Developmental changes in the composition of myofibrillar proteins in the swimming muscles of Atlantic herring,Clupea harengus

Changes in myofibrillar protein composition during development have been investigated in the swimming muscles of the Atlantic herring Clupea harengus L. using a range of electrophoretic techniques. The main muscle-fibre type of larvae, and the fast- and slow-muscle fibres of adult fish were found to contain distinct isoforms of myosin heavy chain (MHC) and myosin light chain 2 (LC2). Larval LC2 was present as a minor component of adult fast-muscle myosin. In contrast, larval and adult fast-muscle myosin appeared to contain identical alkali light chains. Tropomyosin and troponin C were also identical in larval and in adult fast-muscle. All three muscle-fibre types contained unique isoforms of troponin T (TNT) and troponin I (TNI). Larval muscle had multiple isoforms of TNT, some of which may correspond to embryonic forms. It was concluded that although the main muscle-fibre type in larvae shares some myofibrillar proteins with adult fast muscle, it also contains characteristic isoforms of MHC, TNI, TNT and LC2 and therefore represents a distinct fibre type. The particular combination of myofibrillar proteins present at any developmental stage was found to be dependent on the rearing temperature. For example, a higher proportion of embryonic TNT isoforms were present at hatching in larvae reared at 5°C than at either 10 or 15°C. Over a period of 7 d, there was a gradual reduction in the number of TNT isoforms, but the pattern in 5°C larvae after 7 d still did not resemble that in 1 d-old larvae reared at 15°C.     

Temperature influences muscle differentiation and the relative timing of organogenesis in herring (Clupea harengus) larvae

Eggs from spring spawning stocks of herring (Clupea harengus L.) were fertilized and reared at either 5, 8 or 12°C in 1991 and 1992. The differentiation of myotomal muscle fibres was investigated in relation to the development of other organs and tissues using light and electron microscopy. The gut, notochord, eyes and haemocoel appeared at the same relative point in development between fertilization and hatching at all temperatures. In contrast, the formation of the spinal cord, pronephros, pectoral fin buds and muscle fibres was relatively retarded at 5°C compared with 8 and 12°C. Myogenesis in the presumptive inner muscle mass occurred after 12 to 16 d at 5°C, 7 to 10 d at 8°C and 3.5 to 6 d at 12°C. Myoblasts aligned in orderly rows running from myosept to myosept prior to fusion to form myotubes. Actin and myosin filaments were synthesised throughout the cytoplasm in associated with presumptive Z-lines at the periphery of myotubes and immature muscle fibres. Differentiation of the superficial and inner muscle fibres types of larvae occurred at around the same time. Following this initial period of myogenesis, the number of myotomal muscle fibres remained constant until after hatching, so that increases in muscle bulk in the late embryo were entirely due to fibre hypertrophy. At hatching, the number of superficial muscle fibres present in myotomes just posterior to the yolk-sac was significantly less at 5°C (108±12) than at either 8°C (132±10) or 12°C (140±10) (mean±SD, 12 fish/temperature). In contrast, there were around 280 inner muscle fibres/myotome, comprising 90% of the trunk cross-sectional area, at all three temperatures. Myofibrillargenesis occurred relatively slowly at low temperatures, so that the volume density of myofibrils in the inner muscle fibres of larvae at hatching was significantly less at 5°C (39.2±9.0) than at either 8°C (49.6±8.8) or 12°C (50.2±9.8) (mean ±SD, 20 fibres/temperature from total of 5 fish). Undifferentiated myoblasts remained at hatching to form a population of presumptive myosatellite cells. The number of presumptive myosatellite cells per mm² cross-sectional area of muscle fibre was more than two times higher at 8°C (1493±335) than at either 5°C (478±102) or 12°C (924±233) (mean±SD, 5 fish/temperature). The results suggest that temperature can influence the commitment of myoblasts to differentiation at a critical stage in embryogenesis, thereby providing a potential mechanism for influencing future growth characteristics. Correspondence to: I.A. Johnston at Gatty Marine Laboratory     

Influence of temperature on muscle-fibre development in larvae of the herringClupea harengus

The development of swimming (myotomal) muscles was studied in herring larvae (Clupea harengus L.) caught in the Firth of Clyde, Scotland, in spring 1990 and reared at either 5, 10 or 15°C. Two muscle-fibre types can be distinguished in the myotomes of herring larvae using ultrastructural criteria. A single layer of small-diameter muscle fibres, packed with mitochondria, is present beneath the entire surface of the skin (superficial muscle fibres). The remaining bulk of the muscle is composed of larger diameter fibres (inner muscle fibres) containing significantly more myofibrils than the superficial fibres. In 1 d-old larvae, the number of inner muscle fibres in myotomes immediately posterior to the yolk-sac was 311±41 at 15°C, 257±22 at 10°C and 187±22 at 5°C (mean±SD,n=6). The average diameter of inner muscle fibres increased with decreasing temperature, so that the total cross-sectional area of muscle was similar at each temperature. After 6 to 7 d, the number of muscle fibres had significantly increased at 15°C (383±25), but not at 10°C (281±32) or 5°C (192±17). In contrast, the average cross-sectional area of inner muscle fibres had increased by 19% at 15°C, 34% at 10°C, and 26% at 5°C. Temperature also influenced the relative proportions and spatial distributions of muscle-fibre organelles. For example, in 1 d-old larvae, the fraction of muscle-fibre volume (volume density) occupied by mitochondria in the superficial fibres was significantly higher at 15°C (46.0%) than at either 5°C (37.6%) or 10°C (38.8%). In the inner muscle fibres, the volume density of mitochondria was 26.1% at 15°C, 20.5% at 10°C and 15.9% at 5°C, whereas the volume density of myofibrils was similar at the three temperatures (33 to 38%). Typically, inner muscle fibres from 10°C larvae, but not from 5 or 15°C larvae contained a large central mitochondrion.     

Temperature and development in larvae of the turbot Scophthalmus maximus

Turbot (Scophthalmus maximus L.) were reared at 12 and 16°C until 26 d after hatching. At both temperatures, starting at the neural plate stage, somites were initially formed every 75 min. Expressed as a percentage of development time (DT, fertilisation to 90% larvae hatching) somite formation occurred relatively earlier during embryogenesis at 12°C (45% DT) than at 16°C (55% DT). At 12°C, after the 32-somite stage the rate of somite formation decreased to one every 300 min. The larvae hatched after 6 d at 12°C and 3 d at 16°C at a relatively primitive stage of development, prior to the opening of the mouth and anus, with unpigmented eyes, and a straight gut. Temperature altered the relative timing of organogenesis in the larval stages. At 12°C, the following characters appeared (in this order): swimbladder>loop in the gut (at the time of yolk exhaustion)>caudal fin. In contrast, at 16°C, the caudal fin appeared at the same time as the loop in the gut. At 16°C, spines formed on the head in the region of the otic capsule at the time the swimbladder formed and the yolk was exhausted, but were absent in 12°C larvae. At both temperatures, in 1 d-old larvae the myotomes just behind the yolk-sac contained 200 inner muscle fibres (presumptive white muscle). The initial growth of inner muscle was largely due to hypertrophy, but by 26 d at 12°C and 11 d at 16°C hyperplastic growth became important, as evidenced by a significant increase in the number of small fibres (<10 m²). By 26 d the average number of inner muscle fibres had increased to 341 at 12°C and 988 at 16°C. New muscle fibres were added in distinct germinal zones at the dorsal and ventral apices of the myotomes. Metamorphosis was associated with a thickening of the superficial (presumptive red) muscle layer and the appearance of tonic muscle fibres.     

Muscle structure and differentiation in pelagic and demersal stages of the Antarctic teleost Notothenia neglecta

The distribution and fine structure of muscle-fibre types has been determined for the pelagic fingerling and demersal adult stages of the antarctic teleost Notothenia neglecta Nybelin, collected from Signy Island, Antarctica, between January and February 1984. In both stages, the pectoral fin adductor muscle (m.ad.p) is largely composed of slow fibres which contain abundant mitochondria (34 to 36%). During development, the ratio of capillaries to fibres increases less than does fibre diameter, so that capillary density is significantly lower in the m.ad.p of adults (498 mm^-2) than fingerlings (1 727 mm^-2). The secondary metamorphosis from a pelagic to a demersal mode of life is associated with the disappearance of subcutaneous lipid sacs and major changes in the distribution and structure of muscle fibres in the myotomes. The trunk cross-section of adult fish is almost entirely composed of poorly vascularised fast-muscle fibres (100 capillaries mm^-2), which contain densely packed myofibrils (86.3%), and have few mitochondria (1.4%). Slow-muscle fibres in adults are restricted to a thin wedge adjacent to the lateral line canal. In contrast, slow fibres occur around the entire circumference of the trunk in fingerlings representing 24% of the total cross-sectional area at the post-anal level. Volume densities (%) of mitochondria, intracellular lipid and myofibrils in this tissue are respectively 37.0, 7.9, 38.6 for fingerlings and 13.1, 0, 70.3 for adults. Slow-muscle fibres in adult fish are of unusually large diameter (50 to 120 m) and have relatively low capillary densities (266 mm^-2). These morphological changes reflect a general decrease in activity and a shift from a sub-carangiform to a labriform mode of swimming following transition from the fingerling to adult stage of the life cycle. The results are briefly discussed in relation to the physiology and ecology of antarctic fish.Please address all correspondence and requests for reprints to Dr. I.A. Johnston     

Nitrogen balance in marine fish larvae: influence of developmental stage and prey density

The utilization and fate of nitrogen in larvae of plaice (Pleuronectes platessa), blenny (Blennius pavo) and herring (Clupea harengus), from the stage of first-feeding to metamorphosis, was examined under laboratory conditions. Rates of ammonia excretion, primary amine defaecation, and growth in terms of protein-nitrogen were monitored throughout larval life. Data were used to calculate daily ration, the coefficient of nitrogen utilization (absorption efficiency), and gross and net growth efficiencies. The developmental pattern of nitrogen balance was similar for plaice and blenny larvae. These species showed increasing growth efficiency (k₁: 55 to 80%) with decreasing weight-specific waste nitrogen losses with age. Absorption efficiencies. were high (83 to 98%) in plaice and blenny larvae, and tended to increase with development in the former species. Ration relative to body weight decreased with growth in both species. Herring larval development, although at a slower rate than blenny and plaice, appeared normal up to 33 d, after which high mortality occurred. Absorption efficiency in this species tended to decline (83 to 43%) with age, until metabolic costs exceeded the absorbed ration and growth ceased. Artemia sp. nauplii proved a suitable food source for the rearing of plaice and blenny larvae, but this diet may have long-term toxicity or deficiency effects on herring. Availability and density of food affected nitrogen balance in the larvae of all three species. Feeding stimulated the output of wastes in excretion and defaecation by a factor of up to ten times the 12-h non-feeding basal rates. Waste nitrogen output reached a peak some 2 to 3 h after commencement of feeding and returned slowly to the baseline in 5 to 10 h after cessation of feeding. There was an asymptotic increase in ration, ammonia output and growth of larvae as prey density increased. Ration saturated at a higher prey density (>4 prey ml^-1) than either growth or excretion rate (1 prey ml^-1). Thus the efficiency with which food is absorbed and utilized for growth must eventually decline in response to high prey density. The idea that larval fish are adapted to maximize ingestion and growth rate, rather than optimize growth efficiency and thus to respond to prey occurring in either low density or in occasional patches, is supported by these results.     

Asymptotic growth trajectories of larval sardine (Sardinops melanostictus ) in the coastal waters off western Japan

Growth trajectories of individual larvae of Japanese sardine, Sardinops melanostictus, caught in the coastal waters off western Japan were back-calculated from the first feeding stage up to date of capture (approximate size of 20 to 35 mm total length; TL) based on individually determined allometric relationships between otolith daily ring radii and fish total lengths. The larvae in January-, February-, and March-hatched cohorts in the coastal waters grew faster and more uniformly than those in the oceanic waters offshore of the Kuroshio current. Growth trajectories of the three hatch-month cohorts were similar and could be expressed by the Gompertz model. The inflection points of the growth curves were reached at 9 to 11 d after hatching, when larvae were 10.8 to 11.8 mm TL. Maximum growth rates at these points were 0.80 to 0.85 mm d⁻¹. Growth rates gradually declined after the inflection points, and larval TLs converged into the infinite length of 29 to 32 mm, the sizes at which metamorphosis from larvae to juveniles is initiated. This asymptotic growth pattern in the larval stage resulted in the narrow ranges in TLs in spite of the wide range of ages of the larvae caught by boat seiners in the coastal waters. Slow growth and therefore long duration of the metamorphosing stage could be influential in determining the cumulative total mortality in the early life stages of the Japanese sardine. Received: 14 July 1996 / Accepted: 20 August 1996     

Delayed growth of mussel (Mytilus edulis) and scallop (Pecten maximus) veligers at low temperatures

Veliger larvae of Mytilus edulis (L.) from Menai Straits, North Wales, were maintained for up to 2 mo during 1981 at 5°C and then grown on to metamorphosis at 17°C. Larvae so treated showed similar low mortality and equivalent spat production to control larvae. Growth rate at 17°C was less in treated larvae than in controls, but treated larvae grew a little during the period at low temperature. Larvae of Pecten maximus (L.) from the Irish Sea suffered high mortality at low temperature but larvae surviving 2 wk at 8°C could be grown on to spat at 17°C. The longevity of M. edulis larvae is discussed in relation to the genetic homogeneity of adult populations around the UK.     

Chemical changes during growth and starvation of larval Pleuronectes platessa

Plaice (Pleuronectes platessa L.) were sampled during periods of growth and starvation, from the end of the yolk-sac stage through metamorphosis, for changes in water, triglyceride, carbohydrate, total nitrogen, total carbon, and ash. The percentage of water in larvae decreased continuously during development. During post-hatching growth (up to late Stage 2) nitrogen and carbohydrate were laid down faster than triglyceride. The pattern changed during later larval development. The early deposition of protein in preference to neutral fat suggests that conversion of food during growth, without simultaneously laying down fatty energy stores, may be advantageous to pelagic marine fish larvae. During starvation the percentage of water in plaice larvae increased. Triglyceride, carbohydrate, nitrogen and carbon (as a percentage of the dry body weight) decreased during starvation, but ash increased sharply. The continuous use of nitrogen during starvation may be a catabolic adaptation to the marine environment.     

Active pelagic migrations of the bivalve Macoma balthica are dangerous

The bivalve Macoma balthica migrates twice during the benthic part of its life cycle. During the spring migration (May-June), the newly settled spat (0-group) migrates to the nurseries in the high intertidal. Seven to nine months later, the bivalves migrate back to the low tidal flats and the subtidal (winter migration, 1-group). Both 0- and 1-group M. balthica use byssus threads for active pelagic migrations. As many M. balthica disappear during these migrations, we examined experimentally the importance of predation on 0- and 1-group M. balthica. Laboratory experiments using a circular aquarium determined predation rates on buried (no current) and drifting (current) 0- and 1-group M. balthica by several fish species (plaice, flounder, goby and whiting) and the shore crab. Under illuminated conditions, more M. balthica were consumed when migrating than when buried, whereas there was no difference between experiments in conditions of darkness. For the 0-group, predation rates on migrating and buried M. balthica in the dark were lower than in the light. The stomachs of pelagic fish in the Wadden Sea and Oosterschelde estuary did not contain M. balthica during winter migration. In the Wadden Sea, 1-group M. balthica primarily migrated at night. In conclusion, enhanced predation on drifting, as compared to buried, M. balthica may be the mechanism that explains enhanced mortality during migration in light, and may explain why M. balthica mainly migrates at night in the field. As we found no M. balthica in stomachs of pelagic fish, we do not know whether predation on byssus drifting M. balthica exists in the field. There are, however, some indications for fish predation on infaunal polychaetes during pelagic migrations.     

The food of four species of pleuronectiform larvae in the eastern English Channel and southern North Sea

An examination was made of the stomach contents of the larvae of the plaice Pleuronectes platessa Linnaeus, 1758; the flounder Platichthys flesus (Linnaeus, 1758), the dab Limanda limanda (Linnaeus, 1758), and the sole Solea solea (Linnaeus, 1758) collected in the eastern English Channel and in the Southern Bight during the winter and spring of 1971. These 4 species of flat fish have distinct diets, and competition for food between them is largely avoided. Plaice larvae fed almost exclusively on Oikopleura dioica; flounder larvae also ate O. dioica, but in addition a wide range of planktonic organisms including phytoplankton, polychaete larvae, lamellibranch larvae, and copepod nauplii. Dab larvae fed mainly on the nauplii and copepodite stages of a variety of copepods, but particularly of Temora longicornis. Some T. longicornis copepodites and polychaete larvae were eaten by sole larvae, but the principal prey of these was lamellibranch larvae. The larvae of all the species began to feed in the yolk-sac stage; the initial food of all except plaice consisted of dino-flagellates, followed by tintinnids and copepod nauplii. Feeding began at dawn and the number of feeding fish and the number of food organisms in their stomachs increased throughout the day to a maximum near sunset. There were no consistent differences between the two areas in the diets of any of the species.     

Shadow cinematography of fish larvae

A simple system of shadow cinematography, consisting of a small tungsten halogen lamp, 2 large biconvex lenses and a 16 mm camera, is described for recording the swimming and feeding behaviour of larval fish. The system can be used either with infra-red film to record swimming behaviour independently of ambient light intensity, or with high-resolution film to record food organisms and feeding behaviour. Small plankton organisms of 0.2 mm width can be resolved using high-resolution film. The technique has been used to record the behaviour of plaice larvae (Pleuronectes platessa L.) feeding on the nauplii of Artemia salina L. The perceptive field of the larvae extends to approximately ±60° in azimuth, ±40° in elevation and 1.5 body lengths in range.     

Effects of acute changes in temperature and salinity on the oxygen uptake of larvae of herring (Clupea harengus) and plaice (Pleuronectes platessa)

Routine oxygen uptake (QO₂) by yolk-sac and firstfeeding larvae of herring (Clupea harengus L.) and plaice (Pleuronectes platessa L.) was studied after acute change of temperature (8°, 13°, 18°C) and salinity (5, 12.7, 32, 40). In both species, QO₂ (l mg^-1 dry wt h^-1) of both larval stages increased with increasing temperature. Salinity effect on QO₂ varied: for yolk-sac larvae of both species a lower QO₂ was found at lower combined salinities (5 and 12.7); for feeding larvae a lower QO₂ was observed at 12.7 for both species, possibly due to the relatively smaller size of larvae used at this salinity. For both species, oxygen uptake increased as larvae grew and weight regression coefficients were between 0.74 and 1.33. At 32 S, no difference was found in oxygen consumption between species as a function of temperature.Based on a dissertation submitted in partial fulfillment of the requirements for the degree of Master of Science at the University of Stirling, Stirling, Scotland. The work was performed at the Dunstaffnage Marine Research Laboratory, Oban, Scotland     

Scaling relationships,individual variation and the influence of temperature on maximum swimming speed in early settled stages of the turbot Scophthalmus maximus

Escape-swimming speeds (U _max) were studied in settled turbot (Scophthalmus maximus L.) reared at 18°C. Metamorphosis was complete at 4.0 cm total length (TL). U _max scaled in proportion to TL^0.74 in fish of 0.88 5o 8.00 cm TL at 18C. The scaling relationship for U _max was similar for temperatures between 13 and 23°C and could be fitted by the model: . U _max temperature-dependent, with a Q₁₀ of 1.77 over the temperature range studied. Analysis of covariance showed that U _max for farmed turbot was 14% lower than for wild fish filmed within 2 wk of capture; 3 mo after capture the average differences in escape performance were no longer significant, which suggests that the lower escape speeds of farmed fish are due to acclimation effects and not genetic stock differences. In order to assess the individual variability of U _max, 18 wild juvenile turbot [TL=6.2±0.4 cm (Week 1) to 7.5±0.5 cm (Week 17); means±SD] were maintained in individual containers at 18°C. U _max was determined weekly for 6 wk, standardised for fish length using the scaling relationship U _max=1.46 TL ^0.74, and individuals were ranked in order of performance. Temperature was reduced after 6 wk to 13°C, resulting in a significant decline in U _max from 104.0±14.4 to 87.8±12.5 cm s^-1 (means±SD). After 3 wk at 13°C U _max had increased to a level not significantly different from that at 18°C. Kendall's coefficient of concordance showed that repeatability of ranking of the experimental U _max of individuals was maintained over a 13 wk period and through temperature change. The results demonstrate that escape-swimming speeds in juvenile turbot are repeatable, individually variable, and can be modified in response to temperature acclination.     

Effects of temperature and salinity acclimation of adults on larval survival,physiology, and early development of Lytechinus variegatus (Echinodermata: Echinoidea)

Larval survival and developmental rates of Lytechinus variegatus (Lamarck) were determined as a function of temperature and salinity in two experiments by: (1) directly transferring fertilized eggs to 35, 30, 27.5, 25, 20, 15, and 10S seawater at 18 and 23°C, and (2) acclimation of adult sea urchins to the conditions described above for 1 to 4 wk prior to spawning. Developmental rates and percent survival of larvae prior to metamorphosis decreased at salinities below 35 (Q₁₀ values for metamorphosis=0.380 to 0.384). Temperature and salinity significantly (P<0.05) affected metabolic rates of L. variegatus plutei. These results show that L. variegatus larvae are stenohaline when compared to larvae of other echinoderm species. LC₅₀ values (S), developmental rates, and survival to metamorphosis indicate that acclimation of adult sea urchins to lower salinity prior to spawing and fertilization does not enhance development or survival of embryos exposed to low salinity.     

Einfluß von Benetzungsspannung und lonen auf die Substratbesiedlung und das Einsetzen der Metamorphose bei Bryozoenlarven (Bowerbankia gracilis)

Settlement and metamorphosis in larvae of Bowerbankia gracilis depend on the wettability of the substratum. The wettability of a solid can be characterized by it's specific contact angle, . Larvae settle on solids with angles >17°, but not on surfaces with high wettability properties. In an attempt to explain this phenomenon, we consider the larvae as a second liquid and their attachment as a second wetting. In such case, water and larvae would compete in wetting the substratum. Adhesion can be accounted for by assuming the wetting tension of solid to water to be lower than that of the larval surface (|_solid/water| < |_solid/larvae|). Adhesion would thus be favoured energetically. The rate of settlement and that of metamorphosis are demonstrated as depending on the degree of contact angle, , in an all-or-none fashion. Settlement of larvae on high-wetting solids is prevented by high capillary pressure accruing at the point of contact. Metamorphosis is initiated by contractions occurring after adhesion. Application of CsCl or KCl induces such contractions and metamorphosis in free-swimming larvae. MgCl₂ prevents the onset of metamorphosis but not adhesion. Two mechanisms are considered as possibly accounting for induction of metamorphosis: (1) mechanical stimulation by shearing the cilia at the point of contact, and (2) a chemical stimulation of the cell membrane after application of K⁺ or Cs⁺.     

New insights into temperature-induced white muscle growth plasticity during Dicentrarchus labrax early life: a developmental and allometric study

Dicentrarchus labrax is a major finfish of interest in Mediterranean aquaculture. As the development of its hatchery production had gone with an increase of its larval rearing temperature, we studied the effect of a constant high (20°C) and two lower (13 and 15°C) temperatures on its early white muscle growth, with developmental and allometric approaches. D. labrax, sampled at hatching and at three developmental stages corresponding to main events in fish early life (first exogenous feeding, notochord flexion and completion of fin ray counts), were histologically processed in order to follow changes in the white myotomal muscle size, cellularity and localisation of proliferative nuclei. Morphometric analyses showed that, for 13, 15 and 20°C incubated/reared D. labrax, the total cross-sectional area of white muscle increased slowly in eleutheroembryos shorter than 5.2 mm, and four times faster in longer fish (free-swimming larvae). White muscle growth occurred both by hypertrophy (increase in muscle fibre size) and hyperplasia (increase in total number of muscle fibres) in all sampled fish, but both processes were higher in free-swimming larvae than in eleutheroembryos. The morphometric establishment of a higher hyperplastic growth of white muscle in free-swimming larvae was confirmed at immunological level by a high occurrence of proliferative nuclei. Early thermal environment was demonstrated to affect the growth process of white muscle on a different way in eleutheroembryos and free-swimming larvae: in the former, white muscle hypertrophy was stimulated at 20°C and hyperplasia at lower temperature, whereas in the latter, both white muscle hypertrophy and hyperplasia were more stimulated at 20°C than at lower temperatures. This was verified at once when the fish length and the white muscle total cross-sectional area were used as explicative variables.     

Recruitment in the intertidal limpet Lottia digitalis (Patellogastropoda: Lottiidae) may be driven by settlement cues associated with adult habitat

The ribbed limpet, Lottia digitalis, is found high in rocky intertidal habitat throughout its geographic range. In order to identify likely natural settlement locations for larvae of this species, laboratory-reared larvae were settled onto substrata collected from within and near an adult L. digitalis habitat. Of larvae exposed to rock chiseled from within high-intertidal adult habitat, 31.0% and 23.3% underwent metamorphosis during two separate experiments. Similarly, an unidentified filamentous green alga that was isolated from this rock induced metamorphosis in 26.6% and 8.7% of larvae during additional experiments. In contrast, larvae did not metamorphose upon bare rocks or upon rocks encrusted with a crustose corraline alga (CCA) that were collected from lower intertidal zones, nor did they metamorphose upon the macroalgae Ulva sp., Enteromorpha contorta, Alaria marginata, or Polysiphonia sp. The presence of mucus from adult conspecifics during these experiments did not enhance metamorphosis onto rock taken from adult habitat, but it did induce metamorphosis in an average of 13.5% and 7.0% of larvae introduced to the mid-intertidal bare rock and CCA substrata, respectively. Finally, 38.0% and 34.4% of larvae from two experiments underwent metamorphosis when exposed to the high-intertidal barnacle Pollicipes polymerus. These results suggest that larval settlement, rather than differential post-settlement mortality and/or migration, drives recruitment of L. digitalis in high-intertidal habitats.     

Feeding behaviour and prey size selection of gilthead seabream,Sparus aurata,larvae fed on inert and live food

Prey selection shortly after the onset of feeding by laboratory-reared gilthead seabream, Sparus aurata L., larvae was studied using larvae fed on two types of microcapsule (hard- and soft-walled) having diameters ranging from 25 to 300 m. Preferences between inert food and live prey (rotifers and Artemia sp. nauplii) were also studied. Seabream larvae were able to ingest inert food from first feeding. Larvae of all size classes ingested hard microcapsules with diameters in the range 25 to 250 m. However, larvae with a total length (TL) below 4 mm preferentially selected particles 25 to 50 m in diameter, larvae of TL 4 and 5 mm preferred particles 51 to 100 m in diameter, while larvae above 5 mm TL preferred particles 101 to 150 m in diameter. With soft microcapsules, larvae always preferred particles larger than in the previous case, and above 4.5 mm TL they preferentially selected particles 201 to 250 m in diameter. In addition, the gradual increase of preferred diameters with increasing TL was more pronounced when larvae were increasing TL was more pronounced when larvae were fed on soft particles. Mean values for prey width/mouth width ratios were approximately 0.24 and 0.30 when larvae were fed on hard-walled and soft-walled microcapsules, respectively, irrespective of the absolute value of larval length. When a mixed diet of live and inert food items was offered, live prey were always preferentially selected, even if the prey width/mouth width ratio was apparently not favourable. Only a physical constraint such as excessive prey width could counter this preference for living prey vs inert microcapsules. These results contribute to our knowledge in larval feeding behaviour, especially in the presence of inert food, and represent a fundamental step in developing prepared food for marine fish larvae.     

Chemical defense and aposematic coloration in larvae of the ascidian Ecteinascidia turbinata

In the coastal waters of Florida (USA) tadpole larvae of the colonial ascidian Ecteinascidia turbinata contain chemicals which make them unpalatable to planktivorous juvenile pinfish Lagodon rhomboides. Experiments demonstrate that the bright organe color of E. turbinata tadpoles is aposematic. Fish that have recently tasted larvae of E. turbinata will not attack the palatable tadpoles of Clavelina oblonga when the latter are dyed organe to resemble larvae of E. turbinata. Tadpoles of E. turbinata that have been mouthed and rejected by fish generally survive to complete a normal metamorphosis. Individual selection explains the evolution of aposematic coloration in E. turbinata better than kin selection. The identity of the defensive chemical is unknown. The unpalatable substance in larvae of E. turbinata is removed by dialysis, indicating that it has a molecular weight less than 14000 d. Larvae are not acidic, nor is the active substance denatured by doiling.