Development of digestive system and swim bladder of larval nase (Chondrostoma nasus L.)

Digestive tract and swim bladder differentiation during nase (Chondrostoma nasus L.) larval development was evaluated using histological methods. The mouth, pharynx and oesophagus were impervious at the time of hatching. The straight tubular intestine consisted of undifferentiated cells and was situated over the yolk sac. The intestine, lined with a single layer of columnar epithelial cells, opened on 2 day posthatching (dph). First evidence of protein digestion and absorption by the enterocytes was observed on the sixth day, while lipid digestion and absorption began on the ninth day of larval development. Intestinal mucous cells developed and became active by the fourth day. The posterior chamber of the swim bladder inflated on the third day, while the anterior one filled 12 dph. Complete yolk sac resorption took place on the ninth day.     

Embryonic and larval development of honeycomb grouper Epinephelus merra Bloch

Spawning and successful rearing of larvae of honeycomb grouper Epinephelus merra Bloch 1793 upto juvenile stage was accomplished at the finfish hatchery of Mandapam Regional Centre of Central Marine Fisheries Research Institute during 2004. The fertilized eggs were free, spherical and buoyant with size ranging from 710 to 730 μm. Complete early embryonic development took place within 24–27 h and hatching occurred. The hatchlings measured 1.5 mm. Mouth opening (115 μm) appeared at 72 h when the larvae were 2.2 mm in size. Pectoral fin developed on the fifth day. Complete metamorphosis took place and by the 60th day the larvae transformed into juveniles (45 mm) and attained skin colouration and honeycomb pattern.     

Effect of incubation temperature on the embryonic development and yolk‐sac larvae of the Pacific red snapper Lutjanus peru (Nichols & Murphy, 1922)

The effect of incubation temperature on embryonic development and yolk‐sac larva of the Pacific red snapper Lutjanus peru were evaluated by testing the effect of 26, 28 and 30°C, as this is the natural thermal interval reported during the spawning season of Pacific red snapper in the Gulf of California, Mexico. Sixteen developmental stages were observed. The incubation temperature affected the rate of development and time to hatching, being shorter at 30 than at 26°C, but no significant effect (P < 0.05) on larval length at hatching was registered. The depletion rate of yolk sac and oil globule was affected by incubation temperature particularly during the first 12 h post hatching (hph). At the end of the experiment (48 hph), significantly (P < 0.05) larger larvae were recorded at 26°C (TL = 3.22 ± 0.01 mm) than at 28° (TL = 3.01 ± 0.02 mm) and 30°C (TL = 2.97 ± 0.05 mm). Incubation of newly fertilized eggs at 26°C produces larger larvae, which may help to improve feeding efficiency and survival during first feeding.     

Embryonic and larval development of serpae tetra Hyphessobrycon eques (Steindachner, 1882)

In this study, the embryonic and larval development stages of one of the most important ornamental fish serpae tetra (Hyphessobrycon eques) are described. The early life stage is documented from fertilization until the beginning of the juvenile period. The fertilized eggs (the average diameter = 938.55 ± 35.20 µm) were incubated at a water temperature of 26 ± 0.5°C. The cleavage finished in 1:10 hr (=h) and the early blastula stage occurred at 1:26 hr post fertilization (hpf). The gastrulation started at 3:05 hpf, and 50% epiboly was observed at 3:25 hpf. Segmentation stage was monitored at 7:26 hpf. Embryonic developmental stage was completed and hatching occurred 20–21 hpf. The total length (TL) of newly hatched larvae was 2.64 ± 0.21 mm. The larval development of serpae tetra was divided into four different periods: Yolk‐sac larva (1–4 DAH, TL = 2.77 ± 0.09 mm ‐ 3.85 ± 0.11 mm), preflexion larva (5–12 DAH), flexion larva (13–15 DAH, TL = 5.78 ± 0.46 mm on the 15th day) and post‐flexion larva (16–30 DAH, TL = 10.7 ± 0.27 mm on the 28th–30th days). The mouth and anus are closed at 1 DAH. The mouth and anus opened at 4 DAH. Exogenous feeding started on the 4th day. The first gulping of the swim bladder was on days 3. The larva begins to swim freely, and the yolk sac was completely consumed at 4 DAH. Histological structures of the eye and brain of new hatched larva were clearly identified at 1 day after hatching (DAH). According to histological findings, the digestive system (stomach, intestine) started to develop and the liver could be seen on the ventral side of the swim bladder at 5 DAH. No histological difference was observed between the anterior intestine and the posterior intestine at 15–16 DAH. The larval metamorphosis was completed, and the larvae transformed into juveniles at 28–30 DAH.     

First results of embryonic development,spawning and larval rearing of the Mediterranean spider crab Maja squinado (Herbst ) under laboratory conditions,a candidate species for a restocking program

The Mediterranean spider crab, Maja squinado, is depleted due to overfishing. The crab has virtually disappeared from areas where it was abundant, such as the Balearic Islands and the Catalan coast. Maja squinado, is economically and ecologically very valuable, and it is essential to obtain information on its biology and rearing conditions to attempt to repopulate the damaged stocks of the species in the Mediterranean basin. Herein, we describe the first successful rearing of M. squinado under laboratory conditions. Our results show that M. squinado is an excellent candidate for restocking using cultured juveniles. Two consecutive broods with a 1–4 day interbrood period were observed in the laboratory in wild‐caught females, the maximum observed duration of embryonic development of the egg mass being 32 days at 18.4 ± 0.9°C, and went through four different stages. The complete larval and first juvenile development was studied in laboratory cultures fed enriched Artemia metanauplius. At 19.6 ± 0.6°C, development from hatching to first crab moult took 17 days, and it comprised two zoeae stages and one megalopa stage. The survival rate at the different stages was monitored, and 7.13 ± 2.3% was achieved at the first crab instar.     

The Larval Development,Metamorphosis and Juvenile Growth of the Turban Snail Lithopoma (Astraea) undosa (Wood, 1828) (Gastropoda:Turbinidae)

This work presents the embryonic, larval, and postlarval development of Lithopoma undosa (L. undosa) (20 ± 2 C and the effect of natural inducers on the larval metamorphosis. Embryonic development until hatching of the trochophore larva took 15 h. Early veliger larvae with well‐developed shell were after 24 h. At 48 h, the operculum was completely formed. Cephalic tentacles were visible after 72 h. Larvae reached competence 6 to 7 d after fertilization, and was characterized by the presence of branched cephalic tentacles. The effect of natural inducers was evaluated. The percentage of metamorphosis with diatoms and adult mucus was 94% and 58% respectively, differing from the 25% without inducer. When was repeated, the metamorphosis was 100%, and 16% with benthic diatoms. The initial mean size of the postlarvae was 0.316 ± 0.02 mm. After 264 d the juveniles had attained a mean size of 5.98 ± 0.16 mm. These results will not only be of use in the evaluation of the culture of L. undosa, but also in studies on the characterization of inducers.     

Larval development in European hake (Merluccius merluccius L.) reared in a semi-intensive culture system

Eggs of European hake (Merluccius merluccius L.) were stripped from fish caught at sea. Larvae were kept under semi‐intensive conditions at around 12°C. In addition, eggs were incubated in single wells at 9.2, 12.7 and 14.5°C, where hatching, development and survival were closely examined. During the larval stage, a total of 299 larvae were sampled to follow development and growth. In addition a small number of juveniles were sampled. Larvae hatched approximately 4 days after fertilization, and were 2.9 mm in total length (TL). At 6‐day post hatching (dph), the larvae were 4.1 mm (TL), the jaw apparatus was developed, and the larvae had started to feed. Most of the growth during the early larval period is restricted to the head, and there is almost no increase in length for the first 3–4 weeks post hatching. Teeth and pelvic fins appear at 25 dph. Development of unpaired fins at approximately 30 dph marks the start of the larval–juvenile transition. Weaning to formulated feed was accomplished 50 dph, when external morphology was similar to that of adult hake.     

Breeding and larval rearing of the camouflage grouper Epinephelus polyphekadion (Bleeker) in the hypersaline waters of the Red Sea coast of Saudi Arabia

This is the first report on the successful year-round natural spawning and larval rearing of Epinephelus polyphekadion (Bleeker) in captivity and under hypersaline water conditions of 42-43%₀ salinity in the Red Sea coast of Saudi Arabia. Although the fish spawned naturally once or twice a year during 1992-94 culture period, incorporation of cod-liver oil in the broodstock diet during the 1995 culture period enabled the fish to spawn continuously for 2-3 days in each month during March, April, May and August. The egg fertilization and hatching rates also increased during the 1995 spawning period. The egg fertilization rate varied from 90 to 100% with a mean of 96.5 ± 3.38%. The egg hatching rate varied from 70 to 95% with a mean of 83.1 ± 10.12%. The fertilized egg diameter averaged 757.3 ± 37.36 μm. There was a linear relation between the fertilized egg size and the egg hatching rate. The increase in the hatching rate relevant to the egg size was statistically significant (P < 0.01). The egg development time until hatching lasted for 19 h at 29°C. The newly hatched larval size ranged from 1.55-1.71 mm with a mean of 1.65 ± 0.052 mm in total length. The larval growth was slow in the early stages and the growth curve until metamorphosis showed a curvilinear pattern. Wide variations in larval size, range 22-47 mm with a mean of 33.40 ± 7.01 mm, were observed during the metamorphosis stage at day 50. No significant difference (P > 0.05) in growth and survival was observed between the larvae reared using white and grey coloured tanks. The larval survival up to metamorphosis was 1.6-4.7% with a mean of 2.98 ± 1.56% in the grey coloured tanks and 1.6-1.9% with a mean of 1.73 ± 0.16% in the white tanks. The results demonstrated the possibility of breeding E. polyphekadion under captive culture conditions. However, methods to improve the larval survival have to be pursued further for commercial farming of this species.     

Effects of embryonic exposure to α‐lipoic acid or ascorbic acid on hatching rate and development of zebrafish (Danio rerio)

This study investigates the effects of embryonic exposure to two different antioxidants on growth and development in fish. Zebrafish (Danio rerio) embryos (100 per group) were exposed to lipoic acid (LA, 6–12 μM) or ascorbic acid (AA, 100–200 μM) and the hatching rate, standard lengths (SL) at hatching, development and growth post‐hatching monitored. The SLs at hatching were increased (P<0.05) in both antioxidant‐exposed groups relative to the controls, with no effect on yolk reserves. This enhanced development persisted up to 15 days post hatching. At hatching, cell proliferation rates (P<0.0005) and basic fibroblast growth factor (P<0.001), were greater in the antioxidant‐exposed fish than in the controls (0 μM antioxidant); no oxidative DNA damage was detected (P>0.05). Activity of the endogenous antioxidant enzyme superoxide dismutase was greater (P<0.001) in LA‐treated fish than in the controls. The results suggest that embryonic treatment of zebrafish with LA or AA during embryogenesis enhanced cell proliferation, leading to increased somatic growth in the larval stages, persisting into the juvenile stage. The findings support the treatment of embryonic fish with antioxidants for enhanced results in aquaculture.     

Morphological development and growth patterns of the leopard grouper Mycteroperca rosacea during larval development

At 25 °C, metamorphosis in leopard grouper Mycteroperca rosacea larvae took 60 days after hatching. The total length at day 1 was 1.95±0.22 mm and juveniles reached a length of 30.64±0.23 mm at day 60; the increase was approximately linear. We describe eight stages of development during this period. Larvae with the yolk sac attached occur from days 1 to 4 (Stages 1 and 2). The preflexion occurs on days 5–20 (Stages 3 and 4). Bending notochord occurred at day 25 (Stage 5). The other morphological changes that precede the juvenile phase occurred progressively until day 60 (Stages 6–8). Allometric growth in the height and length of the head, trunk length, height and length of the tail and the diameter of the eye compared with the total length showed two distinct stages of growth. Inflexion point, where growth is positive, occurred when larvae reached between 18.75 and 21.59 mm, which corresponds to larvae at days 35–40.     

Studies on the broodstock production and larval rearing of Coral demoiselle Neopomacentrus nemurus (Bleeker 1857)

Steps taken for broodstock development of Neopomacentrus nemurus, Coral demoiselle (Bleeker, 1857) in a hatchery, its spawning and development of egg to the juvenile stage are described. Among the three treatments tried, only the trial using Parvocalanus crassirostris nauplii as the first diet helped in the development of larvae. Larval and post‐larval growth was studied for a period of 65 days post hatch. The larva measured 2–2.4 mm in total length at the time of hatching. The mouth size of larva at the time of hatching measured 237.92 µm. By the 5th day, the width of the larval body had significantly increased. All the fins were conjoined, except for the caudal fin which had begun its appearance on the 2nd day itself. The digestive system was functional by the 5th day and the copepod remains were seen in the digestive tract. The larva metamorphosed by the 15th day with fully developed fins and fin rays. On the 20th day, the larval body had begun to display signs of fin pigmentation. The larva assumed adult pigmentation by the 35th day. By the 65th day, the larva grew to a total length of about 27 mm and weighed about 0.56 g.     

雅鲁藏布江尖裸鲤胚胎和仔稚鱼发育的研究

尖裸鲤(Oxygymnocypris stewartii)成熟卵呈圆形,黄色,沉性,卵径(2.57±0.07)mm,受精卵吸水充分膨胀后卵径为(3.22±0.07)mm。水温9.5～11.8℃,尖裸鲤胚胎发育历时265h,根据胚胎发育的外部形态及典型特征,将其分为受精卵、胚盘形成、卵裂、囊胚、原肠、神经胚、器官分化和孵化共8个阶段、32个时期。出膜仔鱼全长(10.27±0.15)mm,出膜20d后卵黄耗尽,全长(15.22±0.27)mm。出膜后80d,鱼苗全长(23.65±0.59)mm,臀鳞出现,各鳍均已出现,鱼苗侧线全部形成且清晰,体色和体型接近成鱼。尖裸鲤早期发育的特点反映了其对雅鲁藏布江水域的适应性。报道了尖裸鲤胚胎发育和仔稚鱼发育的生物学特性,为雅鲁藏布江特有鱼类资源保护利用提供科学依据和理论基础。     

Is batch variability in hatching time related to size heterogeneity and cannibalism in pikeperch (Sander lucioperca)?

Larval size heterogeneity is known to induce cannibalism, and procedures to avoid larval size differences are consequently implemented already during egg incubation and hatching. We investigated the relation between larval development variability, size heterogeneity and cannibalism in pikeperch. Larvae were sorted into five groups according to the time of hatching during a 65‐h period. The larvae with different times of hatch were then reared separately or together during an 18‐day period. Late hatched larvae were longer (P=0.003) and had less yolk remaining (P<0.001) than early hatched individuals at the time of hatching. However, on 11 days post fertilization, the late‐hatching larvae tended to have larger yolk reserves than earlier hatched individuals (P=0.07). Furthermore, the next day, a lower proportion in the late fraction had switched to exogenous feeding (P=0.024). That larvae with a late hatching time developed slower suggests a positive relationship between the hatching time and the embryonic developmental rate. However, differences in the length and available yolk reserves at hatching between larval fractions with different hatching times suggest that hatching is not strictly associated with a specific developmental stage, and that factors other than the development rate of the embryo are involved in the timing of hatching.     

Mesocosm: A Reliable Technology for Larval Rearing of Diplodus puntazzo and Diplodus sargus sargus

Species diversification is today considered as a major issue for the sustainable development of the Mediterranean aquaculture. For successful propagation of any species however, larval rearing is considered a bottleneck and therefore the development of appropriate tools is essential. Mesocosm is a semi-intensive technology that facilitates larval rearing of several species integrating principles of both intensive and extensive aquaculture, which solves biological problems and many of their technical, human and economical consequences. The extensive (and now even the semi-extensive) strategy is used in the most critical segments of the rearing process during the early developmental stages, when larvae are still extremely weak, sensitive to intensive environment, easily stressed and difficult to feed. The intensive strategy is used as soon as larvae are considered mature enough to be reared easily using classical methods. The technology was used for the rearing of two species, with potential for aquaculture, the sharpsnout seabream (Diplodus puntazzo) and the white seabream (Diplodus sargus sargus). Three groups of each species were monitored for a period of 50–70 days post hatching. Survival for both species was about 54% at the end of the trials. Sharpsnout seabream larvae reached 19.6 ± 0.9 mm total length and 107.2 ± 31.9 mg body weight 50 days post hatching. White seabream larvae 60 days post hatching reached 32.7 ± 2.7 mm total length and 450 ± 70 mg body weight. In order to verify the economical viability of the technology, the individual production cost for each species was estimated and reached ¢0.027 for white seabream and ¢0.043 for sharpsnout seabream. Results indicate the reliability of the technology for the larval rearing of the two species.     

Post‐hatching development of the ornamental ‘Red Cherry Shrimp’ Neocaridina davidi (Bouvier, 1904) (Crustacea,Caridea, Atyidae) under laboratorial conditions

The ‘Red Cherry shrimp’, Neocaridina davidi is a small freshwater caridean shrimp living, originally, in various kinds of inland water bodies around Asian countries. This shrimp has reached several countries for ornamental use; however, basic information on the biology of the species is still scarce in the literature. Its early post embryonic development morphology has not yet been described. This paper focused on the production and the development of early post‐hatching stages of N. davidi, its male secondary sexual features, observation of the gonads and the presence of females with embryos. The larval development of N. davidi was almost suppressed as noted by the presence of relatively large‐sized eggs, first stage hatching as a decapodid; and the tail fans were present only from the 2nd post‐hatching stage. A biological important consequence of the presence of this Red Cherry shrimp species in the Neotropics is its potential release into nature, which could cause its rapid dispersion affecting populations of other indigenous caridean freshwater shrimps.     

Feeding rhythm and growth of the tongue sole, Cynoglossus semilaevis Günther, during its early life stages

The feeding rhythm and growth characteristics of the early life stages of the tongue sole, Cynoglossus semilaevis Günther, were studied. Larvae began to prey on rotifers about 2–3 days after hatching at 23°C. About 12 days after hatching, larvae had grown to 8–9 mm in total length and were able to prey on Artemia larvae. As the larvae grew, they showed an increasing feeding capacity and a distinct feeding rhythm. Feeding intensity for day‐6 larvae was highest at 12:00 and 18:00 hours, about 2–3 h after the maximum feeding incidence. The highest levels of feeding intensity for day‐16 larvae occurred between 09:00 and 18:00 hours. By day 26, when the larvae had metamorphosed, feeding capacity had again increased considerably and, in contrast to the earlier stages, feeding intensity peaked at 18:00 and 24:00 hours concurrently with feeding incidence. Thus, tongue sole were found to have different feeding rhythms in the pre‐ and post‐metamorphosis stages, with the highest feeding activity in the daytime during the larval planktonic stage, and at night during the juvenile benthic stage. The estimated maximum daily feeding rates were 65%, 40% and 11% of body weight on days 6, 16 (larvae) and 26 (juveniles) respectively. Size variation increased markedly with development.     

Effects of different step‐wise temperature increment regimes during egg incubation of Atlantic cod (Gadus morhua L.) on egg viability and newly hatched larval quality

We carried out an experiment to determine how rapidly the early incubation temperature of Atlantic cod eggs can be increased without affecting normal embryonic development and hatching. Atlantic cod eggs were incubated at a constant low temperature (4.5 ± 0.5°C; T5 – control) and four temperature increment treatments where the temperatures were increased stepwise from 4.5°C at zygote stage to 9.5 ± 05°C (T1‐8 h, T2‐32 h, T3‐64 h and T4‐96 h). Embryonic cell symmetry, embryonic mortality, hatching success and larval skeletal abnormalities, length and yolk sac volume were recorded. Larval samples were also taken at hatch for histological analysis. Except for higher egg mortality and lower hatching success in the T1, the differences among experimental groups were minor. Cell asymmetries and embryo mortalities were not significantly different between the control and T2–T4 treatment groups. Control larvae were significantly longer and had smaller yolk reserves at hatch than T1–T4 larvae and larvae from T2 had the largest yolk reserves. Tissue and organ histology of hatched larvae were similar. Considering embryonic cleavage pattern, hatching success and larval morphology and histology, a gradual increment of temperature in 32 h seems to be the better choice for future developmental programming studies in Atlantic cod.     

Embryonic and larval development in the semelparous Nereid polychaete Hediste diversicolor (OF Müller, 1776) in Norway: Challenges and perspectives

Knowledge of broodstock manipulation, gametogenesis, artificial fertilization and larval nutrition is a prerequisite to reach a large‐scale production of the polychaete Hediste diversicolor. In this study, the characteristics of oogenesis and spermatogenesis and embryonic and larval development were observed. Moreover, the effect of temperature on embryonic development and effect of diets and sediment on earlier juvenile development were tested. Mature oocytes were characterized by a spherical shape with a diameter of about 200 μm. They showed radial symmetry with a large centric nucleus surrounded by four distinctive layers. Embryo cleavage started between 4 and 10 hr after fertilization, and developed to the swimming trochophore stage after 5–6 days after fertilization at 11°C. The rate of embryonic development increased with a temperature increase from 6.1 to 21.2°C. The trochophore larvae increased in length from day 9 and commenced differentiation into 3‐setiger stage larvae at day 12. The length growth of larvae until 7‐setiger stage was 0.43 mm/day when fed with fish feed, while those fed shellfish diet and smolt sludge both grew around 0.21–0.23 mm/day. The results suggest that fish feed is a superior diet compared to shellfish diets and smolt sludge for the early larval stage of H. diversicolor.     

The effects of cadmium and copper on embryonic and larval development of ide Leuciscus idus L.

The effects of Cd and Cu on embryos and larvae of the ide Leuciscus idus were evaluated. The ide is an European cyprinid fish, natural populations of which tend to decrease. The ide is also used as a bioindicator organism to evaluate water quality. However, sensitivity of ide early developmental stages to heavy metal intoxication is not known. Fish were exposed to Cd or Cu (100 μg/L) during embryonic, larval or both developmental periods. Survival of the embryos, time of hatching, size and quality of newly hatched larvae were evaluated at the end of embryonic period. Correctly developed larvae from the control and Cd or Cu-exposed groups were transferred to clean water, Cd or Cu solutions (100 μg/L) immediately after hatching. Larval development was observed, and the larvae were photographed. Time of yolk sac resorption, onset of active feeding and swim bladder inflation were evaluated, and the measurements were done on body and swim bladder size. The results showed that exposure of embryos to Cd and Cu significantly reduced embryonic survival and increased frequency of body malformations and death in newly hatched larvae and delayed hatching. Exposure to Cd and Cu during larval period reduced larval survival, growth and delayed development (yolk utilization, beginning of active feeding and swim bladder inflation). Cadmium was more toxic to the ide embryos and larvae than copper. Exposures to metals during embryonic period alone caused adverse impact on larval performance even when larval development took place in clean water. However, exposure of embryos to Cu reduced toxic impact of metal on larvae in continuous Cu exposure compared to the non-preexposed fish, but no such an effect occurred in case of Cd exposure. The results show that even a short-term exposure to Cd or Cu during early development of ide may adversely affect recruitment of this species. Among the measured endpoints, quality of newly hatched larvae (frequency of body malformations and larvae dead immediately after hatching) and swim bladder size were the most sensitive to intoxication with both metals. Embryos were more sensitive to Cu intoxication than larvae, while in case of Cd, sensitivity of both stages was similar.