Availability of two forms of dissolved nitrogen to the coral Pocillopora damicornis and its symbiotic zooxanthellae

The relative contribution of dissolved nitrogen (ammonium and dissolved free amino acids DFAAs) to the nitrogen budget of the reef-building coral Pocillopora damicornis was assessed for colonies growing on control and ammonium-enriched reefs at One Tree Island (southern Great Barrier Reef) during the ENCORE (Enrichment of Nutrient on Coral Reef; 1993 to 1996) project. P. damicornis acquired ammonium at rates of between 5.1 and 91.8 nmol N cm⁻² h⁻¹ which were not affected by nutrient treatment except in the case of one morph. In this case, uptake rates decreased from 80.5 to 42.8 nmol cm⁻² h⁻¹ (P < 0.05) on exposure to elevated ammonium over 12 mo. The presence or absence of light during measurement did not influence the uptake of ammonium ions. Nitrogen budgets revealed that the uptake of ammonium from concentrations of 0.11 to 0.13 μM could completely satisfy the demand of growing P. damicornis for new nitrogen. P. damicornis also took up DFAAs at rates ranging from 4.9 to 9.8 nmol N cm⁻² h⁻¹. These rates were higher in the dark than in the light (9.0 vs 5.1 nmol m⁻² h⁻¹, P < 0.001). Uptake rates were highest for the amino acids serine, arginine and alanine, and lowest for tyrosine. DFAA concentrations within the ENCORE microatolls that received ammonium were undetectable, whereas they ranged up to 100 nM within the control microatolls. The contribution of DFAAs to the nitrogen budget of P. damicornis constituted only a small fraction of the nitrogen potentially contributed by ammonium under field conditions. Even at the highest field concentrations measured during this study, DFAAs could contribute only ≃11.3% of the nitrogen demand of P.␣damicornis. This contribution, however, may be an important source of nitrogen when other sources such as ammonium are scarce or during periods when high concentrations of DFAAs become sporadically available (e.g. cell breakage during fish-grazing). Received: 22 April 1998 / Accepted: 3 November 1998     

Modification of shell formation in the giant clam Tridacna gigas at elevated nutrient levels in sea water

The effect on shell formation of Tridacna gigas by sea water supplemented for 3 mo with ammonium (5, 10 M, N) and phosphate (2, 5, 10 M, P), separately or in combination, was examined. Exposure to N and N+P significantly enhanced shell-extension rates, but significantly reduced shell weights at equivalent size. Scanning-electron microscopy further revealed structural alterations in the outer shell layer, such as misshapen aragonite crystals, irregular crossed-lamellar orientation, and relatively porous shell microstructure. These observations are consistent with results of X-ray diffractometry on the shells which show distinct shifts in the positions of reflections from the (012) and (200) crystal planes relative to the control, indicating changes in crystal lattice parameters following addition of nutrients.     

No genetic differentiation of giant clam (Tridacna gigas) populations in the Great Barrier Reef,Australia

Six Tridacna gigas populations were sampled in 1990 from locations throughout the central and northern Great Barrier Reef (GBR). Despite separations in excess of 1000 km, mean Nei's unbiased genetic distances among the populations was 0.0007. The complete lack of spatial variation observed among populations did not results from lack of genetic variability. Genetic variation within populations was high, with mean heterozygosities from 0.221 to 0.250. Gene frequencies were consistent with expectations under conditions of Hardy-Weinberg equilibrium. These data suggest panmixis, or random mating, throughout the highly connected reef system of the central and northern GBR. The large gene exchange among the giant clam populations has important implications for conservation management of one of the few large populations of T. gigas in the world. Small local effects are likely to be overcome in time by inputs from other sources. However, large genetic perturbations, particularly from up-current sources, may spread rapidly through the population.Contribution No. 561 from the Australian Institute of Marine Science     

Characterization of two glutamate dehydrogenases from the symbiotic microalga Symbiodinium microadriaticum isolated from the coral Acropora formosa

In view of their possible involvement in ammonium assimilation in the coral/algal symbiosis, we have purified two distinct glutamate dehydrogenase isoenzymes from the symbiotic dinoflagellate Symbiodinium microadriaticum (Freudenthal) extracted from the staghorn coral Acropora formosa collected from Magnetic Island, North Queensland, Australia, in 1986–1987. An NADPH-specific glutamate dehydrogenase (GDH) displayed biphasic kinetics with respect to ammonium as the variable substrate; at low substrate concentrations the apparent K _m was below 1 mM, whereas at high substrate concentrations the corresponding value was approximately 200 mM. The NADPH-GDH displayed extremely low activity in the direction of glutamate oxidation; together with the kinetic data this suggests a probable role in ammonium assimilation. A second (NADH-specific) GDH was found to have both amination and deamination activities, and presumably functions in vivo in glutamate oxidation. Kinetic constants are reported for both GDH isoenzymes.     

Illumination and the coral community beneath tabular Acropora species

Light profiles beneath the tabular coral species Acropora cytherea and A. hyacinthus were examined at Rib and Broadhurst Reefs, central Great Barrier Reef, in November 1980. They show a strongly decreasing illumination gradient towards the central stem. Beneath tables at 10m deep, which receive about 7.0 mW cm^–2 at solar noon, substrate irradiance falls to a minimum of about 5% ambient or to about 0.4 mW cm^–2. Thus, from previously reported compensation values, most sub-table substrate may receive above-compensation irradiance. Illumination beneath tables screened by foil is significantly lower. The coral communities beneath tables of these two species plus those of A. subulata and A. clathrata support an average 26 colonies m^–2, at a cover of 40%, nearly identical to adjacent, unshaded quadrats. Species richness is likewise very similar. Species richness and colony density beneath the tables increase inwards from the perimeter of the shaded areas, decreasing only near the central stems. Dark adaptation and reduced competitive and grazing pressure are suggested explanations for the latter. It is concluded that shading does not provide a significant competitive advantage for Acropora spp. tables at 10 m deep.     

Nutrient enrichment and the ultrastructure of zooxanthellae from the giant clam Tridacna maxima

The separate and combined effects of ammonium (10M) and phosphate (2M) on the ultrastructure of zooxanthellae (Symbiodinium sp.) from giant clams, Tridacna maxima, were examined in the field. Nitrogen addition significantly changed the ultrastructure of the zooxanthellae inhabiting the clams. After 9 mo exposure, the cross-sectional area of zooxanthellae from N-treated clams was significantly lower than that from other treatments [N=39.3 m²; C=47.9 m²; P=43.2m²; N+P=44.5 m²; (P=0.001)]. There was also a significant decrease in the size of starch bodies, especially around the pyrenoid of the zooxanthellae from N and N+P treatments [N=1.2 m²; C=2.0 m²; P=1.8 m²; N+P=1.2 m²; (P=2.08E-11)]. This presumably occurs as a result of the mobilization of organic carbon stores in response to stimulated amino acid synthesis under enriched nutrient conditions. These data strongly suggest that the symbiotic zooxanthellae of clams are limited to some extent by the availability of inorganic nitrogen, and that relatively minor changes to the nutrient loading of the water column can have substantial effects on the biochemistry of symbioses such as that which exists between clams and zooxanthellae.     

Influence of field-based nutrient enrichment on the photobiology of the giant clam Tridacna maxima

Nutrients were added to 12 microatolls in One Tree Island lagoon every low tide for 13 mo to an initial concentration of 10 μM (ammonium, N) and 2 M (phosphate, P). These concentrations remained above background for 2 to 3 h after addition. The addition of ammonium (N and N+P but not P alone) significantly increased P _g (gross photosynthesis) P _n (net photosynthesis) and R (respiration) per unit wet-tissue weight and α (photosynthetic efficiency) in Tridacna maxima after 3 mo nutrient enrichment. These responses to small and transient changes in ammonium concentrations suggest that symbiotic clams are not nutrient-replete, and that even subtle changes in nutrients can have a measurable effect on photosynthesis. The same clams did not show significant differences in photosynthetic parameters 6 mo after the beginning of nutrient enrichment, suggesting that their previous responses had either been seasonal or that symbiotic clams such as T. maxima are able to adjust their photophysiology following external changes in nutrient concentrations. Received: 26 August 1997 / Accepted: 11 December 1998     

Neoplasia,regeneration and growth in the reef-building coral Acropora palmata

Abnormal processes of calcification, such as regenerating lesions and neoplasia, situated near the tips(<25 cm) of colonies of Acropora palmata (Lamarck) suppressed normal linear growth. Branches having neoplasia at a larger distance from the tip do not grow significantly differently from controls. This indicates a functional minimal area in terms of energy supply. Neoplasia are pure aragonite and have the same coenosteal structure as regenerative skeletal material. Regeneration of tissue as well as tissue+skeleton lesions involves the simultaneous formation of tissue and regenerative skeleton, trapping foreign material under the regenerated surface. Recovery of a damaged surface slows down with time and this may, in other coral species, result in permanent lesions. A. palmata recovered from all lesions (n=32) within 80 d and appears to be a superior regenerator among Caribbean corals. This is consistent with other life-history characteristics of this highly specialized coral species.     

Diurnal lipid and mucus production in the staghorn coral Acropora acuminata

Net ¹⁴C-accumulation into lipids of Acropora acuminata was rapid and increased with light intensity. Dark ¹⁴C-incorporation was less than 1% noon maximum. Structural lipids were the first radioactively labelled lipid types showing linear ¹⁴C-uptake kinetics. Storage lipids showed non-linear, power-curve kinetics for ¹⁴C-uptake. The rate of ¹⁴C-incorporation into triglycerides and wax esters was maximal during early afternoon and at midday, respectively. Electron microscopic evidence is given for zooxanthellae being primary sites for synthesis of lipids which are exuded from chloroplasts and transferred to animal tissues. Free lipid droplets and crystalline inclusions (wax ester) were common in animal tissues, the inclusions being often associated with mucus-producing cells. The diurnal rate of mucus production was constant. However, ¹⁴C-mucus-lipid production showed a light-dependent diurnal pattern and accounted for 60 to 90% total ¹⁴C of mucus during periods of photosynthetically-saturating light. Here, ¹⁴C was primarily associated with wax esters which were always present in the mucus-lipid. ¹⁴C-triglycerides occur in mucus released only during the day. Lipid and mucus synthesis is discussed in relation to the carbon budget of A. acuminata, in which mucus represented a loss of 40% net C fixation.     

Gas-exchange studies with the staghorn coral Acropora acuminata and its zooxanthellae

Compensation point and light-saturation values were determined from oxygen-exchange experiments with branches and isolated zooxanthellae from the staghorn coral Acropora acuminata. Branches and dense suspensions of zooxanthellae showed similar lightresponse curves for oxygen exchange, with light saturation at about 23 Klux (300 W. m^-2) and compensation point occurring between 4 and 6 Klux (60–80 W. m^-2). Zooxanthellae appear to be mutually shaded in dense suspensions and coral tissues. The effects of metabolic inhibitors, including photosynthetic and respiratory inhibitiors, on oxygen exchange in coral branches and isolated zooxanthellae are presented. Bubbles formed on coral tissues and on several macroalgae under conditions of high illumination contained large amounts of oxygen, suggesting that a high oxygen tension may occur in coral tissues during the day. Photorespiration and dissolved organic carbon production by suspensions of zooxanthellae are discussed in relation to a high oxygen tension which probably occurs in coral tissues during daylight.     

Compartmentation and turnover of organic carbon in the Staghorn coral Acropora formosa

Four colonies of Acropora formosa were incubate with Na₂ ¹⁴CO₃ for separate 2 h periods within a 24 h period, and then returned to the reef from which they were collected. Terminal branches were collected at intervals over the following 5 d and analysed for radioactivity associated with the skeleton and certain organic pools. Colonies incubated at night showed little or no loss of fixed radioactivity during the 5 d on the reef. However, 50–60% of photosynthetically-fixed ¹⁴C was lost from the terminal branches during the first 40 h on the reef. This loss of radioactivity probably resulted from release of mucus and dissolved organic carbon from the coral tissues. Most of the loss of photosynthetically-fixed ¹⁴C was due to decrease in the radioactivity of lipids (80% of the total ¹⁴C loss) and methanol-water soluble compounds. Determination of any sequencing in metabolic compartments was made difficult by the rapidity with which ¹⁴C dissappeared from most of the metabolic pools measured. ¹⁴C was incorporated into the skeleton throughout the 5 d on the reef, although the rate of incorporation was very low in colonies which had been incubated with Na₂ ¹⁴CO₃ at night.     

Genetic structure of giant clam (Tridacna derasa) populations from reefs in the Indo-Pacific

Large genetic differences were observed among the Great Barrier Reef (GBR), Fiji and Philippine populations of Tridacna derasa (Roding) sampled in 1989 and 1990 (Nei's unbiased genetic distance, D,=0.137 to 0.341). This result contrasted strongly with the low genetic distance (D=0.032) reported previously for the giant clam T. maxima over similar geographical scales. No significant genetic differentiation was observed among most populations from the GBR (mean D=0.007), consistent with the high gene exchange expected in this highly connected reef system. However, significant differentiation resulting from differences in the frequencies of less common alleles between the North-Central GBR and South GBR (Swain region) were observed. Historical isolation of blocks of the Central Indo-West Pacific from the GBR and present-day restrictions to gene exchange between the GBR, Fiji and the Philippines as a result of oceanographic current patterns, were thought to be responsible for the high degree of genetic differentiation of T. derasa populations. The relevance of these findings to clam mariculture and reef restocking are briefly discussed.Contribution No. 197 from the Marine Science Institute, University of the Philippines and Contribution No. 562 from the Australian Institute of Marine Science     

Reproduction,recruitment and fragmentation in nine sympatric species of the coral genus Acropora

Multispecies assemblages of the coral genus Acropora occur commonly throughout the Indo-Pacific Ocean. Nine species from such an assemblage comprising 41 species of Acropora, at Big Broadhurst Reef on the Great Barrier Reef, were studied during 1981–1983. Similarities and differences in reproductive modes and timing, oocyte dimensions and fecundity, recruitment by larvae and by fragments, and mortality were recorded. All species had an annual gametogenic cycle, were simultaneous hermaphrodites, and had the same arrangement of gonads in polyps. In six species, most colonies released gametes on the same night of the year, in early summer, during a mass spawning event involving many coral genera. A seventh species had colonies spawning at this as well as other times of the year. Another species spawned in late summer, and gametes were not observed to mature in the last species. Eggs were very large (601 to 728 m geometric mean diameter) and fecundity of polyps low, compared with other corals; no reduction in oocyte numbers occurred during oogenesis. Reef-flat species had slightly bigger and fewer eggs than reef-slope species. All species recruited by larvae, but four also multiplied by fragmentation, either year-round or during occasional rough weather. Yearround fragmenters had few larval recruits; non-fragmenters had many, and a rough-weather fragmenter had an intermediate number of larval recruits. It was concluded that larval recruitment largely determined species composition, and that reduced larval recruitment was responsible for sparse distribution of fragmenting species. Subsequent mortality in some species and increase by fragmentation in others probably determined relative abundances.     

Stable isotopic investigation of physiological and environmental changes recorded in shell carbonate from the giant clam Tridacna maxima

The aragonitic shell of the photosymbiont-bearing bivalve Tridacna maxima contains a record of the physiological and environmental changes the organism has experienced during its lifetime. This record is preserved as chemical and microstructural variations throughout the shell. Stable isotopic analyses of oxygen (¹⁸O/¹⁶O) and carbon (¹³C/¹²C) in shell carbonate were combined with growth increment studies to interpret the shell record of specimens collected from the Rose Atoll (Lat. 14°31S; Long. 168°10W) in April 1982. The seasonal water temperature cycle is recorded in the oxygen isotopic signature of the clams, permitting the recognition of annual cycles in the ¹⁸O profile. The total number of these cycles corresponds to the age of a specimen, while the cycle length is a measure of the yearly growth rate. Large-amplitude cycles, reflecting year-round calcification, characterize the early portion of the growth record. With the onset of sexual maturity and slower growth at an age of approximately ten years, the cycles decrease in amplitude and become more erratic. During this later growth phase calcification is limited to the cooler months of the year, perhaps in response to a re-ordering of energy priorities between growth and gametogenesis. A growth curve developed from the ¹⁸O profile indicates rapid juvenile shell growth followed by slower growth thereafter producing a lifespan of several decades. Carbon isotopic analyses of T. maxima were compared to analyses of the symbiont-barren gastropod Terebra areolata collected from the same locality in April 1984. A 2 depletion in the ¹³C composition of T. maxima shell carbonate is attributed to a symbiontenhanced metabolic rate and an increased flow of isotopically light, respired CO₂ into the carbon pool used in calcification. Such a depletion may prove useful in identifying the presence of photosymbionts in extinct species of fossil mollusks.     

Seasonal growth in Laminaria longicruris: Relations with dissolved inorganic nutrients and internal reserves of nitrogen

Observations have been made on seasonal fluctuations in dissolved inorganic nutrients, internal reserves of nitrogen and growth rates in Laminaria longicruris. The onset of winter growth in shallow-water stations (6 and 9 m) correlated well with improved dissolved nitrate conditions in the sea. During the winter, reserves of NO ₃ ^- were accumulated by the plants and reached maximum values of 150 moles per g fresh weight in March. This represents a concentration factor of approximately 28,000 over the ambient levels, or an internal nitrogen reserve of 2.1% of the dry weight of the tissue. Depletion of this nitrogen pool followed the disappearance of the external NO ₃ ^- with a lag period of up to 2 months. Rapid kelp growth was measured during this period. Reserves of organic nitrogen also reached maximum values in March and declined slowly throughout the summer into autumn. It is suggested that the combined inorganic and organic nitrogen reserves sustain the rapid growth rates into July and at reduced rate through the late summer. Fertilization of an experimental perimental kelp bed with NaNO₃ increased the internal plant reserves of NO ₃ ^- and produced a much improved summer growth rate. The enriched plants developed very small reserves of carbohydrate during the rapid summer growth phase.NRCC No. 15549.     

Growth of the staghorn coral Acropora formosa at Houtman Abrolhos, Western Australia

The growth (extension rate, number of radial branches, skeletal mass, branch diameter) of the␣staghorn coral Acropora formosa (Dana, 1846) was examined at four sites on the Beacon Island platform at Houtman Abrolhos, in subtropical Western Australia (28°S). Sites were at depths of 7 to 11 m, with variable exposure to weather and swell conditions. Two sites on the western reef slope were partly exposed to the oceanic swell, and two sites in the lagoon were largely protected from wave action. Linear extension rate between 1994 and 1995 varied significantly between sites, with greater linear extension at the more protected lagoonal sites. However, accumulation of skeletal mass per branch and number of newly initiated radial branches did not vary significantly between the sites. Carbonate was deposited in similar amounts, but either as porous, rapidly extending branches, or as denser branches which extended more slowly. Branch extension rate over 11.5 mo ranged from a mean of 50.3 mm (range=13 to 93 mm) at a reef slope site to a mean of 76.0 mm (range=31 to 115 mm) at a sheltered lagoonal site. Mean extension rates were almost twice that previously reported for this species in Houtman Abrolhos (37 to 43 mm yr⁻¹) from a shallower site where environmental conditions were apparently sub-optimal. Growth was within the range reported for A. formosa from tropical sites, which is consistent with the relatively high calcification and reef-accretion rates recorded for Houtman Abrolhos in geological and metabolic studies. The role of reduced coral growth-rate in limiting coral reef formation at high latitudes remains equivocal. Received: 19 November 1997 / Accepted: 5 May 1998     

Effect of nutrient enrichment in the field on the biomass, growth and calcification of the giant clam Tridacna maxima

Nutrients were added separately and combined to an initial concentration of 10 μM (ammonium) and/or 2 μM (phosphate) in a series of experiments carried out with the giant clam Tridacna maxima at 12 microatolls in One Tree Island lagoon, Great Barrier Reef, Australia (ENCORE Project). These nutrient concentrations remained for 2 to 3 h before returning to natural levels. The additions were made every low tide (twice per day) over 13 and 12 mo periods for the first and second phase of the experiment, respectively. The nutrients did not change the wet tissue weight of the clams, host C:N ratio, protein content of the mantle, calcification rates or growth rates. However, ammonium (N) enrichment alone significantly increased the total population density of the algal symbiont (Symbiodinium sp.: C = 3.6 · 10⁸ cell clam⁻¹, N = 6.6 · 10⁸ cell clam⁻¹, P = 5.7 · 10⁸ cell clam⁻¹, N + P = 5.7 · 10⁸ cell clam⁻¹; and C = 4.1 · 10⁸ cell clam⁻¹, N = 5.1 · 10⁸ cell clam⁻¹, P = 4.7 · 10⁸ cell clam⁻¹, N + P = 4.5 · 10⁸ cell clam⁻¹, at the end of the first and second phases of the experiment, respectively), although no differences in the mitotic index of these populations were detected. The total chlorophyll a (chl a) content per clam but not chlorophyll a per cell also increased with ammonium addition (C = 7.0 mg chl a clam⁻¹, N = 13.1 mg chl a clam⁻¹, P = 12.9 mg chl a clam⁻¹, N + P = 11.8 mg chl a clam⁻¹; and C = 8.8 mg chl a clam⁻¹, N = 12.8 mg chl a clam⁻¹; P = 11.2 mg chl a clam⁻¹, N + P = 11.3 mg chl a clam⁻¹, at the end of the first and second phases of the experiment, respectively). The response of clams to nutrient enrichment was quantitatively small, but indicated that small changes in inorganic nutrient levels affect the clam–zooxanthellae association. Received: 2 June 1997 / Accepted: 9 June 1997     

Ammonium, but not nitrate, stimulates an increase in glutamine concentration in the haemolymph of Tridacna gigas

The concentration of glutamine in Tridacna gigas haemolymph increased >35-fold following exposure to sea water supplemented with ammonium (20 μM), but no increase was observed with nitrate (20 μM). Lack of a diel cycle, no decrease in haemolymph glucose levels, the expression patterns of glutamine synthetase in zooxanthellae and host, and the lack of glutamine release in response to nitrate supplementation all support the proposition that the increase in haemolymph glutamine is a product of the host and not the zooxanthellae. Unlike ammonium, nitrate accumulates rapidly in the haemolymph. It has no effect on the concentration of glutamine in the haemolymph, but there is an increase in arginine, histidine and lysine in the haemolymph, suggesting the release of these essential amino acids from zooxanthellae. Glutamine synthetase (GS) activity decreased markedly in the gill and less so in the mantle over a period of 6 d exposure to elevated ammonium (20 μM). In contrast, GS activity in zooxan- thellae doubled. The response of zooxanthellae in situ was confirmed by incubating freshly isolated zooxanthellae for 4 d in ammonium, which resulted in a ten-fold increase in GS activity. Comparison of the in situ response of zooxanthellae with that obtained in vitro indicates that the symbionts are likely to be exposed to ammonium concentrations lower than that found in the haemolymph. Received: 14 November 1997 / Accepted: 28 April 1998     

Light harvesting by wavelength transformation in a symbiotic coral of the Red Sea twilight zone

We report an extraordinary depth range for Leptoseris fragilis (Milne Edwards and Haime), a reef building coral of the Red Sea living in cytosymbiosis with zooxanthellae. The coral harbours an as yet unknown pigment system. We suggest that the heterotrophic host — the coral — provides its photoautotrophic symbionts with additional light. The supplementary light is provided by host pigments which transform light of short wavelengths into suitable wavelengths for photosynthesis, thus amplifying and increasing the transfer of photoassimilates from the zooxanthellae to the host.