X-ray diffraction study of the first larval shell of Ostrea edulis

X-ray powder diffraction was used to study the calcification of the first larval shell of Ostrea edulis (sampled in Limski kanal, Istria, Adriatic Sea in April 1986) from the trochophore stage to the veliger larvae (prodissoconch I), and development of the latter up to several days postfertilization (prodissoconch II). In the first stage, only the amorphous component is present (periostracum and organic matrix). The beginning of shell formation is manifested by the appearance of calcite (up to 1–4% of the total vol.) and then aragonite (2 to 7%). In a later stage of the veliger larvae the fraction of calcite decreases, as well as the fraction of the amorphous component, while the fraction of aragonite rapidly increases. In the prodissoconch II stage, aragonite is dominant, with a very small amount of amorphous component and traces of calcite. In contrast, the valves of the adult O. edulis are composed mainly of calcite, with traces of aragonite.     

Differential timing of larval starvation effects on filtration rate and growth in juvenile Crepidula onyx

This study demonstrates that the timing of larval starvation did not only determine the larval quality (shell length, lipid content, and RNA:DNA ratio) and the juvenile performance (growth and filtration rates), but also determine how the latent effects of larval starvation were mediated in Crepidula onyx. The juveniles developed from larvae that had experienced starvation in the first two days of larval life had reduced growth and lower filtration rates than those developed from larvae that had not been starved. Lower filtration rates explained the observed latent effects of early larval starvation on reduced juvenile growth. Starvation late in larval life caused a reduction in shell length, lipid content, and RNA:DNA ratio of larvae at metamorphosis; juveniles developed from these larvae performed poorly in terms of growth in shell length and total organic carbon content because of “depletion of energy reserves” at metamorphosis. Results of this study indicate that even exposure to the same kind of larval stress (starvation) for the same period of time (2 days) can cause different juvenile responses through different mechanisms if larvae are exposed to the stress at different stages of the larval life.     

Quantitative genetic analysis of growth in larval scallops (Placopecten magellanicus)

We estimated the broad-sense heritability of larval size in 20 full-sib families of the giant scallop, Placopecten magellanicus (Gmelin, 1791) grown in laboratory culture in August and September 1991. The goal was to compare scallops with other bivalves which have been shown to have significant heritabilities for larval growth. Secondly, we estimated the lipid content of occytes from female parents, since this is hypothesized to affect larval growth and survival. Finally, we estimated the among-family variation in mortality from 4 to 21 d as a test of possible genetic variation for viability among larval scallops. Significant genetic variation (h²=1.10 to 1.24) was estimated for larval shell length at 4, 14, and 21 d. There was a significant correlation (r=0.66) between larval size at 4 d and lipid content of oocytes, but only when two females with high levels of lipid oocyte^–1 were excluded as outliers. There was no correlation between larval size at 14 and 21 d and lipid content of oocytes. Mortality among families from 4 to 21 d was high (69 to 97%), and was significantly different among families. These results indicate that there is significant heritability for larval growth which is largely independent of the lipid content of the oocytes. A high heritability for larval growth may indicate that this trait is only weakly correlated with fitness.     

Comparative effects of two antifouling paints on the oyster Crassostrea gigas

The effects of two antifouling paints, one containing organotin compounds, the other copper oxide, were studied from September 1983 to October 1984 on Crassostrea gigas (Thunberg) grown under natural field conditions in the Bay of Arcachon, France. The organotin paint reduced growth rate expressed as weight, length and width, but did not affect shell height; it drastically decreased the dry condition factor and shell density, but did not affect the viability of embryos and larvae from exposed oysters. However, some decrease in larval growth rate was observed. The copper paint had no effect on oyster growth, but lowered the condition factor compared to controls. Neither viability of embryos or larvae nor larval growth were affected by this paint.     

Prodissoconch morphology is environmentally modified in the brooding bivalve Lasaea subviridis

Embryonic shell sculpture of intertidal and laboratory brooded individuals of the direct developer Lasaea subviridis was studied using scanning electron microscopy. Intertidally brooded individuals develop a distinct prodissoconch I (PI) and prodissoconch II (PII) of unusual morphologies. The PI is relatively small (50–100 m in length), oval in outline, lacks a radial sculpture and is restricted to the initial pitted zone of the prejuvenile shell. A larger PII is formed (510–680 m in length) and it is dominated by pronouced commarginal striae but also includes a belt of radial ridges extending from the PI/PII boundary. Laboratory brooded individuals kept constantly submerged do not develop the pronounced commarginal striae characteristic of PII. This implies that PII formation in L. subviridis is not a direct result of the brooding habit, and can be modified by environmental factors, which possibly include low-tide exposure. There is no single feature of external prodissoconch morphology that unambiguously indicates a direct development mode combined with mantle cavity brood protection. General guidelines are presented to help recognize this developmental mode, based on prodissoconch morphology. In comparing prodissoconch morphologies of brooding bivalves, the habitats as well as the phylogenies of species should be considered, especially in comparisons of intertidal and subtidal species.     

Measurement of calcium carbonate deposition in molluscs by controlled etching of radioactively labeled shells

Rates of calcium carbonate removal from shell pieces of Argopecten irradians (Say) and Mercenaria mercenaria (L.) were dependent on the type of etching fluid used and not shell origin. Etching was uniform over the entire shell surface, but surface morphology differed with etching fluids. Peak radioactivity was in early eluant fractions of shells etched immediately after radioactive labeling, and in later fractions when individuals were placed in isotope-free seawater after labeling. The etching technique can measure growth during the labeling period and subsequently by estimating the amount of calcium in fractions prior to the radioactive peak. Geometry of shell layers influenced the pattern of radioactivity seen in fractions. Peak location varied inside and outside the pallial line of individual M. mercenaria. A significant portion of the inorganic carbon used in shell formation by M. mercenaria was derived from metabolic CO₂.     

X-ray diffraction study of calcification processes in embryos and larvae of the brooding oyster Ostrea edulis

X-ray powder diffraction was used to study shell calcifications of the oyster Ostrea edulis, sampled in the Limski Kanal, Istria (Adriatic Sea), in May 1992. All the developmental stages were followed, from the embryonic stage through the transition between the trochophore and veliger larva (prodissoconch I and II) and later, after swarming, the pelagic free-swimming larval stages, up to their settlement and attachment (from the D-shaped to the fully formed pediveliger larva), and finally during metamorphosis and juvenile stages (dissoconch). In the first gastrula stage, only an amorphous tissue is present (a periostracum and organic matrix). The beginning of shell formation (at the end of gastrulation) in early trochophores is manifested by the appearance of calcite (up to 1–7% of total volume) and then aragonite (about 1%). In the later stage of the veliger larva the fraction of calcite decreases as well as the amorphous fraction, while the fraction of aragonite rapidly increases. In the prodissoconch II stage and during the whole pelagic period aragonite is dominant, accompanied by a very small amorphous fraction and traces of calcite. The shell mineral composition does not change until metamorphosis, whereupon the fraction of calcite rapidly increases and the fraction of aragonite decreases. The postmetamorphic valves of the juvenile and adult oyster consist mainly of calcite, except the resilium and myostracum which remain aragonitic, possibly as a continuation of the inner layer of the larval shell. Received: 28 May 1997 / Accepted: 1 July 1997     

Heritability of juvenile growth for the hard clamMercenaria mercenaria

The heritability of juvenile growth in the hard clamMercenaria mercenaria (L.) sampled from Wellfleet Harbor, USA, in 1987, was analyzed in two experiments using standard quantitative genetic methods. In the first experiment, culture density was not controlled and was found to have a significant impact upon the non-additive genetic component of growth, possibly through genotype-environment correlation. Variable clam density appeared to have little impact on the additive genetic variance component. Estimates of the narrow sense heritability, calculated from the additive genetic component, ranged from 0.72 (±0.32) to 0.91 (±0.17). Culture density was controlled in the second experiment. The narrow sense heritability estimate from this experiment was 0.37 (±0.13). These estimates suggest that selection, either natural or artificial could bring about rapid change in the mean phenotype(s) of juvenile growth characteristics in hard clams.Contribution no. 793 from the Belle W. Baruch Institute for Marine Biology and Coastal Research     

Genetics of larvae and spat growth rate in the oyster Crassostrea virginica

The heritability of oyster (Crassostrea virginica) larval growth rate was estimated to be in the range of 0.25 to 0.50 and a significant part of this genetic variation is of the additive type. Larval growth rate and spat growth rate were found to be highly correlated. These results suggest that a selection program for faster growing larvae and spat would be successful.     

Food limitation stimulates metamorphosis of competent larvae and alters postmetamorphic growth rate in the marine prosobranch gastropodCrepidula fornicata

The effects of food limitation on growth rates and survival of marine invertebrate larvae have been studied for many years. Far less is known about how food limitation during the larval stage influences length of larval life or postmetamorphic performance. This paper documents the effects of food limitation during larval development (1) on how long the larvae ofCrepidula fornicata (L.) can delay metamorphosis in the laboratory after they have become competent to metamorphose and (2) on postmetamorphic growth rate. To assess the magnitude of nutritional stress imposed by different food concentrations, we measured growth rates (as changes in shell length and ash-free dry weight) for larvae reared in either 0.45-m filtered seawater or at phytoplankton concentrations (Isoehrysis galbana, clone T-ISO) of 1 × l0³, 1 × 10⁴, or 1.8 × 10⁵ cells ml^–1. Larvae increased both shell length and biomass at 1 × 10⁴ cells ml^–1, although significantly more slowly than at the highest food concentration. Larvae did not significantly increase (p > 0.10) mean shell length in filtered seawater or at a phytoplankton concentration of only 1 × 10³ cells ml^–1, and in fact lost weight under these conditions. To assess the influence of food limitation on the ability of competent individuals to postpone metamorphosis, larvae were first reared to metamorphic competence on a high food concentration ofI. galbana (1.8 × 10⁵ cells ml^–1). When at least 80% of subsampled larvae were competent to metamorphose, as assessed by the numbers of indlviduals metamorphosing in response to elevated K⁺ concentration in seawater, remaining larvae were transferred either to 0.45-m filtered seawater or to suspensions of reduced phytoplankton concentration (1 × 10³, 1 × 10⁴, or 5 × 10⁴ cells ml^–1), or were maintained at 1.8 × 10⁵ cells ml^–1. All larvae were monitored daily for metamorphosis. Individuals that metamorphosed in each food treatment were transferred to high ration conditions (1.8 × 10⁵ tells ml^–1) for four additional days to monitor postmetamorphic growth. Competent larvae responded to all food-limiting conditions by metamorphosing precociously, typically 1 wk or more before larvae metamorphosed when maintained at the highest food ration. Surprisingly, juveniles reared at full ration grew more slowly if they had spent 2 or 3 d under food-limiting conditions as competent larvae. The data show that a rapid decline in phytoplankton concentration during the larval development ofC. fornicata stimulates metamorphosis, foreshortening the larval dispersal period, and may also reduce the ability of postmetamorphic individuals to grow rapidly even when food concentrations increase.     

Accumulation and loss of biomass inLiocarcinus holsatus larvae during growth and exuviation

Liocarcinus holsatus (Fabricius) larvae, of females originating from the Elbe Estuary, FRG, were reared in the laboratory at constant 15°C in May 1988. For each larval stage, developmental time was measured by individual cultures (Zoea I: 6.7±0.7d; Zoea II: 5.0±0.6d; Zoea III: 4.8±0.7 d; Zoea IV: 5.3±0.6d; Zoea V: 6.1±1.1d; Megalopa: 10.45±0.7d). During the entire period of development, dry weight (W), carbon (C), nitrogen (N), and hydrogen (H) were measured daily (Zoea I to V) or every second day (Megalopa). The energy content (E) was estimated from C. Biomass and energy (per individual) increased in each larval stage as a parabolic function of age and is described by power functions. C, H, and E exhibit a higher percentage gain (relative to initial values at the time of hatching) than W and N. It is suggested that proportionally more lipid than protein is accumulated during larval development. Cyclical changes in the relative biomass (% W) correspond to the larval moult cycle, indicating a rapid uptake of water and minerals immediately after hatching and a later increase in tissue growth. Changes in the C:N ratio suggest that during the first period more lipid than protein is accumulated. These patterns of growth and elemental composition are compared with literature data and a high degree of similarity in the growth characteristics of decapod larvae is seen. In addition W, C, N, and H values as well as E were measured for the exuviae of Zoea I to V and Megalopa. The percentage loss of growth rate by exuviae for each larval instar were higher in W (12 to 16%) and C (8 to 12%), and varied between 5 and 10% for N, H, and E.     

Development of the pericalymma larva of Solemya reidi (Bivalvia: Cryptodonta: Solemyidae) as revealed by light and electron microscopy

Solemya reidi Bernard 1980 is a gutless protobranch bivalve known to possess intracellular chemoautotrophic bacterial symbionts in its gill. A light and electron microscope study on the embryology and larval development of S. reidi provides data for the bivalve Subclass Cryptodonta. S. reidi spontaneously spawned large eggs (271 m in diameter), which developed within individual gelatious egg capsules. The first several cleavages were equal and a distinct molluscan cross was formed at the animal pole of the embryo, features previously unreported in bivalve development. Lecithotrophic pericalymma larvae (similar to the larvae of paleotaxodont protobranch bivalves and aplacophoran molluscs) hatched at 18 to 24 h and remained in the water column for a further 5 d at 10°C. At hatching, larvae measured from 360 to 440 m in length and from 225 to 265 m in cross-sectional diameter. Definitive adult structures developed within an epithelial locomotory test entirely covered with compound cilia. The test histolysed at metamorphosis and was ingested throught the mouth into the perivisceral cavity. Length and height of the shell following metamorphosis was 433 m (±42 m, n=16) and 282 m (± 29 m, n=13), respectively. Primary data and data from the literature show that the type of larval development in both paleotaxodont and cryptodont bivalves cannot be reliably estimated from egg or prodissoconch sizes.     

Relationship between larval and juvenile growth rates in two marine gastropods,Crepidula plana and C. fornicata

Previous studies on various marine mollusc species have shown that both larval and juvenile growth rates are substantially heritable, but few workers have examined the extent to which larval and juvenile growth rates covary. We examined the relationship between larval and juvenile growth rates in seven laboratory experiments conducted between 1986 and 1993, using the prosobranch gastropods Crepidula plana Say and C. fornicata (L.). In most experiments larvae were reared individually, measured twice nondestructively to determine larval grwoth rate, allowed or stimulated (daily 5-h exposure to 20 mM excess K⁺ in seawater) to metamophose, and then measured at least twice after metamorphosis to determine juvenile growth rates. Generally, there was no significant (p >0.10) relationship between larval and juvenile growth rates, suggesting that in these two species selection can act independently on the two stages of development. A positive correlation (p=0.007) between larval and juvenile growth rates was observed for C. fornicata in one experiment, but only for offspring from females maturing the most rapidly in laboratory culture. Even for these larvae, however, variation in larval growth rate explained<2% of the variation in juvenile growth rate, so that larval and juvenile growth rates are at most only weakly associated in this species.     

Electrophoretic identification and genetic analysis of bivalve larvae

Taxonomic identification and genetic analysis of larval marine invertebrates have been vexing problems. We describe a polyacrylamide mini-gel electrophoresis technique for resolving proteins from individual larval bivalves (shell length 250 to 350 m) and apply this technique to three species of laboratory-cultured larval oysters [Ostrea edulis L., 1758, Crassostrea gigas (Thunberg, 1793) and c. virginica (Gmelin, 1791)] reared during summer 1989. Electrophoretic patterns of proteins clearly discriminate among the three species and allow genetic analysis of a polymorphic allozyme locus (Pgi) in field-collected larvae and juveniles of C. virginica. This technique provides an economical tool for largescale taxonomic, ecologic, and genetic studies of meroplanktonic stages of various species.     

Identification of settling and early postlarval stages of mussels (Mytilus spp.) from the Pacific coast of North America, using prodissoconch morphology and genomic DNA

 Detailed inventories of the benthos and field studies of the settlement and recruitment processes of marine benthic invertebrates require accurate identification of newly settled larvae and early juvenile stages. We provide morphological criteria, visible under a good quality dissecting stereomicroscope, by which to discriminate between species of the settling larval and early postlarval stages (∼250 to 700 μm shell length) of mussels of the genus Mytilus on the west coast of Vancouver Island and Southern California. Compared to the bay mussel (M. trossulus), the sea mussel (M. californianus) has: (i) a shallower and flatter umbo, the latter corresponding to a significantly less pronounced prodissoconch I (PI) curvature and (ii) a greater PI length; as well as (iii) a wider separation between the provincular lateral teeth (PLT). The PLT distance is a new term denoting the separation between the midpoint of two reddish pigment spots of the provinculum (larval hinge apparatus) region of settling larvae and early postlarvae of Mytilus spp. from the East Pacific Coast. These spots mark the larger provincular lateral teeth, situated at either end of the provinculum. We confirmed the validity of morphological criteria by comparing PCR products of genomic DNA of provisionally identified postlarvae. Furthermore, measurements of PI lengths and PLT distance from well-preserved postlarvae of sea mussels (M. californianus) and of bay mussels (M. galloprovincialis) from Southern California indicate that the PI morphology and morphometry, and PLT distance criterion apply for that region as well. The criteria presented here can also apply to the advanced (competent) veliger stages, as the latter may settle (i.e. become the “settling” stage) upon encountering a suitable substrate. Our present and previously published work provide economical and effective identification methods that can be used to discriminate among early life history stages (∼250 μm to 5.0 mm shell length) of Mytilus spp. along the west coast of North America. Received: 10 November 1999 / Accepted: 6 September 2000     

General and specific combining abilities of larval and juvenile growth and viability estimated from natural oyster populations

Diallel crosses of oysters from three geographically isolated natural populations were produced to evaluate the relative importance of genetic, maternal, and environmental effects on larval and juvenile growth and viability. Significant additive genetic effects were observed only in larval viability at Day 12 and larval shell length at Day 2. The presence of significant male and female mean square for larval viability (suggesting non-additive genetic variance) is consistent with fitness related characters. Important maternal effects were observed for the larval and juvenile shell length and viability characters. These female mean squares are probably affected by both real and spurious maternal effects and potential contributing influences are discussed. The performance of the crosses can be largely explained by two factors: parental performance and the heterotic gene effects. This is based on an apparent positive correlation of mean values between the parental populations and their crosses. The crosses' mean viability at the end of the larval phase was 14.0% lower than the pure matings and support a previous observation of lower heterozygote viability in the larval phase (Mallet and Haley, 1983b).     

Relationship between growth rate and age at recruitment ofAnguilla japonica elvers in a Taiwan estuary as inferred from otolith growth increments

The growth history and recruitment dynamics of eel (Anguilla japonica) elvers were studied. Observations were based on growth increments in sagittal otoliths of elvers collected from Shuang-Chi River estuary off northeastern Taiwan, from November 1985 to February 1986. Total lengths of elvers upon arrival at the estuary were similar in most case; mean total lengths were from 55.99 to 59.06 mm. Daily ages of elvers at arrival ranged from 112.8±9.4 (±SD) to 156.5±13.5 d, indicating that migration of eel larvae from their oceanic spawning ground to the estuary requires 4 to 5 mo. Elver hatching dates, back-calculated from estimated daily ages, indicated that the spawning season lasted 5 mo (from late June to early October). Furthermore, the earlier eels spawned, the earlier elvers reached the estuary. The transition in growth history during the larval stage was obvious, as indicated from the change in increment width in elver otoliths. The inverse correlation between daily age and mean daily growth rates of fish length and otolith indicated that the age of elvers upon arrival at the estuary was susceptible to larval growth rate. In other words, the time taken on migration from oceanic spawning ground to the estuary was shorter for fast-growing larvae than for slowgrowing ones.     

Growth and differentiation during delayed metamorphosis of feeding gastropod larvae: signatures of ancestry and innovation

Extent of larval growth among marine invertebrates has potentially profound implications for performance by benthic recruits because body size influences many biological processes. Among gastropods, feeding larvae often attain larger size at metamorphic competence than non-feeding larvae of basal gastropod clades. Delay of metamorphosis can further influence size at recruitment if larvae continue to grow during the delay. Some caenogastopod larvae grow during delayed metamorphosis, but opisthobranch larvae do not. Data on larval growth of neritimorph gastropods are needed to help determine which of these growth patterns for planktotrophic gastropod larvae is more derived. We cultured planktotrophic larvae from all three major gastropod clades with feeding larvae through delays of metamorphosis of 3–10 weeks. Larvae of the caenogastropod Euspira lewisii and the euthyneurans Haminoea vesicula (Opisthobranchia) and Siphonaria denticulata (Pulmonata) conformed to previously described growth patterns for their respective major clades. Furthermore, the caenogastropod continued to lengthen the prototroch (ciliary band for swimming and feeding) and to differentiate prospective post-metamorphic structures (gill filaments and radular teeth) during delayed metamorphosis. Larvae of the neritimorph Nerita atramentosa arrested shell growth during delayed metamorphosis but the radula continued to elongate, a pattern most similar to that of non-feeding larvae of Haliotis, a vetigastropod genus. Character mapping on a phylogenetic hypothesis suggests that large larval size and capacity for continued growth during delayed metamorphosis, as exhibited by some caenogastropods, is a derived innovation among feeding gastropod larvae. This novelty may have facilitated post-metamorphic evolution of predatory feeding using a long proboscis.     

Growth and mortality in young larval herring (Clupea harengus); effects of repetitive changes in food availability

Following yolk resorption, laboratory-reared larval Baltic herring (Clupea harengus L.) were exposed to two sequences of food restriction for 5 d and re-alimentation for 10 d. Comparisons regarding larval growth (standard length and content of water-soluble protein), mortality and content of the sum of trypsin and trypsinogen were made with larvae at a continuous high ration. Larvae exposed to varying prey abundance grew less in length than the control, and during the second high-ration period (Day 22 to 32) growth in length ceased. From the first low-ration period onwards, the content of water-soluble protein in these larvae was lower than that of the control larvae, and the survival rate of the low-high ration group was 59% compared to 77% in the larvae at a continuous high ration. In contrast, the effects of varying food availability were minor on larval content of trypsin and trypsinogen. Results are compared with previous findings in larval Clyde herring, and the effects of larval stock and timing and duration of food restriction on larval growth performance are discussed.     

Carbon content and nutritional condition of sardine larvae (Sardina pilchardus) off the Atlantic coast of Spain

As part of a “European Sardine/Anchovy Recruitment Program” (SARP), sardine larvae (Sardina pilchardus) were sampled off the Atlantic coast of Spain through the spawning season from March to June. The larvae were analysed for carbon and nitrogen content as a measure of nutritional condition and survival potential. There was no significant diel variation in larval carbon content, but there was a small significant diel variation in nitrogen; the absence of a strong diel signal in elemental composition was ascribed to the overnight retention of the gut contents. There was an increase in carbon content with increase in body length which reached an asymptote at ∼40% carbon content at a larval length of 20 mm. It is argued that larvae with a carbon content of <25% of body weight were nutritionally stressed, with the smaller larvae (<10 mm in length) appearing to be more vulnerable to food limitation. Although larvae with the lowest age-specific carbon content (poorest condition) occurred on the cruise with the lowest food availability, there was no consistent relationship between carbon content and food availability. While the successive monthly estimates of carbon content revealed differences in potential recruitment between months, these were not related to the birth-date distribution of the surviving juveniles. Received: 23 November 1998 / Accepted: 3 March 1999