UV absorption by mycosporine-like amino acids in Phaeocystis antarctica Karsten induced by photosynthetically available radiation

Mycosporine-like amino acids (MAAs), which occur in diverse taxonomic groups, exhibit in vivo absorption maxima between 310 nm and 360 nm and may play a photoprotective role against ultraviolet (UV) exposure. Using cultures of colonial Phaeocystis antarctica, we examined the relationship between MAA concentration, in vivo UV absorption, photoprotective (carotenoid) and photosynthetic pigments, and photosynthetically available radiation (PAR, 350–700 nm). UV absorption was high; chlorophyll-specific absorption, a ^* _ph, at 330 nm ranged from 0.06 to 0.41 m²/mg chlorophyll a. Values of a ^* _ph (330) were 4–13 times greater than a ^* _ph (676). Mycosporine-glycine, shinorine, and mycosporine-glycine valine are responsible for the strong in vivo UV absorption. The sum of all MAAs increased with irradiance when normalized to chlorophyll a or carbon concentrations, whereas individual MAAs varied independently from each other. Mycosporine-glycine concentrations showed no statistically significant change over the range of light intensities, whereas mycosporine-glycine and shinorine concentrations increased at higher irradiances. The relative fluorescence yield for chlorophyll a was low in the UV region compared to the visible region, implying that absorbed UV radiation (<375 nm) is transferred inefficiently to chlorophyll a in the reaction center. Quantitative estimates of UV screening by MAAs are attributed to elevated MAA concentrations and increased diameter at high light. Received: 31 March 1999 / Accepted: 13 July 2000     

Selective accumulation of mycosporine-like amino acids in ovaries of the green sea urchin Strongylocentrotus droebachiensis is not affected by ultraviolet radiation

Field sampling and laboratory experiments examined whether ultraviolet radiation (UVR) affects the reproduction or the accumulation of mycosporine-like amino acids (MAAs) and ascorbic acid in ovaries of the green sea urchin Strongylocentrotus droebachiensis (Müller). Ovaries of sea urchins sampled across a depth gradient (0.5–10 m) in March 1998 did not differ in their gonadal index, or in concentrations of MAAs, or ascorbic acid. Concentrations of MAAs and ascorbic acid in ovaries were higher in sea urchins collected from a kelp bed compared with those collected from a community of crustose coralline algae. The concentrations of MAAs in ovaries varied seasonally, peaking in March, when sea urchins had high gonadal indices just before spawning. Ovaries of sea urchins maintained on controlled diets from October 1997 to April 1998 accumulated significantly higher concentrations of MAAs when fed a diet rich in MAAs than did ovaries of sea urchins fed an alga lacking MAAs, but the gonadal indices did not differ between diets. Sea urchins accumulated principally one MAA, shinorine, but not others that were available in high concentrations in their diet. Neither the gonadal index nor the ovarian concentrations of MAAs were affected by daily exposure of adult urchins to UVR for 6 months. Concentrations of ascorbic acid in ovaries did not differ among diets or UV-treatments. The percentages of nutritive phagocytes and gametic cells were not affected by diet or UVR, and did not co-vary with concentrations of MAAs or ascorbic acid in ovaries. These data support previous demonstrations that female sea urchins accumulate MAAs from their diet of macroalgae, but further show that the accumulation is selective for specific MAAs, particularly shinorine, and that adult S. droebachiensis do not accumulate MAAs in their ovaries or eggs in response to UV-exposure. These are also the first experimental studies to address whether MAAs are affected by or regulate gametogenesis, and indicate that they do not. Received: 5 May 2000 / Accepted: 29 September 2000     

Mycosporine-like amino acids prevent UVB-induced abnormalities during early development of the green sea urchin Strongylocentrotus droebachiensis

Experiments were performed to determine how ultraviolet radiation (UVR) in the environmentally relevant range affects development of the sea urchin Strongylocentrotus droebachiensis (Müller) and whether mycosporine-like amino acids (MAAs), present in the early life stages, reduce UV-induced damage. Eggs, embryos, and larvae contained five MAAs having absorption maxima ranging from 320 to 334 nm. Eggs contained principally shinorine and porphyra-334, which absorb maximally at 334 nm and half-maximally at 312 and 348 nm, spanning much of the environmental range of biologically effective UVR. Concentrations of MAAs remained constant in unirradiated embryos through the gastrula stage, but decreased significantly in two-armed pluteus larvae. Daily exposure to combined photosynthetically active radiation (PAR, 400–700 nm) and UVR did not affect the concentration of MAAs in these embryos up to the two-armed pluteus stage. Prism larvae of sea urchins and the sand dollar Echinarachnius parma (Lamarck) did not accumulate shinorine from the surrounding seawater. Daily exposure of embryos to UVA (320–400 nm) and UVB (295–320 nm) radiation in the presence of PAR induced delays and abnormalities during development, and removing UVB eliminated this effect. Abnormalities in embryos included thickening of the blastoderm wall, filling of the blastocoel by abnormal cells, exogastrulation, and formation of abnormal spicules. The percentage of embryos that developed normally was lower in batches of embryos exposed to PAR + UVA + UVB, except in embryos from urchins maintained on MAA-rich diets. In all cases, the percentage of PAR + UVA + UVB-exposed embryos that developed normally was positively related to the concentration of MAAs in eggs from which the embryos developed. Thus, the MAAs found in S. droebachiensis embryos protect them against UVB-induced abnormalities during their development to at least the four-armed pluteus larval stage. Received: 8 May 2000 / Accepted: 29 September 2000     

Temporal and spatial occurrence of UV-absorbing mycosporine-like amino acids in tissues of the antarctic sea urchin Sterechinusneumayeri during springtime ozone-depletion

From September to November 1991, UV-absorbing mycosporine-like amino acids (MAAs) were monitored in a natural population of the sea urchin Sterechinusneumayeri from a coastal area of Anvers Island (Antarctic Peninsula). MAA concentrations were determined for specific tissues (gonad, digestive tract and body wall) from adults collected at four depths (intertidal, 8, 15 and 24 m). Four MAAs were identified: mycosporine-glycine, shinorine, porphyra-334 and paly-thine. Concentrations of MAAs among replicate individuals varied considerably. Ovaries had high concentrations of MAAs (84 to 1389 μg g⁻¹ dry wt), while testes had non-detectable levels. The relative abundance of specific MAAs in ovaries appeared to be related to the spawning cycle. Digestive-tract samples had MAA concentrations as high as 3000 μg g⁻¹ dry wt, but the mean MAA content in intertidal individuals decreased by 70% over 3 mo during spring. The body walls of sea urchins had very low amounts of MAAs (≤ 0.08 μg g⁻¹ dry wt). There were significant depth differences in the␣total MAA content of the ovary ( p <0.001), ( p <0.015), digestive tract ( p <0.001), and body wall with organisms from the intertidal and 8 m depth having the highest concentrations of MAAs. Biological dosimetry indicated that UV-B (280 to 320 nm) wavelengths penetrated 3 to 7 m below the sea ice during the study period. The total MAA content in ovaries decreased with depth on all sample dates; however, the MAA content of the digestive tract and body wall did not exhibit a consistent pattern of change with depth. The MAA content of tissues did not change significantly with the temporal gradient of light exposure that was established by both ozone depletion and increasing photoperiod, except in the digestive tract sampled from intertidal specimens. Adult urchins are probably well-protected from UV exposure by the water column and a calcareous test; however, the results of this study suggest that, even under ice cover, depth of habitation is a determinant of MAA content in S. neumayeri. Large daily and seasonal fluctuations in the light regime, which are characteristic of Antarctic coastal environments, apparently do not provide reliable cues to elicit a detectable, temporal, biochemical response. Received: 19 February 1997 / Accepted: 26 March 1997     

Mycosporine-like amino acids: possible UV protection in eggs of the sea hare Aplysia dactylomela

We investigated mycosporine amino acid (MAA) involvement as protective sunscreens in spawn of the sea hare Aplysia dactylomela to determine if adult diet and ultraviolet (UV) exposure affected the UV sensitivity of developing embryos. Adults were fed a red alga rich in MAAs (Acanthophora spicifera) or a green alga poor in MAAs (Ulva lactuca). Adults on each diet were exposed for 2 wk to ambient solar irradiance with two types of acrylic filters; one allowed exposure to wavelengths >275 nm (designated UV) and one to wavelengths only >410 nm (designated NOUV). Spawn from each adult group was likewise treated with UV or NOUV and monitored during development for differences in mortality and metabolic rate (measured as oxygen consumption: V˙ _O2). Also recorded were number of eggs or embryos per capsule, times to hatching, hatching success, size at hatching, and V˙ _O2 of adults. Spawn from adults eating red algae was almost twice as rich in MAAs as spawn from adults eating green algae, suggesting that MAA content is diet-related. Although overall quantities of MAAs in the spawn reflected MAA contents of the adult diet, specific MAAs were differentially sequestered in the spawn. Thus, porphyra-334, found in high concentration in Aplysia dactylomela's preferred red algal food, was present in only low concentration in the spawn. Conversely, mycosporine-glycine, in low concentration in red algal food, was the most abundant MAA in the spawn. UV treatment of adults had no effect on quantities of MAAs in the spawn. Adults exposed to UV had significantly higher V˙ _O2s and spawned twice as often. The UV-treated adults produced spawn with significantly higher V˙ _O2s and their embryos developed to hatching sooner. The only significant effect of UV exposure of the spawn was to reduce the percentage of veligers hatching from 71 to 50%. There was no significant effect on hatching time or size of the veligers at hatching, nor on number of eggs per capsule. Received: 13 May 1997 / Accepted: 27 June 1997     

UV-absorbing substances in zooxanthellate and azooxanthellate clams

The effects of UV-A and UV-B radiation on photosynthesis of zooxanthellae within the siphonal mantle of the giant clam, Tridacna crocea, and in isolation were studied. While UV-B irradiation (2.4 W m⁻², 20 min) completely suppressed photosynthesis of the isolated zooxanthellae, it had little effect on their photosynthetic ability if they were irradiated while within the siphonal mantle of the host tissue. Chemical analysis of the siphonal mantle of T. crocea showed the presence of significant amounts of mycosporine-like amino acids (MAAs), which absorb UV-A and -B light. However, no MAA was detected in the isolated zooxanthellae. MAAs were concentrated in the siphonal mantle and kidney tissues in comparison with other tissues. In the siphonal mantle, MAA concentrations were the highest in the outermost surface layer where most of the zooxanthella cells resided. This indicates that the zooxanthellae are protected from UV radiation by a screen of concentrated MAAs in the host clam. Aside from T. crocea, significant amounts of MAAs were found not only in other zooxanthellate clams, such as T. derasa, Hippopus hippopus, Colculum cardissa and Fragum unedo, but also in a closely related azooxanthellate clam, Vasticardium subrugosum. On the other hand, no MAA was detected in any of the zooxanthellae from these zooxanthellate clams. No MAA was detected in the tissues of a deep-sea bivalve, Calyptogena soyoae. Although MAAs seem to block strong UV radiation in the shallow-water clam, they are probably not essential for the clam's life in the dark. MAAs in shallow-water clams may be derived from food and accumulated in their tissues, especially in the siphonal mantle and kidney. Received: 29 November 1996 / Accepted: 13 January 1997     

Diel fluctuations of mycosporine-like amino acids in shallow-water scleractinian corals

Reports of bathymetric decrease in the total mycosporine-like amino acid (MAA) concentration of benthic invertebrates suggest that light gradients may be important determinants of MAA content. With the pronounced diel light changes, distinct temporal variations in MAA concentrations might also be expected. We examined the changes in the abundance of MAA in three shallow-water scleractinian corals, Pavona divaricata, Galaxea fascicularis and Montipora digitata from Okinawa, Japan, in relation to daily cycles in solar radiation and tested whether the species have different capabilities for protection against UVR depending on their MAA composition. The results show that symbiotic algae freshly isolated from the investigated coral species do not contain MAAs and that distribution of these compounds resided only within the animal tissue. Total MAA content in the tissue of P. divaricata, G. fascicularis and M. digitata rose rapidly at midday and significantly dropped at night. The observed variations were by a factor of two and, thus, very dramatic. For all the investigated coral species, total MAA concentrations were significantly correlated with the diurnal cycle in solar radiation, during both winter and summer seasons. Seawater temperature was significantly correlated with MAA levels only in the June experiment, but represented no more than 20% of the MAA variation in all three coral species, whereas solar radiation explained 60–70% of the MAA fluctuations. This suggests that MAAs are an integral component of the hard corals biochemical defense system against high solar irradiance stress. The diurnal increase in total MAA concentrations was due to an increase in the concentration of imino-MAA species of up to 2–2.5-fold of their pre-dawn values. In contrast, the oxocarbonyl-MAA mycosporine-glycine (Myc-Gly) showed the lowest (Tukey–Kramer HSD test: P<0.05) values at midday, compared to afternoon and night hours. Analysis of diel changes in chlorophyll fluorescence and chlorophyll a content of the investigated species revealed that P. divaricata and G. fascicularis were less sensitive to the high levels of ambient irradiance compared to M. digitata. In P. divaricata and G. fascicularis, Myc-Gly, an MAA with an antioxidant function, is the most abundant MAA, contributing about 70% to the total MAA pool, whereas the major MAA factions in M. digitata were represented by oxidatively robust imino-MAAs. We speculate that MAAs furnish scleractinian corals with protection from biologically damaging ultraviolet radiation through both the direct sunscreening activity of imino-MAAs and the antioxidant properties of oxocarbonyl-MAAs and suggest that the predominance, in the host tissue, of MAA species with an antioxidant ability may render corals more tolerant to high photosynthetically active and ultraviolet radiation.Communicated by T. Ikeda, Hakodate     

Interplay of host morphology and symbiont microhabitat in coral aggregations

The Belizean reef coral Agaricia tenuifolia Dana forms aggregations in which rows of thin, upright blades line up behind each other. On average, the spacing between blades increases with depth and hence with decreasing ambient irradiance. We designed and built a small, inexpensive light meter and used it to quantify the effect of branch spacing on light levels within colonies at varying distances from branch tips. Concurrently, we measured photosynthetic pigment concentrations and population densities of symbiotic dinoflagellates (zooxanthellae) extracted from coral branches of colonies with tight (≤3 cm) vs wide (≥6 cm) branch spacing, collected at 15 to 17 m and from colonies with tight branch spacing collected at 1 to 2 m. Light levels decreased significantly with tighter branch spacing and with distance from the branch tips. Total cellular pigment concentrations (chlorophylls a, c ₂ and peridinin) as well as chlorophyll a:c ₂ and chlorophyll a: peridinin ratios all increased significantly with distance from the branch tip, indicating very localized differences in photoacclimation within individual branches. Zooxanthellae from colonies with widely-spaced branches displayed significantly lower chlorophyll a:c ₂ and chlorophyll a:peridinin ratios, and were present at significantly higher population densities than those from colonies with tightly-spaced branches collected at the same depth (15 m). Tightly-spaced colonies collected from shallow environments (1 to 2 m) displayed pigment ratios similar to those from widely-spaced colonies from deeper water (15 m), but maintained zooxanthellae populations at levels similar to those in tightly-branched colonies from deeper water. Thus, variation in colony morphology (branch spacing and distance from branch tip) can affect symbiont physiology in a manner comparable to an increase of over 15 m of water depth. These results show that a host's morphology can strongly determine the microhabitat of its symbionts over very small spatial scales, and that zooxanthellae can in turn display steep gradients in concordance with these altered physical conditions. Received: 12 June 1997 / Accepted: 24 June 1997     

Dietary accumulation of UV-absorbing mycosporine-like amino acids (MAAs) by the green sea urchin (Strongylocentrotus droebachiensis)

The bioaccumulation of ultraviolet radiation-absorbing mycosporine-like amino acids (MAAs) by Strongylocentrotus droebachiensis (Müller) was determined in specimens maintained on MAA-rich and MAA-deficient diets. Individuals were fed either the red alga Mastocarpus stellatus (Stackhouse), which has a high concentration of the MAA shinorine (6.98 nmolmg^–1 dry wt), and trace amounts of the MAAs porphyra-334, asterina-330 and palythine, or the brown alga Laminaria saccharina (Lamouroux), which contains no detectable MAAs. Reproductively spent urchins were fed ad libitum during a 9 mo period in 1992 to 1993 until they were once again gravid. To monitor accumulation, samples of urchin tissues and ingested food were taken throughout the 9 mo period from males and females maintained on each diet, and the concentrations of MAAs determined using high performance liquid chromatography. Urchins maintained on a diet of M. stellatus showed an ovarian shinorine concentration (8.33 nmol mg^–1 dry wt) 25 times higher than those fed L. saccharina. There was no change in MAA content of testes or somatic tissues in either feeding group. More than 99% of the MAAs present in an experimental meal of M. stellatus were removed during passage through the gut. Previous studies have inferred dietary accumulation of MAAs by comparing MAAs present in animal tissues with the consumer's potential diet in the field. This is the first study to have monitored MAA accumulation in animal tissues in conjunction with a controlled diet having a known MAA composition. Concentrating MAAs in the ovaries may provide eggs released into the water column with protection from damaging solar ultraviolet radiation.     

Role of secondary metabolites and pigments in the epidermal tissues, ripe ovaries, viscera, gut contents and diet of the sea cucumber Holothuria atra

The epidermal tissues, ovaries, viscera, gut contents, and the diet of Holothuria atra (Jaeger), collected from the Great Barrier Reef (GBR), Australia, just prior to spawning, contained carotenoid pigments, mycosporine-like amino acids (MAAs) and three other metabolites in varying proportions. Mycosporine-glycine (mycosporine-gly), palythine, asterina-330, shinorine, mycosporine-2-gly, porphyra-334, and palythinol were present in the epidermal tissues. Mycosporine-gly was the major MAA in the epidermal tissues, gut contents and the diet, and the only MAA present in the ripe ovaries and the viscera. Gadusol, a metabolite with antioxidant properties, which co-occurs with MAAs, and in certain instances with trace amounts of 6-deoxygadusol in unfertilised and fertilised eggs and developing larvae of some marine invertebrates and vertebrates, was absent from H. atra. However, 6-deoxygadusol, with similar physical and chemical properties to gadusol, and the proposed biosynthetic precursor of MAAs, was present, but only in the ovaries. This is first report of the presence of this proposed biogenetic precursor of MAAs in significant amounts in a marine invertebrate. Approximately 90% of the total carotenoids of the epidermal tissues, ovaries and viscera of H. atra are highly oxidised, the main component being astaxanthin followed by canthaxanthin. These were the major carotenoids present in the ovaries and the total carotenoid content was highest in the ovaries. β-carotene, a common egg carotenoid, was present in trace quantities in the ovaries but, along with the xanthophylls lutein and zeaxanthin, it occurred in significant amounts in the gut contents and the diet of H. atra. Carotenoid patterns in the epidermal tissues and viscera were strikingly similar, containing β-carotene, canthaxanthin, astaxanthin, zeaxanthin, lutein, phoenicoxanthin and echinenone. The alkaloids homarine and isomeric trigonelline, (structurally unrelated to MAAs) and 6-deoxygadusol were present only in the ovaries and the viscera. It is suggested that these metabolites and pigments are either involved with photoprotection or reproduction, or associated with both processes. The origin, biogenetic relationships and the roles of these metabolites and pigments in H. atra are discussed. Received: 21 November 1997 / Accepted: 14 September 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     

Short-term variation and long-term changes in the oceanographic environment and zooplankton community in the vicinity of a sub-Antarctic archipelago

 Mesozooplankton community structure in the vicinity of the Prince Edward Islands (PEIs) was investigated during six surveys conducted in late austral summer (April/May) from 1996 to 1999. Zooplankton samples were collected by oblique tows using a Bongo net fitted with 300-μm mesh. Surface temperature, average temperature and chlorophyll a were measured in conjunction with each net tow. The positions of the Sub-Antarctic Front (SAF) and the Antarctic Polar Front (APF), in relation to the islands, were determined by CTD and/or XBT transects to the west of the islands (upstream). Both fronts were characterized by a high degree of latitudinal variation. Changes in position of the fronts occurred rapidly, the SAF moving up to ∼120 km in a 2-week period. Consequently, the oceanographic environment in the vicinity of the PEIs was subject to a high degree of intra- and inter-survey variation. The positions of the SAF and APF appeared to have a significant impact on phytoplankton biomass in the vicinity of the PEIs, possibly through the alteration of local oceanographic flow dynamics. Water retention over the island shelf in 1996, associated with location of the SAF far to the north of the PEIs, corresponded to enhanced chlorophyll-a concentrations (∼1.54 mg m⁻³). Conversely, when the fronts were close to the islands, as in 1997 and 1999, higher current velocity limited water retention and chlorophyll-a concentrations in the inter-island region were relatively low (∼0.4 mg m⁻³). Cluster analyses showed that, in many instances, there was greater similarity among zooplankton communities from different surveys than among communities within surveys, indicating that short-term variability exceeded inter-annual variability. The population structure of the copepod Calanus simillimus indicated that there was inter-annual variation in the timing of the biological season. Differences in the population structure of species, and consequently their contribution to abundance and biomass, may therefore have been an important contributor to inter-annual variation in community structure. Evidence is provided of a long-term southward shift in the position of the SAF. It is postulated that this may affect the PEIs by increasing the proportion of allochthonous energy input, because the PEIs now lie in the path of the front, altering the tropho-dynamics of the island ecosystem. Lower mesozooplankton biomass associated with warmer sub-Antarctic water may have important negative consequences for higher trophic levels that depend on mesozooplankton for food. Received: 10 June 2000 / Accepted: 22 September 2000     

Availability of ammonium influences photosynthesis and the accumulation of mycosporine-like amino acids in two<Emphasis Type="Italic"> Porphyra</Emphasis> species (Bangiales,Rhodophyta)

The effect of ammonium concentration on photosynthetic activity estimated as in vivo chlorophyll fluorescence, i.e. maximal quantum yield (F_v/F_m) and electron transport rate (ETR) and on the accumulation of mycosporine-like amino acids (MAAs), chlorophyll a (chl a), biliproteins (BP) and soluble proteins (SP) in the red algae Porphyra leucosticta Thuret in Le Jolis collected from Lagos (Málaga, Spain) and Porphyra umbilicalis (Linnaeus) J. Agardh from Helgoland (Germany) was evaluated. Discs of both species were incubated with three ammonium concentrations (0, 100 and 300 µM) under artificial PAR and UV radiation for 7 days. Photosynthetic activity decreased under the culture conditions due to UV radiation and ammonium availability. The decrease of both F_v/F_m and maximal ETR was related to ammonium supply, i.e. the lowest decrease occurred in algae growing with the highest concentration of ammonium. In both species, after 7 days of culture, the content of chl a, BP and SP was higher under 300 µM than that under 0 and 100 µM ammonium. In both species, the content of MAAs was increased under 300 µM ammonium compared to the initial value, whereas a decrease under 0 and 100 µM ammonium was observed only in P. leucosticta. The content of MAAs in P. umbilicalis did not present significant differences compared to the initial value, probably because of the high initial content of MAAs. In both Porphyra species, four MAAs were identified: shinorine, porphyra-334, palythine and asterina-330. However, P. leucosticta modified its MAA pattern during the incubation time, reaching the same percentages found for P. umbilicalis, which did not show any change during the experimental period. P. leucosticta exhibited a decrease in BP/SP and BP/chl a ratios through the incubation time and an increase in MAAs/BP. The ratio MAAs/chl a did not show any variation with time or treatment, as was also true for all ratios in P. umbilicalis. In summary, ammonium supply diminished the decrease of F_v/F_m, increased the content of photosynthetic pigments (chlorophyll and biliprotein) and soluble protein, and stimulated of the accumulation of MAAs in the red algae P. leucosticta and P. umbilicalis.Communicated by O. Kinne, Oldendorf/Luhe     

The oxygen microenvironment adjacent to the tissue of the scleractinian Dichocoenia stokesii and its effects on symbiont metabolism

The biology of symbiotic scleractinians is profoundly influenced by their intracellular zooxanthellae, and many studies have focused on the mechanistic basis of this influence. This has usually been accomplished by examining the metabolism of zooxanthellae under physical conditions measured in the open reef and assumed to be similar to conditions in hospite. Recent advances in the measurement of conditions near and within coral tissue suggests that this assumption may result in substantial errors. To address this possibility, the role of water flow in determining oxygen saturation adjacent to the tissue of Dichocoenia stokesii was investigated, and the effect of these measured oxygen saturations on the respiration and photosynthesis of zooxanthellae isolated from the same species was quantified. Using a microelectrode (700 μm diam), we measured oxygen saturations above (≤4 mm) the tissue in two flow speeds over 24 h periods in a flume receiving sunlight at in situ levels. The results were used as a proxy for ecologically relevant intracellular oxygen saturations, which were applied to zooxanthellae in vitro to assess their effect on symbiont metabolism. Microenvironment oxygen saturations (% air saturation) ranged from 74–159% in slow flow (2.7 cm s⁻¹) to 88–110% in faster flow (7.5 cm s⁻¹) over day–night cycles. Therefore, the metabolic rates of zooxanthellae were measured at 50 to 54% (hypoxia), 98 to 102% (normoxia) and 146 to 150% (hyperoxia) oxygen saturation. Oxygen saturation significantly affected the metabolism of zooxanthellae, with gross photosynthesis increasing 1.2-fold and dark respiration increasing 2-fold under hyperoxia compared to hypoxia. These results suggest that the metabolism of zooxanthellae in hospite is affected markedly by their microenvironment which, in turn, is influenced by flow-mediated mass transfer. Received: 13 July 1998 / Accepted: 30 April 1999     

Effect of sub-lethal damage to juvenile colonies of massive Porites spp. under contrasting regimes of temperature and water flow

In this study, juvenile colonies of massive Porites spp. (a combination of P. lutea and P. lobata) from the lagoon of Moorea (W 149°50′, S 17°30′) were damaged and exposed to contrasting conditions of temperature and flow to evaluate how damage and abiotic conditions interact to affect growth, physiological performance, and recovery. The experiment was conducted in April and May 2008 and consisted of two treatments in which corals were either undamaged (controls) or damaged through gouging of tissue and skeleton in a discrete spot mimicking the effects of corallivorous fishes that utilize an excavating feeding mode. The two groups of corals were incubated for 10 days in microcosms that crossed levels of temperature (26.7 and 29.6°C) and flow (6 and 21 cm s⁻¹), and the response assessed as overall colony growth (change in weight), dark-adapted quantum yield of PSII (F _v/F _m), and healing of the gouged areas. The influence of damage on growth was affected by temperature, but not by flow. When averaged across flow treatments, damage promoted growth by 25% at 26.7°C, but caused a 25% inhibition at 29.6°C. The damage also affected F _v/F _m in a pattern that differed between flow speeds, with a 10% reduction at 6 cm s⁻¹, but a 4% increase at 21 cm s⁻¹. Regardless of damage, F _v/F _m at 21 cm s⁻¹ was 11% lower at 26.7°C than at 29.6°C, but was unaffected by temperature at 6 cm s⁻¹. The lesions declined in area at similar rates (4–5% day⁻¹) under all conditions, although the tissue within them regained a normal appearance most rapidly at 26.7°C and 6 cm s⁻¹. These findings show that the response of poritid corals to sub-lethal damage is dependent partly on abiotic conditions, and they are consistent with the hypothesis that following damage, calcification and photosynthesis can compete for metabolites necessary for repair, with the outcome affected by flow-mediated mass transfer. These results may shed light upon the ways in which poritid corals respond to biting by certain corallivorous fishes.     

Effects of dissolved ammonium addition and host feeding with Artemia salina on photoacclimation of the hermatypic coral Stylophora pistillata

 Effects of nutrient treatments on photoacclimation of the hermatypic coral Stylophora pistillata (Esper) were studied. Studies on photoacclimation of colonies from different light regimes in the field were evaluated and used to design laboratory experiments. Coral colonies were collected in the Gulf of Eilat (Israel) from January to March 1993. Exterior branches of colonies from different depths (1 to 40 m) displayed different trends in production characteristics at reduced and very low levels of illumination. From 24 ± 3% to 12 ± 2% of incident surface photosynthetic active radiation (PAR_o), zooxanthella population density and chlorophyll a+c per 10⁶ zooxanthellae increased, a trend seen in the range of light levels optimal for coral growth (90 to 30% PAR_o). The P _max of CO₂ per 10⁶ zooxanthellae decreased, while P _max of CO₂ per 10³ polyps increased, indicating an increase in zooxanthella population density at low light levels. Proliferous zooxanthella frequency (PZF, a measure of zooxanthella division) declined significantly at light levels <18 ± 3% PAR_o. At the lowest levels of illumination (<5% PAR_o), zooxanthella population density decreased, as did the PZF; chl a+c per 10⁶ zooxanthellae was unchanged. In 28-d experiments, exterior coral branches from the upper surfaces of colonies from 3 m depth (65 ± 4% PAR_o) were incubated in aquaria under bright (80 to 90% PAR_o), reduced (20 to 30% PAR_o), and extremely low (2 to 4% PAR_o) light intensities. At each light intensity, the corals were maintained in three feeding treatments: sea water (SW); ammonium enriched SW (SW + N); SW with Artemia salina nauplii (SW + A). An increase in P _max of CO₂ per 10³ polyps was found in corals acclimated to reduced light (20 to 30% PAR_o) in nutrient-enriched SW, while in SW, where the increase in zooxanthella population density was smaller, it did not occur. Nutrient enrichments (SW + N at 2 to 4% PAR_o and SW + A at 20 to 30% PAR_o) increased zooxanthella population density, but had no effect on chl a+c per 10⁶ zooxanthellae. Acclimation for 14 d to reduced (10 to 20% PAR_o) and extremely low (1 to 3% PAR_o) light intensities shifted ¹⁴C photoassimilation into glycerol and other compounds (probably glycerides), rather than sugars. Both ammonium addition and feeding with Artemia salina nauplii resulted in an increase in photosynthetic assimilation of ¹⁴C into amino acids. We conclude that acclimation to reduced light consists of two processes: an increase in photosynthetic pigments and in zooxanthella population density. Both processes require nitrogen, the increase in zooxanthella population density needing more; this adaptation is therefore limited in nitrogen-poor sea water. Received: 19 June 1998 / Accepted: 13 June 2000     

Variability of stable isotopes and maximum linear extension in reef-coral skeletons at Kaneohe Bay, Hawaii

Stable-isotope and growth records of coral skeletons are often used to reconstruct tropical paleoclimate, yet few surveys have systematically examined the natural variability in coral skeletal ¹³C, ¹⁸O and maximum linear skeletal extension (MLSE) across depth. Here, interspecific, intraspecific, and geographical variations in coral skeletal ¹³C, ¹⁸O, and MLSE were examined in the corals Porites compressa, P. lobata, and Montipora verrucosa grown at 1.7, 5.0, and 8.3 m depth from August 1996 to March 1997 at The Point Reef and Patch Reef #41 field sites in Kaneohe Bay, Hawaii. Coral skeletal ¹³C values significantly decreased with depth and differed between species, but did not vary between field sites. ¹⁸O values were not significantly different across depth within a species, but did differ among species and field sites. High-resolution analysis of the intra-annual variation in skeletal ¹³C and ¹⁸O in P. compressa at 2.0 m depth confirms that these isotopes reflect changes in solar irradiance and temperature, respectively. Changes in MLSE across depth were consistent within, but highly variable among, species. Peak MLSE occurred at 1.7, 5.0, and 8.3 m for P. lobata, P. compressa, and M. verrucosa, respectively. Such interspecific variation in MLSE patterns may be attributable to one or more of the following: increases in zooplankton in the diet, changes in metabolic processes, or changes in growth form with depth. Overall, these results imply that natural inter- and intraspecific variability in coral skeletal ¹³C, ¹⁸O, and MLSE should be considered when interpreting and comparing coral-based tropical paleoclimate data from various coral species, depths, and field sites. Received: 6 October 1998 / Accepted: 8 July 1999     

Feeding and bacteriolytic responses of the deposit-feeder Abarenicola pacifica (Polychaeta: Arenicolidae) to changes in temperature and sediment food concentration

Deposit-feeders can respond to seasonal fluctuations in food concentration both functionally (e.g. by adjusting feeding rates) and physiologically (e.g. by changing the concentration of bacteriolytic agents in gut fluids). Laboratory feeding experiments were carried out (11 to 21 July 1997) with the arenicolid polychaete worm Abarenicola pacifica (Healy and Wells). Objectives were to test for separate and interactive effects of sediment food concentration and temperature (6, 11, and 16 °C) on deposit-feeder functional (feeding rates) and physiological (bacteriolytic activity of gut fluids) responses. Food concentration was varied experimentally using sieved (1 mm) natural sediments (Md φ=2.00; 0.6% organic) mixed with combusted (500 °C, 8 h) sediments for final concentrations of 25, 50, and 100% natural sediment. Sediment food quality was measured as: (1) bioavailable amino acids (EHAA), (2) chlorophyll a (chl a), and (3) bacterial abundance. Feeding rates were inferred from egestion rates (ER, g h⁻¹) and analyzed with respect to worm size. Bacteriolytic activity of midgut fluids was assayed turbidimetrically against two bacterial isolates, after worms had fed on experimental sediments for 15 d. Temperature and food concentration both significantly affected feeding rates, with maxima occurring at 50 and 100% natural sediment mixtures, and at high (16 °C) temperature. ER was positively, but not significantly correlated with EHAA and chl a; a positive, significant correlation was detected between ER and sediment bacterial abundance. Overall, functional responses agreed with earlier compensatory intake models for deposit-feeders. However, the size and direction of these responses was temperature-sensitive, suggesting that these models need to be adjusted for changes in absorption rates. No effects of ambient temperature or food concentration on bacteriolytic rates were observed, possibly due to compensatory mechanisms or the presence of multiple bacteriolytic agents in gut fluids. Received: 28 June 1999 / Accepted: 14 March 2000     

Population structure, growth and recruitment of the euryalinid brittle-star Astrobrachion constrictum (Echinodermata: Ophiuroidea) in Doubtful Sound, Fiordland, New Zealand

The euryalinid brittle-star (snake star) Astrobrachion constrictum (Farquhar) lives coiled around the branches of black coral (Antipathes fiordensis) colonies. Twenty-two vertical transects, 10 m wide by 30 m deep, were swum in Doubtful Sound over a 2.5 yr period from 1993 to 1995. Numbers, disc diameters and colour morphotypes of brittle-stars inhabiting coral colonies were recorded. 36.3% of the coral colonies >200 mm tall (n = 292) hosted ≥1 Astrobrachion constrictum (range 0 to 12). Overall, the population was patchily distributed on the available coral habitat. The dark red colour morphotype of A. constrictum was most common (87%, n = 279) followed by the yellow, striped, and then spotted varieties. The population was comprised mainly of large (≥10 mm disc diam) individuals, and juveniles were rarely encountered, indicating low rates of recruitment or a high mortality of recruits. Disc-diameter data gathered from this and previous studies indicated that growth in A. constrictum is initially rapid, with individuals reaching a disc diameter of 15 mm in ≃2.5 yr; growth decreases with age, as in other deep-sea ophiuroids. Growth rate within years, however, was not constant, with faster growth in the spring/summer. Maximum size for A. constrictum is reached in ≃8 yr at ˜23 mm disc diam. Anecdotal evidence indicates that A. constrictum may not be confined solely to black coral colonies. Received: 25 September 1996 / Accepted: 16 October 1996     

A novel symbiosis between a cyanobacterium, Synechococcus sp., an aplastidic protist, Solenicola setigera, and a diatom, Leptocylindrus mediterraneus, in the open ocean

The abundance, biomass and distribution of Solenicola setigera, a colonial heterotrophic protist found only with the centric chain-forming diatom Leptocylindusmediterraneus, are reported for four major ocean basins. The distribution is cosmopolitan, and abundances and biomass are usually low (<500 colonies l⁻¹); however, in the summer of 1993, we observed a major biomass component (range = 5 to 31 μg C l⁻¹) in the surface waters of the North Atlantic attributable to S. setigera. These colonies of S. setigera were exceptionally large, and unusual in possessing high abundances of Synechococcus sp., a normally solitary cyanobacterium, embedded in the matrix covering the cells. We hypothesize that this relationship was mutually beneficial for both Solenicola setigera and Synechococcus sp. Received: 5 January 1998 / Accepted: 22 May 1998