Organogenesis and histogenesis in the planktotrophic veliger of Doridella steinbergae (Opisthobranchia: Nudibranchia)

Development of the planktotrophic veliger of the dorid nudibranch Doridella steinbergae (Lance) was studied by histological examination of 4, arbitrarily defined larval stages. Following an embryonic period of 7 1/2 to 8 days (12° to 15°C), the newly hatched veligers possess a functional digestive tract, a pair of nephrocysts, a secondary kidney, a pair of cerebral ganglia, a larval shell consisting of a two-thirds whorl, and the metapodial component of the foot. Development during Stage I mainly involves growth of the larval shell and the visceral organs. Stage II is marked by the retraction of the mantle fold from the shell aperture and the appearance of the eyespots, gonadal rudiment, larval heart, and the optic, pedal, and pleural ganglia. At Stage III the radular sac rudiment evaginates from the esophageal wall, the buccal ganglia differentiate, and the propodial rudiment begins to develop on the ventral surface of the metapodium. Stage IV veligers, which are competent to metamorphose, possess 6 pairs of radular teeth, lipid deposits in the left digestive gland, rudiments of the adult kidney and the oral lip glands, an hypertrophied mantle fold, a propodium, and densely packed cilia over the entire ventral surface of the foot. The length of the obligatory larval period, from hatching of the veliger until the attainment of metamorphic competence, is 25 to 26 days under laboratory culture conditions and the larval shell grows from 142 to 168 m in length. The sequence of morphogenetic events and the structure of the competent veliger of D. steinbergae is compared to that of other opisthobranch veligers. It is suggested that the relatively small maximal shell size attained by D. steinbergae results from precocious retraction of the mantle fold. It is further suggested that interspecific differences in the kinds of structures that develop during the veliger phase of opisthobranchs may relate to variations in the requirements of the juvenile phase. The functional adaptations of the gut of planktotrophic veligers are discussed and compared to those of lecithotrophic veligers.     

A method for rearing Pecten maximus larvae in the laboratory

Development of the scallop Pecten maximus (L.) from egg to metamorphosis takes 33 to 38 days at 16°C. The shelled veliger first appears 2 days after fertilisation, and crawling pediveligers at around 2 days before metamorphosis. The pediveliger can attach temporarily to filamentous objects by means of a byssus thread and, even after metamorphosis, periods of attachment alternate with periods of crawling. The larvae are reared in polythene bins containing 30 l of full salinity sea water (pre-filtered through 0.2 filters), which is changed every 2 days. Mixtures of algal foods (Isochrysis galbana, Chaetoceros calcitrans, Pyramimonas obovata and Tetraselmis suecica), concentrated by centrifugation, are added at each change from the third day onwards to give a concentration of 50 cells/l. Larval density is maintained below 10/ml. Antibiotics are added at each change, but very careful sterilisation of all equipment is necessary to guard against the introduction of diseases.     

Reproduction and development of Paedoclione doliiformis,and a comparison with Clione limacina (Opisthobranchia: Gymnosomata)

Observations on breeding season, copulation, spawning, and development are described for Paedoclione doliiformis, the only gymnosomatous pteropod which consistently retains external larval features throughout its life. The number of eggs spawned per individual is dependent upon temperature and availability of Spiratella retroversa, its thecosome prey. At 17° to 19°C, planktotrophic veligers hatch from floating egg masses within 3 days, and cast their shells approximately 11 days later. The resultant shell-less veligers could complete metamorphosis to the polytrochous larval stage, in which the body is encircled by 3 ciliary bands, within 12 h. Some polytrochous larvae begin feeding on veligers of S. retroversa within 2 days. Unlike other gymnosome species, there is no further change in external appearance except for growth. A comparison of reproduction and development has been made with Clione limacina, another gymnosome which competes for food with P. doliiformis in Nova Scotian (Canada) coastal waters.     

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.     

Feeding,growth, and survival of Engraulis mordax larvae reared in the laboratory

Methods are described for the successful rearing of northern anchovy larvae (Engraulis mordax Girard) on cultured foods. Larvae were fed successively on the unarmored dinoflagellate Gymnodinium splendens, the veliger of the gastropod Bulla gouldiana, and nauplii of the brine shrimp Artemia salina. Rearing containers ranging in capacity from 4.5 to 510 l were tested; the smaller ones were found to be most useful for laboratory experimentation. Irreversible starvation occurred when E. mordax were denied food for more than 1.5 days after yolk absorption. Growth rates of larval anchovies fed different diets were compared. Larvae fed G. splendens grew for 1 week at the same rate as animals fed wild plankton, but did not maintain this rate. Laboratory survival of E. mordax larvae on a diet of G. splendens alone, did not differ significantly when veligers supplemented the diet. However, when G. splendens and veligers were fed simultaneously to E. mordax larvae, growth rate was greatly improved, although still not matching the growth attained on a diet of wild plankton. Length (L) versus weight (W) analyses were made for all larvae at all diets. The results showed that weight could be calculated most accurately from length by the relationship log W=3.3237 log L-3.8205, regardless of diet.     

A response-surface approach to the combined effects of temperature and salinity on the larval development of Adula californiensis (Pelecypoda: Mytilidae). II. Long-term Larval survival and growth in relation to respiration

Larvae of the bivalve molluso Adula californiensis (Phillippi, 1847) were reared for 3 days, from fertilization to veliger stage, at optimum conditions (15°C, 32.2 S), and then transferred to experimental temperatures and salinities for 22 more days to determine the effects of these factors on survival and growth. For larvae surviving to 25 days, maximum survival was estimated, by response-surface techniques, to occur at temperatures below 10°C and at salinities above 25. A comparison of 60% survival response contours for 3, 15 and 25-day old larvae indicated a progressive shift in temperature and salinity tolerance with age of larvae. The older larvae became more tolerant to reduced salinity, but less tolerant to high temperatures. Growth of the larvae over 25 days of culture was slight, and relatively independent of temperature and salinity conditions found in the environment. Oxygen consumption of 3-day old veliger larvae measured at various combinations of temperature and salinity generally increased from 7° to 18°C, and then sharply decreased from 18° to 21°C. A plateau of oxygen consumption from 9° to 15°C at 32.9 S indicated that the larvae are adapted to oceanic rather than estuarine conditions. A comparison of 25-day larval survival, mean length, and growth, with oxygen consumption of 3-day old veliger larvae indicated that high temperatures (15°C, and above) coupled with reduced salinities (26.1, and below) were unfavorable for prolonged larval life. Because of the lack of larval adaptations to estuarine conditions, larva survival and, hence, successful recruitment of this species within Yaquina Bay (Oregon, USA) depends upon the essentially oceanic conditions found only during the summer in the lower part of the Bay.     

Tisbe pori n. sp. (Copepoda: Harpacticoida) from the Mediterranean coast of Israel and its cultivation in the laboratory

Tisbe pori n. sp., from the Mediterranean coast of Mikhmoret, Israel, was cultivated in the laboratory and is described. The relative food values of 9 species of marine algae were evaluated according to the copepods' development time (length of period from hatching to reproductive maturity). Individuals fed on Dunaliella tertiolecta or D. primolecta developed the fastest. The rate of feeding of the adults on Dunaliella primolacta was determined. In the temperature range 17° to 31.5°C, the shortest development time (7 days) was at 26° to 28°C. The range of salinity tolerated by adults was 26 to 48.     

Encapsulated development of the marine prosobranch gastropod Nucella lapillus

Nucella (=Thais) lapillus (L.) develops to the juvenile stage within an egg capsule. This study was undertaken to investigate nutritive and bacteriostatic properties of the intracapsular fluid for this species. Capsules were collected intertidally at Nahant, Massachusetts, USA, in the summers of 1982 and 1983. The average capsule contained only 1.1 l of intracapsular fluid per embryo. A statistically significant correlation was observed between capsule height and number of developing embryos contained therein, although the number of embryos per capsule was more closely related to capsule volume. Total dry weight of encapsulated, shelled embryos increased exponentially as a function of shell length. However, weight increases may be entirely due to shell calcification; the average dry tissue weight of shelled embryos was significantly below that for pre-shelled individuals, suggesting a net loss of biomass during development. Aliquots of intracapsular fluid failed to inhibit the growth of the 13 bacterial strains tested. Despite the above results, pre-shelled embryos generally survived only several days following their premature removal from egg capsules at 12°C. Shelled veligers were reared outside of the capsules with no apparent ill effects. An explanation for these survivorship results is elusive.     

Establishment of the photosymbiosis in the early ontogeny of three giant clams

Distribution and morphology of zooxanthellae were investigated histologically and ultrastructurally in veligers and juveniles of three giant clam species, Tridacna crocea, T. derassa, and T. squamosa. No zooxanthellal cells were associated with gametes. In veliger larvae, zooxanthellae were ingested and digested in the stomach. Within several days after metamorphosis from veliger to a juvenile clam, the zooxanthellal tube, in which zooxanthellae were packed, elongated from the stomach toward the mantle. Zooxanthellae in the tube appeared in a line, and we designated the appearance of the lined zooxanthellae in the mantle of juvenile clams as the first sign of the establishment of symbiosis. The zooxanthellal tubular system developed as the clams grew, particularly in the mantle margin, and then hypertrophied siphonal tissue formed. In zooxanthellal tubes, zooxanthellae usually had intact ultrastructures suggesting that they were photosynthetically active, while the stomach always contained degraded zooxanthellae that were probably discharged from the zooxanthellal tube. Giant clams probably digest zooxanthellae directly, and ingest the secreted photosynthates from zooxanthellae. There may be a selection mechanism to discharge unhealthy zooxanthellae from the mantle into the stomach. In addition to zooxanthellae, digested diatoms and other unidentified digested materials in the stomach suggest that filter-feeding also contributes to giant clam nutrition.     

Egg laying and embryonic-larval development in the snail<Emphasis Type="Italic"> Thais (Stramonita) chocolata</Emphasis> (Duclos, 1832) with observations on its evolutionary relationships within the Muricidae

Oviposition and embryonic-larval development are described for the muricacean snail Thais (Stramonita) chocolata from the Southeast Pacific coast. As with numerous other muricacean snails, this species engages in communal egg laying, with females depositing egg capsules in clusters on subtidal rocks. Each cluster of capsules contains 100–150 pedunculate, ampulliform egg capsules, with each capsule containing an average of 2,600 small (130 m) eggs. Intracapsular development was followed using light and scanning electron microscopy to describe the successive embryonic stages of the species. Free-swimming veliger larvae of about 225 m length were released from capsules after 49 days incubation at 13.6°C. The planktotrophic larvae were cultured in seawater aquaria by feeding with pure cultures of phytoplankton, recording growth and form of the larvae. Larvae reached competence after 4 months at 22°C, at 1,450–1,740 m in size, and a few larvae were observed through metamorphosis and early definitive growth. The embryonic-larval development of T. chocolata coincides with the general characteristics of the ontogeny observed in other Thais species as well as of other genera of the Rapaninae such as Concholepas. This lent support to grouping these genera into a single clade. The lack of knowledge of the development of free larvae of Thais spp. means that we do not know whether these similarities also include an extensive larval phase as generally characteristic of other members of the clade. The mode of development may be useful in characterizing some clades of this family. Thus for example, the transference of some Thais to the genus Nucella (Subfamily Ocenebrinae) is supported by differences in the mode of embryonic development, which differentiates these subfamilies. Paleobiological data reported for Neogastropoda allow postulation of primitiveness in planktotrophic larval development compared to more recent developmental strategies such as direct development of different types, which characterize various clades of this family.Communicated by O. Kinne, Oldendorf/Luhe     

Laboratory rearing of the clupeid fish Harengula pensacolae from fertilized eggs

Pelagic eggs of the scaled sardine Harengula pensacolae (Goode and Bean), have been hatched and reared in the laboratory for the first time. Larvae were reared in two 75 l aquaria under constant illumination, at an average temperature of 26.2°C. Zooplankton collected in a 35 mesh net was fed to the newly hatched larvae, and the diet was supplemented later with Artemia salina nauplii and a pelleted food. Larvae hatched at 4 mm TL (total length), and metamorphosed about 25 days later at 25 to 30 mm TL. Survivors averaged 76 mm TL 100 days after hatching. Of the 500 incubated eggs, 2.8% survived until 20 days, after which no significant natural mortality occurred. Sources of natural mortality included starvation, a copepod parasite (Caligus sp.), and injuries from contact with the sides of the tank. Larvae began feeding at 4.5 mm TL on copepod nauplii averaging 62 in body width. Scaled sardines were photopositive throughout the larval stage.Contribution No. 149, Bureau of Commercial Fisheries Tropical Atlantic Biological Laboratory, Miami, Florida 33149, USA.     

The effect of temperature-salinity combinations on survival and growth of juvenile Patiriella pseudoexigua (Echinodermata: Asteroidea)

Adult Patiriella pseudoexigua were collected in October 1989 from Wanlitung, Taiwan and then induced to spawn in the laboratory. Post-metamorphosed juvenile P. pseudoexigua were reared on a diet of benthic algae Navicula sp. at 25°C and salinity (34). Six weeks after metamorphosis, juvenile P. pseudoexigua at ca. 400 m in radius were reared on a diet of benthic algae Navicula sp. at different combinations of temperatures (20, 25, 30°C) and salinities (26, 30, 34) for 40 d. Both temperature and salinity had a significant effect on juvenile survival and growth. Juveniles survived best (>90%) at 25°C and 34 and grew best (to ca. 750 m in radius) at 30°C and 34. Variation in juvenile size was small immediately after metamorphosis and increased with time.     

Some observations on gametogenesis,larval development and substratum selection of the sea pen Ptilosarcus guerneyi

Ptilosarcus (Leioptilus) guerneyi (Gray) maintained in the laboratory, were observed to spawn in late March, 1972. Gametes, developed in the leaf proper, discharged through the mouths of feeding polyps and were fertilized externally in the sea water. The sea pen's eggs are 500 to 600 in diameter; a large female is capable of producing over 200,000 eggs in one season. A pear-shaped and free-swimming planula larva developed 4 days after fertilization, at a temperature of 12 °C. The larvae were ready to settle and metamorphose when 7 days old if favorable substratum was available, but would remain as planulae for at least 30 days if kept in glass dishes only. The 30-day-old larvae would metamorphose if a suitable substratum (coarse sand, for example) was presented. The larvae do not feed and, hence, development is lecithotrophic. Studies of histogenesis showed that metamorphosis greatly enhanced the rate of cellular differentiation. The high fecundity, lecithotrophic development, and the ability of substratum selection by the larvae explain the success of this species in maintaining a high-density population in many areas of sandy substratum in the shallow waters of Puget Sound (USA), despite the fact that it is preyed upon by 7 species of predators.     

Food selection by laboratory-reared larvae of the scaled sardine Harengula pensacolae (Pisces,Clupeidae) and the bay anchovy Anchoa mitchilli (Pisces,Engraulidae)

Food selection by laboratory-reared larvae of scaled sardines Harengula pensacolae, and bay anchovies Anchoa mitchilli, was compared. Natural plankton was fed to the larvae during the 22 days following hatching. Food levels in the rearing tanks were maintained at an average of 1,600 to 1,800 potential food organisms per liter. Larvae of both species selected as food copepod nauplii, copepodites, and copepods; initial feeding was on organisms of 50 to 75 body width. Larvae of H. pensacolae averaged 4.2 mm in total length at hatching and those of A. mitchilli about 2.0 mm. H. pensacolae larvae grew about 1.0 mm per day and A. mitchilli 0.70 mm per day. The mean number of food organisms in each digestive tract was greater in H. pensacolae than in A. mitchilli, and the difference in number increased as the larvae grew. Average size of food organisms eaten increased for both species with growth, because of selection by the larvae; the average size of copepodites and copepods in digestive tracts increased at a faste rate in H. pensacolae than A. mitchilli. A. mitchilli longer than 8 mm did not eat copepod nauplii.Contribution No. 170, Bureau of Commercial Fisheries Tropical Atlantic Biological laboratory, Miami, Florida 33149, USA.     

Effects of high temperature on larval development and metamorphosis of Arachnoides placenta (Echinodermata: Echinoidea)

Adults of the sea urchin Arachnoides placenta (L.) were induced to spawn, and eggs were fertilized at 28°C in September 1989. After 5 min, eggs were transferred to 28, 31, 34, or 37°C and reared to metamorphosis. Embryos were observed at 20-min intervals during the first 2 h; larvae were observed daily. The cleavage was higher at higher temperatures. Embryos reared at 28°C were still at the 4th cleavage (16-cell stage) after 100 min, while those at 34°C had reached the 5th cleavage (32-cell stage). All embryos reared at 37°C died on the second day. Incidence of abnormality was 20 to 30% at 28 and 31°C, 48% at 34°C, and 77% at 37°C. The 8-arm stage was reached after 4 d at 28°C, 3 d at 31°C and 2 d at 34°C. Larvae displayed decreasing body length and arm length with increasing temperature. Larvae at 31°C have relatively long arms, as a result of a decrease in body length, not because of increased arm length. Incidence of metamorphosis was 43.9±1.7% (mean/plusmn;SD) at 28°C, 24.5±1.9% at 31°C, and 5.3% at 34°C. The size of metamorphosed juveniles was significantly larger at 28°C than at 31 and 34°C. Temperatures of 31°C negatively affect larvae and juveniles of the sand dollar.     

Procedures adopted for the laboratory cultivation of Trichodesmium erythraeum

The present paper describes the procedures adopted for axenic or pure cultivation of the marine blue-green alga Trichodesmium erythraeum (Ehr.)¹ in the laboratory. Strains of T. erythraeum were isolated from the nearshore waters of the Bay of Bengal (11°29 N 79°49 E) during the pre-bloom period in March, 1965, at a depth of 9 to 12m. Modified, enriched, Erdschreiber medium was used for laboratory cultivation of T. erythraeum. A differential concentration of streptomycin 6 mg/100 ml, tetracycline 5 mg/100 ml and sulfadiazine 4 mg/100 ml was used to inhibit the microbial growth in cultures. Difficulties experienced in the initial culture stages of T. erythraeum and the necessary specific instructions employed are described in detail.     

Effect of temperature,salinity and food level on sexual and asexual reproduction in Brachionus plicatilis (Rotifera)

The reproductive response of sexual and asexual female Brachionus plicatilis (Muller) was examined over temperatures ranging from 20° to 40°C, salinities from 5 to 40 S, and food levels from 0.25 to 20 g Chlorella vulgaris dry-weight per ml. Reduced food levels, as well as temperature and salinity extremes, reduced reproduction of both sexual and asexual females, but did so differentially. Reproduction by sexual females was reduced to a greater extent at environmental extremes than asexual females. The broad, flat reproductive response curve of asexual females extended beyond the limits of the narrower, more sharply peaked curve of sexual females. Thus zones of exclusively asexual reproduction exist at environmental extremes where sexual reproduction is physiologically restricted. These results are corroborated by a comparison of the lifetime fecundity of individual sexual and asexual females over a 20°C temperature range. No differences in lifetime fecundity occurred between sexual and asexual females at 18° and 28°C. At 38°C, however, asexual female fecundity reached its highest level, while sexual female fecundity declined 15%. The appearance of sexual females in rotifer populations in the result of both inducible and repressible factors.     

Sexual and asexual reproduction of Anthopleura dixoniana (Anthozoa: Actiniaria): periodicity and regulation

Reproduction of the sea anemone Anthopleura dixoniana (Haddon and Shackleton) from the high intertidal zone of southern Taiwan (120°41 E; 22°01N) was studied from April 1987 through March 1989. A. dixoniana spawns once a year, in July, and divides asexually by longitudinal fission throughout the year, with a peak in July. During the spawning season, sea anemones>3 mm pedal dise diameter can be sexed, and display a 1:1 sex ratio. Dividing sea anemones are significantly larger than non-dividing individuals, and increase in body size before fission. Under laboratory conditions, individuals kept at 28 C and fed had larger oocytes and a higher division rate than those kept at 18, 22, 25 or 32°C or starved. The division rate significantly influenced the oocyte diameter. The present study revealed for the first time, that a long photoperiod (14 h hight:10 h dark) significantly enhances the growth of oocytes in A. dixoniana under laboratory conditions.     

Evidence for a water-borne cue inducing metamorphosis in the dorid nudibranch mollusc Adalaria proxima (Gastropoda: Nudibranchia)

The dorid nudibranch Adalaria proxima (Alder & Hancock) is a specialist predator of the cheilostome bryozoan Electra pilosa (L.). Natural induction of metamorphosis of the pelagic lecithotrophic larva of A. proxima was assessed in response to solutions from sonicated prey tissue and (live) E. pilosa-conditioned seawater (Electra-CSW). We exploited the tendency of larvae to become entrapped (rafted) at the air-water interface in cultures to examine whether larvae require direct contact with the live prey for metamorphosis to proceed. Larvae metamorphosed when rafted above colonies of live E. pilosa, above plankton mesh bags isolating live E. pilosa, and in choline chloride controls; there was no metamorphosis of larvae that were rafted in filtered seawater controls. Entrapped veliger shells remained rafted throughout the experimental period in all cases. No metamorphosis occurred in treatments containing either the supernatants or pelleted particulates obtained from sonicated colonies of E. pilosa. Both one-colony and three-colony Electra-CSW induced metamorphosis of larvae. These data are at variance with previous results in showing that direct contact with the live prey is not necessary for metamorphosis to proceed. Furthermore, the fact that competent larvae metamorphosed in response to Electra-CSW in the absence of any other cue strongly suggests that the inductive cue is water-borne.     

Comparison of larval development and growth of the sea cucumberActinopyga echinites: Ovary-induced ova and DTT-induced ova

AdultActinopyga echinites (Jaeger) were collected from northern Taiwan in September 1989. Oocytes were induced to mature by bathing them in ovary juice (ovary-induced ova) or in 10^–2 M dithiothreitol (DTT-induced ova). The percentage of germinal vesicle breakdown (GVBD) increased from 0.4 to 6.4% in the former treatment and to 84% in the latter. After artificial fertilization, the embryos were cultured in seawater (35 S) at 25 to 28°C. Larvae were fed with the algaIsochysis aff.galbana at 10^4–5 cells/ml. Larvae from ovary-induced ova developed faster (18 d to the doliolaria stage) and grew to a larger size (1.13 mm length) than those from DTT-induced ova (20 d to the doliolaria stage and 0.62 mm in length). On the twelfth day, larvae from ovary-induced ova bear lipid spheres. The number of spheres is positively correlated with larval size. Lipid spheres may provide nutrient reserves for larvae during metamorphosis.