Osmotic and structural changes during early development of eggs and larvae of the cod, Gadus morhua L.

Newly extruded cod eggs have a similar osmoconcentration ( c. 400 mosmol kg^-1) to maternal blood, maternal ovarian fluid and paternal semen. After extrusion, average egg osmolarities rise rapidly to about 520 mosmol kg^-1 but this is entirely due to the cortical reaction resulting in the formation of a perivitelline space filled with fluid isosmotic with the external medium. The ovoplasm remains at around 400 mosmol throughout development; free swimming cod larvae also possess body fluids of this concentration. Over the salinity range 10.2-37.4%₀, salinity had no effect on egg dimensions and average egg osmolarities were consistent with a model assuming changes in perivitellic fluid concentration only.     

Effects of temperature and salinity on yolk utilization in Bairdiella icistia (Jordan & Gilbert) (Pisces: sciaenidae)

The effects of nine different combinations of temperature and salinity on yolk utilization in the eggs and larvae of the sciaenid fish, Bairdiella icistia (Jordan & Gilbert) have been investigated. The rate of yolk absorption was accelerated at higher temperatures, but salinity had only a slight effect. Increased water content resulted in larger yolk sacs at hatching in low salinities and at low temperatures. In all treatments, functional eyes and jaws were formed before the yolk had been completely exhausted. Yolk was utilized most efficiently at the intermediate temperature (24 °C) and lowest salinity (20 %.).     

The metabolism of neutral and polar lipids in eleuthero-embryos and starving larvae of the African catfish Clarias gariepinw

Lipid class analysis was carried out on developing eggs, eleuthero-embryos (yolk sac larvae) and starving larvae of the freshwater species Clarias gariepinus , using thin layer chromatography. Samples were taken at fixed intervals from a large pool of fertilized eggs obtained through induced reproduction of several parent fish. The total lipid content of fertilized eggs fluctuated around 22% of the dry weight and decreased from 21% at hatching to about 12–5% at yolk absorption. In starving larvae, the amount of total lipid per individual remained relatively constant. Polar lipids [phosphatidylcholine (PC) and phosphatidylethanolamine (PE)] together accounted for 73·6 to 80% of total lipid. PC was by far the most abundant lipid class during the entire experimental period (70–75% of total lipid). PC was catabolized proportionally to total lipid, demonstrating its role as the main energy supplier. All yolk PE was converted to body tissue. The neutral Hpids consisted of triglycerides (TAG), cholesterol and cholesteryl esters (respectively 12·5, 10 and 3% of total lipid in newly fertilized eggs). All TAG were depleted before complete yolk absorption.     

The effect of parental acclimation to spawning salinity on the survival of larval Cynoscion nebulosus

The yolk and oil depletion of eggs and larvae of spotted seatrout Cynoscion nebulosus , produced by fish collected from two bays with historically different salinity regimes (Matagorda Bay (MB; 18-24%) and Upper Laguna Madre (ULM; 40–50%), Texas, U.S.A. and spawned in salinities of 20, 30 and 40%, differed in their response to both salinity and history. Time to 90% yolk depletion was significantly longer for low salinity bay fish (MB) kept at 20%, but not for high salinity bay fish (ULM) at 20%. The neutral buoyancy salinity of 1 and 2 day old MB 20% larvae was significantly lower than that of MB larvae spawned in 30 or 40%. Overall, eggs and larvae spawned by MB fish were able to hatch out and survive to 3 days post-hatch in lower salinities than those from ULM. Furthermore, the tolerance of eggs and larvae to very low salinities increased with decreasing spawning salinity. The ability of 1–9 day old ULM, but not MB, larvae to survive 18 h exposure to salinities above or below that of spawning exhibited an age-dependent pattern with day 3 being the most sensitive. This study shows that the response of spotted seatrout eggs and larvae to changes in salinity is dependent upon the spawning salinity of the adults and the prevailing salinity regime within the bay.     

Egg formation and the pre-laying period of the Common guillemot Una aalge

The patterns of colony attendance of male and female Common guillemots, relative to calendar date and relative to the female's laying date during the three weeks prior to egg laying, are presented. The probability of a male being at the colony was consistently higher than that for females. Male attendance peaked in the three days before his mate laid: female attendance was lowest at this time. Examination of the yolk showed that the egg was formed over 14–15 days, with yolk deposition (of first eggs) taking 11–5 days, and a lag period (between the end of yolk deposition and laying) of 3–4 days. Yolk deposition occurred over a shorter period (9–3 days) in replacement eggs, and followed a different pattern from first eggs.     

Early ontogeny of coal grunter from hormone-induced spawnings and laboratory-reared embryos and larvae

Spawning of coal grunter Hephaestus carbo was successfully induced using doses of human chorionic gonadotrophin (hCG) between 500 and 3000 IU kg (body weight)⁻¹. Water hardened eggs are telolecithal, amber in colour, spherical, transparent, demersal and slightly adhesive with a single large oil droplet and perivitelline space 47% of total egg volume. Cleavage begins 10–15 min after fertilization. Epiboly begins 6h after fertilization and continues for 4 h. Invagination of the neural tube is apparent 11·5 h after fertilization, followed by progressive organogenesis up to hatching 60–80 h after fertilization. An invagination in the yolk, consistent in shape, position and time of appearance among embryos spawned from numerous brood stock pairs, was visible in all fertilized eggs between neurulation (11-5 h) and early organogenesis (20 h). The functional significance of this yolk invagination is unknown. Newly-hatched larvae (4·2 mm L _T) are elongate and possess well developed eyes, a functional mouth, and a large yolk sac. Yolk is fully resorbed and first feeding occurs at 6 days posthatching. The sequence of fin formation is caudal, second dorsal and anal, first dorsal, pectoral and pelvic. The prefiexion larval stage lasts for c. 8 days and flexion of the notochord is complete within a further 8–9 days. Squamation commences at 30 days posthatching and transition to the juvenile life stage is complete by 35–40 days posthatching.     

Lipovitellins and phosvitins of the fertilized eggs during embryo growth in the oviparous lizard Podarcis sicula

In the lizard Podarcis sicula, the major vitellogenin (VTG)-derived yolk proteins, lipovitellins and phosvitins, were extracted from the yolk globules of laid and fertilized eggs at different periods of incubation up to 44 days close to hatching. Embryonic development was almost over at this time. Yolk proteins were isolated by precipitation in saturated (NH(4))(2)SO(4), separated on SDS-PAGE and detected by Western blotting with homologous polyclonal anti/VTG antibody. Two lipovitellins of 110 and 116 kDa were always present in the yolk of laid eggs after 1, 10, 18, and 44 days from oviposition. Both these proteins were glycosylated and were recognized by the anti/VTG antibody; their N-terminal sequences were analyzed. Four phosvitins were detected in freshly laid eggs, but their number decreased during incubation, and after 44 days only a single protein of approximately 6.5 kDa was present. The results indicated that, in this lizard, during embryonic development, lipovitellins remain unchanged, whereas the phosphorylated components of yolk undergo continuous degradation.     

Reproductive behaviour and early development of the Drysdale hardyhead, Craterocephalus sp. nov. (Pisces: Atherinidae), from the Alligator Rivers System, Northern Territory, Australia

Craterocephalus sp. nov. showed sexual dimorphism in body shape during the breeding season. Pairing occurred during daylight with a single release of eggs amongst submerged vegetation between sunrise and the early afternoon on the same day. The eggs were demersal, adhesive, spherical (0.87–0.96 mm diameter) and had 12 adhesive filaments (0.5–1.5 mm long) at the animal pole. Approximately 24 oil droplets (0.02–0.08 mm diameter) persisted throughout egg and larval development. Hatching occurred 155–160 h after spawning at 25–27° C.
The yolk-sac larvae were 3.85–3.95 mm notochord length at hatching and began feeding at the surface after absorption of the yolk (3–12 h after hatching). All fin rays were developed in 9.4 mm standard length fry, which moved from midwater to feed on the substrate. Aquarium reared fish first spawned at 30 mm s.L. when 165 days of age.
Features of Craterocephalus reproduction, as they relate to a specific survival strategy, are briefly discussed.     

Growth and respiration of embryos and larvae of the rabbittish Siganus randalli (Pisces, Siganidae)

Growth and respiration of larval rabbitfish from Guam were examined. Larvae were reared from eggs in 2- to 10-ton tanks and were fed rotifers, Anemia , and artificial feed in succession as development proceeded through metamorphosis. Growth in length was rapid during the 12 h after hatching, then slowed until the larvae began to feed. The yolk sac was usually absorbed by 36 h after hatching. Rates of respiration of larvae and eggs were determined with a dissolved oxygen electrode at various times through development. Larval metabolism increased steadily during the embryonic stages culminating in a metabolic burst immediately after hatching. Respiration rates remained relatively stable from shortly after hatching until the onset of exogenous feeding, after which respiration rates increased with larval size. The respiration rates of post-yolk-sac larvae scaled isometrically with larval dry mass. Daily growth of feeding larvae was 27 to 28% of larval dry mass.     

Post‐hatch dispersal of lake sturgeon (Acipenser fulvescens,Rafinesque, 1817) yolk‐sac larvae in relation to substrate in an artificial stream

Knowledge of the effects of environment and genotype on behavior during early ontogenetic stages of many fish species including lake sturgeon (Acipenser fulvescens) is generally lacking. Understanding these effects is particularly important at a time when human activities are fundamentally altering habitats and seasonal and diel physical and biotic stream features. Artificial stream channels were used in a controlled experiment to quantify lake sturgeon yolk‐sac larvae dispersal distance and stream substrate preference from different females (N = 2) whose eggs were incubated at different temperatures (10 and 18°C) that simulated stream conditions during early and late spawning and incubation periods in the Black River, Michigan. Data revealed that yolk‐sac larvae exhibited considerable variability in dispersal distance as a function of family (genotype), temperature experienced during previous (embryonic) ontogenetic stages, and environmental ‘grain’. Yolk‐sac larvae dispersal distance varied as a function of the juxtaposition of substrate to location of egg hatch. Lake sturgeon yolk‐sac larvae dispersed from mesh screens attached to bricks and settled exclusively in gravel substrate. Dispersal distance also varied as a function of family and egg incubation temperatures, reflecting differences in offspring body size and levels of endogenous yolk reserves (yolk sac area) at hatch. Expression of plasticity in dispersal behavior may be particularly important to individual survival and population levels of recruitment contingent upon the location, size, and degree of fragmentation of suitable (gravel) habitats between adult spawning and yolk‐sac larvae rearing areas.     

Yolk Degradation in Tick Eggs: III. Developmentally Regulated Acidification of the Yolk Spheres

Yolk spheres in tick eggs contain a latent procathepsin L, which is activated in vivo , in parallel with yolk degradation, and in vitro by acid treatment (Fagotto, F., Arch. Insect Biochem. Physiol., 1990b in press). Mature cathepsin L hydrolyzes vitellin at acidic pH (Fagotto, F., Arch. Insect Biochem. Physiol., 1990a in press). Here, yolk spheres' pH has been estimated using acridine orange. In the early development, all yolk spheres are neutral, then an increasing number acidify, until hatching, where general acidification seems to occur. This fits well with vitellin utilized slowly during embryogenesis, more intensely at hatching (Chinzei, Y. and I. Yano, Experientia 41 , 948, 1985), and can be related to sequential degradation of individual spheres observed during embryonic development, then extensive yolk liquefaction in the larva. Different yolk spheres populations have been separaded on Percoll density gradients. In freshly laid eggs, yolk spheres are dense, neutral, undegrated and contain exclusively the precursor of cathepsin L. In later stages, yolk spheres are progressively recovered in lower density fractions, displaying acidic interior and cytological signs of degradation. They cosediment with mature cathepsin L. It is concluded that acidification initiates yolk degradation through procathepsin L activation.     

Egg formation and the pre-laying period of Black-browed and Grey-headed Albatrosses Diomedea melanophris and D. chrysostoma at Bird Island, South Georgia

Egg composition and factors influencing egg formation were studied in Black-browed and Grey-headed Albatrosses Diomedea melanophris and D. chrysostoma at Bird Island, South Georgia. At nests where eggs were laid, females arrived 6–7 days after males, stayed one day during which 96% of observed copulations occurred, then departed to sea for c. 16 days in D. chrysostoma, c. 10 days in D. melanophris , returning c. two days before laying. Yolk deposition, however, lasted 21 and 20 days, and started 32 and 29 days before laying, in D. chrysostoma and D. melanophris respectively. Therefore, it is probably initiated by environmental factors not by copulation. Egg, albumen and yolk mass are significantly greater in D. chrysostoma but the proportionate composition of the species' eggs is nearly identical. Reduced differences in chick mass at hatching may reflect the longer incubation period in D. chrysostoma or relate to subsequent differences in chick growth rate.     

A study of the assimilation of fluorescent pigments of microalgae Isochrysis galbana by the early larval stages of turbot and herring

The appearance of large supranuclear vacuoles in the enterocytes of 1- and 4-days post-hatch larvae of turbot and herring, respectively, revealed by the pinocytotic uptake of a fluorescent marker (FITCdextran), indicates a potential for the absorption of dissolved nutrients by the endotrophic stages of marine fish larvae. Ingestion of algal cells by turbot larvae was observed soon after hatching, but low level pinocytotic absorption of algal material was first seen during the second day. More extensive lysis of algal cells and pinocytotic absorption occurred 24 h later. Although lysis of I. galbana was shown to occur at low external osmolarities, it is unlikely that sufficiently low osmolarities present in the hind gut of turbot larvae explained the observed rupture of algae. Other mechanisms for digestion are discussed. In newly hatched herring larvae, algal cells were unable to pass beyond the constriction in the mid-gut caused by the yolk sac. When algal cells were eventually seen to pass into the hind gut, there was no evidence of algal digestion or absorption throughout the remaining endotrophic and early exotrophic stages of herring larvae, although pinocytosis was observed to occur mid-way through the endotrophic stage.     

The combined effects of temperature and salinity on hatching success and the survival,growth, and development of the larval stages of Metapenaeus bennettae (Racek & Dall)

During the spawning season of the estuarine prawn Metapenaeus bennettae (Racek & Dall), laboratory and field experiments were conducted to examine the combined effects of temperature and salinity on hatching success of eggs and the survival, growth and development of larvae. Response surface analysis showed that optimal levels of temperature and salinity for maximum hatching success varied depending on conditions during spawning. Similarly, temperature and salinity conditions that produced maximum survival and growth of larvae depended on conditions during rearing prior to experimental temperature/salinity treatments. At the onset of feeding, larvae showed the lowest tolerance to changes in temperature and salinity. Supplementary feeding experiments in the laboratory, and survival rates in field experiments indicated that starvation was a more potent factor than the effects of temperature and salinity in determining survival through the protozoeal larval stages. Late larval stages were relatively indifferent to the effects of temperature and salinity. It is suggested that, during early development, adaptive response to the prevailing physical conditions enhances survival in an estuarine environment.     

Effects of development, temperature and salinity on metabolism in eggs and yolk-sac larvae of milkfish, Chanos chanos (Forsskål)

Oxygen uptake rates and yolk-inclusive dry weiGhts were measured during the egg and yolk-sac larval stages of milkfish, Chanos chanos (Forsskal). Oxygen uptake by eggs and yolk-sac larvae was measured to assess the effects of four salinities (20,25,30,35 ppt) at 28°C. The effects of three temperatures (23,28,33°C) on oxygen uptake by yolk-sac larvae were determined at a salinity of 35 ppt. Dry weights were measured throughout embryonic development at 28°C and the yolk-sac stage at 23.28 and 33°C.
Oxygen uptake rates of eggs increased more than fivefold during embryogenesis (0.07±0.03 to 0.40 ± 03 μl O₂ egg ⁻¹ h ⁻¹;blastula to prehatch stage). Larval oxygen uptake did not change with age but was affected by rearing temperature (0.33 ± 0.08, 0.44 ± 0.07 and 0.63 ± 0.13 μl O₂ larva ⁻¹ h⁻¹ at 23, 28 and 33°C, respectively; Q₁₀= 1.93). Acute temperature changes from 28 to 33°C caused significant increases in oxygen uptake by embryos (Q ₁₀= 1.69–3.58) and yolk-sac larvae (Q ₁₀=2.55). Salinity did not affect metabolic rates.
Dry weight of eggs incubated at 28°C decreased 13% from fertilization to hatching. Incubation temperatures from 23–33°C did not affect dry weights at hatching. Rearing temperatures significantly affected the rate of larval yolk absorption (Q ₁₀= 2.25).     

Yolk androgens reduce offspring survival

Females may favour some offspring over others by differential deposition of yolk hormones. In American kestrels (Falco sparverius), we found that yolks of eggs laid late in the sequence of a clutch had more testosterone (T) and androstenedione (A4) than yolks of first-laid eggs. To investigate the effects of these yolk androgens on nestling 'fitness', we injected both T and A4 into the yolks of first-laid eggs and compared their hatching time, nestling growth and nestling survival with those of first-laid eggs in which we injected vehicle as a control. Compared to controls, injection of T and A4 at a dose intended to increase their levels to those of later-laid eggs delayed hatching and reduced nestling growth and survival rates. Yolk androgen treatment of egg 1 had no effect on survival of siblings hatching from subsequently laid eggs. The adverse actions of yolk androgen treatment in the kestrel are in contrast to the favourable actions of yolk T treatment found previously in canaries (Serinus canaria). Additional studies are necessary in order to determine whether the deposition of yolk androgens is an adaptive form of parental favouritism or an adverse by-product of endocrine processes during egg formation. Despite its adaptive significance, such 'transgenerational' effects of steroid hormones may have helped to evolutionarily shape the hormonal mechanisms regulating reproduction.     

Correlation of yolk phosphatase expression with the programmed proteolysis of vitellin in Blattella germanica during embryonic development

Proteolytic processing of the vitellin in Blattella germanica eggs occurs 4 days postovulation and is correlated with both the onset of its utilization and the major portion of the embryo's growth. Yolk phosphatase is also expressed coincident with this event, and some aspects of its activation have been investigated. The enzyme is absent from the ooplasm (yolk) during the first 2 days following ovulation but increases approximately 20-fold in specific activity between days 3 and 4, when assayed at pH 3.9 or 4.8 and 9-fold at pH 6.5. No activation is observed for yolk-bound α-mannosidase, its specific activity remains elevated through the first 6 days following ovulation. This suggests that expression of the phosphatase is regulated independently of that of α-mannosidase in the yolk. Yolk with active phosphatase can dephosphorylate native vitellin, delipidated vitellin, and phosphocasein. Sucrose density gradient centrifugation of yolk obtained from eggs 4 days postovulation, revealed that phosphatase activity cosediments with material which reacts with antivitellin antibodies, while α-mannosidase and β-N-acetyl glucosaminidase are found near the top of the gradient. Oothecae derived from crossing certain translocational heterozygote males and wild-type females contain some eggs with severely depressed levels of yolk phosphatase in which embryos do not grow. Vitellin in these eggs fails to undergo proteolytic processing as late as day 5 postovulation and retains the subunit composition of freshly ovulated vitellin.     

Developmental fate of the yolk protein lipovitellin in embryos and larvae of winter flounder, Pleuronectes americanus.

The developmental fate of the vitellogenin-derived yolk protein, lipovitellin (Lv), was investigated in winter flounder embryos and yolk-sac larvae. Since Lv is present as only one major polypeptide in ovulated winter flounder eggs, unlike the multiple yolk polypeptides found in the mature eggs of most teleosts, this system is presented as a simpler model of yolk protein structure and utilization during teleostean development. Winter flounder Lv is cleaved during embryogenesis from a 94 kD polypeptide at fertilization to 67 kD and 26 kD polypeptides at hatching. The rate of this proteolytic processing is slow during early embryonic development, but enters a more rapid phase between days 8 and 12 post-fertilization in embryos reared at 4-5 degrees C, and approaches 50% completion at day 10. Lv processing is essentially complete 3 days before hatching; nevertheless, major degradation of the Lv peptide by the developing winter flounder does not occur until after hatching. The Stokes radius of Lv changes only moderately following processing, from 4.50 nm in unfertilized eggs to 4.19 nm in late embryos and newly hatched larvae, whereas the processed Lv retains its heat stability relative to other yolk polypeptides. Nearly 50% of its lipid content, however, is released from the Lv particle during embryogenesis, concomitant with cleavage of the Lv 94 kD polypeptide. Lv processing may thus render a portion of the yolk protein-associated lipid more accessible to the developing embryo, whereas other yolk components are retained for later use by the winter flounder larva. Alternately, removal of lipid may lead to proteolytic vulnerability of the Lv polypeptide. In either case, only a portion of the lipid moiety of the Lv particle appears to play a significant nutritive role for the embryo, whereas its protein component is reserved for larval use. J. Exp. Zool. 284:686-695, 1999.     

Direct Effects of Microalgae and Protists on Herring (Clupea harengus) Yolk Sac Larvae

This study investigated effects of microalgae (Rhodomonas baltica) and heterotrophic protists (Oxyrrhis marina) on the daily growth, activity, condition and feeding success of Atlantic herring (Clupea harengus) larvae from hatch, through the end of the endogenous (yolk sac) period. Yolk sac larvae were reared in the presence and absence of microplankton and, each day, groups of larvae were provided access to copepods. Larvae reared with microalgae and protists exhibited precocious (2 days earlier) and ≥ 60% increased feeding incidence on copepods compared to larvae reared in only seawater (SW). In the absence and presence of microalgae and protists, life span and growth trajectories of yolk sac larvae were similar and digestive enzyme activity (trypsin) and nutritional condition (RNA-DNA ratio) markedly declined in all larvae directly after yolk sac depletion. Thus, microplankton promoted early feeding but was not sufficient to alter life span and growth during the yolk sac phase. Given the importance of early feeding, field programs should place greater emphasis on the protozooplankton-ichthyoplankton link to better understand match-mismatch dynamics and bottom-up drivers of year class success in marine fish.