Influence of salinity on embryonic development and the distribution ofSepia officinalis in the Delta Area (South Western part of The Netherlands)

The effect of salinity on embryonic development ofSepia officinalis (cuttlefish) in the Delta Area (South Western part of The Netherlands) was investigated in 1988/1989, and compared with data concerning the distribution ofS. officinalis in the three main parts of this area: Oosterschelde, Westerschelde and Grevelingen. Embryos hatched in water collected at Yerseke (Oosterschelde), Vlissingen (Western part of the Westerschelde) and Bommenede (Grevelingen), i.e., at salinity values above 28.1, but not in water sampled at Hoedekenskerke and Hansweert (Middle and Eastern part of the Westerschelde; salinities below 22.0). Under laboratory conditions, using diluted Oosterschelde water, the highest hatching percentages ofS. officinalis were found at salinities above 29.8. Some embryos hatched at a salinity value of 26.5 but no hatching occurred at salinities below 23.9. In embryos exposed to salinity changes during late embryonic development, the developmental rate decreased at salinity values of 28.7 or less. Below 22.4 embryos with morphological malformations were found. It can be concluded that salinity is an important factor limiting the distribution ofS. officinalis in most parts of the Delta Area, with the exception of the Western part of the Westerschelde and the Grevelingen.Contribution no. 489 of the Library of the Delta Institute for Hydrobiological Research     

Respiration of Sepia officinalis during embryonic and early juvenile life

Adult Sepia officinalis L. were caught in June 1984, in the coastal waters of Wimereux (France). Deposition of the eggs took place in the seawater aquaria of the Station Marine. The oxygen consumption of S. officinalis was measured during embryonic and juvenile development. Aerobic metabolism occurs as soon as the early embryonic Stage 21. Oxygen diffuses through the initially thick egg shell; the oxygen level in the perivitelline liquid reaches a maximal value just before hatching (116.7±6.9 mm Hg). Hatchings display only a slight increase in oxygen consumption compared to embryos in the last stage of development. Respiration experiments with 40 d old juveniles showed that oxygen consumption increases with temperature, but is not affected by photoperiod. Experiments under increasing hypoxia revealed that S. officinalis juveniles are good regulators and maintain a constant oxygen consumption in the range of 4 to 7 mg O₂l^-1. Juveniles successfully recover from an hypoxic stress of 2 mg O₂l^-1 maintained for 1 h. This suggests that the respiratory pigments (pre-hemocyanins) of 40 d-old juveniles have a high oxygen affinity and/or that these juveniles have the ability to adapt to anaerobic conditions.     

Effects of zinc on epidermal ultrastructure in the larva of Clupea harengus

In March, 1983 changes in epidermal ultrastructure were examined in Clupea harengus L. larvae hatched from eggs incubated in four zinc concentrations (0.5 2.0, 6.0 and 12.0 ppm). In addition to the outer and inner epidermal cell types described previously, a third type of cell is present. Ultrastructurally this resembles the epidermal chloride cells of Sardinops caerulea Girard, characterised by numerous mitochondria, extensive smooth endoplasmic reticulum and a free surface exposed to the environment. These cells occur on the yolk sac, head and in the regions of the trunk just above the yolk sac. In larvae treated in 12.0 ppm zinc, the cells contain fewer mitochondria with fewer cristae than those in the controls. Smooth endoplasmic reticulum is much reduced forming nodular masses and contains granular osmiophilic inclusions. In larvae hatching from eggs previously incubated in 6.0 and 12.0 ppm zinc, the epidermal cells contain more vesicles, intracellular spaces, and swollen mitochondria and show signs of necrosis. The viability of these larvae is discussed.     

Lipid biochemistry and reproductive biology in two species of Gammaridae (Crustacea: Amphipoda)

Gammarus oceanicus Segerstråle, 1947 and Echinogammarus marinus (Leach, 1815) were sampled during the breeding season from Oslofjord in 1984, and their lipid composition examined in relation to reproductive condition. In G. oceanicus, female lipid content increased as the ovary matured. Both the amount of lipid stored and the rate of accumulation were greater in spring than in winter. Spring eggs contained 12.4 g lipid, of which 63% was triacylglycerol and 27% phospholipid. Both fractions decreased steadily during embryonic development. Winter eggs contained 19.2g lipid, of which 52% was triacylglycerol and 43% phospholipid. During the early stages of embryonic development the amount of phospholipid decreased sharply, whereas that of triacylglycerol increased, suggesting that some of the fatty acid released from phospholipid was sequestered temporarily as triacylglycerol. When newly spawned, both winter and spring eggs were richer in monoenoic fatty acids than adult amphipods and these acids were the major fuel consumed during development. 6 fatty acids were utilised more slowly than 3 acids, and egg carotenoid pigment content remained constant. Female E. marinus increased in lipid content as the ovary matured. Spring eggs contained 14.7 g lipid when newly spawned and this increased to 16.6 g during the early stages of development. This increase was entirely triacylglycerol, which declined in later stages; the source of the extra lipid was unclear. Eggs contained very little phospholipid or sterol, and both of these components remained at a steady low level during development. E. marinus eggs were not significantly rich in thonoenoic acids compared with adults, and saturated, monoenoic and polyenoic acids were utilised about equally during development. Both adults and eggs were rich in 20.46, which was utilised at a slower rate than the 3 polyunsaturated acids during embryonic development; again, egg carotenoid pigment content remained constant. In both species there was a decrease in the size of the egg (and as a result, of the newly hatched juvenile), but an increase in total reproductive output (i.e., the total weight of the egg clutch) per female as the breeding season proceeded. The reproductive output of an individual female is probably related to food availability during the period of ovarian maturation, whereas the size of an individual egg is dictated largely by feeding conditions for the juveniles once they are independent of the female. The different patterns of lipid utilisation during development found in this study emphasize the flexibility of response in the reproductive biology of gammarid amphipods. It is not yet possible, however, to relate the differing patterns in a simple way either to egg size or total female reproductive output. Two outstanding problems are the source of extra triacylglycerol during the early stages of development of E. marinus and the metabolic cost of brooding eggs.     

Embryo development and maternal–embryo nutritional relationships of piked spurdog (<Emphasis Type="Italic">Squalus</Emphasis><Emphasis Type="Italic"> megalops</Emphasis>)

The maternal–embryo relationship was determined for the piked spurdog (Squalus megalops). In addition, the increase in offspring size with maternal size was studied and the embryonic development was described. Wet weight of in utero eggs and offspring size was correlated with maternal size; larger females produced larger embryos which would have higher survival rate and reproductive value. All embryos present in a female were at a similar stage of development. The external yolk sac is reabsorbed late in gestation, suggesting that embryos are mostly nourished by yolk sac reserves. Embryo size-at-birth varied considerably (180–244 mm total length) as a result of the significant variability in ova size at ovulation. The amounts of water, organic and inorganic matter of embryos at different stages of development were measured to determine possible maternal contributions during embryonic development. Total wet weight from smallest and largest in utero eggs to smallest and largest term embryos changed by +46 and +58%, respectively. This pattern was due to a change in water content by +137 and +154%, and inorganic matter by +100 and +156%. Organic matter of smallest and largest in utero eggs changed by −23 and −17%, respectively. The uterus of pregnant females became specialised for water and mineral transport, not nutrient provision. These results indicate that S. megalops is a strict yolk-sac viviparous species with no maternal contribution of organic matter during development.     

Distribution of copper and iron in some central organs of Sepia officinalis (Cephalopoda). A comparative study by flameless atomic absorption and electron microscopy

In juvenile and adult Sepia officinalis (L.) of the Bassin d'Arcachon, France, the copper and iron contents of blood, hepatopancreas, branchial gland, branchial heart, branchial heart appendages, heart ventricle, and pancreatic appendages were determined by flameless atomic absorption. Despite the significant differences between the absolute values of the copper and iron concentrations in the different organs, a certain parallelism can be demonstrated in the ability to concentrate the two metals. The hepatopancreas, as the most important storage organ, shows the highest concentrations of copper and iron, followed by the branchial heart system with distinctly lower values. The cytolysosome-like dense bodies in these tissues of s. officinalis and of Octopus vulgaris (Lam.) can be characterized as the cellular sites of storage by histochemistry and electron microscopy. The low copper content of the branchial gland does not disprove the conception of a hemocyanin synthesis in this organ; biochemical experiments indicate a subsequent incorporation of copper released by the endoplasmic reticulum (ER) of the gland.This study was supported by the Deutsche Forschungsgemeinschaft.     

Hypoosmoregulatory ability of eyed-stage embryos of chum salmon

To evaluate the osmoregulatory ability of eyedstage embryos of laboratory-reared chum salmon, Oncorhynchus keta, we examined changes in osmolality of the perivitelline fluid and blood following transfer to 50 and 100% seawater (SW), together with morphological changes in chloride cells present in the yolk sac membrane. Transfer to SW did not cause any significant change in the whole egg weight. However, the embryos shrank when the eggs were transferred to SW, whereas the perivitelline space increased at the expense of the embryo. Osmolality of the perivitelline fluid increased rapidly to reach environmental levels 3 h after transfer, indicating that the egg shell is permeable to ions and water. Blood osmolality increased after transfer to SW, reached a peak level at 3 h, and then decreased gradually. The chloride cells in the yolk sac membrane became activated following transfer, as shown by increased cell size and frequent appearance of apical openings. These results indicate that the eyed-stage embryos of chum salmon possess hypoosmoregulatory ability and that chloride cells in the yolk sac membrane may be involved in salt extrusion, in place of gill chloride cells, during the late embryonic stage.     

Yolk utilization and hatching time in the Canadian lobster Homarus americanus

During the course of its embryonic development, the Canadian lobster Homarus americanus Milne-Edwards exhibits steady increases in water content (56.2 to 86.8%) and ash (5.8 to 21.2%), and a progressive decrease in energy content from 6636 to 4292 cal/g dry weight. Mean dry weight of a single egg is 965 g, equivalent to 6.4 cal; a freshly hatched egg. The lobster hatches about 1,500 larvae per night over a period of 4 to 5 days. Dry weight, ash and calorific contents of larvae hatched on different days show considerable variations. After larvae hatch on the first day, continuous salt absorption by eggs to hatch on subsequent days leads to a steady increase in ash content from 143 g/larva hatched on the first day to 255 g/larva hatched on the fourth day, and consequently, to an increase in dry weight from 854 to 956 g/larva. Metabolism of embryos (0.1 cal/day), which are yet to be hatched on subsequent days, depletes the calorific content per unit weight (from 4637 to 3837 cal/g dry weight) as well as per larva (from 3.98 to 3.67 cal).Dedicated to my colleague K.-H. Schumann (Biologische Anstalt Helgoland) who died on December 12, 1969 due to a diving accident.     

Studies on sharks. I. Reproduction in the female sumitsuki shark Carcharhinus dussumieri

During the last 5 years, the T.S. Nagasaki Maru, Nagasaki University, Japan, carried out trawling operations in the waters off Kuching, northwest Borneo, for 1 week in July of each year. During this period, 119 specimens of the sumitsuki shark Carcharhinus dussumieri Valenciennes (Müller et Henle, 1841) were caught and their reproductive system investigated. In these waters, C. dussumieri has no definite reproductive seasons; it was, therefore, possible to obtain sufficient suitable material to examine all stages of the sexual cycle. The left ovary is rudimentary throughout the life cycle, and only the right ovary is functional. The ova in the ovary are already developing for the succeeding ovulation while the embryo already present in the uterus is growing; the 2 mature eggs from the right ovary are ovulated in the peritoneal cavity soon after parturition. The developing ova which remain in the ovary after ovulation are reabsorbed. The 2 mature eggs ovulated enter a common ostium and each oviduct receives one egg. In the nidamental gland, the fertilized egg is enclosed by an embryonic membrane and descends into the uterus where it grows into an embryo (simple pregnancy: 1 embryo in each uterus). During earlier embryonic development, nourishment is obtained from the yolk stored in the yolk sac. The placenta is established when the embryo attains a total length of about 150 mm; the embryo is thenceforth nourished by the placenta through the umbilical stalk until birth.     

Spawning behaviour,egg and larval development of the myctophid fish Benthosema pterotum

The distribution of eggs and early larval stages of the myctophid fish Benthosema pterotum was studied during three cruises to the Gulf of Oman during January to February 1981 and February 1983. The eggs and yolk sac larvae were identified and described after artificial fertilization on board the Research Vessel Dr. Fridtjof Nansen. The mature fish spawn in the Gulf at 300 to 100 m depths during early night. The eggs and smallest larvae were found at 300 to 200 m depths. The eggs hatch within 12 h at 21°C. Hatching occurs before the slightly bouyant eggs reach the upper 50 m of the water column.     

Regulation of respiration during juvenile development of Sepia officinalis (Mollusca: Cephalopoda)

A culture of juvenile Sepia officinalis L. was kept during summer 1985 in the aquaria of the Station Marine, Wimereux, France. During the first four months of juvenile development, oxygen consumption under increasing hypoxia was measured with a closed respirometer. The experiments revealed a high regulatory capacity of juvenile S. officinalis. The critical oxygen concentrations were calculated and their ontogenetical evolution was studied. The critical oxygen concentration increased with increasing development. A linear relationship emerged between the critical oxygen concentration and the logarithm of the wet weight [CO_c (mg O₂ l^-1)=-0.393+0.893×log₁₀(W _w )]. The decreasing regulatory capacity of growing S. officinalis is most probably related to adaptations to a changing ecological environment during development. Another possibility is a physiological change, most probably related to the shift from embryonic to adult hemocyanin.     

Reproductive biology of shovelnose guitarfish <Emphasis Type="Italic">Rhinobatos productus</Emphasis> from the eastern Gulf of California México

Guitarfishes are a primary component of artisanal elasmobranch fisheries and are commonly taken as trawl fishery bycatch throughout the Gulf of California. However, little is known of the life history of this species. To address this lack of critical biological information, the reproductive biology of Rhinobatos productus was investigated in the eastern Gulf of California. Development of claspers and testes indicated that males reach maturity at 53 cm total length (TL). Measurements of oviducal gland, largest ovum diameter, and uterus width indicated that females >57 cm TL are mature. This species possesses two functional ovaries: the ovarian cycle and gestation run concurrently. Histological analysis of oviducal glands did not provide evidence of sperm storage, but females carrying uterine capsules were observed over an extended period, suggesting the possibility of diapause in the early embryonic development. Following 4–5 months of embryonic growth, pups were typically born from late June to October after a gestation period of approximately 11–12 months. Width of yolk sac was inversely related to embryo length. The rate of reduction of yolk sac width suggests that embryos depend on the sac until birth. Mean fecundity was estimated to be 5 (range 1–10, s.d. = 2.24) with a 1:1 sex ratio. Average size at birth was 175 mm TL. Seasonally, gravid females enter shallow waters for parturition, becoming extremely vulnerable to gill nets used in the artisanal ray fishery.     

吡唑醚菌酯对不同阶段斑马鱼的毒性效应评价

为评价吡唑醚菌酯对鱼类不同生命阶段的毒性效应,以斑马鱼(Brachydanio rerio)为测试生物,测定了其对斑马鱼胚胎、仔鱼(孵化3 d和孵化12 d)和成鱼的急性毒性及对胚胎和仔鱼(孵化3 d)的发育毒性。结果表明,吡唑醚菌酯对斑马鱼胚胎、仔鱼(孵化3 d)、仔鱼(孵化12 d)和成鱼的96 h-LC50分别为0.0578(0.0540~0.0631)、0.0299(0.0282~0.0320)、0.0330(0.0310~0.0350)和0.0756(0.0692~0.0862)mg·L-1,其毒性大小依次为仔鱼(孵化3 d)仔鱼(孵化12 d)胚胎成鱼。与对照组相比,吡唑醚菌酯0.101 mg·L-1处理,可对斑马鱼胚胎24 h自主运动产生显著抑制作用;0.0609、0.0777和0.101 mg·L-1处理,可对斑马鱼胚胎48h心跳速率产生显著抑制作用;0.0609和0.0777 mg·L-1处理,可对斑马鱼胚胎96 h孵化仔鱼体长产生显著抑制作用。在供试浓度下吡唑醚菌酯暴露96 h后,均可对斑马鱼仔鱼(孵化3 d)体重和体长产生明显的抑制效应,对斑马鱼仔鱼(孵化3 d)心跳未产生明显影响。在高浓度下,吡唑醚菌酯可诱导斑马鱼胚胎产生畸形,主要包括心包水肿、卵黄囊水肿等;而对仔鱼(孵化3 d)未观察到明显的致畸效应。     

Yolk testosterone organizes behavior and male plumage coloration in house sparrows (<Emphasis Type="Italic">Passer domesticus</Emphasis>)

Vertebrate eggs contain maternal steroid hormones in their yolks; and in avian species the concentration of these steroids vary within and among clutches. The organizational actions of these variable doses of maternal steroids in the avian egg, analogous to those regulating the development of sexually dimorphic traits, are little explored. In this study, we examined the organizational effects of yolk testosterone in the house sparrow, Passer domesticus, a sexually dichromatic passerine bird in which males are characterized by the presence of a throat patch or badge of black feathers that varies in size among males. We injected a physiological dose of testosterone or vehicle into eggs in the field, removed nestlings, and hand-raised them in the laboratory. At 5 months of age, we investigated treatment effects on plumage and behavior. Males that hatched from testosterone-injected eggs developed a larger badge than males that hatched from vehicle-injected (control) eggs. However, testosterone did not induce the expression of a badge in females. In staged dyadic encounters, both males and females hatched from testosterone-injected eggs were more successful at obtaining and defending a food source than individuals of the same sex hatched from control eggs. The results suggest that variable concentrations of maternal testosterone in the eggs of the house sparrow organize the expression of a plumage trait in males and behavior in both sexes.Communicated by J. Graves     

Evaluation of developmental toxicity and apoptotic induction of the aqueous extract of Millettia pachycarpa using zebrafish as model organism

Extensive and indiscriminate use of synthetic compounds and natural compounds obtained from plant sources have resulted in serious threats to the aquatic ecosystem and human health. Aqueous extract of the root of the plant, Milletia pachycarpa Benth, is currently used for killing fish in the state of Manipur, India. Moreover, this plant is also used as traditional medicine in this region. Although it is widely used in traditional medicine, there is limited information available regarding the adverse effects and mechanism underlying its toxicity. This study examined the effects of exposure to aqueous extract of M. pachycarpa (AEMP) on early embryonic development of zebrafish embryos and mechanisms underlying toxicity. Zebrafish embryos treated with different concentrations of the AEMP produced embryonic lethality and developmental defects. The 96-hr-LC₅₀ of AEMP was found to be 4.276 µg/mL. Further, multiple developmental abnormalities such as pericardial edema, yolk sac edema, spinal curvature, swim bladder deflation, decreased heart rate, and delayed hatching were also observed in a dose-dependent manner. Zebrafish embryo showing moderate-to-severe developmental defects following AEMP exposure cannot swim properly. Further, this study examined oxidative stress and apoptosis in embryos exposed to AEMP. Enhanced production of ROS and apoptosis was found in brain, trunk, and tail of zebrafish embryos treated with AEMP. Data suggest that oxidative stress and apoptosis are associated with AEMP-induced embryonic lethality and developmental toxicity in zebrafish embryos.     

Changes in yolk total proteins and lipid components and embryonic growth rates during lobster (<Emphasis Type="Italic">Homarus americanus</Emphasis>) egg development under a simulated seasonal temperature cycle

From August 2000 to June 2001, seven egg-carrying female lobster (Homarus americanus) from the Îles de la Madeleine population (Gulf of St. Lawrence, Canada) were held under a simulated seasonal temperature cycle to monitor egg development from extrusion to hatching. For the first time, changes in the yolk components (total lipids and major lipid classes, total proteins) and embryo growth of single eggs were monitored separately over the entire development period. Under the controlled temperature conditions, egg development proceeded in three phases. (1) Autumn, from extrusion to early December, was marked by a rapid increase in the Perkinss eye index and rapid declines in yolk total proteins and triacylglycerols (TAG). Embryo daily growth rate was estimated between 1 and 2 µg proteins day^–1. (2) Winter, from late December to early April (temperature stable at ca. 1°C) was characterized by a stationary phase in the evolution of the eye index and yolk lipid use, and embryo growth slowed significantly. (3) Spring, from late April to hatching in June was the period with the most rapid changes in yolk TAG and embryo growth rates >6 µg proteins day^–1 were recorded. Almost 65% of the live biomass (total proteins) of the hatching larvae was accumulated during the last few weeks of development. An index of embryo growth efficiency was estimated as the slope of the relationship between embryo total proteins and yolk TAG during egg development. A relationship was found between the initial mean egg dry weight and the embryo growth efficiency index suggesting that under the same experimental conditions bigger eggs used yolk lipids more efficiently and sustained faster embryonic growth than smaller eggs. The relationship may also explain why larger larvae originate from larger eggs.Communicated by R.J.Thompson, St Johns     

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.     

Depth distribution of developing Euphausia superba embryos,predicted from sinking rates

Sinking rates of the embryos of Euphausia superba were measured during development in the laboratory at Palmer Station. Antarctic Peninsula, during austral summer 1982. Two densities of seawater were used in these experiments; the lower density (1.027 g cm^-3) is characteristic of the upper 300 m of the water column, the higher (1.029 g cm^-3) is characteristic of the deeper circumpolar deep water. During development, the eggs sank at an initial average rate of 175 m d^-1, declining to 51 m d^-1 during the gastrula to early limb bud stages and increasing before hatching to near initial rates. The larvae hatched within 6 d. We predict that the majority of the embryos of E. superba should hatch at a depth of about 850 m.     

Diapause embryos in the neustonic copepodAnomalocera patersoni

The neustonic copepodAnomalocera patersoni Templeton, collected from the Gulf of Naples from January to May 1988, laid subitaneous and diapause eggs that were of equal size and dark and smooth in appearance. In January and February, most eggs were subitaneous and hatched within 2 to 3 d at room temperature. Conspicuous numbers of diapause eggs were first obtained in February when several clutches contained a second egg type which did not hatch in a subitaneous manner. By March and April, most clutches contained only diapause eggs. Transmission electron microscope studies of subitaneous and diapause eggs revealed striking morphological differences. Subitaneous eggs had a thin vitelline coat covering the plasma membrane, whereas diapause eggs were enveloped by a complex four-layer structure which is assembled after spawning and which probably serves as a protective shell during dormancy. No major morphological differences were discernible in the oogenic cycle of females spawning either of the two egg types. Diapause eggs were shown to be embryos in early stages of development, and segmentation of these embryos was arrested for the entire 6-mo period of investigation. Diapause eggs hatched after 7 mo of dormancy.     

The importance of free amino acids to the energy metabolism of eggs and larvae of turbot (Scophthalmus maximus)

A study was undertaken to establish the role of free amino acids (FAA) in aerobic energy dissipation in embryos of turbot (Scophthalmus maximus) which contain an oil globule in the egg. Laboratory-reared developing eggs and larvae (15°C, 34 salinity) were measured for oxygen uptake, ammonia excretion, contents of FAA, protein, and ammonium, and volumes of yolksac and oil globule. Newly spawned eggs from different batches contained 55 to 90 nmol egg^–1 of FAA. Resorption of FAA occurred in parallel with the consumption of yolk. Resorption of the oil globule, however, occurred predominantly after hatching and mainly after yolk resorption. The combined data suggest that approximately 70% of the FAA are utilized as an energy substrate, while the rest are polymerized into body proteins. FAA become a significant energy substrate in the early egg stage and account for 100% of the aerobic energy dissipation 2 d after Fertilization then decrease to ca. 60% at the time of hatching. Lipids derived from the oil globule seem to be the main fuel after hatching and account for ca. 90% of the energy dissipation at the onset of first-feeding. Thus, the energetics of fish embryos which contain an oil globule seems to be different from those that depend exclusively on the nutritional reserves of the yolk.