The larval settling response of the lined chiton Tonicella lineata

The lined chiton Tonicella lineata (Wood, 1815) is found on enerusting coralline algae in the lower rocky intertidal zone of Oregon and San Juan Island, Washington, USA. An encrusting coralline alga is the major food item of this chiton. Experiments were performed to test the settling response of T. lineata larvae to various algae and other substrata. In these experiments, the larvae would settle only on pieces of encrusting coralline algae and piecesof ceramic roofing tile soaked in a coralline algal extract. The settling stimulus is probably chemical in nature, and is inactivated by boiling. In laboratory cultures, normal development stops at the trochophore stage (110 to 160 h, depending on the temperature). Metamorphosis and further development will take place only after stttlement on encrusting coralline algae.     

Waterborne chemical compounds in tropical macroalgae: positive and negative cues for larval settlement

Settlement sites of marine invertebrate larvae are frequently influenced by positive or negative cues, many of which are chemical in nature. Following from the observation that many shallow-water, Hawai'ian marine macroalgae are free of fouling by sessile invertebrates, we predicted that the algae are chemically protected and dependent on either surface-bound or continuously released soluble compounds to deter settling invertebrate larvae. To address the importance of waterborne algal compounds, we experimentally determined whether larvae of two of Hawai'i's dominant hard-surface fouling organisms, the polychaete tube worm Hydroides elegans and the bryozoan Bugula neritina, would settle in the presence of waters conditioned by 12 species of common Hawai'ian macroalgae (representing the Phaeophyta, Chlorophyta, Rhodophyta and Cyanophyta). The results included a full spectrum of biological responses by each larval species to waterborne algal compounds. Larval responses to conditioned water were consistent for each algal species, but the outcomes were not predictable based on the taxonomic relationships of the algae. For example, among the species of Phaeophyta examined, different conditioned waters were: (1) toxic, (2) inhibited settlement, (3) simulated settlement, or (4) had no effect, compared to larvae in control dishes containing filtered seawater. Additionally, larval responses to aged (24 h) conditioned waters could not be predicted from the results of assays run with conditioned waters utilized immediately after preparation. Finally, settlement by larvae of one species did not predict outcomes of tests for the other species. Four of 12 shallow-reef Hawai'ian macroalgae tested released compounds into surrounding waters that immediately killed or inhibited settlement by both H. elegans and B. neritina (toxic: Dictyota sandvicensis; inhibitory: Halimeda discoidea, Sphacelaria tribuloides, Ulva reticulata); the remaining 8 algal species prevented settlement by one of these fouling organisms but for the other had no effect or, in some cases, even stimulated settlement     

Chemoautotrophic symbiosis in the tropical clamSolemya occidentalis (Bivalvia: Protobranchia): ultrastructural and phylogenetic analysis

Patterns of algal seasonality, and their effect on the diet and feeding preferences of the herbivorous crab Grapsus albolineatus were investigated over an 18-mo period from March 1993 to August 1994 on an exposed tropical rocky shore (Hok Tsui Peninsula at Cape d' Aguilar, Hong Kong). Algal cover was greatest in the winter months, and lowest in the summer. Foliose algae such as Ulva fasciata, Porphyra suborbiculata, and Dermonema frappieri were dominant in the winter, but died off in the summer. During the hot summer months, perennial encrusting algae e.g., Ralfsia expansa, Hildenbrandia rubra, H. occidentalis, coralline crusts and the encrusting cyanobacteria Kyrtuthrix maculans, were the dominant algal species. Seasonal variation in algal abundance influenced the dietary selectivity of the herbivorous crab G. albolineatus. In the winter, the crab fed selectively on filamentous algae (e.g. Hincksia spp., Cladophora spp., Enteromorpha spp., and the cyanobacteria Lyngbya sp.). Foliose algae (e.g. U. fasciata, P. suborbiculata, Pterocladia tenuis) formed a small part of the diet, despite being the dominant species on the shore. Foliose and filamentous algae were virtually absent from the shore in the summer, and the crabs switched to feeding solely on encrusting algae. Electivity indices revealed preferences for green and brown turf species, and avoidance of foliose algae. Faecal analysis revealed that a greater proportion of the food is digested in the winter, suggesting that G. albolineatus is able to digest filamentous algae more efficiently than encrusting algae. Feeding preferences of G. albolineatus appear to be influenced by a number of factors, including the availability, digestibility and morphology of algae. The foraging behaviour and cheliped morphology of the crab also affect food choice. The monsoonal nature of Hong Kong's climate controls the diversity and abundance of intertidal algae and, therefore, indirectly influences the diet and subsequent growth and reproductive success of the herbivorous crab G. albolineatus.     

Dissolved histamine: a potential habitat marker promoting settlement and metamorphosis in sea urchin larvae

Many species of marine invertebrate larvae settle and metamorphose in response to chemicals produced by organisms associated with the adult habitat, and histamine is a cue for larvae of the sea urchin Holopneustes purpurascens. This study investigated the effect of histamine on larval metamorphosis of six sea urchin species. Histamine induced metamorphosis in larvae of three lecithotrophic species (H. purpurascens, Holopneustes inflatus and Heliocidaris erythrogramma) and in one planktotrophic species (Centrostephanus rodgersii). Direct comparisons of metamorphic rates of lecithotrophic and planktotrophic larvae in assays cannot be made due to different proportions of larvae being competent. Histamine (10 μM) induced metamorphosis in 95% of larvae of H. purpurascens and H. inflatus after 1 h, while the coralline alga Amphiroa anceps induced metamorphosis in 40–50% of these larvae. Histamine (10 μM) and A. anceps induced 40 and 80% metamorphosis, respectively, in the larvae of H. erythrogramma after 24 h. Histamine (10 μM) and the coralline alga Corallina sp. induced 30 and 70% metamorphosis, respectively, in the larvae of C. rodgersii after 24 h. No metamorphosis of any larval species occurred in seawater controls. Larvae of two planktotrophic species (Tripneustes gratilla and Heliocidaris tuberculata) did not metamorphose in response to histamine. Seagrasses, the host plants of H. inflatus, induced rapid metamorphosis in larvae of the two Holopneustes species, and several algae induced metamorphosis in C. rodgersii larvae. Histamine leaching from algae and seagrasses may act as a habitat marker and metamorphic cue for larvae of several ecologically important sea urchin species.     

Inhibition of larval settlement to a soft bottom benthic community by drifting algal mats: An experimental test

The hypothesis that drifting red algal mats inhibit settlement of planktonic larvae was tested in a field experiment in 1986. Substratum free of algae (caged boxes) was compared with substratum covered with algae (natural substratum and open boxes). Whereas settling densities of 1500 to 5500 ind. m^-2 of the bivalves Macoma balthica, Cardium glaucum and Mya arenaria were observed in sediment without algal cover, no recruits of the same species were found beneath the algae during the period of peak settlement (June to July). The same difference was observed for the polychaete Nereis diversicolor, although in this case occasional individuals were found beneath the algae. The results demonstrate that algae mats may be efficient inhibitors of larval settlement to benthic soft-substratum communities. It is concluded that algae act as a larval filter.     

Bacteria on the surface of crustose coralline algae induce metamorphosis of the crown-of-thorns starfish Acanthaster planci

The crustose coralline alga Lithothamnium pseudosorum induces high rates of settlement and metamorphosis of larvae of the coral-eating crown-of-thorns starfish (Acanthaster planci). In cases where crustose coralline algae (CCA) induce metamorphosis of marine invertebrate larvae it is normally assumed that the inductive molecules are produced by the alga, but an alternative is that they originate from bacteria on the plant surface. Bioassays using shards of L. pseudosorum treated with several antibiotics, whereby some shards were reinfected with bacteria from the alga, showed that if bacteria populations are depleted then settlement and metamorphosis of larvae of A. planci are inhibited. This demonstrates that bacteria are necessary for induction and suggests that morphogenic substances are produced by bacteria on the surface of the alga and not directly by the alga itself. However, surface bacteria are not inductive if they are isolated from soluble algal compounds, suggesting either that they require a substrate from the alga to produce the inductive agents or, alternatively but less likely, that compounds from both the alga and bacteria are required. There is no evidence that inductive compounds derive from the alga, since algal cell debris and soluble extracts prepared from the alga do not induce metamorphosis of A. planci. This is the first time that induction of metamorphosis in a marine invertebrate by CCA has been shown to be mediated by bacteria associated with the alga.     

Effects on larval crabs of exposure to algal toxin via ingestion of heterotrophic prey

We tested whether ingesting toxic algae by heterotrophic prey affected their nutritional value to crab larval predators, using toxic algal strains that are either ingested directly by larval crabs or rejected by them. Ingestion of toxic strains of the dinoflagellates Alexandrium andersoni and A. fundyense by the rotifer Brachionus plicatilis was confirmed. Rotifers having ingested either algal type for five days were fed to freshly hatched larvae of three crab species, with larval survival and stage durations determined. For both algal/rotifer treatments in all three crab species, larvae fed algae directly died during the first zoeal stage, while those fed rotifers that had been fed either algal strain survived to the experiment’s end (zoeal stage 3). Survival was lower, and stage duration longer, for larvae fed rotifers cultured on toxic algae when compared to those fed non-toxic algae. The role of toxic algae in the planktonic food web may be influenced by its direct or indirect ingestion by larval crabs.     

Settlement and recruitment of sea urchins (Strongylocentrotus spp.) in a sea-urchin barren ground and a kelp bed: are populations regulated by settlement or post-settlement processes?

I sampled recruitment of very small sea urchins (Strongylocentrotus spp.) by using the anesthetic magnesium chloride to remove individuals from substrata collected in sea-urchin barren grounds (barrens) and kelp beds at Naples Reef near Santa Barbara, California, USA. Preliminary sampling found low numbers of newly settled individuals(<0.6 mm test diam) from April–July in 1984 and 1985, and in April, 1986. In early May, 1986, I found many newly settled seaurchins (0.3 to 0.6 mm, 5 to 17 d old), and I compared the densities of the cohort on several types of natural substrata in barrens and kelp-bed habitats. Newly settled individuals of both purple sea urchins (S. purpuratus) and red sea urchins (S. franciscanus) were present in similar, high densities (1 000 S. purpuratus m^-2) on foliose red algal turf, a dominant substratum ofthe kelp bed, and on crustose coralline algae, the dominant substratum of an adjacent barrens. Larvae of S. purpuratus reared and tested in the laboratory showed high rates of settlement on both red algal turf and on crustose coralline algae, but significantly lower rates on rock. Larvae also settled in response to a partiallypurified extract of coralline algae. The reduced settlement on natural rock surfaces relative to either algal treatment and the significant settlement in response to the extract of coralline algae indicate that larvae discriminate between natural substrata and probably respond to a settlement cue other than, or in addition to, a simple microbial (bacterial) film. The similar densities of young recruits of S. purpuratus on dominant substrata of barrens and kelp bed show that, at least in this case, differential settlement cannot explain the high densities of sea urchins in the barrens habitat. Movement between barrens and kelp bed is unlikely given the small sizes of the newly recruited sea urchins relative to the large distances often involved. Reduced post-settlement mortality of newly settled individuals in the barrens remains the most likely mechanism leading to the higher densities of sea urchins in barrens relative to kelp-bed habitats.     

Epifluorescence microscopy,a technique for the study of feeding in Crassostrea virginica veliger larvae

Epifluorescence microscopy was applied as a new technique to observe the response of Crassostrea virginica veliger larvae to the algal foods Monochrysis lutheri or Chlorella autotrophica. Uptake, lysis, and digestion or rejection of these algae by oyster larvae were observed microscopically through the autofluorescence of chlorophyll a and its derivatives. Both species of algae were taken up by large numbers of larvae within 5 min after being added to larval cultures. The good growth of larvae obtained with M. lutheri as a food source was related to rapid uptake, lysis, and digestion of this alga. Larvae fed with C. autotrophica did not grow. Althogh C. autotrophica was also taken up rapidly by the larvae, there was no lysis or digestion of these algae and cells were eventually voided.     

Effects of gamma-aminobutyric acid on the settlement of larvae of the black chiton Katharina tunicata

Laboratory-cultured larvae of the black chiton Katharina tunicata (Wood), collected from central California in May and June of 1979 and 1980, settled preferentially on the encrusting coralline alga Lithothamnium sp. Metamorphosis, which involves the loss of the prototrochal ciliary band, occurred within a period of 2.5 h after larvae began crawling upon the alga. In response to gamma-aminobutyric acid (GABA) at a concentration of 10^-6 M, the lecithotrophic trochophore larvae ceased swimming and settled rapidly. Developmental metamorphosis did not follow settlement in the absence of encrusting coralline alga. However, when both GABA and Lithothamnium sp. were present, larval metamorphosis occurred within 2 h of settlement. Significantly different rates of settlement exist for sibling larvae of different ages; 12 d-old larvae responded more rapily and in greater numbers to treatment with GABA than 10 d-old larvae. Differences in settling rates are attributed to the onset of metamorphic competence.     

Selective consumption and facilitation by mesograzers in adult and colonizing macroalgal assemblages

Grazing pressure on macroalgae in littoral communities may vary with algal species, the age of an algal individual and grazer identity. Previous studies on alga–grazer interactions have shown that grazer preference for an algal species may release another one from interspecific competition. We measured the impacts of four common grazer taxa and the natural grazer guild on macroalgal communities at both their colonization and adult stages, and compared the impacts to grazer exclosures. The grazer effects were stronger on colonizing than on adult macroalgae; grazers did not reduce the total density of adult algae. Grazers both feed on propagules and indirectly facilitate other algae, depending on the grazer or algal species. Hydrobia species increased the settlement of spores of the red alga Ceramium tenuicorne. Similarly, the gastropod Theodoxus fluviatilis tended to facilitate one crustose algal species, but decreased the propagule density of annual filamentous algae, suggesting a preference for one species to the advantage of another. Effects of crustacean mesograzers on the studied macroalgae were weak. These results indicate that northern Baltic macroalgae are limited to grazing mainly during their colonization stage.     

The induction of larval settlement and metamorphosis of two sea urchins,Pseudocentrotus depressus and Anthocidaris crassispina,by free fatty acids extracted from the coralline red alga Corallina pilulifera

Lipophilic inducers of larval settlement and metamorphosis of Pseudocentrotus depressus and Anthocidaris crassispina, two commercially important sea urchin species in Japan, were isolated from the foliose coralline red alga Corallina pilulifera (collected in 1990 near Saga, Japan) and identified. Larval assays of the fractions obtained by silica gel column chromatography of the total lipids showed that non-polar groups of lipids were effective at inducing larval settlement and metamorphosis. The effective fractions were further subjected to gel filtration (Sephadex LH-20) and also to silica gel column chromatography, and the effective components isolated as single spots by thin-layer chromatography. The components at a concentration of ca. 0.4 mg paper^-1 (sample was adsorbed on a paper with 20 cm²) induced high rates of larval settlement of both P. depressus and A. crassispina. Chemical analyses of the components revealed a mixture of free fatty acids (FFAs), dominated by eicosapentaenoic acid (20:5, 41 to 50%), palmitic acid (16:0, 11 to 17%), arachidonic acid (20:4, 9 to 15%), and palmitoleic acid (16:1, 4 to 5%). In assays with the four standard FFAs, only 20:4 and 20:5 induced larval settlement and metamorphosis of the two species, while 16:0 and 16:1 were ineffective. The larvae underwent significant rates of settlement and metamorphosis in response to the two former FFAs at levels as low as 0.18 mg paper^-1. Amongst the free fatty acid components of the alga, 20:5 was isolated as the chemical inducer of larval settlement and metamorphosis of the sea urchins in the laboratory.     

Phenology and demography of a marine specialist herbivore: Placida dendritica (Gastropoda: Opisthobranchia) on the central coast of Oregon

The occurrence of small specialist herbivores in marine rocky intertidal communities has been largely ignored. From 1984 to 1989, the phenology and demography of the common oligophagous ascoglossan opisthobranch Placida dendritica A.& H. was documented at several sites along the central coast of Oregon, USA. P. dendritica was found on the low intertidal green algae Codium setchellii Gardn., C. fragile (Sur.) Har. and Bryopsis corticulans Setch. during spring and summer. Ascoglossan abundance on Codium spp. varied spatially and temporally. Peak density usually occurred in May or June. Maximum ascoglossan live mass was 4 to 6 mg on Codium spp. and 100 mg on B. corticulans. Larval recruitment of P. dendritica to algal hosts was continuous during spring and summer. Rapid recruitment (200 to 400 sea slugs thallus^-1mo^-1) of newly metamorphosed ascoglossans to C. setchellii in wave-protected areas suggests that high densities of competent larvae occurred in nearshore waters and/or that larvae were efficient at locating algal hosts. Ascoglossan growth was rapid, with less than one month elapsing from larval settlement to adult sexual maturity. Therefore, populations of P. dendritica were composed of many overlapping generations. Movement of adult ascoglossans among thalli of C. setchellii was rapid. Continuous larval settlement and adult movement enabled P. dendritica not only to locate and colonize its algal hosts quickly but also to maintain a persistent association with algal hosts during spring and summer.     

Larval settlement and metamorphosis of the mussel <Emphasis Type="Italic">Mytilus galloprovincialis</Emphasis> on different macroalgae

Settlement of mussels is commonly associated with macroalgae. The effects of 19 macroalgal species on the settlement and metamorphosis of pediveliger larvae of the mussel Mytilus galloprovincialis were investigated in the laboratory. Settlement and metamorphosis inducing activities of macroalgae Chlorodesmis fastigiata and Ceramium tenerrimum collected each month during the period between January 2005 and April 2006 were also investigated. Furthermore, C. fastigiata and C. tenerrimum were subjected to various treatments to investigate the roles of bacteria and diatoms on the algal surface in the induction of larval settlement and metamorphosis of M. galloprovincialis and the characteristics of the cues in these two macroalgae. Pediveliger larvae of M. galloprovincialis settled and metamorphosed in high percentages on Cladophora sp., Chlorodesmis fastigiata, Centroceras clavulatum, and Ceramium tenerrimum, all of which were filamentous in morphology. Macroalgae that were cylindrical, phylloid, flabellate, palmate and pinnate all showed low (<8%) percentages of post-larvae but four other filamentous macroalgae also had low mussel larval settlement, suggesting that chemical factors may also be involved. Seasonal variation had no effect on inductive activities of C. fastigiata and C. tenerrimum. Treatment of C. fastigiata and C. tenerrimum with formalin, ethanol and heat resulted in the significant decrease or loss of their inductive activities. Survival of algal cells within treated macroalgae also decreased significantly. Treatment of the two macroalgae with antibiotics and GeO₂ reduced the numbers of bacteria and diatoms on their surface but did not affect their inductive activities, indicating that the cue was produced by macroalgae and not by coexisting bacteria and diatoms. However, conditioned water and crude extracts of these two macroalgae did not induce larval settlement and metamorphosis. Thus, larvae of M. galloprovincialis settled and metamorphosed on specific filamentous macroalgae. The chemical cues produced by C. fastigiata and C. tenerrimum were susceptible to ethanol and heat treatments and were not recovered in the conditioned water nor in the extracts. The finding that inactive C. tenerrimum can be produced from culturing its apical segments provides a new tool to elucidate the chemical cue(s) from macroalgae through manipulation of their culture conditions.     

Community development on subtidal temperate reefs: the influences of wave energy and the stochastic recruitment of a dominant kelp

Patterns of community development on subtidal rocky reefs in Marmion Lagoon, southwest Australia, were investigated with a settlement panel experiment. We tested the hypothesis that community development would differ between outer and inner reefs lines, because exposure to swell and wave energy was significantly greater on outer reefs. Following a 14-month deployment, we recorded pronounced variability between panels and sites, but did not detect any effect of wave exposure on the structure of panel assemblages. Subsequent data exploration suggested the importance of the presence of kelp recruits (Ecklonia radiata) in structuring the overall assemblage. Panel assemblages with kelp recruits were significantly different in structure to those without, principally because of greater space coverage of encrusting coralline algae and less coverage of red turfing algae, spirorbids, and bryozoans. Mature E. radiata act as ecosystem engineers in subtidal rocky reefs in southwest Australia. Our results suggested the importance of young, recruiting kelps in determining patterns of early community development on newly available hard substrata.     

Use of artificial holes in studying community development in cryptic marine habitats in a tropical rocky intertidal region

Previous studies on the rocky intertidal shores of the Bay of Panama indicate that for many sessile and mobile organisms holes and crevices are important refuges from consumers. To study the dynamics of these hole-dwelling species assemblages, we devised a method allowing repetitive, non-destructive sampling and species manipulations in artificial holes. These are fiberglass sleeves with flanges at the outer edge that are inserted into receptacles made of quick-setting concrete. Predator and herbivore manipulations include: fences excluding slow-moving benthic consumers, bars across hole mouths restricting entry of large fishes and homogeneous (shelterless) surfaces created around a treatment decreasing abundance of small crabs. This design has withstood the rigors of two wet seasons in Panama. Results from two sites in Panama indicate that colonization occurs rapidly (6–7 months) in holes established in mid dry season. Encrusting fleshy algae (Ralfsia sp.) and ephemeral green algae (Cladophora spp.) colonize first and are succeeded by encrusting species of coralline algae, bryozoans and colonial tunicates and by erect fleshy red algae. Where consumers are present, encrusting corallines predominate; without consumers, bryozoans, tunicates and erect algae are more abundant. Zonation patterns develop within the holes, with the desiccation/heattolerant Ralfsia sp. dominating at the outer edges. With increasing deph, encrusting coralline algae, bryozoans and colonial tunicates reach their respective peak abundances. In contrast to the high variability observed among naturally occurring holes, replicates of each treatment tend to be similar. Between-treatment and between-site comparisons are less so. The naturally-occurring high level of small-scale patchiness is thus presumably due to variation in recruitment and in the local (microspatial) consumer regime. Substratum heterogeneity is therefore directly and indirectly important in maintaining a high local diversity in this community. As in other studies, consumers or disturbances are key factors in regulating patterns of community structure. More experiments of longer duration are necessary to ascertain the relative importances of consumers/disturbance and competition in controlling such assemblages.     

Substratum preferences in planula larvae of two species of scleractinian corals, Goniastrea retiformis and Stylaraea punctata

To test whether coral planulae recruit randomly to different coral reef habitats or have specific substratum preferences, the settling behavior of planulae from two shallow water coral species from Pago Bay, Guam (13°25.02N, 144°47.30E) were examined in the laboratory in June and July of 1995. Goniastrea retiformis is generally restricted to the shallow reef front (<10 m depth) in areas dominated by crustose coralline algae (CCA), while Stylaraea punctata is abundant on inner reef flats were CCA coverage is low and sand and carbonate rubble covered by biofilms is common. When presented with four substrata (1) carbonate rock scrubbed free of biofilm and dried as a control, (2) the CCA Hydrolithon reinboldii, (3) the CCA Peyssonelia sp., and (4) naturally conditioned carbonate rubble covered by a biofilm, G. retiformis larvae showed a significant preference for H. reinboldii, and S. punctata larvae for the carbonate biofilm treatment. The preference shown by S. punctata larvae for biofilmed surfaces did not diminish with increasing larval age up to 11 days. These results suggest that the larvae of both species are capable of habitat selection, and that the preferred substrata among those tested bears a relationship to the habitats in which adult colonies were found. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

The response of encrusting coralline algae to canopy loss: an independent test of predictions on an Antarctic coast

We assessed whether published observations of the ecology of encrusting coralline algae (Rhodophyta) from tropical and temperate coasts could be used to predict patterns and responses on a polar coast where such knowledge does not exist. On subtidal rocky coasts near Casey, East Antarctica, we detected a strong positive association of understorey encrusting coralline algae with canopies formed by the endemic alga Himantothallus grandifolius. The experimental removal of H. grandifolius caused corallines to bleach from red to pink/white concomitant with a decline in their photosynthetic activity. The magnitude of this decline (mean ± SE = 56.85±8.43%) was remarkably similar to that observed on temperate coasts (45.98±5.91%). Positive effects of nutrient enrichment of the surrounding water, hypothesized to alleviate the negative effects of canopy loss on encrusting corallines, were not detected. Removing H. grandifolius increased the intensity of photosynthetically active radiation and ultra-violet radiation reaching the substratum by three orders of magnitude, providing the basis for models invoking enhanced irradiance as the primary cause of the negative effects of canopy loss. Striking similarities among our results and those from tropical and temperate coasts suggest that responses of encrusting corallines to loss of canopies may have predictive properties across large distances and environmental gradients (tropical–temperate–polar).     

Inhibitory effects of red algal extracts on larval settlement of the barnacle Balanus improvisus

We examined the chemical antifouling properties of four sublittoral red algae, Chondrus crispus, Delesseria sanguinea, Osmundea ramosissima, and Polyides rotundus, which are all rarely fouled in the field. Two different approaches were used. Firstly, we tested the effects of lipophilic crude extracts on the settlement behaviour of cyprid larvae of the co-existing barnacle Balanus improvisus. Secondly, in a settlement preference experiment, we tested whether B. improvisus cyprid larvae settle on living algae when given a choice between natural algal surfaces and control surfaces. With this procedure, we were able to test both if the algae inhibit recruitment of cyprids, and if this inhibition is a result of chemistry. The settlement of B. improvisus larvae was strongly inhibited at concentrations estimated to be potentially ecologically relevant for all of the tested extracts. However, only C. crispus significantly inhibited settlement in the preference experiment, even though there was also a tendency for settlement inhibition on P. rotundus and O. ramosissima. In contrast, D. sanguinea seemed to stimulate settlement. This contradiction probably resulted from an extraction of metabolites that naturally occur only inside the alga. However, as this study shows, a combination of settlement assays with whole-cell extracts and preference tests of ecologically relevant fouling organisms on natural algal and control surfaces may be a useful procedure to avoid erroneous conclusions regarding natural antifouling roles of compounds based on settlement assays with only whole-cell extracts. Furthermore, this study also shows that production of inhibitory metabolites may explain the low degree of fouling, especially by B. improvisus, on C. crispus.Communicated by L. Hagerman, Helsingør     

Characteristics of feeding on dinoflagellates by newly hatched larval crabs

The zoeal larvae of brachyuran crabs must feed soon after hatching on a diet that includes large micro- and mesozooplankton in order to satisfy nutritional requirements. However, newly hatched larvae have been shown to ingest a variety of dinoflagellates, perhaps using microbial carbon sources to sustain them until they encounter more favored prey. Ingestion of dinoflagellates by larval crabs has been documented previously under conditions in which the larvae were exposed to algae provided in monoculture or in defined mixtures of cells. We report here on experiments conducted on the hatching stage of five crab species to determine if ingestion of dinoflagellates occurred when they were provided in combination with Artemia sp. nauplii or after a period of feeding on mesozooplankton. Quantitative measurements of chl a in the larval guts provided evidence of ingestion of algal cells. Active ingestion of the dinoflagellate Prorocentrum micans at specified intervals during an extended feeding period was determined on larvae of two crab species using fluorescently labeled cells provided for brief periods at prescribed time intervals. Stage 1 larvae of four of the five crab species ingested dinoflagellates when they were provided in combination with nauplii and larvae of all five species ingested cells after feeding solely on nauplii for 24 h. Ingestion of algal cells was first evident in the larval guts after 6 h of feeding at both low (200 cell ml⁻¹) and high (1,000 cells ml⁻¹) prey densities. Higher prey densities resulted in higher gut chl a. Larvae continuously exposed to dinoflagellates actively ingested cells at every 3 h interval tested over a 36 h period. Results confirm previous studies that larvae will ingest dinoflagellates even when they are encountered in a mixed prey field or when having previously fed. Ingestion of cells may occur on a continual basis over time.