Validation of a technique for reconstructing daily patterns in the recruitment of coral reef damselfish

Counting growth increments in otoliths recently has become an accepted method of ageing tropical fishes, however, verification is essential for each new species. In this study, growth increments in otoliths of the juveniles of several coral reef damselfishes (Pomacentridae) were deposited daily and a distinct transition from wide to narrow increments coincided with settlement from the pelagic larval phase into the demersal habitat. Thus, the data of settlement for each individual fish could be calculated with acceptable accuracy. The daily pattern of recruitment to a coral reef was successfully reconstructed using the otoliths from a large sample of juvenile fish collected at the end of the breeding season. This was because the original pattern of settlement was preserved in the age distribution for at least 4 to 5 months. This application of otolith ageing techniques may be extended to reveal the temporal patterns of recruitment to many localities encompassing spatial scales larger than would be logistically possible using visual censuses.     

Transcriptional signature of accessory cells in the lateral line, using the Tnk1bp1:EGFP transgenic zebrafish line

Background

A metamorphic life-history is present in the majority of animal phyla. This developmental mode is particularly prominent among marine invertebrates with a bentho-planktonic life cycle, where a pelagic larval form transforms into a benthic adult. Metamorphic competence (the stage at which a larva is capable to undergo the metamorphic transformation and settlement) is an important adaptation both ecologically and physiologically. The competence period maintains the larval state until suitable settlement sites are encountered, at which point the larvae settle in response to settlement cues. The mechanistic basis for metamorphosis (the morphogenetic transition from a larva to a juvenile including settlement), i.e. the molecular and cellular processes underlying metamorphosis in marine invertebrate species, is poorly understood. Histamine (HA), a neurotransmitter used for various physiological and developmental functions among animals, has a critical role in sea urchin fertilization and in the induction of metamorphosis. Here we test the premise that HA functions as a developmental modulator of metamorphic competence in the sea urchin Strongylocentrotus purpuratus.

Results

Our results provide strong evidence that HA leads to the acquisition of metamorphic competence in S. purpuratus larvae. Pharmacological analysis of several HA receptor antagonists and an inhibitor of HA synthesis indicates a function of HA in metamorphic competence as well as programmed cell death (PCD) during arm retraction. Furthermore we identified an extensive network of histaminergic neurons in pre-metamorphic and metamorphically competent larvae. Analysis of this network throughout larval development indicates that the maturation of specific neuronal clusters correlates with the acquisition of metamorphic competence. Moreover, histamine receptor antagonist treatment leads to the induction of caspase mediated apoptosis in competent larvae.

Conclusions

We conclude that HA is a modulator of metamorphic competence in S. purpuratus development and hypothesize that HA may have played an important role in the evolution of settlement strategies in echinoids. Our findings provide novel insights into the evolution of HA signalling and its function in one of the most important and widespread life history transitions in the animal kingdom - metamorphosis.     

Nutritional condition of metamorphosing sole: spatial and temporal analyses

Sole Solea solea larvae were collected during five cruises in the Bay of Biscay in 1992 and 1993. The larval growth rate, triacylgycerol (TAG) and sterol (ST) content, and the width of the marginal otolith increments were determined on the same individual larva as indicators of nutritional condition. The TAG content increased during the ontogenetic larval development up to an elevated value corresponding to the end of metamorphosis. During metamorphosis, sole larvae continued to accumulate TAG and to increase in length. Seasonal and yearly variations of growth and nutritional condition of sole larvae at the onset of metamorphosis were more important than spatial variations. Analysis of larval growth showed that, during each cruise, the faster growing larvae were generally those with the highest TAG content (and TAG : ST ratio). In 1993, larvae began to metamorphose earlier and their growth (both larval growth and recent growth) were significantly higher than in 1992. Their relatively low nutritional condition, however, seemed to be the result of a low accumulation of TAG. Sole metamorphosis occurs without growth arrest and the majority of the larvae during this phase are judged to be in good nutritional condition (TAG : ST ratio > 1). In the Bay of Biscay, metamorphosis does not seem to be a critical period in the life cycle of the sole. It is suggested that larvae originating from different cohorts probably have the same chance of survival during or just after metamorphosis when they enter the benthic environment.     

Short-term and latent post-settlement effects associated with elevated temperature and oxidative stress on larvae from the coral Porites astreoides

Coral reefs across the Caribbean are undergoing unprecedented rates of decline in coral cover during the last three decades, and coral recruitment is one potential process that could aid the recovery of coral populations. To better understand the effects of climate change on coral larval ecology, the larvae of Porites astreoides were studied to determine the immediate and post-settlement effects of elevated temperature and associated oxidative stress. Larvae of Porites astreoides were exposed to 27 °C (ambient) and +3.0 °C (elevated temperature) seawater for a short duration of 24 h; then, a suite of physiological parameters were measured to determine the extent of sublethal stress. Following the +3.0 °C treatment, larvae did not show a significant difference in maximum quantum yield of PSII (F _v/F _m) or respiratory demand when compared to controls maintained at 27 °C. The addition of micromolar concentrations of hydrogen peroxide did not impact respiration or photochemical efficiency. Catalase activity in the larvae increased (>60 %) following exposure to elevated temperature when compared to the controls. Short-term larval survival and settlement and metamorphosis were not affected by increased temperature or the H₂O₂ treatment. However, the settled spat that were exposed to elevated temperature underwent a 99 % reduction in survival compared to 90 % reduction for the control spat when examined 24 days following the deployment of 4-day-old settled spat on settlement tiles in the field. These results show that short-term exposure to some stressors might have small impacts on coral physiology, and no effects on larval survival, settlement and metamorphosis. However, due to post-settlement mortality, these stressors can cause a significant reduction in coral recruitment.     

Growth during the early life history of the Pacific tarpon,Megalops cyprinoides

The early growth of the Pacific tarpon,Megalops cyprinoides, was studied by larval otolith analysis and rearing of larvae and juveniles in the laboratory. Morphology of the sagitta, validation of sagittal daily increments, age at the start of metamorphosis, decrement of standard length in early metamorphosis, and growth under rearing conditions are described. The sagitta of fully-grown Pacific tarpon leptocephali were transparent and circular, with regular intervals between the neighboring rings becoming wider at the onset of metamorphosis. Alizarin complexone treatment of larvae confirmed the daily formation of the sagittal rings. Metamorphosis was estimated to start about one month after hatching. After drastic shrinkage during the first several days of metamorphosis, the body length more or less stabilized for one month and then resumed rapid growth. The early growth of Pacific tarpon was divided into four phases as follows: A) leptocephalus positive growth phase; B) leptocephalus negative growth phase; C) sluggish growth phase; and D) juvenile growth phase.     

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.     

Larval settlement preferences of Acropora palmata and Montastraea faveolata in response to diverse red algae

Settlement specificity can regulate recruitment but remains poorly understood for coral larvae. We studied larvae of the corals, Acropora palmata and Montastraea faveolata, to determine their rates of settlement and metamorphosis in the presence of ten species of red algae, including eight species of crustose coralline algae, one geniculated coralline and one encrusting peyssonnelid. Twenty to forty percent of larvae of A. palmata settled on coralline surfaces of Hydrolithon boergesenii, Lithoporella atlantica, Neogoniolithon affine, and Titanoderma prototypum, whereas none settled and metamorphosed on Neogoniolithon mamillare. Larvae of M. faveolata had 13–25 % settlement onto the surface of Amphiroa tribulus, H. boergesenii, N. affine, N. munitum, and T. prototypum, but had no settlement on the surface of N. mamillare, Porolithon pachydermum, and a noncoralline crust Peyssonnelia sp. Some of these algal species were common on Belizean reefs, but the species that induced the highest rates of larval settlement and metamorphosis tended to be rare and primarily found in low-light environments. The shallow coral, A. palmata, and the deeper coral, M. faveolata, both had increased larval settlement rates in the presence of only a few species of red algae found at deeper depths suggesting that patterns of coral distribution can only sometimes be related to the distribution of red algae species.     

Effects of temperature on survival, development, growth and feeding of larvae of Yellowtail clownfish Amphiprion clarkii (Pisces: Perciformes)

To understand the physiological and ecological responses of marine fishes to the change of water temperature, newly-hatched larvae of Yellowtail clownfish Amphiprion clarkii were reared in captivity at water temperatures of 23, 26 and 29 °C till they completed the metamorphosis to juvenile phase, and larval survival, development, growth and feeding were evaluated during the experimental period. The results showed that water temperature influenced the physiological performance of larvae of A. clarkii significantly. The survival and growth rates of larvae of A. clarkii increased significantly with the increase of water temperature from 23 to 29 °C (P < 0.05). Water temperature also influenced larval development of A. clarkii significantly and larvae reared at 23 °C took longer time for post-larval development and metamorphosis compared to 26 and 29 °C (P < 0.05). Total length and body weight for post-larval development and metamorphosis decreased with the increase of water temperature from 23 to 29 °C (P < 0.05). Q₁₀ in developmental rate was higher than in daily growth rate at the same rearing temperature, indicating that at water temperature had greater influence on larval development than on growth. Water temperature also influenced larval feeding of A. clarkii significantly with feed ration (FR) and feed conversion efficiency (FCE) increased with the increase of water temperature from 23 to 29 °C (P < 0.05). There was a positive correlation between FR and specific growth rate (SGR) (P < 0.05) but not between FCE and SGR (P > 0.05), indicating that FR influenced growth rate significantly in larvae of A. clarkii. This study demonstrated that the physiological responses of larvae of A. clarkii to the change of water temperature and confirmed that water temperature influenced larval survival, development, growth and feeding significantly. This study suggests that the decline of larval survival and growth rates, extension of pelagic larval duration and reduction of larval feeding at lower temperature have ecological impacts on larval dispersal and metamorphosis, juvenile settlement and population replenishment in A. clarkii in the wild.     

Otolith microstructure during the pelagic, settlement and benthic phases in burbot

During the pelagic larval phase of burbot Lota lota L., the pattern of otolith increments changes, showing three, clearly distinguishable growth sectors: a first sector with faint increments, difficult to enumerate, comprising an average (±S.D.) of 17˙5±6˙7 increments, a second sector with distinct increments comprising an average of 33˙1±7˙6 increments and a third sector where increments again become faint and difficult to enumerate. Laboratory experiments conducted in parallel to the field investigation showed that settlement occurs after the formation of this third, faint sector and is marked by the formation of numerous accessory growth centres within the range of three to five daily increments. There was a strong linear relationship between sagittal width and total length of the burbot (r²=0˙928) over the range examined. Significantly different growth rates were calculated for the three otolith sectors (faint, distinct, faint) in burbot larvae, indicating large environmental changes during their pelagic larval phase in Lake Constance. These results suggest that inshore migration of burbot larvae does not take place in the warm epilimnetic surface waters but via an alternative pathway, the cold hypolimnion or profundal zone.     

Ecological regulation of development: induction of marine invertebrate metamorphosis

In the marine environment a wide range of invertebrates have a pelagobenthic lifecycle that includes planktonic larval and benthic adult phases. Transition between these morphologically and ecologically distinct phases typically occurs when the developmentally competent larva comes into contact with a species-specific environmental cue. This cue acts as a morphogenetic signal that induces the completion of the postlarval/juvenile/adult developmental program at metamorphosis. The development of competence often occurs hours to days after the larva is morphologically mature. In the non-feeding--lecithotrophic--larvae of the ascidian Herdmania curvata and the gastropod mollusc Haliotis asinina, gene expression patterns in pre-competent and competent stages are markedly different, reflecting the different developmental states of these larval stages. For example, the expression of Hemps, an EGF-like signalling peptide required for the induction of Herdmania metamorphosis, increases in competent larvae. Induction of settlement and metamorphosis results in further changes in developmental gene expression, which apparently is necessary for the complete transformation of the larval body plan into the adult form.     

Effect of delayed metamorphosis on larval competence, and post-larval survival and growth, in the abalone Haliotis iris Gmelin

Marine invertebrate species vary in their ability to delay metamorphosis, and in the degree to which delayed metamorphosis compromises juvenile performance. Abalone (Haliotis iris) larvae were deprived of metamorphosis cues and the effects of delayed metamorphosis on larval competence, and post-larval growth and survival were quantified. Larvae were exposed to a metamorphosis inducer (the coralline alga Phymatolithon repandum (Foslie) Wilks and Woelkerling) on Days 11, 18, 22, 26, 30 and 34 post-fertilisation (temperature 16-17 degrees C). Post-larvae were reared on diatoms (Nitzschia longissima Grunow) for 3-4 weeks post-metamorphosis. Delayed metamorphosis caused progressive negative effects on post-larval performance. Virtually all larvae initiated metamorphosis in response to P. repandum, regardless of larval age. The proportion of post-larvae that developed post-larval shell growth within 2 days of metamorphosis induction dropped only approximately 20% from Day 11 to Day 26 (P>0.05), but was significantly lower by Day 30 and Day 34 (P<0.001). Larvae that metamorphosed on Days 11, 18 and 22 showed high survival (>80%) and growth rates (means of 20-22 μm shell length per day). In contrast, larvae that metamorphosed on Day 26 and Day 30 had poor survival (30-40%) and lower (P<0.05) growth rates (15-16 μm/day). Of the larvae that metamorphosed on Day 34, only 7 (30%) survived their first week post-metamorphosis, and they grew only 2 μm/day on average. Only one of these post-larvae (4%) survived the second week. The visible yolk supply diminished over the life of the larvae and was near zero by Day 34. Nearly all larvae had died by Day 38. H. iris larvae remained competent to metamorphose for at least 3 weeks after they attained competence. Post-larval growth and survival were not reduced if metamorphosis occurred within 3 weeks of fertilisation. This extended period of larval competence implies that H. iris larvae can potentially disperse for up to several weeks before successful metamorphosis.     

Role of habitat in mediating mortality during the post-settlement transition phase of temperate marine fishes

The transition phase describes a distinct post-settlement stage associated with the recruitment to benthic habitats by pelagic life stages. The habitat shift is often accompanied by feeding shifts and metamorphosis from larval to juvenile phases. Density-dependent settlement, growth and mortality are often the major factors controlling recruitment success of this phase. Habitat use also becomes more pronounced after settlement. The role of habitat-mediated post-settlement mortality is elucidated by focusing on the early life history of Atlantic cod ( Gadus morhua ) and cunner ( Tautogolabrus adspersus ) in the north-west Atlantic. In these species, settlement can occur over all bottom types, but habitat-specific differences in post-settlement mortality rates combined with size and priority at settlement effects on growth and survival determine recruitment and eventual year-class strength. These results and those from other temperate marine fish species along with work on tropical reef species emphasize the generality of habitat-based density-dependent mortality during the transition phase and its potential for population regulation. These results have implications for fisheries management and can be used to outline a procedure to assist managers in identifying and managing essential transitional habitats including the potential role of marine protected areas in habitat conservation.     

Coral larvae under ocean acidification: survival, metabolism, and metamorphosis

Ocean acidification may negatively impact the early life stages of some marine invertebrates including corals. Although reduced growth of juvenile corals in acidified seawater has been reported, coral larvae have been reported to demonstrate some level of tolerance to reduced pH. We hypothesize that the observed tolerance of coral larvae to low pH may be partly explained by reduced metabolic rates in acidified seawater because both calcifying and non-calcifying marine invertebrates could show metabolic depression under reduced pH in order to enhance their survival. In this study, after 3-d and 7-d exposure to three different pH levels (8.0, 7.6, and 7.3), we found that the oxygen consumption of Acropora digitifera larvae tended to be suppressed with reduced pH, although a statistically significant difference was not observed between pH conditions. Larval metamorphosis was also observed, confirming that successful recruitment is impaired when metamorphosis is disrupted, despite larval survival. Results also showed that the metamorphosis rate significantly decreased under acidified seawater conditions after both short (2 h) and long (7 d) term exposure. These results imply that acidified seawater impacts larval physiology, suggesting that suppressed metabolism and metamorphosis may alter the dispersal potential of larvae and subsequently reduce the resilience of coral communities in the near future as the ocean pH decreases.     

Promoting larval settlement of coral Pocillopora damicornis by calcium

Larval settlement is a critical bottleneck in the process of coral sexual propagation. Promoting coral larval settlement by inducers is a promising strategy in coral reef restoration engineering. In this study, the settlement-promoting effect of Ca²⁺ on larvae of the brooding coral Pocillopora damicornis was investigated for the first time. Treatment with 40 mM CaCl₂ for 24 h effectively promoted coral larval settlement (~ 80%). Moreover, CaCl₂ is comparable with the natural inducer, crustose coralline algae (CCA), in both promoting coral larval settlement and post-settlement growth. CaCl₂ showed toxic effects on larval survival and growth at high concentrations, and this could be minimized by optimizing CaCl₂ concentration and shortening the exposure period. Our study suggests that applying Ca²⁺ to effectively and efficiently induce coral larval settlement is viable for laboratory research and small-scale aquaculture systems, and it might become a useful tool in future coral reef restoration engineering.

     

Larval settlement preferences and post-settlement survival of the threatened Caribbean corals Acropora palmata and A. cervicornis

The settlement specificity of two threatened Caribbean corals, Acropora palmata and A. cervicornis, was tested by measuring their rates of larval metamorphosis in response to crustose coralline algae (CCA) and other substrata. In the no-choice experiments, the coral larvae were placed in six treatments: filtered seawater (FSW), a fragment of biofilmed dead skeleton of A. palmata, or a fragment of one of four species of CCA (Hydrolithon boergesenii, Porolithon pachydermum, Paragoniolithon solubile, and Titanoderma prototypum). Within each CCA treatment, there were three different substrata on which to settle and metamorphose: (1) the CCA surface, (2) the rock under the CCA, or (3) the plastic dish. The 5-day-old larvae of both A. palmata and A. cervicornis had similar rates of total metamorphosis (all substrata combined) in every treatment (excluding FSW) even in the absence of CCA. However, there were differences in larval behavior among the CCA species since the larvae settled and metamorphosed on different substrata in the presence of different CCA species. In the no-choice experiments the larvae of both corals had higher rates of metamorphosis on the top surfaces of H. boergesenii and/or T. prototypum than on P. pachydermum. In the choice experiments, the coral larvae were offered two species of CCA in the same dish. When given a choice, both species of coral larvae had more settlement and metamorphosis on the surface of H. boergesenii or T. prototypum or clean rock than onto the surface of P. solubile. After 6 weeks in the field, transplanted A. palmata recruits had approximately 15% survival on both T. prototypum and H. boergesenii, but A. cervicornis recruits only survived on T. prototypum (13%). Some, but not all, CCA species facilitated the larval settlement and post-settlement survival of these two threatened corals, highlighting the importance of benthic community composition for successful coral recruitment.     

Microstructural growth in otoliths of black rockfish,sebastes schlegeli, from prenatal larval to early juvenile stages

Microstructural growth in the sagittae ofSeastes schlegeli, a viviparous scorpaenid, is described from prenatal larval to early juvenile stages, and related to morphological changes. Embryos and prenatal larvae were extruded from a gravid female from 21 d prior to birth onwards, and released larvae reared and sampled up to 58 d after birth. Eggs hatched in the ovary 14 d prior to birth. At this time, otoliths consisted of a core surrounded by a prominent check, similar to the otolith structure seen in oviparous fishes. Fourteen growth increments had been deposited by birth. The parturition mark it-self comprised a prominent check and narrow growth increment Growth increments were deposited daily from hatching up to 58 d after birth, whereas accessory primordia first appeared in otoliths by ca. 32 d after birth, at a specimen total length of ca. 13 mm. This corresponded to the period during which the larvae metamorphosed into juveniles. Otoliths grew exponentially during the larval stage and linearly during the juvenile stage. when plotted against total length. Growth in total length from hatching to 58 d after birth could be represented by the Gompertz curve.     

Growth of North Alboran Sea sardine larvae estimated by otolith microstructure, nucleic acids and protein content

Wet mass and DNA, RNA and protein content increased significantly with standard length ( L _S) of sardine Sardina pilchardus larvae, collected in January 1995, in the Bay of Málaga, North Alboran Sea. L _S, wet mass and DNA, RNA and protein content were closely related allometrically to otolith radius ( R ). Larval daily length increments decreased but DNA, protein and wet mass daily increments increased with larval age. Daily length increments showed a negative and poor relationship with long-term otolith growth. In contrast, DNA, protein and wet mass daily increments were positively correlated. Differences between observed and back-calculated otolith radius-at-age indicated that larvae with slow otolith growth were under represented in older age groups, suggesting the existence of growth-selective mortality. Recent otolith growth, estimated from the mean widths of the last six increments, increased with age and R . Individual RNA: DNA and protein: DNA ratios were correlated significantly, although weakly, with L _S and larval growth.     

Otolith microstructure of Japanese sea bass larvae and juveniles: interpretation and utility for ageing

This paper interprets and discusses the usefulness of otolith microstructure for ageing Japanese sea bass ( Lateolabrax japonicus ) larvae and juveniles. Samples were collected from the Tango Sea along the Japan Sea coast, January–March 2007. Known-age (0-day and 10-day-old) larvae were obtained from the Ibaragi Prefectural Hatchery, Japan. Sagittal and lapillar otolith were processed and read using an otolith reading system. Clearly discernible hatch- and first-feeding marks were evident on sagitta, and development of accessory premordia (AP) appeared to be associated with larva-juvenile transition; however, no other marks indicating metamorphosis or settlement were evident. In lapillus, no discernible check mark was found. Known-age larvae showed that deposition of the first daily increment (DI) corresponded to first-feeding, which occurred at day-4 post-hatch. However, mean increment counts were significantly lower in lapillus than in sagitta, caused by poorly expressed increments around the centrum as well as relatively unclear centrum of the lapillus. The authors suggest that the use of lapillus can cause significant underestimation of age. Therefore, the sagitta is recommended for age and growth estimations of larvae and juveniles, although the presence of numerous subdaily increments warrants careful preparation and interpretation of the microstructure. A test for asymmetry showed the right and left otoliths to be quite symmetrical and their DI counts not significantly different, suggesting that either otolith can be used for studying age and growth of Japanese sea bass larvae and juveniles.     

Influence of delayed metamorphosis on postsettlement survival and growth in the sipunculan Apionsoma misakianum

Abstract. Certain stresses experienced by marine larvae from many groups can dramatically reduce aspects of juvenile performance. This study reports the effects of delayed metamorphosis and nutritional stress on survival and growth of the deposit-feeding sipunculan Apionsoma (= Golfingia ) misakianum . Approximately 600 larvae collected from the Florida Current plankton were distributed among 3 treatment groups. Ninety larvae (controls) were offered sediment and adult-conditioned seawater 4 d after collection, to induce metamorphosis; larvae of this species could not be induced to metamorphose by increasing the K⁺ concentration of seawater. The remaining 500 larvae were kept swimming for either 2 or 4 weeks, with or without phyto-plankton (clone T-ISO). At the end of the periods of prolonged larval swimming, subsampled larvae (360) were induced to metamorphose as in the controls. Surviving individuals were retrieved 6 weeks after the addition of excess sediment in all treatments, and weighed to document growth. Neither delayed metamorphosis nor starvation influenced juvenile survival. However, starving larvae for 2 weeks significantly reduced mean juvenile growth rates relative to the mean growth rate of control individuals (p<0.0001), while prolonging larval life by 4 weeks significantly reduced mean juvenile growth rates (p<0.05) whether or not larvae were fed. Reduced juvenile growth rates may have been caused by nutritional stress experienced by larvae in both the starved and fed treatments. The rapid response of freshly collected larvae to sediment indicates that competent larvae of this species routinely delay metamorphosis in the field. The extent to which they also experience food limitation is not yet clear. If competent larvae are food limited while delaying metamorphosis in the field, our results suggest that juveniles will grow more slowly and may thus exhibit reduced fitness.