Less inhibited with age? Larval age modifies responses to natural settlement inhibitors

As larvae of marine invertebrates age, their response to settlement cues can change. This change can have significant consequences to both the ecology of these organisms, and to their response to antifouling coatings. This study examines how larval age affects the settlement response of larvae to two naturally derived settlement inhibitors, non-polar extracts from the algae Delisea pulchra and Dilophus marginatus, the former of which contains compounds that are in commercial development as antifoulants. Two species of marine invertebrates with non-feeding larvae were investigated: the bryozoans Watersipora subtorquata and Bugula neritina. Larval age strongly affected larval settlement, with older larvae settling at much higher rates than younger larvae. Despite having strong, inhibitory effects on young larvae, the non-polar extracts did not inhibit the settlement of older larvae to the same degree for both species studied. The results show that the effects of ecologically realistic settlement inhibitors are highly dependent on larval age. Given that the age of settling larvae is likely to be variable in the field, such age specific variation in settlement response of larvae may have important consequences for host-epibiont interactions in natural communities.     

Interaction of conspecific cues in Balanus amphitrite Darwin (Cirripedia) settlement assays: continued argument for the single-larva assay

Gregariousness in marine invertebrate larvae is an important regulator of benthic community structure. Previous laboratory settlement assays employing Balanus amphitrite Darwin cyprids found gregarious effects with as few as 3 larvae well(-1), together with modulation of such effects by chemical cues. Here, the relationship between settlement rate and larval density was rigorously tested through a fully randomised design. Seawater conditioned with adult B.amphitrite was tested alongside unconditioned seawater to determine the effect of a conspecific cue on gregarious interactions. Gregarious effects were detected in both conditioned and unconditioned seawater at < or =4 larvae well(-1). In untreated seawater, settlement rate increased linearly with larval density, levelling off at densities of > or =10 larvae well(-1). In conditioned seawater, settlement induction was observed at < or =4 larvae well(-1), switching to inhibition at 6, 8 and 10 larvae well(-1), before asymptoting at the highest densities tested. These results advocate the use of individual larvae in laboratory assays that investigate factors stimulating barnacle settlement.     

Crustose coralline algae and a cnidarian neuropeptide trigger larval settlement in two coral reef sponges

In sessile marine invertebrates, larval settlement is fundamental to population maintenance and persistence. Cues contributing to the settlement choices and metamorphosis of larvae have important implications for the success of individuals and populations, but cues mediating larval settlement for many marine invertebrates are largely unknown. This study assessed larval settlement in two common Great Barrier Reef sponges, Coscinoderma matthewsi and Rhopaloeides odorabile, to cues that enhance settlement and metamorphosis in various species of scleractinian coral larvae. Methanol extracts of the crustose coralline algae (CCA), Porolithon onkodes, corresponding to a range of concentrations, were used to determine the settlement responses of sponge larvae. Cnidarian neuropeptides (GLW-amide neuropeptides) were also tested as a settlement cue. Settlement in both sponge species was approximately two-fold higher in response to live chips of CCA and optimum concentrations of CCA extract compared to 0.2 μm filtered sea water controls. Metamorphosis also increased when larvae were exposed to GLW-amide neuropeptides; R. odorabile mean metamorphosis reached 42.0±5.8% compared to 16.0±2.4% in seawater controls and in C. matthewsi mean metamorphosis reached 68.3±5.4% compared to 36.7±3.3% in seawater controls. These results demonstrate the contributing role chemosensory communication plays in the ability of sponge larvae to identify suitable habitat for successful recruitment. It also raises the possibility that larvae from distinct phyla may share signal transduction pathways involved in metamorphosis.     

Studies on the factors influencing larval settlement in Balanus amphitrite and Mytilus galloprovincialis

Understanding of factors influencing settlement(attachment and metamorphosis) of marine invertebratelarvae is of great importance in aquaculture andcontrol of biofouling. The influence of two factors onsettlement of larvae was assessed from two separateinvestigations: 1, the influence of age (endogenousfactor) on cyprids of the barnacle Balanusamphitrite; and 2, the influence of a microbial film(exogenous factor) on pediveligers of the mussel Mytilus galloprovincialis.The settlement response of cypris larvae of B.amphitrite was found to be age-dependent. Oldercyprids responded more readily to settlement factorsthan newly molted ones. In M.galloprovincialis, competent pediveligers settled inresponse to a microbial filmed surface but not toan unfilmed surface. Moreover, a factor with MW of lessthan 5000 dalton, derived from culture medium of abacterial strain C1.1 (Pseudomonas-Alteromonasgroup), induced the settlement of M. galloprovincialis larvae.Thus, marine invertebrate larvae may require a periodof competence acquisition, during which they arepoorly responsive to settlement inducers. Uponacquisition of competence, larvae readily respondto external cues (e.g. microbial film, bacterialextracellular products).     

Effect of meiofauna on macrofauna recruitment: settlement inhibition of the polychaete Hydroides elegans by the harpacticoid copepod Tisbe japonica

Various aspects of marine macroinvertebrate ecology cannot be understood without detailed knowledge of larval settlement processes. An important effect underscored during the settlement process is the disturbance of marine invertebrate larvae by predators. We demonstrated that biotic disturbance, comprising physical elimination and mortality due to predation and the behavioral irritation of larvae by the harpacticoid copepod Tisbe japonica, prevent a significant portion of larvae of the polychaete Hydroides elegans from settlement on otherwise suitable substrata. Experiments were performed both in the laboratory and the field showing reproducible significant differences in larval settlement and mortality rates between gender-specific copepod treatments and the control. The trend of decreased larval settlement in the presence of copepods coincided with increased larval mortality in these treatments. In the corresponding field experiments, larval settlement and mortality were similar to the ones obtained under laboratory conditions.     

The influence of resident adults on larval settlement: experiments with four species of ascidians

Residents within any community can affect the larval settlement of both their own and other species. In marine sessile communities resident adults can affect larval settlement by preying on settling larvae, removing or adding space for the larvae to colonize, or stimulating or prohibiting larval settlement on available substratum nearby. To examine those processes by which residents affect settlement, we exposed experimental substrata with three densities of adults of a single species at a site in eastern Long Island Sound, USA for a 24-h period. Four species of common ascidians, Botryllus schlosseri (Pallas), Botrylloides diegensis Ritter and Forsyth, Diplosoma macdonaldi Herdman, and Molgula manhattensis (De Kay), were used in 11 separate experiments. Few individuals of any species settling attached to the surfaces of these species and this resulted in the main effect of these residents being the usurpation of space and the restricting of settlement to unoccupied areas. A model is also presented to explain the apparent aggregated settlement of several species in open areas adjacent to the resident ascidians. From this model we suggest that the aggregated settlement can result from limited larval mobility such that some larvae that contact and reject the resident species as settlement sites may subsequently contact open surfaces of the same substratum and increase settlement densities there over those observed on control substrata. Finally, settlement data for several species indicate that Molgula may influence settlement by preying on larvae.     

Induction of settlement in mussel (Perna canaliculus) larvae by vessel noise

Underwater sound plays an important role in the settlement behaviour of many coastal organisms. Large steel-hulled vessels are known to be a major source of underwater sound in the marine environment. The possibility that underwater sound from vessels may promote biofouling of hulls through triggering natural larval settlement cues was investigated for the mussel, Perna canaliculus. The mussel larvae showed significantly faster settlement when exposed to the underwater noise produced by a 125-m long steel-hulled passenger and freight ferry. Median time to attachment on the substrata (ie settlement) was reduced by 22% and the time taken for all experimental larvae to settle was reduced by 40% relative to a silent control. There was no difference in the survival of the mussel larvae among the various noise treatments. The decrease in settlement time of the mussel larvae appeared to correlate with the intensity of the vessel sound, suggesting that underwater sound emanating from vessels may be an important factor in exacerbating hull fouling by mussels.     

The importance of flow and settlement cues to larvae of the abalone, Haliotis rufescens Swainson

In flow tank experiments, I tested the relative importance of active and passive processes to larvae settling on manufactured casts that were hydrodynamically rough at a small scale (mm to <1 cm). I predefined two distinct regions of small-scale flow that I used to manipulate larval settlement behaviour of the red abalone Haliotis rufescens Swainson. The larvae show a stringent settlement response associated with coralline red algae. Haliotis rufescens larvae settled preferentially to an inducer regardless of the flow conditions, as expected. However, the ability of H. rufescens larvae to show this stringent behaviour was altered by changing the small-scale flow. When the free-stream velocity was low, the larvae responded to a settlement cue regardless of the small-scale hydrodynamics. When free-stream velocity was higher, the larvae acted increasingly as passive particles in their deposition, but settled only in response to an inducer. The results were consistent in two flow tanks, across 2 years and between different batches of larvae.     

Sediment-associated cues for larval settlement of the infaunal spionid polychaetes Polydora cornuta and Streblospio benedicti

Do patchy distribution patterns of infaunal polychaetes result from active site selection of larvae influenced by sediment-associated microbial cues? This hypothesis was tested with still-water laboratory settlement assays revealing the acceptance or rejection of polychaete larvae to qualitatively different sediments. Laboratory brood cultures of the spionid polychaetes Polydora cornuta and Streblospio benedicti yielded a sufficient number of larvae with planktotrophic development for bioassays. High settlement rates (75-95%) of test larvae were observed in response to natural sediment. Sterilization of natural sediment significantly decreased settlement of P. cornuta (25-55%) while combustion of sediment significantly decreased the settlement rate in both species (5-50%). Differences in settlement responses to sediments treated by sterilization or combustion most likely resulted from a variety of factors such as modified sediment fabric, grain size distribution and quantity of adsorbed organic matter. To experimentally address the potential role of microorganisms and microbial metabolites as mediators of larval settlement, ashed sediment was inoculated with viable microorganisms obtained from natural sediment. In both polychaete species, this treatment significantly increased larval settlement in comparison to the control of ashed sediment indicating that larval settlement was at least partially mediated by the presence of microorganisms associated with sediment.     

Effects of delayed settlement on post-settlement growth and survival of scleractinian coral larvae

Demographic connectivity requires both the dispersal of individuals between sub-populations, and their subsequent contribution to population dynamics. For planktonic, non-feeding marine larvae, the capacity to delay settlement enables greater dispersal distances, but the energetic cost of delayed settlement has been shown to adversely impact post-settlement fitness in several taxa. Here, we assess whether delayed settlement influences mortality rates or growth rates for the first 6 weeks following settlement of the scleractinian coral, Acropora tenuis. Coral larvae that were settled at 2, 4, and 6 weeks after spawning, and then deployed in the field, showed negligible effects of delayed settlement on post-settlement survival and time to initial budding for colony formation. Between-cohort differences in budding rate appeared to be explained by temporal variation in the post-settlement acquisition of zooxanthellae. The potential for coral larvae to remain in the pelagic zone for increased periods of time with little to no effect on post-settlement survival and growth suggests that the capacity for delayed settlement is likely to have meaningful demographic consequences for broadcast-spawning reef-building corals, and that the predicted trade-off between delayed settlement and post-settlement fitness is less applicable to reef-building scleractinian corals than other taxa with non-feeding larvae.     

In situ observation of settlement behaviour in larvae of coral reef fishes at night

The swimming behaviour of 534 coral reef fish larvae from 27 species was explored at Moorea Island (French Polynesia) while they searched for a suitable settlement habitat, on the first night of their lagoon life. Most larvae swam actively (74%) and avoided the bottom (77%). A significant relationship was highlighted between the vertical position of larvae in the water column and the distance they travelled from lagoon entrance to settlement habitat: larvae swimming close to the surface settled farther away on the reef than bottom-dwelling larvae.     

Adding coral rubble to substrata enhances settlement of Pocillopora?damicornis larvae

Settlement preferences of Pocillopora damicornis larvae were examined on artificial substrata. Planulation of P. damicornis followed a lunar cycle and the release of larvae occurred after new moon. P. damicornis larvae had the highest rates of settlement within 3 days of being presented settlement substrata. Cumulative settlement gradually increased from 3 to 8 days, and post-settlement mortality was most frequent after 8 days. Settlement experiments showed greatest settlement preference to cement tiles containing 10% coral rubble. This study suggests that physical cues are important in the settlement process, which may be useful for coral reef rehabilitation projects.     

Depth and timing of settlement of veligers from different populations of giant scallop, Placopecten magellanicus (Gmelin), in thermally stratified mesocosms

The present study was conducted to evaluate the relative roles that water column stratification intensity and possible inter-population behaviour differences play in determining depth of larval settlement of giant scallops, Placopecten magellanicus, in relation to thermoclines. Differences in timing of settlement of larvae from various populations were also examined. Two separate experiments were conducted in a 10.5-m-deep, 3.7-m-diameter, thermally stratified tank with larvae spawned from scallops collected from several adult beds located in areas with differing oceanographic regimes (Georges Bank, Mahone Bay, Passamaquoddy Bay). Previous mesocosm experiments had shown that veligers from these various populations differ in their vertical migration patterns. In the first experiment (December 1992-February 1993), larvae from all three populations were held in separate 9.5-m-deep tubes and exposed to a 1.5 °C temperature differential established over a depth interval of 1 m. The number of settled juveniles (spat) of each population collected at the end of the experiment increased with depth and showed no peak at or above the thermocline. This depth distribution of spat was most likely driven by preferential larval settlement. In the second experiment (February-May 1994), larvae spawned from Georges Bank and Passamaquoddy Bay stocks were held in 9.0-m-deep tubes and exposed to a 5 °C temperature differential established over a depth interval of 1 m. The number of settled spat of both populations was greater above this thermal boundary and increased with decreasing depth. This depth distribution was most likely driven by preferential settlement above the thermocline followed by upward post-settlement migration. The results from the two experiments indicate that larvae from various populations show similar trends in settlement patterns in response to similar thermal stratifications. Stratification intensity, however, does affect depth of larval settlement. In the second experiment, both populations of larvae settled throughout the time interval of collector deployment (larvae 32-82 days old). These results extend the range of planktonic developmental times generally reported in the literature and may be more indicative of natural planktonic development in the field. While Georges Bank larvae settled in consistent numbers through time, Passamaquoddy Bay larvae showed peaks in settlement at certain time periods, indicating that pulses of larval settlement may occur even from an individual spawning event.     

Rapid settlement in broadcast spawning corals: implications for larval dispersal

Broadcast spawning of gametes with planktonic development of larvae is the most common reproductive mode in tropical corals, and is generally thought to optimize the dispersal potential of larvae. To this end, many previous studies of coral larval dispersal have focused on the maximum time larvae can remain competent to settle and consequently how far they might disperse. However, dispersal ability of broadcast-spawned coral larvae will be linked, at least in part, to the minimum time to settlement competency as well as the length of the planktonic period—although estimates of minimum time to competency remain largely anecdotal, with few rigorous studies of the pre-competent period. To determine the minimum time to larval settlement in two species of broadcast-spawning coral ( Platygyra daedalea and Goniastrea favulus), we monitored larval settlement rates in aquaria every 6 h from the time larvae commenced swimming (i.e. were ciliated, fully developed larvae) for a period of approximately 10 days. For P. daedalea, peak settlement occurred between 60 and 66 h following fertilization (2.5 and 2.75 days), which is markedly earlier than the 4- to 6-day time period commonly cited as the minimum time before broadcast-spawned coral larvae are competent to settle. Surprisingly, it was also clear from our experimental results that settlement in P. daedalea occurred as a distinct pulse during the 60- to 66-h period, rather than continuously throughout the study period. G. favulus larvae also appear to be able to settle quickly (from 54 h following fertilization). We argue, on the basis of these short competency times and apparently rapid settlement, that dispersal in broadcast-spawning coral larvae may not be as great as has previously been assumed.     

Effect of temperature on the settlement choice and photophysiology of larvae from the reef coral Stylophora pistillata

To better understand the consequences of climate change for scleractinian corals, Stylophora pistillata was used to test the effects of temperature on the settlement and physiology of coral larvae. Freshly released larvae were exposed to temperatures of 23 degrees C, 25 degrees C (ambient), and 29 degrees C at light intensities of approximately 150 micromol photons m(-2) s(-1). The effects were assessed after 12 h as settlement to various substrata (including a choice between crustose coralline algae [CCA] and limestone) and as maximum quantum yield of PSII (F(v)/F(m)) in the larvae versus in their parents. Regardless of temperature, 50%-73% of the larvae metamorphosed onto the plastic of the incubation trays or in a few cases were drifting in the water, and 14% settled on limestone. However, elevated temperature (29 degrees C) reduced the percentage of larvae swimming by 81%, and increased the percentage choosing CCA nearly 7-fold, both relative to the outcomes at 23 degrees C. Because temperature did not affect settlement on limestone or plastic, increased settlement on CCA reflected temperature-mediated choices by larvae that otherwise would have remained swimming. Interestingly, F(v)/F(m) was unaffected by temperature, but it was 4% lower in the larvae than in the parents. These results are important because they show that temperature can affect the settlement of coral larvae and because they reveal photophysiological differences between life stages that might provide insights into the events associated with larval development.     

Loss and gain of the juvenile rudiment and metamorphic competence during starvation and feeding of bryozoan larvae

SUMMARY In many animals, larval structures and juvenile rudiments develop independently. One advantage of this independence is that juvenile rudiments can be expended as a nutrient reserve or for energy conservation. When bryozoan cyphonautes larvae were starved, structures required for settlement and metamorphosis shrank. When the larvae were again fed, these structures grew back. Starvation reduced the size of both the internal sac, a rudiment of postlarval juvenile structures, and the pyriform organ, which functions in sensing and crawling on the substratum at settlement. In contrast, starvation affected neither the size of the larval shell nor the lengths of the ciliary bands used in swimming and feeding. Starved larvae that had reduced the pyriform organ and internal sac did not metamorphose in response to stimuli from a laminarian alga. The laminarian alga did stimulate metamorphosis of the same larvae after renewed feeding, when the larvae had regrown these structures. Thus starved larvae expended body parts needed for settlement and metamorphosis when food was scarce while retaining structures for feeding, swimming, and defense. Starved larvae thereby retained the capacity to regrow structures needed for settlement and metamorphosis when they again encountered food. Advantages from expendable juvenile rudiments may enhance selection for their being developmentally distinct from structures for larval swimming and feeding.     

Adrenoceptor compounds prevent the settlement of marine invertebrate larvae: Balanus amphitrite (Cirripedia), Bugula neritina (Bryozoa) and Hydroides elegans (Polychaeta)

The effects of the neurotransmitter blockers idazoxan and phentolamine on the larval settlement of three marine invertebrate species belonging to three different phyla were investigated by using in vitro concentration-response bioassays. Since neurotransmitters are known to influence metamorphic transitions in invertebrate larvae, neurotransmitter blockers were tested to evaluated their sublethal effects on larvae. The alpha-adrenergic antagonists idazoxan and phentolamine inhibited settlement of Balanus amphitrite (Cirripedia), Bugula neritina (Bryozoa) larvae, and larvae of the polychaete Hydroides elegans (Polychaeta) in a concentration-and taxon-dependent manner. At concentrations of 10(-3) M of both agents, larvae of all three species became immobile and subsequently died within 24 h. While cumulative settlement rates were observed after 48 h for B. amphitrite and H. elegans, and after 5 h for B. neritina, >90% of the larvae that settled did so within 24 h for the first two species and within 1 h for B. neritina. The tendency of the hydrophobic idazoxan and phentolamine to accumulate at solid surfaces most probably contributes to their successful inhibition of larval settlement. This ability makes them particularly attractive as candidates for the development of slow-release carriers in antifouling paints.     

Coral settlement onto Halimeda opuntia: a fatal attraction to an ephemeral substrate?

Degraded reefs with a high abundance of macroalgae usually also have low densities of coral recruits. Few studies, however, have examined whether these algae affect coral larval settlement. This study demonstrates, experimentally, that larvae of the Caribbean coral Favia fragrum can settle on the green alga Halimeda opuntia even when another substrate more suitable for settlement is present. Larval settlement onto experimental substrates was quantified under three treatments: rubble only, rubble plus plastic algal mimic, and rubble plus live H. opuntia. Similar total larval settlement was observed in all treatments. No larvae settled on the algal mimic, but total settlement was similar on the rubble in the first two treatments, showing that the rubble alone offered sufficient substrate for high settlement success. About half the larvae in the live algal treatment settled on H. opuntia instead of on the rubble, showing that larvae did not reject this substrate as they did the algal mimic. This result raises the possibility that corals will settle on some macroalgae when their abundance is high. Most macroalgae, including H. opuntia, are ephemeral substrates unsuitable for post-settlement survival. Such unexpected settlement may therefore have significant consequences for coral recruitment success on algal-dominated reefs.     

Developmental dimorphism: consequences for larval behavior and dispersal potential in a marine gastropod

Specific effects of alternative developmental programs on swimming and settlement behavior for marine larvae have not been identified experimentally. A major impediment to this research has been the rarity of species with variable development. Here, we compared traits related to movement and habitat selection for different ontogenetic stages of long-lived, feeding larvae (planktotrophic) and short-lived, nonfeeding larvae (lecithotrophic) of the herbivorous gastropod Alderia modesta. Newly hatched planktotrophic larvae swam in meandering paths with equal rates of upward and downward movement. As planktotrophic larvae developed towards competence (physiological ability to metamorphose), their swimming paths became straighter, faster, and increasingly directed towards the bottom, traits shared by newly hatched lecithotrophic larvae. Despite differing in developmental history, competent planktotrophic (32-d-old) and lecithotrophic larvae (competent upon hatching) exhibited qualitatively similar swimming behaviors and substrate specificity. However, lecithotrophic larvae moved downward at twice the speed of competent planktotrophic larvae, potentially producing a 5-fold higher rate of contact with the bottom in natural flows. Competent larvae swam downwards rather than passively sinking, even though sinking rates were faster than swimming speeds; active swimming may allow larvae to keep the velum extended, permitting rapid response to chemical settlement cues and promoting successful habitat colonization. Differences between larvae of the two development modes may reflect fine-tuning by selection of traits important for dispersal and settlement into patchy adult habitats.     

Habitat differentiation in the early life stages of simultaneously mass-spawning corals

The settlement process of coral larvae following simultaneous mass-spawning remains poorly understood, particularly in terms of population and community parameters. Here, the larval settlement patterns of Acropora corals, which are the most diverse genera of scleractinian corals at the species (haplotype) level, were investigated within a single subtropical reef. Across a 4-year period (2007–2010), the mitochondrial and nuclear molecular markers of 1,073 larval settlers were analyzed. Of the 11 dominant haplotypes of recruited populations, nine exhibited non-random patterns of settlement distribution. This result suggests that the actual habitat segregation starts during the early swimming larval stages of their life history, rather than by natural selection after random settlement. In addition, the presence of a depth-related settlement pattern supports that species-specific vertical zonation of coral larvae may play a role in the establishment of habitat segregation. Moreover, in some species that showed a preference toward the shoreward area of the bay, the settlement pattern was consistent with that of the adult distribution. This result indicates that the gametes were not mixed between fore and back reefs in the period from fertilization to settlement during the mass-spawning event, even within a single small reef. Another compatible hypothesis of this pattern is that the larvae are able to recognize various types of environmental information, facilitating the selection of optimal micro-habitats. Overall, Acropora coral larvae that are produced from a simultaneous mass-spawning event may have adapted to complex reef topography by means of multi-step habitat selection at settlement, corresponding to different spatial scales.