ALBERT

Notes: Abstract Pocillopora damicornis (Linnaeus) is a ubiquitous branching coral found throughout the Indo-Pacific region. Like many other species of coral, P. damicornis displays a large range of morphologies. At One Tree Island, it occurs as two distinct morphs that are easily distinguished by the presence or absence of pink pigmentation. The two colour morphs of P. damicornis were found to differ in their distribution and abundance in the One Tree Island Lagoon. The brown morph was more abudant than the pink morph in the shallows (〈1 m),whereas the pink morph was more abundant at deeper sites (〉3 m). The two morphs also differed physiologically. The brown morph tended to have a greater calcification rate than the pink morph, regardless of environmental conditions. However, the difference in the calcification rate between the two morphs became non-significant under shaded conditions (5% full sunlight), indicating some degree of physiological plasticity of the morphs. The pink colour in P. damicornis was due to a hydrophilic pigment with a major peak absorbance at 560 nm. The expression of pink pigment had both genetic and phenotypic components. The brown morph has a reduced genetic capacity to express the pigment relative to the pink morph. On the other hand, pigment expression could be induced by light in the pink morph. Although genetic differences ultimately determine the differences between the two morphs of P. damicornis, the extent of pigment expression is under some degree of environmental influence.

Notes: Abstract The life-history of the crown-of thorns starfish (Acanthaster planci) includes a planktotrophic larva that is capable of feeding on particulate food. It has been proposed, however, that particulate food (e.g. microalgae) is scarce in tropical water columns relative to the nutritional requirements of the larvae of A. planci, and that periodic shortages of food play an important role in the biology of this species. It has also been proposed that non-particulate sources of nutrition (e.g. dissolved organic matter, DOM) may fuel part of the nutritional requirements of the larval development of A. planci as well. The present study addresses the ability of A. planci larvae to take up several DOM species and compares rates of DOM uptake to the energy requirements of the larvae. Substrates transported in this study have been previously reported to be transported by larval asteroids from temperate and antarctic waters. Transport rates (per larval A. planci) increased steadily during larval development and some substrates had among the highest mass-specific transport rates ever reported for invertebrate larvae. Maximum transport rates (J max in) for alanine increased from 15.5 pmol larva−1 h−1 (13.2 pmol μg−1 h−1) for gastrulas (J max in=38.7 pmol larva−1 h−1 or 47.4 pmol μg−1 h−1) to 35.0 pmol larva−1 h−1 (13.1 pmol μg−1 h−1) for early brachiolaria (J max in just prior to settlement=350.0 pmol larva−1 h−1 or 161.1 pmol μg−1 h−1) at 1 μM substrate concentrations. The instantaneous metabolic demand for substrates by gastrula, bipinnaria and brachiolaria stage larvae could be completely satisfied by alanine concentrations of 11, 1.6 and 0.8 μM, respectively. Similar rates were measured in this study for the essential amino acid leucine, with rates increasing from 11.0 pmol larva−1 h−1 (or 9.4 pmol μg−1 h−1) for gastrulas (J max in=110.5 pmol larva−1 h−1 or 94.4 pmol μg−1 h−1) to 34.0 pmol larva−1 h−1 (or 13.0 pmol μg−1 h−1) for late brachiolaria (J max in=288.9 pmol larva−1 h−1 or 110.3 pmol μg−1 h−1) at 1 μM substrate concentrations. The essential amino acid histidine was transported at lower rates (1.6 pmol μg−1 h−1 at 1 μM for late brachiolaria). Calculation of the energy contribution of the transported species revealed that larvae of A. planci can potentially satisfy 0.6, 18.7, 29.9 and 3.3% of their total energy requirements (instantaneous energy demand plus energy added to larvae as biomass) during embryonic and larval development from external concentrations of 1 μM of glucose, alanine, leucine and histidine, respectively. These data demonstrate that a relatively minor component of the DOM pool in seawater (dissolved free amino acids, DFAA) can potentially provide significant amounts of energy for the growth and development of A. planci during larval development.

Notes: Abstract The reproductive cycle of the sea urchin Centrostephanus rodgersii (Agassiz) was investigated in two populations, at Clovelly and Little Bay, in Sydney, New South Wales, Australia. C. rodgersii were collected at monthly intervals from February 1992 through January 1993. The reproductive cycle was determined by histological examination of oogenesis and spermatogenesis, monthly measurements of gonad index (GI), and induction of spawning by KCl injection. C. rodgersii has an annual reproductive cycle that was highly synchronous in both populations. From February to June, gametogenesis was accompanied by a decline in the amount of nutritive tissue in the gonads. The urchins were mature from June to September, with peak spawning between July and August, as indicated by a significant drop in GI. The breeding season of C. rodgersii therefore coincides with the lowest sea temperatures and the shortest days of the year. The gonads returned to the recovering condition within a month of spawning, with a substantial thickening of the nutritive layer along the gonad wall, and the GI returned to near pre-spawning levels. As a result, the spent phase was rarely found in C. rodgersii. With the exception of a significant decrease in the GI following spawning of urchins from the Clovelly population, the GI measurements did not show any distinct pattern through time. Specimens spawned in response to KCl injection from mid-May to early October, with the maximum response in July. Although all individuals sampled were at a similar stage of maturity at any one time, inter-site differences were seen with all of the methods used. Gonad indices from Little Bay were consistently higher and less variable than those from Clovelly for most of the year. The Little Bay population could also be induced to spawn for a longer period of time than could the Clovelly population. The breeding season of the Little Bay population appears to be longer than that of the Clovelly population. The relationship between size and sexual maturity was also examined. All C. rodgersii with a test diameter of 〉60 mm could be induced to spawn and produced viable gametes. C. rodgersii has been nominated for commercial exploitation in New South Wales, and the results of this investigation are used to make recommendations on the timing and size limits for a fishery utilising this species.

Notes: Abstract 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).

Notes: Abstract Symbiotic Aiptasia pulchella and freshly isolated zooxanthellae were incubated in NaH14CO3 and NH4Cl for 1 to 240 min, and samples were analysed by reverse-phase high-performance liquid chromatography (HPLC) and an online radiochemical detector. NH4 + was first assimilated into 14C-glutamate and 14C-glutamine in the zooxanthellae residing in A. pulchella. The specific activities (dpm nmol−1) of 14C-glutamate and 14C-glutamine in vivo, were far greater in the zooxanthellae than in the host tissue, indicating that NH4 + was principally incorporated into the glutamate and glutamine pools of the zooxanthellae. 14C-α-ketoglutarate was taken up from the medium by intact A. pulchella and assimilated into a small amount of 14C-glutamate in the host tissue, but no 14C-glutamine was detected in the host fraction. The 14C-glutamate that was synthesized was most likely produced from transamination reactions as opposed to the direct assimilation of NH4 +. The free amino acid composition of the host tissue and zooxanthellae of A. pulchella was also measured. The results presented here demonstrate that NH4 + was initially assimilated by the zooxanthellae of A. pulchella.

Notes: Abstract 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.

Notes: Abstract The separate and combined effects of ammonium (10μM) and phosphate (2μM) 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 μm2; C=47.9 μm2; P=43.2μm2; N+P=44.5 μm2; (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 μm2; C=2.0 μm2; P=1.8 μm2; N+P=1.2 μm2; (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.

Notes: Abstract 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.

Notes: The photochemical efficiency of symbiotic dinoflagellates within the tissues of two reef-building corals in response to normal and excess irradiance at water temperatures 〈 30 °C were investigated using pulse amplitude modulated (PAM) chlorophyll fluorescence techniques. Dark-adapted Fv/Fm showed clear diurnal changes, decreasing to a low at solar noon and increasing in the afternoon. However, Fv/Fm also drifted downwards at night or in prolonged darkness, and increased rapidly during the early morning twilight. This parameter also increased when the oxygen concentration of the water holding the corals was increased. Such changes have not been described previously, and most probably reflect state transitions associated with PQ pool reduction via chlororespiration. These unusual characteristics may be a feature of an endosymbiotic environment, reflective of the well-documented night-time tissue hypoxia that occurs in corals. Fv/Fm decreased to 0·25 in response to full sunlight in shade-acclimated (shade) colonies of Stylophora pistillata, which is considerably lower than in light-acclimated (sun) colonies. In sun colonies, the reversible decrease in Fv/Fm was caused by a lowering of Fm and Fo suggesting photoprotection and no lasting damage. The decrease in Fv/Fm, however, was caused by a decrease in Fm and an increase in Fo in shade colonies suggesting photoinactivation and long-term cumulative photoinhibition. Shade colonies rapidly lost their symbiotic algae (bleached) during exposure to full sunlight. This study is consistent with the hypothesis that excess light leads to chronic damage of symbiotic dinoflagellates and their eventual removal from reef-building corals. It is significant that this can occur with high light conditions alone.

Notes: The early effects of heat stress on the photosynthesis of symbiotic dinoflagellates (zooxanthellae) within the tissues of a reef-building coral were examined using pulse-amplitude-modulated (PAM) chlorophyll fluorescence and photorespirometry. Exposure of Stylophora pistillata to 33 and 34 °C for 4 h resulted in (1) the development of strong non-photochemical quenching (qN) of the chlorophyll fluorescence signal, (2) marked decreases in photosynthetic oxygen evolution, and (3) decreases in optimal quantum yield (Fv/Fm) of photosystem II (PSII). Quantum yield decreased to a greater extent on the illuminated surfaces of coral branches than on lower (shaded) surfaces, and also when high irradiance intensities were combined with elevated temperature (33 °C as opposed to 28 °C). qN collapsed in heat-stressed samples when quenching analysis was conducted in the absence of oxygen. Collectively, these observations are interpreted as the initiation of photoprotective dissipation of excess absorbed energy as heat (qN) and O2-dependent electron flow through the Mehler-Ascorbate-Peroxidase cycle (MAP-cycle) following the point at which the rate of light-driven electron transport exceeds the capacity of the Calvin cycle. A model for coral bleaching is proposed whereby the primary site of heat damage in S. pistillata is carboxylation within the Calvin cycle, as has been observed during heat damage in higher plants. Damage to PSII and a reduction in Fv/Fm (i.e. photoinhibition) are secondary effects following the overwhelming of photoprotective mechanisms by light. This secondary factor increases the effect of the primary variable, temperature. Potential restrictions of electron flow in heat-stressed zooxanthellae are discussed with respect to Calvin cycle enzymes and the unusual status of the dinoflagellate Rubisco. Significant features of our model are that (1) damage to PSII is not the initial step in the sequence of heat stress in zooxanthellae, and (2) light plays a key secondary role in the initiation of the bleaching phenomena.