haptophycean Hymenomonas elongata Droop (Braarud) at three concentrations: 9 to 14×10⁵, 24 to 37×10⁵, 43 to 61×10⁵ μm³ cells ml^-1 d^-1. Optimum growth took place at a density of 24 to 37×10⁵ μm³ algal cells. Growth of the plutei was estimated in terms of weight increases in protein, carbohydrate and lipid; growth equations are given. The relationship between growth and the food ingested was calculated for the different larval stages. Earlier field data for the bay of Villefranche have shown the mean biovolume of nanoflagellates to be 0.65×10⁵ μm³ ml^-1; at such in situ food concentrations, P. lividus larvae would metamorphose only after one month of planktotrophic life. Chemoreception by larvae could lead to prey selection, thereby altering the amounts of protein, carbohydrate and lipid ingested, and hence the duration of larval life.     

Co-occurrence of copepods and dissolved free amino acids in shelf sea waters

The vertical distribution of chlorophylla, copepods, dissolved free amino acid concentration and the fixation of¹⁴C by phytoplankton were monitored in the springs of 1983, 1987 and 1988 in the Ushant front region, shelf edge of the Celtic Sea and central Irish Sea, respectively. In each area, two stations characterized by mixed and stratified water conditions were compared. Vertical distributions of amino acids coincided with the distribution of copepods. A positive and significant correlation was found between the abudance of copepods and the concentration of amino acids dissolved in seawater. A negative and significant correlation was found between chlorophylla and the concentration of amino acids. Enrichment of amino acids ( 20 to 500 nM l^–1 at specific depths) due to aspartic and glutamic acids, glutamine and ornithine, was assumed to reflect copepod feeding activity and faecal production. At these depths, the natural concentration and diversity of amino acids, including aspartic acid, glutamic acid, asparagine, serine, histidine, glutamine, arginine, threonine, glycine, alanine, tyrosine, valine, phenylalanine, ornithine and lysine, were high enough and in the correct proportions for triggering feeding and swimming and swarming behavior of copepods, as well as their remote detection of food at the micro- and meso-scales (1 to 10 m). This accumulation of amino acids also constitutes a potential additional source of organic nitrogen for bacteria and phytoplankton.     

Primary site and initial products of ammonium assimilation in the symbiotic sea anemone Anemonia viridis

Invertebrates containing endosymbiotic dinoflagellate algae (zooxanthellae) retain excretory nitrogen, and many are able to take up ammonium from the surrounding seawater. However, the site of assimilation and role of nitrogen recycling between symbiont and host remains unclear. In the present study, ammonium uptake by the symbiotic sea anemone Anemonia viridis (Forskål) was examined by following the pathway of assimilation using ¹⁵N-enriched ammonium. Since zooxanthellae became enriched with ¹⁵N from ammonium at up to 17 times the rate of the host, they appear to be the primary site of assimilation. In the light, the rate of zooxanthellae enrichment at 20?M was twice that at 10?M, whereas the rate of host enrichment was not significantly affected by ammonium concentration. When anemones were incubated with [¹⁵N]ammonium in the dark, after 12?h without light the rate of enrichment was lowered in both zooxanthellae and host. However, while the enrichment of the host was significantly reduced when the light level was lowered from 300 to 150?μmol photons m^?2?s^?1, zooxanthellae enrichment was unchanged. Low molecular weight material from the zooxanthellae became enriched at 20 times the rate of that from the host, and enrichment was detected in the amino acids glutamate, glutamine, aspartate, alanine, glycine, phenylalanine, threonine, valine, tyrosine, and leucine from zooxanthellae. In the zooxanthellae, amino acids accounted for 65% of the total enrichment of low molecular weight material. Of the amino acids detected in zooxanthellae, over 90% of the enrichment was accounted for by glutamate, glutamine and aspartate. The enrichment of the amide group of glutamine was greater than that of the amine group of glutamate or glutamine, consistent with the glutamine synthetase/glutamine 2-oxoglutarate amidotransferase cycle as the mechanism of ammonium assimilation. To examine the flux of ¹⁵N from zooxanthellae to host, anemones were pulse-labelled with [¹⁵N]ammonium and then transferred to an unlabelled chase. Over a 2?h period there was no evidence for a flux of nitrogen from zooxanthellae to host. However, during the chase period, the enrichment of low molecular weight material declined and that of high molecular weight material increased in both zooxanthellae and host, indicating that protein was synthesized using ¹⁵N from ammonium in both components of the symbiosis. Again by using a pulse-chase system, it was found that glutamate was metabolised most rapidly by zooxanthellae, followed by (in order of decreasing rate of turnover) aspartate, alanine, glycine and valine (no data are available for glutamine). Unlike these amino acids, nitrogen was transferred to the essential amino acids phenylalanine and threonine, increasing their enrichment during the chase period. While recycled nitrogen is clearly important to this symbiosis, the mechanism by which it is cycled remains to be resolved.     

Effect of nitrogen supply on nitrogen uptake,accumulation and assimilation in Porphyra perforata (Rhodophyta)

Porphyra perforata J. Ag. was collected from a rocky land-fill site near Kitsilano Beach, Vancouver, British Columbia, Canada and was grown for 4 d in media with one of the following forms of inorganic nitrogen: NO ₃ ^- , NH ₄ ⁺ and NO ₃ ^- plus NH ₄ ⁺ and for 10 d in nitrogen-free media. Internal nitrogen accumulation (nitrate, ammonium, amino acids and soluble protein), nitrate and ammonium uptake rates, and nitrate reductase activity were measured daily. Short initial periods (10 to 20 min) of rapid ammonium uptake were common in nitrogen-deficient plants. In the case of nitrate uptake, initial uptake rates were low, increasing after 10 to 20 min. Ammonium inhibited nitrate uptake for only the first 10 to 20 min and then nitrate uptake rates were independent of ammonium concentration. Nitrogen starvation for 8 d overcame this initial suppression of nitrate uptake by ammonium. Nitrogen starvation also resulted in a decrease in soluble internal nitrate content and a transient increase in nitrate reductase activity. Little or no decrease was observed in internal ammonium, total amino acids and soluble protein. The cultures grown on nitrate only, maintained high ammonium uptake rates also. The rate of nitrate reduction may have limited the supply of nitrogen available for further assimilation. Internal nitrate concentrations were inversely correlated with nitrate uptake rates. Except for ammonium-grown cultures, internal total amino acids and soluble protein showed no correlation with uptake rates. Both internal pool concentrations and enzyme activities are required to interpret changes in uptake rate during growth.     

Der Einfluß physikalischer und chemischer Faktoren auf die Aufnahme in Meerwasser gelöster Aminosäuren durch Aktinien

Changes in abiotic environmental factors do not adversely influence the capacity of Anemonia sulcata (Coelenterata, Anthozoa) to take up dissolved amino acids from sea water; e. g., A. sulcata resorbs also under anaerobic conditions. The influence of temperature on amino acid uptake (Q₁₀=2) indicates that the translocation of the organic molecules through membranes of the ectoderm depends on energy. Uptake of amino acids is possible against a gradient of up to 9×10⁶:1. It can be postulated that L-amino acids (also D-isomers) and glucose are resorbed by different “carrier systems”, since glucose does not influence the uptake of amino acids; however, amino acids can interfere with each other, e. g. phenylalanine/glycine. Blocking of anaerobic glycolysis results in uptake reduction; at the same concentrations “oxidative” blockers have a much smaller effect on amino acid uptake. The possibility of exploiting the surrounding water as an energy source by taking up dissolved organic material is discussed with reference to ecological and evolutionary aspects.     

Production of glycine betaine and dimethylsulfoniopropionate in marine phytoplankton. I. Batch cultures

The quantitative significance of the nitrogenous compound glycine betaine (GBT) and its sulfur analog dimethylsulfoniopropionate (DMSP) to intracellular pools in marine phytoplankton is not well known. In a series of experiments conducted in August 1993, we measured these compounds, as well as total organic sulfur, carbon, and nitrogen, over the growth cycle in six isolates of marine phytoplankton, Amphidinium carterae Hulburt, Chrysochromulina sp. Lackey, Emiliania huxleyi Hay et Mohler, Prorocentrum minimum (Pavillard) Schiller, Skeletonema costatum (Greville) Cleve, and Tetraselmis sp. At the same time, we measured cellular concentrations of protein, amino acids, chlorophyll, and inorganic nutrients. All six species produced DMSP, while three produced GBT at lesser levels. In the Chrysochromulina sp. isolate, levels of GBT were greater than DMSP during the exponential phase of growth, but declined sharply as the culture approached stationary phase. This change appeared to coincide with the onset of nitrogen limitation. Other nitrogenous osmolytes were produced in five of the six species but in much smaller quantities. DMSP contributed significantly to cellular sulfur throughout the growth cycle although, in some algae, the proportion of dissolved DMSP increased substantially during stationary growth. When present, GBT formed a sizeable fraction of the cellular nitrogen only during exponential growth. A significant percentage (ca. 50%) of the organic nitrogen could not be accounted for even when cellular pools of protein, amino acids, inorganic nitrogen, and nitrogenous osmolytes were combined. Based on these experiments, there does not appear to be a reciprocal relationship between DMSP and GBT production, although GBT production does appear to be correlated with nitrogen availability. Received: 5 January 1998 / Accepted: 29 June 1999     

Zur physiologie der aminbildung bei der marinen rotalge Polysiphonia urceolata

The ceramiaceaen Polysiphonia urceolata rapidly degrades ¹⁴C(U)-L-leucine, added to sea water at a final concentration of 2.5x10^-5 M/l, to isoamylamine. Under experimental conditions, 22% of the total radioactivity is found in the amine within 160 min. The amino acid decarboxylase responsible for this reaction has been characterized by Hartmann (1972a). No other mechanisms of leucine degradation could be detected, and the rate of ¹⁴C-incorporation into algal proteins is considerably lower than that of decarboxylation. The rate of decarboxylation is optimal at a leucine concentration of about 5x10^-5 M/l. The amine formed is found in almost equal amounts in algal extract and environment. No further degradation of isoamylamine could be detected. The amine is a metabolic end product in P. urceolata. When ¹⁴C-isoamylamine is used as a tracer, relatively high amounts of amine are found in the algal extracts. It is supposed that the amine does not accumulate within the algal cells, but rather is bound to the acid polysaccharides of the cell walls by means of ionic exchange. The results strongly suggest that decarboxylation is the main route by which P. ureolata metabolizes amino acids from the environment which are substrates of the decarboxylase. The endogenous amino acid pool does not seem to be available to the enzyme as a substrate source. A possible ecological significance of amino acid decarboxylations is discussed.     

Decarboxylation of environmental L-leucine by marine red algae

The uptake and metabolism of ¹⁴C-L-leucine by Cystoclonium purpureum (Huds) Batt. (Rhodophyllidaceae) at concentrations of 0.5–50 μM were studied. Leucine was taken up from seawater at rates of 10–160 nmol·g^-1 fresh weight·h^-1. At low leucine levels ( 5 μM) the amino acid was preferably incorporated into cellular protein; amine formation was less than 5%. However at leucine concentrations of 50 μM, more than 30% of the total leucine taken up was decarboxylated to 3-methylbutylamine. This effect is expressed even more in Dumontia increassata (O. F. Müll) Lamour. (Dumontiaceae). In both species some of the amine occurred as a non-volatile derivative (bound amine) from which free amine could quantitatively be released by acid hydrolysis. Leucine and amine uptake were inhibited by CCCP. However, from cells preloaded with both leucine and amine, only free amine is released in the presence of CCCP. The results indicate that decarboxylase activity will be switched on at increased levels of amino acids in seawater which may be expected frequently in habitats of littoral algae (tidal zone). A possible function of amine formation and release within the scope of chemical defence of benthic algae is discussed.     

Decomposition of silicate minerals by Bacillus mucilaginosus in liquid culture

The extraction of K⁺ and SiO₂ from silicate minerals by Bacillus mucilaginosus in liquid culture was studied in incubation experiments. B. mucilaginosus was found to dissolve soil minerals and mica and simultaneously release K⁺ and SiO₂ from the crystal lattices. In contrast, the bacterium did not dissolve feldspar. B. mucilaginosus also produced organic acids and polysaccharides during growth. The polysaccharides strongly adsorbed the organic acids and attached to the surface of the mineral, resulting in an area of high concentration of organic acids near the mineral. The polysaccharides also adsorbed SiO₂ and this affected the equilibrium between the mineral and fluid phases and led to the reaction toward SiO₂ and K⁺ solubilization. These two processes led to the decomposition of silicate minerals by the bacterium.     

Patterns of growth and triacylglycerol content in snow crab Chionoecetes opilio (Brachyura: Majidae) zoeal stages reared in the laboratory

Snow carb Chionoecetes opilio zoea I and zoea II larvae, hatched from females in a controlled mating experiment, were reared in the laboratory at 10.1 °C and 28.0 salinity, to resolve the patterns of growth (dry weight [DW]) and change in energy reserves (triacylglycerols [TAG]) within a given moult cycle. The patterns of growth and change in TAG reserves were similar in each zoeal stage. Following hatching or a moult, the zoeae entered a phase of rapid size increase, i.e. high daily growth rates (5.5 to 12.8% DWd^-1), for 1/3 to 2/5 of the duration of the moult cycle. During the same period, the zoeae accumulated TAG reserves until a maximum (TAG DW^-1) was reached at the end of the phase of rapid growth. The period of high growth rates and of TAG accumulation is interpreted as the required time for the zoeae to reach a point in development [i.e. point of reserves saturation (PRS); Anger and Dawirs (1981)] where sufficient growth and energy reserves allow moulting to the next stage. Following the phase of rapid growth and TAG accumulation, the zoeae entered a phase of low daily growth rates (0 to 1% DWd^-1) during which the TAG reserves decreased to a minimum at the end of the phase. Prior to, and during the moult to zoea II, a phase of negative growth was observed in the zoea I larvae. We conclude that measurement of zoeal size and TAG content, along with morphometric criteria (e.g. epidermal retraction), can be used to assess growth and nutritional condition of C. opilio zoeal stages from the sea.