Crushing predation of the spiny star Marthasterias glacialis upon the sea urchin Paracentrotus lividus

Literature data report that only fish predators are able to crush sea urchin tests in Mediterranean rocky reefs. This experimental study showed that the spiny star Marthasterias glacialis is able to break Paracentrotus lividus tests and that the breaking event is more likely to occur for small-sized sea urchins than for big ones. Our results show that the role of M. glacialis in regulating P. lividus population density can be important in specific locations. They may have important implications, moreover, for the use of tethering techniques aimed at identifying predator types of sea urchins.     

Morpho-functional defences of Mediterranean sea urchins,<Emphasis Type="Italic"> Paracentrotus lividus</Emphasis> and<Emphasis Type="Italic"> Arbacia lixula</Emphasis>, against fish predators

Morpho-functional features potentially involved in defence mechanisms against fish predators (i.e. attachment tenacity, spine length, and test robustness and thickness) have been assessed in two Mediterranean sea urchins, Paracentrotus lividus and Arbacia lixula. All four morpho-functional features were significantly and positively related to individual size for both species of sea urchins. Test robustness (i.e. static load needed to break sea urchin tests) was significantly greater for A. lixula (from 3,450 to 15,000 g depending on size) than for P. lividus (1,180–11,180 g). Attachment tenacity (i.e. force needed to dislodge sea urchins from the rocky substrate) was greater in A. lixula (280–3,300 g) than in P. lividus (110–1,450 g), and the difference tended to decrease in relation to smaller sea urchin size. Spine length was greater in A. lixula (1.5–2.9 cm) than in P. lividus (0.5–2.3 cm), but the difference decreased for larger sea urchin size. Test thickness was slightly greater (but not significantly) in A. lixula (0.35–1.10 mm) than in P. lividus (0.12–0.90 mm). These results provide evidence that morpho-functional features of sea urchins could be involved in affecting predation rates by fishes upon P. lividus and A. lixula, with potential implications for the population structure and distribution patterns of the two sea urchins in shallow rocky reefs.Communicated by R. Cattaneo-Vietti, Genova     

The effects of predator abundance and habitat structural complexity on survival of juvenile sea urchins

We studied the effect of the abundance of predatory fishes and structural complexity of algal assemblages on the survival of juveniles of the sea urchin Paracentrotus lividus on Mediterranean infralittoral rocky bottoms. Post-settlement juveniles (2–10 mm) were placed on four distinct natural substrates with increasing structural complexity (coralline barren, algal turf, erect fleshy algal assemblages and small crevices) inside and outside the Medes Islands Marine Reserve. Predation on these sea urchins increased at greater abundance of predatory fishes, and decreased with greater structural complexity. The refuge provided by structural complexity, however, decreased with increasing size of sea urchin recruits. Predation on the smallest post-settlers was carried out almost exclusively by small fishes (<20 cm), mainly the labrid Coris julis, while the dominant predator of larger juveniles was the sparid Diplodus sargus. Our results demonstrate the cascading effects caused by the prohibition of fishing in marine reserves, and highlight the potential role of small predatory fishes in the control of sea urchin populations.Communicated by O. Kinne, Oldendorf/Luhe     

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     

Predators of juvenile sea urchins and the effect of habitat refuges

We evaluated the effects of potential predators from intertidal habitats on Strongylocentrotus purpuratus survival using laboratory experiments and assessed abundances of main predatory species along the Pacific coast of North America. The interactive effects of urchins’ and predators’ sizes in mediating predation were quantified. Habitat complexity (substrate pits, adult spine canopy) was manipulated to examine its effects on predation of most susceptible individuals (<14 mm). Pachygrapsus crassipes was identified as a major predator of urchins up to ≈30 mm. A positive effect of predator size on consumption of progressively larger urchins was detected, probably due to a mechanical limitation on crabs’ ability to consume large prey. Larger claws of males with respect to females of comparable sizes facilitated the handling of larger prey. Substrate refuges significantly reduced mortality on juvenile urchins. These results show that crab predation may be important in organizing intertidal communities, despite multiple ecological mechanisms promoting sea urchin survival.     

Direct estimation of natural mortality rates for littoral marine fishes using populational data from a marine reserve

 Visual censuses conducted in a marine reserve (Medas Islands) were used to estimate the natural mortality rates (M) for five common fish species in the NW Mediterranean Sea (Coris julis, Diplodus annularis, D. sargus, Serranus cabrilla and Symphodus roissali). Visual censuses of these same five species were also performed at three sites in unprotected areas of the coast where both commercial and sport fishing activity was normal. Censuses were conducted over a 3 year period. Estimates of M in the 3 years displayed scant seasonal or interannual variation, which may mean that the populations were in equilibrium during that period. The results of this study showed that the relationships between M and the growth parameters and maximum life span were unclear, and considerable caution is therefore recommended when using indirect methods of estimating M based on those parameters. For certain species the values of M were equal to or greater than the estimated total mortality in the exploited areas. The virtual absence of piscivorous predators in the unprotected area as a consequence of the high level of fishing in that area contrasts with the high abundance of such predators in the marine reserve. Since predation is the main contributor to M, estimated mortality in the unprotected areas is attributable nearly entirely to fishing. It is suggested that M may vary according to alterations taking place in conditions in the ecosystem inhabited by a species and thus that use of a value of M for a pristine population cannot be extrapolated to exploited areas. Received: 18 January 2000 / Accepted: 14 July 2000     

Role of surface glycoproteins in mate-guarding of the marine harpacticoid Tigriopus japonicus

Surface glycoproteins serve as mate- and gamete-recognition molecules in some marine animals such as rotifers and sea urchins. We investigated the role of contact chemoreception of surface glycoproteins in mate recognition, mate-guarding, and spermatophore transfer in the marine harpacticoid Tigriopus japonicus Mori. Adult males accurately distinguished immature females from a pool of mixed-sex juveniles, and their behavior towards both live and dead females is described. A survey of the binding sites of twelve lectins to the surface of juvenile and adult females, and adult males demonstrated localized lectin-binding at sites considered important in mating behavior of each sex/age class. Treating adult males with the Triticum vulgaris lectin sharply inhibited normal mate recognition and guarding behavior, as did the monosaccharide glucosamine. Treatment of males with protease and detergents destroyed their mate-guarding behavior without affecting swimming behavior. These results suggest that protein receptors on the antennules of the males detect glycoprotein signals on the surface of females, recognizing carbohydrate moieties of glycoproteins to identify appropriate partners for guarding or mating. Surface glycoproteins may function as a common mechanism of sexual communication in many marine zooplankters. Received: 6 May 1997 / Accepted: 14 September 1997     

Interactions between sea urchins (Strongylocentrotus droebachiensis) and their predators in field and laboratory experiments

Field observations and manipulative experiments in a nearshore cobble bed (2 to 3 m below mean low water) at Eagle Head, Nova Scotia, Canada, between 1984 and 1986, showed that small juveniles ofStrongylocentrotus droebachiensis (3 to 6 mm diam) sheltering beneath cobbles had a refuge from predators such as rock crabs, small lobsters, and fish. Sea urchins gradually outgrew these refuges and small adults (25 to 30 mm) required larger rocks as shelter from predators, particularly large cancrid crabs. Small juveniles were usually solitary and well dispersed beneath cobbles, whereas small adults tended to aggregate on the undersides and in the interstices of boulders. These aggregations may develop passively as sea urchins accumulate in suitablysized refuges. Chemotaxis experiments indicate that juvenileS. droebachiensis are repelled by waterborne stimuli from conspecifics. In a factorial experiment, effects of the presence of potential predators (rock crabs and lobsters) and/or food (kelp) on the behaviour of large juvenile (10 to 15 mm) and small adult sea urchins were examined in flowing seawater tanks. Both size classes formed exposed feeding aggregations when kelp was provided as food, irrespective of the presence or absence of predators. In the absence of kelp, each size class responded differently to the presence of a predator: juveniles became more cryptic, whereas adults aggregated on the tank sides. Increased movement to the sides of a tank in the presence of a predator may reflect a flight response, since chemotaxis experiments indicated thatS. droebachiensis is repelled by waterborne chemical stimuli from predators. Observational and experimental data suggest that predation at the late juvenile and early adult stages may influence population structure, distribution and abundance ofS. droebachiensis.     

Leading role of the sea urchin <Emphasis Type="Italic">Arbacia lixula</Emphasis> in maintaining the barren state in southwestern Mediterranean

Sympatric sea urchin species are usually considered to belong to the same grazer guild. Nevertheless, their role in community dynamics may vary due to species-specific morphological traits, feeding preferences and foraging behavior. In the Mediterranean Sea, the two species Paracentrotus lividus and Arbacia lixula co-occur in barren areas. Whereas P. lividus is usually considered responsible for creating a barren ground, the roles of the two sea urchin species in its maintenance are currently unclear. The relative and combined effects of P. lividus and A. lixula on maintaining the benthic community in the barren state were tested experimentally, using orthogonal exclusion of the two species. Results show that exclusion of A. lixula, regardless of the presence of P. lividus, led to a significant decrease in the surface of bare rock and a significant change of the algal assemblages, thus demonstrating the major role of this species in maintaining the barren state.     

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     

Manipulation of food and photoperiod promotes out-of-season gametogenesis in the green sea urchin, Strongylocentrotus droebachiensis: implications for aquaculture

Green sea urchins, Strongylocentrotus droebachiensis OF Müller, collected off the coast of New Hampshire, USA, in late February 1995, were brought into the laboratory and fed an artificial diet ad libitum, and subjected to a photoperiod advanced by 4 months. During this study, temperatures and salinities for experimental urchins mirrored those recorded at the collection site. We examined the effects of changes in feeding regime and photoperiod on gametogenesis and compared the experimental urchins with those from the source population. During the 7-month period, experimental urchins showed no detectable changes in mean test height or diameter. Experimental urchins had a significantly higher gonad index (GI) in March, April and May (18 ± 6%) compared with field urchins in March (11 ± 3%). Subsequently, experimental urchins had a mean monthly GI of 25 to 30%, while the mean GI for field urchins was 11 to 13%. Gonial cell mitosis and gametogenesis occurred earlier in experimental male and female urchins compared with field urchins. Stereological and histological observations and stage–frequency data showed that the ovaries of experimental urchins were large because of the accelerated development of nutritive phagocytes, the volume fraction (V _v) of which was 89 to 90% of the gonad, while new vitellogenic primary oocytes occupied <1% V _v. In males, stereological and histological observations and stage–frequency data suggested a mobilization of materials from the nutritive phagocytes beginning between June and August, i.e. earlier than in females, and, by September, new gametes occupied a V _v of 49 ± 3% of the testes. Oocyte size–frequency distributions demonstrated that most primary oocytes were <80 μm in diameter between March and September, suggesting that cold temperatures may be needed for completion of vitellogenesis. We describe changes in the two principal cell types in the germinal epithelium of urchin gonads and indicate how knowledge of their population dynamics may be useful in aquaculture applications. Received: 21 February 1997 / Accepted: 3 June 1998     

Different antipredator behaviour in two anuran tadpoles: effects of predator diet

Recent investigations have indicated that animals are able to use chemical cues of predators to assess the magnitude of predation risk. One possible source of such cues is predator diet. Chemical cues may also be important in the development of antipredator behaviour, especially in animals that possess chemical alarm substances. Tadpoles of the common toad (Bufo bufo) are unpalatable to most vertebrate predators and have an alarm substance. Tadpoles of the common frog (Rana temporaria) lack both these characters. We experimentally studied how predator diet, previous experience of predators and body size affect antipredator behaviour in these two tadpole species. Late-instar larvae of the dragonfly Aeshna juncea were used as predators. The dragonfly larvae were fed a diet exclusively of insects, R. temporaria tadpoles or B. bufo tadpoles. R. temporaria tadpoles modified their behaviour according to the perceived predation risk. Depending on predator diet, the tadpoles responded with weak antipredatory behaviour (triggered by insect-fed predators) or strong behaviour (triggered by tadpole-fed predators) with distinct spatial avoidance and lowered activity level. The behaviour of B. bufo in predator diet treatments was indistinguishable from that in the control treatment. This lack of antipredator behaviour is probably related to the effective post-encounter defenses and more intense competitive regime experienced by B. bufo. The behaviour of both tadpole species was dependent on body size, but this was not related to predator treatments. Our results also indicate that antipredator behaviour is largely innate in tadpoles of both species and is not modified by a brief exposure to predators. Received: 22 August 1996 / Accepted after revision: 31 January 1997     

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     

Susceptibility of non-indigenous ascidian species in British Columbia (Canada) to invertebrate predation

Non-indigenous ascidians are known to significantly alter the structure and composition of benthic communities and adversely affect shellfish aquaculture by fouling both the cultured species and the infrastructure. The ability of these species to persist in new locations and their current and potential distributions are dependent upon physiological tolerances to environmental factors and biotic resistance to competition and predation. Despite significant data on global invasion patterns, potential biotic resistance to non-indigenous ascidians is poorly understood. We identified potential predators of four non-indigenous ascidians (Styela clava, Botryllus schlosseri, Botrylloides violaceus, and Didemnum vexillum) in British Columbia (BC), Canada in order to: (1) assess the potential for biotic interference to limit the establishment and/or spread of these ascidian species in BC, and (2) identify candidate species to be used as ascidian biofouling control agents in shellfish aquaculture. Using a series of single- and multiple-choice laboratory experiments, potential benthic predators (including various species of molluscs, echinoderms, and arthropods) were offered non-indigenous ascidians as prey. The sea urchins Strongylocentrotus droebachiensis and Strongylocentrotus franciscanus, the sea stars Dermasterias imbricata and Evasterias troschelii, the nudibranch Hermissenda crassicornis, and the crabs Cancer productus and Carcinus maenas were found to consume one or more species of non-indigenous ascidians in single-choice experiments. However, when provided a choice, all predators chose their respective preferred food over ascidians. Thus, predation alone is unlikely to prevent large-scale establishment and spread of non-indigenous ascidians in BC, but it may have the potential to significantly reduce localized populations of ascidians. Green sea urchins, S. droebachiensis, were found to be efficient grazers of all four ascidian species, consuming 12.7 ± 5.14 cm² (mean ± SD) of adult B. violaceus over a 3-day period, 15 ± 3.7 juvenile colonies of B. violaceus over a 2-day period, and 63 ± 28.8 juvenile colonies of B. schlosseri over a 2-day period. Using sea urchins as biological control organisms may significantly reduce ascidian fouling in shellfish aquaculture.     

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     

Experimental evaluation of aggregation behavior in the sea urchin Strongylocentrotus droebachiensis

Defensive aggregation by Strongylocentrotus droebachiensis has been invoked as the fulcrum for the transformation of macroalgal beds into coralline barrens in the northwest Atlantic. We critically examined some of the mechanisms contributing to aggregation behavior by experimentally manipulating sea urchins, purported predators and food during autumn 1983 and spring 1984 both in the laboratory and field. We utilized several approaches to examine a range of sea urchin responses to the presence of food, tethered predators, caged predators, crushed con-specifics in the field and predators in laboratory tanks. Some of the field cages had the property of allowing free passage to sea urchins while retaining lobsters; this allowed distinctions to be made between artifacts caused by cage walls or topographic barriers and unrestricted behavioral responses of sea urchins. The results falsified the hypothesis that aggregations of S. droebachiensis are elicited by predators. Except in the presence of algae, sea urchins always avoided decapod predators (but not sea stars) and fled from the vicinity of injured conspecifics. However, avoidance behavior was subordinate to feeding responses, demonstrated by mass migrations of sea urchins into cages (with lobsters) to feed on algae. We reaffirmed by observation and manipulation previous studies which showed that sea urchins aggregate only in the presence of food. Two types of sea urchin groupings were delineated: (1) surficial two dimensional associations, often caused by topographic or other features which inhibited dispersal and (2) cohesive three-dimensional aggregations induced by food.     

Are mangroves and seagrasses sources of organic carbon for penaeid prawns in a tropical Australian estuary? A multiple stable-isotope study

The vertical distribution of the larvae of shelf-dwelling fish species that spawn in the NW Mediterranean Sea in spring was studied in relation to environmental data. Two sampling cycles were carried out at fixed stations on the continental shelf in May and June 1992. Three patterns of larval vertical distribution for the various taxa represented in the samples were observed. The larvae of most species (e.g. Boops boops, Diplodus sargus) were mainly located in the surface layer (10 m), others (e.g. Arnoglossus sp.) had broader distributions in the upper 40 m of the water column, and but a few (e.g. Gobiidae) were present in large concentrations at greater depths. The vertical distribution patterns of the various species showed no variations, despite high hydrographic variability during the study. The vertical distribution of only a few species (e.g. Arnoglossus sp., Crystallogobius linearis and Engraulis encrasicolus) varied over the diel cycle. The possible influence of the vertical distribution of fish larvae on their horizontal distribution patterns is discussed. Received: 10 March 1997 / Accepted: 4 April 1997     

Juvenile–adult associations in sea urchins ( Strongylocentrotus franciscanus and S. droebachiensis ): protection from predation and hydrodynamics in S. franciscanus

Juvenile red sea urchins, Strongylocentrotus franciscanus, aggregate under adult conspecifics, whereas sympatric juvenile green sea urchins, Strongylocentrotus droebachiensis, are typically more solitary and dispersed. Neither the potential advantage of juvenile sheltering nor the differences in post-settlement behavior between the two species has been demonstrated experimentally, but may be related to protection from predators and/or hydrodynamics. In predation experiments, juvenile vulnerability differed in the two species as the seastar Pycnopodia helianthoides consistently chose juvenile S. franciscanus over S. droebachiensis (100% vs. 0%). When associated with adults, juvenile mortality decreased dramatically in S. franciscanus (90% alone vs. 5% with adults), but very little in S. droebachiensis (85% vs. 75%). Not surprisingly, juvenile behavioral responses in the two species reflect this difference in vulnerability. Juvenile S. franciscanus sheltered under adults when predation risk was high, but not when risk was low (44% vs. 13%), whereas sheltering in S. droebachiensis was infrequent and not related to predation risk (7% for high risk versus 5% for low risk). From a hydrodynamic perspective, the presence of an adult led to the creation of a hydrodynamic refuge for juveniles, where average water velocities were reduced by > 60% around the adult urchin. Again, striking differences in sheltering rate were apparent in S. franciscanus (52% vs. 13% for high flow and low flow, respectively), but not S. droebachiensis (5% for high flow versus 4% for low flow). Sheltering behavior was also species-specific as juveniles did not shelter at high rates under adults of the opposite species (≤ 16%). A field survey confirmed these finding in that juvenile S. franciscanus abundance was associated with both adults and water motion (R ² = 0.80, P = 0.008, best-subsets regression). These results suggest that sheltering confers juvenile S. franciscanus with a degree of protection from predators and water motion, and that species-specific differences in this post-settlement behavior may be related to the differences in the protection afforded by adults.     

Sequence divergence of mitochondrial DNA indicates cryptic biodiversity in Octopus vulgaris and supports the taxonomic distinctiveness of Octopus mimus (Cephalopoda: Octopodidae)

DNA sequence diversity of octopods was investigated using the mitochondrial cytochrome oxidase III gene (mtCOIII). DNA was obtained from ethanol- or formalin-fixed tissue of 15 specimens belonging to Octopus mimus Gould, 1852, Octopus vulgaris Cuvier, 1797 and Scaeurgus unicirrhus d'Orbigny, 1840, from coastal waters of the Mediterranean (France), the southwestern Atlantic (Brazil), the Caribbean (Costa Rica) and the southeastern/tropical Pacific (north Chile/Costa Rica). A 612 bp fragment of the mtCOIII gene was sequenced and aligned to the orthologous sequences available from northeastern Pacific Octopus species. Possible phylogenetic relationships were reconstructed. The mtCOIII gene tree revealed two main clusters, one comprised O. rubescens, O. dofleini and O. californicus, while the other included all O. vulgaris specimens, O. bimaculatus, O. bimaculoides and O. mimus. With one exception all South American haplotypes including O. bimaculatus and O. bimaculoides appeared as the sister group of the Mediterranean haplotype of O. vulgaris, indicating that most of the South American O. vulgaris specimens investigated would not belong to the recently redescribed species O. vulgaris restricted to the Mediterranean and eastern Atlantic. The treatment of O. mimus as a species distinct from Mediterranean O. vulgaris is supported by a high nucleotide divergence of 12.7%. Based on the mtCOIII gene tree the existence of cryptic species among O. vulgaris-like octopods is suggested. Received: 15 January 1999 / Accepted: 15 October 1999     

Water-soluble extracts of the diatom Thalassiosira rotula induce aberrations in embryonic tubulin organisation of the sea urchin Paracentrotus lividus

Eggs and embryos of the sea urchin Paracentrotus lividus were used as a model to study the effect at the cellular level of potential anti-mitotic compounds extracted from the diatom Thalassiosira rotula. Eggs and embryos incubated in a water-soluble diatom extract, corresponding to 5 × 10⁶ and 10⁷ cells ml⁻¹, were totally blocked (i.e. cell division was blocked) at the one-cell stage. At lower concentrations (2.5 and 1.25 × 10⁶ cells ml⁻¹), the first mitotic division was inhibited in 32 ± 26% and 25 ± 3.5% of the zygotes, respectively, demonstrating the dose-dependent effect of diatom extracts on sea urchin development. Immunofluorescence dyes, specific for DNA and α-tubulin subunits, were used to stain nuclei and microtubules in sea urchin embryos during various phases of development. Images with the confocal laser scanning microscope showed that tubulin was not organised in filaments at the sperm aster and cortex levels, and that the pronuclei were not fused in embryos incubated soon after fertilisation with water-soluble diatom extracts corresponding to 10⁷ cells ml⁻¹. At lower diatom-extract concentrations (4 × 10⁶ cells ml ⁻¹), fusion of the pronuclei occurred but the mitotic spindle was not formed. Microtubules were clearly de-polymerised and the chromatin appeared globular and compacted at the centre of the cell. A similar structure was observed for sea urchin embryos incubated with 0.1 mM colchicine, a potent anti-mitotic compound. When sea urchin embryos were incubated in water-soluble diatom extracts at different times prior to the first mitotic division, microtubules appeared de-polymerised at each step, from pronuclear fusion to telophase, and cell division was blocked. At the histological level, embryos incubated with 4 × 10⁶ cells ml⁻¹ diatom extract showed nuclear fragmentation without cytokinesis. The possible use of sea urchin embryos as a bioassay to test for other unknown compounds with cytotoxic activity in phytoplankton species is discussed. Received: 7 May 1998 / Accepted: 9 December 1998