Zooxanthellae providing assimilatory power for the incorporation of exogenous acetate in Heteroxenia fuscescens (Cnidaria: Alcyonaria)

The soft coral Heteroxenia fuscescens (Ehrb.) and its isolated zooxanthellae (endosymbiotic dinoflagellates) were investigated with particular regard to uptake and utilization of exogenously supplied ¹⁴C-acetate in the light and in the dark. The incorporation of ¹⁴C from ¹⁴C-acetate into the host tissue and into the zooxanthellae was consistently much higher in the light than in the dark. The incorporated ¹⁴C-acetate was rapidly metabolized by the host and algae and was recovered from different assimilate fractions. The major proportion of radiocarbon from metabolized ¹⁴C-acetate was located in host tissue. The CHCl₃-soluble fraction composed of diverse lipids showed the strongest ¹⁴C-labelling. Zooxanthellae isolated prior to incubation accounted for about 80% of the acetate incorporation recorded for zooxanthellae in situ (in vivo). It is concluded from a comparison of acetate incorporation and conversion under light and dark conditions that most of the lipid reserve of the host tissue originates from fatty acids, which are synthesized within the algal symbionts and are then translocated to the heterotrophic partner via extrusion. The acetate units needed for lipid synthesis are obtained by absorption of free acetate from dissolved organic matter (DOM) in the seawater as well as by photosynthetic assimilation of inorganic carbon. Thus, in H. fuscescens, lipogenesis is operated as a light-driven process to which the zooxanthellae considerably contribute assimilatory power by performing fatty acid synthesis and translocation of lipid compounds to their intracellular environment (host cell). A metabolic scheme is proposed to account for the different pathways of carbon conversion observed in H. fuscescens. The incubations took place in August 1980 and the analytical part from October 1980 to January 1984.     

Carbon fixation and analysis of assimilates in a coral-dinoflagellate symbiosis

Freshly collected pieces of the hermatypic coral Acropora cf. scandens containing dinoflagellate endosymbionts (presumably Gymnodinium microadriaticum) were allowed to assimilated ¹⁴C from H¹⁴CO ₃ ^- in the light and in the dark. Time-dependent carbon uptake resulted in intense ¹⁴C-labelling of ethanol-soluble as well as of insoluble assimilates. About forty ¹⁴C-labelled assimilates have been identified. Polymeric (ethanol-insoluble) compounds achieve about 30% of total radiocarbon incorporation after 60 min incubation. Kinetics of ¹⁴C-labelling of single assimilates are analyzed. Percentages of typical photosynthates in the soluble fraction undergo characteristic time-dependent changes. Lipids proved to be the main accumulation products of carbon assimilation by incorporating more than 50% of ¹⁴C after 60 min photosynthesis. The data indicate that low-molecular weight photosynthates such as ¹⁴C-glycerol and ¹⁴C-glucose are rapidly converted to constituents of the polymeric fraction(s) of the coral. Besides peptides, polysaccharides, and lipophilic substances, considerable amounts of ¹⁴C are confined to skeletal CaCO₃ of the coral. The results are discussed with respect to trophic and metabolic interrelationships between the autotrophic dinoflagellates and the A. cf. scandens tissues.     

Coral calcification: Sources of error in radioisotope techniques

Isotopic exchange occurs between coral skeleton and ⁴⁵Ca⁺⁺ and H¹⁴CO ₃ ^- in seawater. Exchange of ¹⁴C onto skeletons is more rapid than exchange of ⁴⁵Ca⁺⁺. Exchange of ¹⁴C from skeletons to seawater takes place more slowly than exchange of ⁴⁵Ca⁺⁺ to seawater. When living coral is incubated in the dark with radioisotopes for 1 h, the tissues contain considerably more radioactivity than is associated with the skeleton. The tissue radioactivity reflects permeation of tissues and coelenteron by radioactive compounds from the incubation seawater. Addition of alkalis to cardioactive seawater results in a radioactive precipitate, part of which becomes associated with any coral skeleton present, and part of which forms on the wall of the containing vessel. Strong alkali removes biologically-deposited radioisotope from coral skeletons. Deposition, of ¹⁴C from H¹⁴CO ₃ ^- in skeletons of living coral incubated in the dark is greater than in dead coral. The reverse situation occurs with ⁴⁵Ca⁺⁺.     

Uptake of inorganic carbon and internal carbon cycling in symbiont-bearing benthonic foraminifera

Incorporation rates of inorganic carbon and its distribution between the organic matter and the skeleton have been measured using ¹⁴C tracer techniques on two species of symbiont-bearing benthonic foraminifera in the Gulf of Elat: Amphistegina lobifera (a perforate species) and Amphisorus hemprichii (an imperforate species). Under constant experimental conditions, incorporation rates of the radiotracer become linear with time after several hours in A. hemprichii and after one day in A. lobifera. A. lobifera showed a lag time of 24 h for skeletal incorporation, whereas in A. hemprichii uptake into the skeleton started within 2 h. Pulse-chase incubations in radioactive seawater, followed by unlabelled incubations, demonstrate transfer of photosynthetically acquired ¹⁴C into the skeleton of A. lobifera. No such transfer was found in A. hemprichii. The total ¹⁴C uptake by A. lobifera increased during the first 24 h of cold chase incubation. This increase suggests the existence of an internal inorganic carbon pool that was lost (probably evaporated) during the analysis of pulse incubations. However, during the following chase incubations, the ¹⁴C in this pool was incorporated mainly into the skeleton and retained during analysis, causing the increase in the total uptake. No such increase was found in A. hemprichii. Additional ¹⁴C uptake experiments on other species of the genera Operculina, Heterostegina and Borelis suggest that the differences in pathways for incorporation of carbon between A. lobifera and A. hemprichii can be generalized to the perforate and imperforate foraminiferal groups. In perforate species, respired carbon originally taken up through photosynthesis is partly recycled into the skeleton. In imperforate species such a transfer has not been demonstrated. Perforate species seem to have a large internal inorganic carbon pool which serves mainly for calcification and possibly also for photosynthesis, while imperforate species may take up carbon for calcification directly from seawater or have a very small inorganic carbon pool.     

Decomposition of urea associated with photosynthesis of phytoplankton in coastal waters

Decomposition of urea in seawater was studied in Mikawa Bay, a shallow eutrophic bay on the southern coast of central Japan. The urea concentration in seawater ranged from 1.3 to 5.9 μg-at. N/1 and comprised 12 to 40% of the dissolved organic nitrogen. Using ¹⁴C labelled urea, the rate of CO₂ liberation from urea and the incorporation rate of urea carbon into the particulate organic matter were determined. For the surface samples, high rates of CO₂ liberation from urea as well as the incorporation of urea carbon into the particulate organic matter were observed in the light, while much lower rates were obtained in the dark. Incubation experiments with exposure to different light intensities revealed that the rate of CO₂ liberation from urea and the incorporation of urea carbon into particulate organic matter changed with light intensity, showing a pattern similar to that of photosynthesis. The highest liberation and incorporation rates were observed at 12,000 lux. Incubation in light and in dark produced marked decreases and increases, respectively, in urea and ammonia, while no appreciable changes were observed for nitrate and nitrite. It is suggested that urea decomposition associated with photosynthetic activity of phytoplankton is one of the major processes of urea decomposition, and that it plays a significant role in the nitrogen supply for phytoplankton in coastal waters.     

Effect of nitrogen source on short-term light and dark CO2 uptake by a marine diatom

Based on a series of short-term incubations involving the marine diatom Chaetoceros simplex (Bbsm), precultured in NH ₄ ⁺ -, NO ₃ ^- -and urea-limited continuous cultures at several dilution rates, we found that both the short-term specific rate of ¹⁴CO₂ uptake and the amount of CO₂ fixed after 8- and 16-min incubations were unaffected by enrichment with NH ₄ ⁺ , urea, or NO ₃ ^- when NH ₄ ⁺ or urea were the preconditioning forms of N, but were slightly suppressed when the cells were first grown on NO ₃ ^- . Similar enrichments in the dark, however, led to significant CO₂ uptake under all conditions of NH ₄ ⁺ enrichment and to similarly enhanced CO₂ uptake, but only at high growth rates, when urea was the source of enrichment nitrogen. Our light results are contrary to some contemporary findings, but there does seem to be agreement that photosynthetic rates of rapidly growing phytoplankton will not be affected by exposure to pulses of nitrogen. Enhanced dark uptake, in contrast, appears to be characteristic of phytoplankton under all degrees of N limitation, and, as such, may be useful as an “all or nothing” index of the nitrogen status of natural waters. There is some indication that the index may be useful in determining both the form of and the degree of N limitation as well.     

Coral illumination through an optic glass-fiber: incorporation of 14C photosynthates

The pathways of ¹⁴C incorporation into the three major compartments of the coral body were analysed in colonies of Stylophora pistillata. We used the optic glass-fiber method to carry out two sets of experiments: in the first, 11 different colonies were sampled immediately after incubation; in the second, 3 colonies were returned to the reef at the termination of incubation for a further period of 29 h. Within the tissue compartment, significantly more ¹⁴C labeled products were incorporated into illuminated tips or bases than into unilluminated sections. Tips located above illuminated bases contained amounts of ¹⁴C products similar to unilluminated tips. Within the organic matrix compartment, illumination of tip or base segments again resulted in increased amounts of ¹⁴C fixation, and again unilluminated tips located above the illuminated bases did not accumulate more ¹⁴C photosynthates than other tips on the same branches. The absence of detectable translocation was also confirmed after a post-incubation period of 29 h, and raises questions as to the validity of the widely accepted theory of upward translocation. Within the skeletal carbonate compartment, although the results were associated with a high coefficient of variation, significantly more ¹⁴C accumulated in the tips than in the bases. Illumination of tips or bases did not enhance ¹⁴C uptake. A light-independent carbon assimilation (dark fixation) is significant in S. pistillata within the three tested compartments (the tissue, the organic matrix of the skeleton, and the skeletal carbonate). It is suggested that the dark fixation process in corals in a result of accumulation of respiratory CO₂ and CO₂ from sea water as malate or other titratable acids during the night. During the day these acids are broken down, releasing free CO₂ for C₃ pathway photosynthesis.     

A reevaluation of the role of glycerol in carbon translocation in zooxanthellae-coelenterate symbiosis

Glycerol has been traditionally viewed as the main form of carbon translocated from zooxanthellae to the coelenterate host. Most of this glycerol was postulated to be used by the coelenterate host for lipid synthesis. Recent work suggests that large amounts of photosynthetically fixed carbon is synthesized into lipid in the algae, and then translocated as lipid droplets to the host. These two hypotheses of carbon translocation are not mutually exclusive, but to reconcile them the role of glycerol must be reevaluated. In this study the short term metabolic fate of uniformly labelled ¹⁴C-glycerol, ¹⁴C-bicarbonate, and ¹⁴C-acetate was examined in zooxanthellae and coelenterate host tissue isolated from Condylactis gigantea tentacles. When host and algal triglycerides, synthesized during 90-min light and dark incubations in ¹⁴C-bicarbonate and ¹⁴C-acetate, were deacylated, more than 80% of the activity was found in the fatty acid moiety. In contrast, triglycerides isolated from zooxanthellae and coelenterate host tissue incubated in ¹⁴C-glycerol in the dark for 90 min were found to have more than 95% of their radioactivity in the glycerol moiety. During the 90-min ¹⁴C-glycerol incubations in the light, the percentage of radioactivity in the fatty acid moiety of zooxanthellae triglycerides increased to 37%. The percentage of radioactivity in the host tissue triglycerides fatty acid moiety stayed below 5% during the 90-min ¹⁴C-glycerol incubations in the light. These results show that neither the zooxanthellae nor the host can rapidly convert glycerol to fatty acid. Radioactivity from ¹⁴C-glycerol, which does eventually appear in host lipid, may have been respired to ¹⁴CO₂, then photosynthetically fixed by the zooxanthellae and synthesized into lipid fatty acid.     

Respiratory quotient and calcification of Nautilus macromphalus (Cephalopoda: Nautiloidea)

Respiration and calcification were investigated in the ectocochleate cephalopod Nautilus macromphalus Sowerby. Specimens were collected off New Caledonia, in October 1991, and kept at the Nouméa Aquarium until December 1991. The respiratory quotient and calcification rate of 5 individuals were measured during 14 short term incubations (63 to 363 min). Oxygen uptake was recorded with a polarographic oxygen sensor. CO₂ flux and calcification were calculated from changes in pH and alkalinity (alkalinity-anomaly technique). Several methods were used to compute the respiratory quotient (RQ); a functional regression indicated an RQ of 0.74. CaCO₃ exchange rates were linearly related to respiratory quotient, calcification occurring in individuals with a low RQ. CaCO₃ uptake from the surrounding water was noncontinuous. From the highest CaCO₃ uptake, maximum growth rate was estimated as 7.1 mg shell wt h^- (=61 g yr^-1).     

Carbon flow into the end-products of photosynthesis in short and long incubations of a natural phytoplankton population

During a cruise to the eastern Canadian Arctic (Northern Baffin Bay) in the summer of 1980, we took advantage of the 24-h photoperiod to conduct a 32-h time course experiment of ¹⁴C accumulation under natural solar radiation. The degree of non-linearity in the time course was judged against a time-dependent curve of radioactivity constructed by cumulatively adding the amount of ¹⁴C taken up in sequential short (2 h) incubations of plankton held in a replicate bottle but left unlabelled until removed for assay. Departure from linearity was due first to decreasing rates of ¹⁴C incorporation into polysaccharides and then into lipids. There was a close correspondence between ¹⁴C incorporation into proteins in the 32-h incubation and in the sequence of short incubations. These observations are consistent with patterns in utilization of photosynthetic end-products established from laboratory studies of unicellular algal cultures. Based on parallel or independent control experiments, it was judged that complicating factors such as diel light changes, nitrogenous nutrient exhaustion, bottle size effects or inhibitory conterminants in NaH¹⁴CO₃ stock solutions would not seriously affect our interpretation that non-linearity resulted from catabolic loss of radiocarbon.     

Non-photosynthetic oxygen production and non-respiratory oxygen uptake in the dark: a theory of oxygen dynamics in plankton communities

Plankton respiration is commonly measured in terms of oxygen uptake, usually employing the Winkler method, much less commonly the polarographic method. Both methods produce results that can be misinterpreted when H₂O₂ production and decomposition are ignored. This paper: (1) presents experimental evidence of significant H₂O₂ involvement during plankton incubation in dark bottles, (2) explains how results differ between the Winkler and polarographic methods in the presence of H₂O₂, (3) discusses how this difference is clouded by side issues of variability inherent in the Winkler technique and the use of different bottle sizes, and (4) shows that unexpected and variable results of light-/dark-bottle incubations can all be explained by a theory of H₂O₂ production and decomposition. During an initial period in the dark, when plankton respiration has been poisoned by mercuric chloride or chloroform, O₂ increase can be measured with a polarographic oxy‐gen sensor (POS). The trend in O₂ changes is linear for several days when only respiration is occurring, but curvilinear when there is concurrent O₂ production. O₂ production in the dark and H₂O₂ decomposition are one and the same process. Measurement of oxygen by Winkler analysis and POS produce different results when H₂O₂ is present because the former method measures oxidizing equivalent while a POS measures O₂ pressure. A real difference in results between the two methods is prima facie evidence that H₂O₂ is involved. The synthesis of this new empirical evidence with diverse knowledge from various fields shows that the common practice of estimating gross community primary production from oxygen changes in light and dark bottles is based on untenable assumptions. Received: 15 April 1997 / Accepted: 17 June 1997     

Photosynthesis,photorespiration, and dark respiration in eight species of algae

The rates of photosynthesis and dark respiration for 7 marine algae and 1 fresh-water alga were measured and compared. The dinoflagellates Glenodinium sp. and zooxanthellae have high dark respiration rates relative to photosynthetic rates, which may decrease their net growth rates. Photorespiration in the 8 algal species was studied by examining the effects of the concentration of oxygen on the rates of photosynthesis, on the incorporation of ¹⁴CO₂ into the photorespiratory pathway intermediates glycine and serine, and on the postillumination burst of carbon dioxide production and oxygen consumption. A combination of these results indicates that all the algae tested can photorespire, but that Glenodinium sp., Thalassiosira pseudonana, and zooxanthellae either have a photorespiratory pathway different from that proposed for freshwater algae (Tolbert, 1974), or an additional pathway for glycolate metabolism.     

Skeletal calcification patterns in the sea urchin Tripneustes gratilla elatensis (Echinoidea: Regularia)

A comprehensive study on skeletal calcification in the regular echinoid Tripneustes gratilla elatensis was carried out between 1979 and 1981 in the northern Gulf of Eilat (Red Sea), employing size measurement, allometry and radiotracer techniques. Uptake rates of the isotope ⁴⁵Ca were used to estimate the calcification rate of whole tests. Calcification rates of the different skeletal parts and the ⁴⁵Ca uptake along the sutural margins of individual plates were also measured. Whole-test calcification rates for juvenile individuals, 11 to 13 mm in diameter, relative to their skeletal Ca dry weight, calculated using the allometric relationship between skeletal dry weight and diameter, were 1.5 to 1.8% d^-1 for 4 to 8 h of incubation, while calcification rates obtained from periodical size measurements and allometrical constants amounted to 2.75% d^-1. The apparent discrepancy between these results is explained partly by the short duration of the incubations, during which the internal calcium pools, apparently in the coelom, were not fully saturated with the radioactive tracer. This discrepancy decreased when longer radioactive incubations were used, or when a post-incubational deposition of ⁴⁵Ca from these pools was allowed. The size of these pools is roughly 10 mol Ca per 13-mm individual, and it resides probably in the coelom where Ca concentration is 3 times higher than in seawater. The calcification rates given as percentage of skeleton added per day, both for the radioactive and allometric methods, decreased with the urchins' size. The calcification patterns of various components of the skeleton generally fitted their allometrical trends — a relative decrease in the size of the spines, apical plates and lantern. The teeth calcify rapidly to compensate for the constant erosion of their tips. Calcification per plate decreased exponentially from the apex to the peristome. Plate-edge calcification patterns of both interambulacral and ambulacral plates fitted patterns derived from size increment measurements, using natural growth-lines. Typically, most plates grow more horizontally (latitudinally) than vertically. It is suggested that the vertical vs horizontal calcification ratio (v/h) determines the individual plate growth and affects the whole test morphology.     

Ultrastructure and anaerobic metabolism of mitochondria in the marine oligochaete Tubificoides benedii : effects of hypoxia and sulfide

It has often been suggested that ultrastructural properties of mitochondria are correlated with oxygen and sulfide levels from the environment, although careful analyses of this question are rare. In this study the ultrastructure and distribution of mitochondria in Tubificoides benedii, a marine oligochaete from sulfide-rich sediments, were investigated after a series of oxic, hypoxic and hypoxic–sulfidic (200 μM H₂S) incubations up to 24 h. Succinate, one of the key endproducts of an anaerobic metabolism, was used as an indicator of mitochondrial anaerobiosis. Consistent differences in mitochondrial ultrastructure were not observed in any of the incubations, even after 24 h. Stereological parameters of mitochondria (volume density, surface density of the outer mitochondrial membrane, and specific surface) in epidermal and intestinal tissues of T. benedii were not affected by hypoxia or sulfide either. On the other hand, succinate concentrations increased significantly within 24 h under hypoxic and hypoxic–sulfidic conditions. Thus, experimental hypoxia and sulfide clearly caused mitochondrial anaerobiosis without affecting ultrastructure or distribution of mitochondria in T. benedii. Distinct differences in ultrastructural and stereological parameters were common between different tissues and between individuals, showing that different forms of mitochondria can occur within one species. Our results imply that a mitochondrial ultrastructure specific to thiobiotic animals does not appear to exist. Received: 4 August 1996 / Accepted: 20 September 1996     

Growth characteristics of Parmales (Chrysophyceae) observed in bag cultures

Incubations of natural populations of phytoplankton were carried out in neritic and oceanic areas of the western subarctic Pacific in 1991 and 1992. Algae in the order Parmales, class Chrysophyceae, were observed to increase in number during the incubations. In the light-exposed treatments, the growth rate of Parmales at 5 to 12 °C was 0.012 to 0.016 h^-1 or 0.41 to 0.54 doubling d^-1, which is lower than that of diatoms, but comparable to that of common dinoflagellates. On the other hand, heterotrophic choanoflagellates grew positively in both light and dark at the rate of 0.016 to 0.040 h^-1 or 0.54 to 1.39 doublings d^-1, which is comparable or lower than the reported value at 15 °C. The results obtained demonstrate that the Parmales can grow vegetatively in light and prefer low temperatures.     

Transversal profiles of carbon assimilation in the fronds of three Laminaria species

The brown macroalgae Laminaria digitata (Huds.) Lamour., L. hyperborea (Gunn.) Foslie, and L. saccharina (L.) Lamour. (Laminariales, Phaeophyceae) have been investigated for carbon assimilation and assimilate biosynthesis in different tissues (meristoderm, cortex, medulla) of stipes and blades. Carboxylation via ribulose-1,5-bisphosphate carboxylase (EC 4.1.1.39) and phosphoenolpyruvate carboxykinase (EC 4.1.32) follows a transversal gradient from the outer to the inner tissues with maximum activity of enzymatic CO₂ fixation in the meristoderm. Such transversal profiles are also recorded for photosynthetic and dark carbon assimilation as well as for kinetics of assimilate ¹⁴C-labelling in isolated tissues. Differential chemical constitution of the blade and stipe tissues examined and results from assimilate biosynthesis are discussed with regard to a transversal translocation of photosynthates from meristodermal to cortical and medullar cells.     

Chlorophyll fluorescence as a proxy for microphytobenthic biomass: alternatives to the current methodology

Pulse amplitude modulated (PAM) fluorescence has been used as a proxy of microphytobenthic biomass after a dark adaptation period of 15 min to stabilise the minimum fluorescence yield (F _o ¹⁵). This methodology was investigated for in situ migratory and ex situ engineered non-migratory biofilms, comparing dark adaptation to low (5% ambient) and far-red light treatments over different emersion periods. Far-red and low light reduced potential errors resulting from light history effects, by reversal of non-photochemical quenching after 5 min of treatment, compared to over 10 min required by conventional dark adaptation. An in situ decline of minimum fluorescence yield over 15 min was observed during the dark adaptation for migratory biofilms, but was not observed in the non-migratory biofilms indicating that the major cause of decline was downward vertical migration of cells into the sediment. This pattern occurred in far-red light after 10 min, but not for the low light treatment, indicating that low light maintained the biomass at the surface of the sediment. It is therefore concluded that low light treatment is a better option than conventional dark adaptation for the measurement of minimum fluorescence as a proxy of microphytobenthic biomass.     

Dark CO2 uptake by the diatom Chaetoceros simplex in response to nitrogen pulsing

In a continuing investigation of dark CO₂ uptake by nitrogen-limited cultures of the marine diatom Chaetoceros simplex (Bbsm), we expanded on several of our earlier conclusions regarding the potential application of this physiological response for measuring the degree and type of nitrogen limitation in phytoplankton populations. First, the duration over which the maximal enhancement of dark ¹⁴CO₂ uptake was sustained after NH ₄ ⁺ enrichment was a function both of the concentration of added NH ₄ ⁺ and the standing crop of phytoplankton nitrogen — in effect, the total N demand. Second, pulsing with NH ₄ ⁺ for a given degree of N-limitation always produced the same level of enhanced dark CO₂ uptake regardless of whether the cultures were preconditioned with oxidized or reduced nitrogen. In contrast, urea pulsing led to reduced dark CO₂ uptake, but the effect was most pronounced in cells grown on NO ₃ ^– . And third, the assay could be used to distinguish readily between no, moderate, and severe N limitation. The degree of severe N limitation was quantitatively correlated with the degree of enhanced dark CO₂ uptake, but this relationship was not so clear in the region of moderate N limitation. The main advantage of the assay is that it is a relatively simple and effective alternative to more complicated techniques for gauging the degree and form of N limitation in phytoplankton. Further evaluation will be required, both in the laboratory and field, before the assay can be calibrated for quantitative use.Contribution No. 5982 from the Woods Hole Oceanographic Institution     

Effects of pCO2 on physiology and skeletal mineralogy in a tidal pool coralline alga Corallina elongata

Marine organisms inhabiting environments where pCO₂/pH varies naturally are suggested to be relatively resilient to future ocean acidification. To test this hypothesis, the effect of elevated pCO₂ was investigated in the articulated coralline red alga Corallina elongata from an intertidal rock pool on the north coast of Brittany (France), where pCO₂ naturally varied daily between 70 and 1000 μatm. Metabolism was measured on algae in the laboratory after they had been grown for 3 weeks at pCO₂ concentrations of 380, 550, 750 and 1000 μatm. Net and gross primary production, respiration and calcification rates were assessed by measurements of oxygen and total alkalinity fluxes using incubation chambers in the light and dark. Calcite mol % Mg/Ca (mMg/Ca) was analysed in the tips, branches and basal parts of the fronds, as well as in new skeletal structures produced by the algae in the different pCO₂ treatments. Respiration, gross primary production and calcification in light and dark were not significantly affected by increased pCO₂. Algae grown under elevated pCO₂ (550, 750 and 1000 μatm) formed fewer new structures and produced calcite with a lower mMg/Ca ratio relative to those grown under 380 μatm. This study supports the assumption that C. elongata from a tidal pool, where pCO₂ fluctuates over diel and seasonal cycles, is relatively robust to elevated pCO₂ compared to other recently investigated coralline algae.     

Respiratory impairment by water-borne copper and zinc in the edible crab cancer pagurus (L.) (Crustacea: Decapoda) during hypoxic exposure

Specimens of the edible crab Cancer pagurus (L.) collected in the Skagerrak, Denmark, between June and August 1990 were examined in the laboratory. Impairment of respiratory function after pre-exposure (7 d) to sub-lethal concentrations of Cu and Zn (0.4 mg l^-1) was only detectable during hypoxic exposure [PO₂ (partial pressure of oxygen) =60 torr]. This was indicated by a decrease in the transfer factor (TO₂), due principally to an increase in the PO₂ differential across the gills. Cu and Zn exposure did not cause significant changes in ventilation or perfusion although there was some indication that cardiac output may increase in respiratory-impaired individuals. After 28 d exposure no difference was noted in the respiratory responses to hypoxia of treated and untreated crabs. It is concluded that respiratory impairment was due to an increase in the diffusion barrier thickness at the gills and that this was reversible even during continued exposure to trace metal contamination.