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     

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     

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.     

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     

Effects of ultraviolet radiation and water motion on the reef coral Porites compressa Dana: a flume experiment

The effects of water flow and ultraviolet radiation (UVR, 280–400 nm) on the reef coral Porites compressa Dana were explored in a manipulative flume experiment. The aim of this study was to determine whether this coral responds to changes in the UVR environment by adjusting the tissue concentration of UV-absorbing compounds (mycosporine-like amino acids, MAAs), and to see whether such an acclimation is affected by water flow. Also, calcification rate and chlorophyll-a concentration were measured during the experiment to estimate the potential costs (in the form of slowed growth and/or reduced photosynthetic capacity) to the coral–alga symbiosis of being exposed to UVR and producing MAAs. Branches of P. compressa from a single male colony were exposed to high or low flow (15 cm s⁻¹ and 3 cm s⁻¹, respectively) and ambient or no UVR in an outdoor, continuous-flow seawater system. Chlorophyll-a and MAA concentrations were determined after zero, 3 and 6 weeks of exposure to the experimental conditions. Increase in buoyant weight during the two 3-week periods was used to calculate calcification rate. The presence of UVR had a significant positive effect on total MAA concentration in the P. compressa colonies; however, there were significant interactions present. In colonies exposed to UVR, MAA concentration increased and then decreased to initial levels in high water flow, and increased steadily in low water flow. In colonies receiving no UVR, MAA concentration decreased steadily, declining 23% in 6 weeks. The absence of UVR did not result in higher chlorophyll-a concentrations, but the calcification rate was slightly affected by UVR. This study supports the putative photoprotective role of MAAs in P. compressa, and suggests that the costs of mitigating the effects of ambient UVR are detectable, but they are very small. Received: 29 February 2000 / Accepted: 20 September 2000     

UV induces reactive oxygen species,damages sperm,and impairs fertilisation in the sea urchin <Emphasis Type="Italic">Anthocidaris crassispina</Emphasis>

We demonstrated that environmentally relevant levels of UVA and UVB can reduce sperm motility (UVA: by 38–58%; UVB: by 42–85%; P < 0.05) and subsequently fertilisation success (UVA: by 38–72%; UVB: by 91–98%; P < 0.05) of urchins (Anthocidaris crassispina) in a dose-dependent manner, implicating that recruitment of urchin populations might be reduced by UVR (ultraviolet radiation) prevailing in their natural habitats. Concomitantly, reactive oxygen species (ROS) production was enhanced by UVA and UVB in a dose-dependent manner (UVA: by 1.3-fold; UVB: by 6.6–7.3-fold; P < 0.05), and the increase in ROS resulted in an increase in lipid peroxidation (LPO) in urchin sperm (UVA: by 4.2–7.2-fold; UVB: by 2.3–2.7-fold; P < 0.05). This study demonstrated that ROS production and oxidative damages enhanced by UVR may account for the observed declines in sperm motility and fertilisation, and suggests that levels of UVR prevailing in the environment may pose a significant threat to the reproductive success of natural populations of urchins spawning in shallow waters.     

UVR-induced photoinhibition of summer marine phytoplankton communities from Patagonia

During austral summer 2006, experiments were carried out to evaluate the effects of ultraviolet radiation (UVR, 280–400 nm) on carbon fixation of natural phytoplankton assemblages from Patagonia (Argentina). Surface water samples were collected (ca. 100 m offshore) at mid morning using an acid-cleaned (1 N HCl) dark container. The short-term impact of UVR (measured as radiocarbon incorporation) was immediately assessed by exposing samples to three artificial illumination treatments: PAR (400–700 nm), PAR + UV-A (320–700 nm), and PAR + UV-A + UV-B (280–700 nm). Pico-nanoplankton characterized the assemblages, and taxon-specific pigment fingerprinting combined with CHEMTAX and supplemented with microscopic observations showed varied proportions of diatoms, chlorophytes, and cyanobacteria throughout January–February 2006. Photosynthetic efficiency, as assessed through assimilation numbers, was high [between 4.4 and 10.4 μg C (μg chl-a)⁻¹ h⁻¹], and it was probably favored by the supply of inorganic nutrients from the Chubut River. UVR-induced photoinhibition appeared to be related to the taxonomic composition: in general, higher photoinhibition was observed when diatoms dominated, whereas this was lower when samples were dominated by chlorophytes. Our data suggest that xanthophyll pigments might have provided only limited protection in these already highlighted acclimated assemblages.     

Two new UV-absorbing mycosporine-like amino acids from the sea anemoneAnthopleura elegantissima and the effects of zooxanthellae and spectral irradiance on chemical composition and content

Many tropical cnidarians living in shallow water contain a class of ultraviolet-A (UV-A, 320 to 400 nm) and ultraviolet-B (UV-B, 280 to 320 nm) absorbing compounds known as mycosporine-like amino acids (MAAs). These compounds may provide protection from the deleterious effects of solar UV radiation. Using a novel application of reverse-phase high performance liquid chromatography, we find that the temperate sea anemoneAnthopleura elegantissima (collected in 1988 from Bodega Bay, California, and in 1991 from Santa Barbara, California) contains four major MAAs: shinorine, porphyra-334, and two new compounds, mycosporine-taurine and mycosporine-2 glycine. Analysis of zooxanthellate (containing zooxanthellae) and naturally apozooxanthellate (lacking zooxanthellae) specimens acclimated in the presence and absence of UV for 28 d in the spring of 1988 suggests that this anemone, unlike some other anthozoans, does not regulate the concentration of its MAAs in response to UV radiation. The presence of similar concentration of MAAs in apozooxanthellate and zooxanthellate specimens indicates that symbiosis with algae is not required for these compounds to be present in the anemone. The total concentration of MAAs in the zooxanthellae is only about 12% of that in their host's tissues.     

Ultraviolet radiation effects on the behavior and recruitment of larvae from the reef coral Porites astreoides

We tested the rarely considered hypothesis that the ultraviolet portion (UVR, 280–400 nm) of the light spectrum affects patterns of recruitment in reef-building corals. The premise for this hypothesis rests in the fact that biologically relevant intensities of UVR penetrate to considerable depths (>24 m) in the clear waters surrounding many coral reefs, and that reef organisms allocate substantial resources to prevent and repair UVR damage. The ability of larvae spawned by the brown morph of the Caribbean coral, Porites astreoides, to detect and avoid UVR was assessed in petri dishes where one-half of the dish was shielded from UVR and the other exposed. Observations made every 30 min between 10:30 and 13:30 h showed significantly higher densities of larvae swimming in regions shielded from UVR. To determine how this behavior affects settlement patterns, larvae collected from P. astreoides adults at 18 m depth were released into chambers deployed at 17 m depth where they were given a choice of three different light regions in which to settle: PAR (PAR=400–700 nm), PAR+UVAR (UVAR=320–400 nm), and PAR+UVAR+UVBR (UVBR=280–320 nm). At the end of the experiment, greater numbers of P. astreoides larvae had settled in the region of the tube where UVR was reduced than would be expected if dispersion were random. To our knowledge, this is the first demonstration in any reef-building coral species that planula larvae can detect UVR and that it affects their choice of a settlement site. These results indicate that the capacity to detect and avoid habitats with biologically damaging levels of UVR may be one factor contributing to the successful recruitment of coral larvae.     

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     

Effects of food ration and feeding regime on growth and reproduction of the sea urchin Strongylocentrotus droebachiensis

To determine the effects food ration and feeding regime on growth and reproduction of Strongylocentrotus droebachiensis (Müller), sea urchins in laboratory aquaria were fed kelp (Laminaria longicruris) supplied at either a high (H, ad libidum daily) or a low (L, ad libidum 1 d wk⁻¹) ration in two successive 12-wk intervals during the reproductive period. After 24 wk, urchins fed the high ration continuously (HH) or for the last 12 wk only (LH) had a significantly greater mean gonad index [(gonad weight/total body weight) × 100] and body weight than urchins fed the low ration continuously (LL) or for the last 12 wk only (HL). Urchins in the HL treatment had a significantly greater gonad index than those in the LL treatment; there was no significant difference in gonad index between the LH and HH treatments. Females had a greater gonad index than males in the low ration (LL and HL) treatments at the end of the experiment; there was no significant difference between sexes in the high ration (LH, HH) treatments. Gametogenesis proceeded to maturation in all treatments and some individuals spawned at the end of the experiment. Females in the high ration (HH and LH) treatments had a greater proportion of nutritive phagocytes in their ovaries than females in the low ration treatments, but there was no effect of feeding treatment on oocyte or ovum size. Feeding treatment had no effect on the relative abundance of nutritive phagocytes in the testes, although the proportion of spermatocytes was higher (and that of spermatozoa lower) in the high ration than in the low ration treatments. Urchins in the high ration treatments had a lower mean jaw height index [(jaw height/test diameter) × 100] and greater mean test diameter than those in the low ration treatments at the end of the experiment, although these differences were not statistically significant. Feeding rate on kelp at the end of the experiment was significantly greater for urchins in the low ration than in the high ration treatments. Our experimental results show that even relatively low rations of kelp support somatic and gonadal growth in S. droebachiensis. Increasing the supply of kelp, particularly during the period of active gametogenesis, results in maximal rates of growth and reproduction. These results suggest that populations of S. droebachiensis in barrens may derive a substantial proportion of their nutrition from drift kelp, which may contribute to their persistence in these habitats. They also explain the large body size, high reproductive effort and fecundity of urchins grazing on kelp beds. These findings have important implications for understanding the dynamics of natural populations of S. droebachiensis and for development of effective aquacultural practices. Received: 17 February 1997 / Accepted: 5 March 1997     

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     

Effects of food type and ration on reproductive maturation and growth of the sea urchin Strongylocentrotus droebachiensis

We investigated the effects of food quality and quantity on reproductive maturation and growth of juveniles of Strongylocentrotus droebachiensis (Müller) in a 22 month laboratory experiment in which we fed sea urchins four diets: (1) kelp (Laminaria spp.) for 6 d wk⁻¹ and mussel (Mytilus spp.) flesh for 1 d wk⁻¹ (KM); (2) kelp for 7 d wk⁻¹ (high ration, KH); (3) kelp for 1 d wk⁻¹ (low ration, KL) and (4) no food other than encrusting coralline algae (NF). At their first and second opportunity for reproduction, all sea urchins in the KM and KH treatments, and most in the KL treatment were reproductively mature, whereas all sea urchins in the NF treatment remained immature. Gonad index differed significantly among all fed treatments at first and second reproduction, and was highest in the KM and lowest in the KL treatment. Gonad index was similar in both sexes at first reproduction, but it was higher in females than in males at second reproduction. Diet had little or no effect on the relative abundance of spermatocytes, spermatozoa, or nutritive phagocytes in testes at first and second reproduction. In ovaries, nutritive phagocytes were significantly more abundant in females in the KM and KH treatments than in the KL treatment at first reproduction, and significantly more abundant in unfed (NF) than fed (KM, KH, KL) females at second reproduction. Mean oocyte size was similar in all fed females at first reproduction, but significantly larger in fed than unfed females at second reproduction. Mean ovum size was similar in all fed females in both reproductive periods. Increase in test diameter was greatest in the KM treatment and smallest in the KL treatment; sea urchins in the NF treatment decreased slightly in size. Survival was 95 to 100% in all fed sea urchins but significantly lower in unfed ones. The feeding rate on kelp was significantly greater in the KL than the KM and KH treatments. In the KM treatment, the feeding rate on kelp increased significantly over a 6 d period after mussel flesh was provided. Our results demonstrate that a diet of high food quality and quantity accelerates reproductive maturation and growth rate, and enhances gonad production and survival in juvenile and young adult S. droebachiensis. These findings contribute to our understanding of the reproductive ecology of S. droebachiensis in habitats with differing food supplies (e.g., kelp beds and barrens). Our results also can be used to improve aquacultural practices for sea urchins. Received: 3 June 1998 / Accepted: 2 February 1999     

DNA damage induced by ultraviolet radiation in coral-reef microbial communities

Ultraviolet radiation (UVR) has been implicated in coral-bleaching processes and UVR may create stress to marine organisms by damage to DNA. Absorption of energy from UVB (280 to 320 nm) induces direct damage to DNA via cyclobutane pyrimidine dimer photoproduct-formation. We examined the extent of DNA damage created by UVR in coral reef microbial communities and whether the coral-surface microlayer (CSM) provides protection from UVR to the microorganisms found there. Diel patterns and depth profiles of UVR effects were examined in coral mucus (coral-surface microlayer, CSM) from Montastraea faveolata and Colpophyllia natans, and water-column samples of similar depths. UV-induced photodamage was determined using a radioimmunoassay specific for cyclobutane pyrimidine dimers (thymine dimers). Significant photodamage was detected in water-column and CSM samples, although the level of damage in CSM samples was consistently lower than in water-column samples collected from the same depth, suggesting the presence of photoprotective mechanisms within the CSM. Diel patterns of photodamage were detected in both water-column and CSM samples, but peak damage occurred earlier in the day for the CSM samples, suggesting differences in damage and repair kinetics between the water column and CSM. The results suggest that microorganisms within the CSM are afforded some protection from UVR stress and that changes in the amount of DNA damage in these organisms may be an indicator of changing UVR stress to corals. Received: 10 January 1997 / Accepted: 15 September 1997     

Vertical distribution of fish larvae in the Great Barrier Reef Lagoon,Australia

To investigate the natural defenses of Antarctic marine organisms against exposure to ultraviolet (UV) radiation (280 to 320 nm), 57 species (1 fish, 48 invertebrates, and 8 algae) were collected during austral spring 1988 in the vicinity of Palmer Station (Anvers Island, Antarctic Peninsula) and were analyzed for the presence of mycosporine-like amino acids (MAAs), compounds that absorb UV radiation and may provide shielding from these biologically hazardous wavelengths. Nearly 90% of the 57 species examined contained MAAs, and eight specific MAA compounds were identified. Seven of these (palythine, porphyra-334, shinorine, mycosporineglycine, palythene, asterina-330, and palythinol) have been observed previously in marine organisms from temperate and tropical latitudes. A new MAA, mycosporineglycine: valine, was found in the Antarctic fish and in 38 of the invertebrate species examined. This study confirms widespread occurrence of MAAs in Antarctic marine organisms and suggests that these species have some degree of natural biochemical protection from UV exposure.Contribution No. 502 from the Australian Institute of Marine Science     

A method for the assessment of light-induced oxidative stress in embryos of fucoid algae via confocal laserscan microscopy

A method was developed for measurement of active oxygen production in embryonic stages of the brown seaweed Fucus spiralis, using the label CM-DCFH-DA. Active oxygen species convert the label into the green fluorescent CM-DCF (exc/em 488/530 nm) that is detected via confocal laserscan microscopy and quantitative image analysis. Loading of the label did not harm the embryos; loading efficiency was age-independent, and the esterases needed for conversion to CM-DCFH were not inhibited by the effective UV dose (2 W m⁻²) applied in the experiments. After correction for daily variation of the laser power, and calibration with DCF standard solutions, this automated analysis of confocal images rendered active oxygen concentrations in fucoid embryos (μM DCF). An experiment was designed for the assessment of active oxygen production following irradiance stress in the light-sensitive embryos. Dim-light-acclimated, 1-, 2- and 4-day-old embryos were transferred for 60 min to defined high-light conditions (4π-irradiance 300 μmol photons m⁻² s⁻¹), optionally without UV radiation, including UVA, or including UVA plus UVB. PSII yield measurements (PAM fluorometer) were carried out in order to assess the degree of photoinhibition under these light conditions. The imposed light stress initially caused a rapid decrease of the PSII yields (Φ_P). With increasing embryo age, minimum Φ_P values attained under light stress remained higher. Consequently, electron transport rates (ETR) would increase with embryo age, i.e., with the development of their photosynthetic apparatus. Active oxygen production increased with ETR, and when UVB was included, relatively greater amounts of active oxygen were produced. A slow, second-phase decrease of Φ_P under light stress that was proportional to active oxygen production indicated that some photooxidative damage was caused, in particular during UVB exposure. Recovery from light stress was a rapid process in the absence of UVB; in such cases Φ_P was almost restored to the initial values within 60 min. The relative state of recovery of Φ_P was correlated with both the effective UV dose and active oxygen production rate (DCF). Recovery was slowest in embryos exposed for 60 min to an experimental UVB dose, which was representative of a situation at low tide, on a sunny day. The results suggest that active oxygen may cause an in situ inhibition of growth of the earliest life stages of F. spiralis. Received: 26 January 2000 / Accepted: 4 September 2000     

Biological weighting of ultraviolet (280–400 nm) induced mortality in marine zooplankton and fish. I. Atlantic cod (Gadus morhua) eggs

In the Gulf of St. Lawrence, Canada, productivity-determining biophysical interactions occur in the upper 0 to 30 m of the water column. The eggs and larvae of several commercially important marine invertebrates and fishes (e.g. Gadus morhua L.) are found in this layer. Measurements of the diffuse attenuation coefficients for ultraviolet-B radiation (280 to 320 nm, UV-B) at various locations in this geographic region indicated maximum 10% depths (the depth to which 10% of the surface energy penetrates at a given wavelength) of 3 to 4 m at a wavelength of 310 nm. This represents a significant percentage of the summer mixed-layer water column: organisms residing in this layer are exposed to UV-B radiation. Laboratory experiments using a Xenon-arc-lamp based solar simulator revealed that cod embryos exposed to UV-B exhibited high wavelength-dependent mortality. The strongest effects occurred under exposures to wavelengths below 312 nm. This susceptibility was also dependent upon developmental stage; mortality was particularly high during gastrulation. At the shorter wavelengths (<305 nm) UV-B-induced mortality was strongly dose-dependent, and not significantly influenced by dose-rate. The biological weighting function (BWF) derived for UV-B-induced mortality in cod eggs is similar to that reported for naked DNA – suggesting that the mortality is a direct result of DNA damage. There was no evidence of a detrimental effect of ultraviolet-A radiation (320 to 400 nm). Calculations based upon the BWF indicate that, under current noon surface irradiance, 50% of cod eggs located at or very near (within 10 cm) the ocean surface will be dead after 42 h of exposure. Under solar spectral irradiance simulating a 20% decrease in ozone layer thickness, this time drops to 32 h. These are first-order estimates based upon surface irradiance taken at a time of day during which the values would be maximal. Nonetheless, they illustrate the relative changes in UV-B impacts that will result from ozone layer depletions expected over the coming decades. It is also important to point out that variability in cloud cover, water quality, and vertical distribution and displacement of cod eggs and larvae within the mixed layer, can all have a greater effect on the flux of UV-B radiation to which fish eggs are exposed than will ozone layer depletion at these latitudes. Received: 2 March 1998 / Accepted: 18 December 1998     

Density-related reproductive trade-offs in the green sea urchin, Strongylocentrotus droebachiensis

Green sea urchin, Strongylocentrotus droebachiensis (O.F. Müller), populations are being depleted rapidly in the Gulf of Maine and there is justified concern that the potential of this free-spawner to produce larvae may be severely inhibited. We evaluated the opposing effects of different population densities on gonad development and fertilization success, using population surveys and fertilization experiments. We determined gonad indices (gonad mass/body mass) over a range of population densities (0.1 to 250 ind. m⁻²) at seven sites in coastal Maine, USA, sampled at two depths (5 and 15 m). At shallow sites, we found that gonad indices declined by 50% over the 1500-fold range in adult population density. At 15 m deep locations, gonad mass was consistently low and did not vary significantly with density. Patterns of macroalgal abundance suggest urchins at high density and in deeper water were food limited. Because macroalgal cover co-varies inversely with sea urchin density, we designed field experiments to determine the interaction between sea urchin density and kelp canopy on fertilization success. On square arrays we manipulated the spacing of simulated urchins, but held their numbers constant (five sperm-filled syringes interspersed with four Nitex mesh egg containers permeable to sperm). These experiments, simulating the observed range of natural density, suggested that (1) fertilization rates decreased many times faster than individual gamete production increased over the same range in density, and (2) kelp increased fertilization success at high density when eggs were within 25 cm of a sperm source, but not when spaced 1 m apart. Additional laboratory fertilization experiments at ambient temperatures (3 to 5 °C) indicated that diluted sperm were viable for <1 h, but egg viability was virtually unchanged for >8 h. In short, to the individual the reproductive benefits of aggregating appear to outweigh the costs; and while sperm may be limiting at low population density, eggs may remain viable long enough to be fertilized by sperm from more distant males. Received: 10 February 1998 / Accepted: 22 January 1999     

Mass spawning by the sea urchin Evechinus chloroticus (Echinodermata: Echinoidea) in a New Zealand fiord

A mass synchronous spawning of the sea urchin Evechinus chloroticus (Valenciennes) was observed in situ in Doubtful Sound, a large New Zealand fiord. Spawning occurred between 17:30 hrs and 18:30 hrs on 27 January 1994 and coincided with a full moon, spring tides and a period of decreasing sea temperatures. During spawning, the sea urchins formed a dense spawning aggregation of both sexes, with >90% of the urchins observed spawning at the time. Spawned gametes clouded the water column, and some were eaten by small labrid fish species. The spawning, which may have been as widespread as 40 km, marked a 42 to 50% decrease in gonad indices and resulted in a widespread, dense cohort of  E. chloroticus larvae within the fiord. Received: 25 September 1997 / Accepted: 6 March 1998     

Annual reproductive cycle of the green sea urchin, Strongylocentrotus droebachiensis, in differing habitats in Nova Scotia, Canada

We monitored the reproductive cycle of Strongylocentrotus droebachiensis (OF Müller) between April 1993 and August 1995 in kelp beds, barren grounds and grazing fronts at both a wave-exposed and a sheltered site along the Atlantic coast of Nova Scotia. Gonad index and histological analyses showed that S. droebachiensis has an annual reproductive cycle that is synchronous across sites and habitats, and between females and males. Spawning occurs in March/April of each year but a small proportion of sea urchins in the study populations also spawned in fall 1995. During most of the year, sea urchins in kelp beds and grazing fronts have a higher gonad index than those in barren grounds. Gonad indices also tended to be higher at the wave-exposed than the sheltered site. Interannual variability in peak gonad index was significant in the barren grounds at the wave-exposed site and in the grazing front at the sheltered site. The gametogenic cycle is characterized by six stages based on the abundance of nutritive and germinal/gametic cells. Nutritive phagocytes are abundant after spawning and replaced by increasing numbers of germinal and gametic cells as the gametogenic cycle progresses. The temporal patterns of abundance of each cell type were similar among habitats indicating that the gonads were qualitatively similar despite large differences in gonadal mass. The quantity of gut contents (ratio of food volume to body volume) was similar among habitats, but the quality (percentage of organic material) tended to be higher in kelp beds and grazing fronts than in barren grounds suggesting that differences in gonad index of S. droebachiensis in different habitats are related to differences in diet. The high density of sea urchins in grazing fronts combined with their high fecundity suggests that they make the greatest contribution, per unit area, to the overall larval pool. Received: 20 May 1997 / Accepted: 21 January 1998