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.     

The effects of supplemental dietary cholesterol on growth, development and survival of mud crab, Scylla serrata, megalopa fed semi-purified diets

The effects of varying levels of dietary cholesterol on growth, development time and survival of mud crab, Scylla serrata megalopa were investigated using semi-purified microbound diets (MBD). Five iso-energetic diets containing different level of cholesterol ranging from 0.14% to 1% of dry weight of the diet were tested. Fifteen megalopa were reared individually for each dietary treatment, and development time and survival were recorded on a daily basis. More than 25% of megalopa from all treatments were able to metamorphose into the first crab stage, suggesting that the endogenous level of cholesterol in the basal diet (0.14%) was sufficient to support development of the megalopa stage of this species. Widest mean carapace width (3.53 ± 0.08 mm) and highest mean dry weight (2.11 ± 0.22 mg) were recorded for juveniles that molted from megalopa fed live Artemia, whereas no megalopa in the unfed control treatment metamorphosed into crabs. The average development time from megalopa to the juvenile crab stage varied between the treatments, where megalopa fed live Artemia or MBD containing 0.2%, 0.4% or 0.8% total cholesterol showed the most synchronized molting (between 8.0 and 9.9 days). Longest development time was recorded for the megalopa fed diets containing 0.14% or 1% total cholesterol (both 11 days). Highest survival (74.3%) was recorded for the megalopa fed a diet containing 0.8% cholesterol. The results of this study are valuable in research to develop formulated diets for mud crab larvae as a replacement for live food in hatchery culture.     

Embryonic and larval development of black skirt tetra (Gymnocorymbus ternetzi,Boulenger, 1895) under laboratory conditions

The embryonic and larval development of black skirt tetra, Gymnocorymbus ternetzi, are described under controlled laboratory conditions. In addition, major histomorphological changes and the allometric growth patterns during larval development have been described. The laboratory‐reared broodstock, that is 1 year of age, were spawned. Hatching occurred 20–21 h after spawning at 24 ± 0.5°C. The cleavage was finished in 2 h and the early blastula stage occurred at 2:04 hours after spawning. The gastrulation started at 3:20 hours and 30% epiboly was observed at 3:34 hours after spawning. Eight‐somite stage was observed at 08:33 hours. And embryonic developmental stage was completed at 21 h after spawning. The newly hatched larvae were 1442 ± 14.3 μm in mean total length (TL). The mouth opened at 3 days after hatching (DAH). The yolk sac had been totally absorbed and the larvae started to swim actively within 3–4 days. Notochord flexion began at 11 DAH. The metamorphosis was completed and the larvae transformed into juveniles at 32 DAH. In this paper, the full developmental sequence from egg to juvenile of G. ternetzi is described for the first time.     

Effects of four natural diets on the culture performance and biochemical composition of megalopa of Eriocheir sinensis during desalination period

Frozen copepods, frozen adult Artemia, live rotifers and live Artemia nauplii were used as feed for the megalopa of Eriocheir sinensis during desalination stage to determine the most nutritious and cost‐effective feed. The biochemical composition of the four feeds was analysed. Then, the individual wet weight, individual dry weight, survival rate, the cost and the biochemical composition of the megalopa were measured after the desalination. The resulting effects on the metamorphosis ratio and the duration of the megalopa were also assessed. The results showed that the duration of desalinated megalopa to first stage crab for all treatments was approximately 3.5 days, while the metamorphosis rate of the copepod treatment was significantly higher than the other treatments with the value of 96.33 ± 3.79%. In terms of biochemical composition, after desalinization, the moisture content of megalopa in the adult Artemia treatment was the highest, and the total lipid in the Artemia nauplii treatment and rotifer treatment was significantly higher than the other two treatments. The desalinated megalopa from copepod treatment had the highest percentages of DHA (22:6n‐3) and EPA (20:5n‐3) among the four treatments. The desalinated megalopa from copepod treatment and rotifer treatment had the higher contents of protein. In conclusion, frozen copepods have higher nutritional value for the megalopa during the desalination period. Therefore, feeding the megalopa with copepods can improve the culture performance and megalopa quality, and can also reduce the feed cost during desalination stage.     

Effect of starvation during late megalopa stage of Mithraculus forceps (Brachyura: Majidae) on larval duration, synchronism of metamorphosis, survival to juvenile, and newly metamorphosed juvenile size

Larval resistance to temporary starvation is considered a key factor for successful development in the wild. Subjecting larvae to temporary starvation during early and/or late development is occasionally used in larviculture to reduce production costs.Mithraculus forceps is a popular species in the marine aquarium industry for their ability to control nuisance algae in aquarium tanks; a larval culture methodology was previously proposed in order to avoid collection from the wild. In an attempt to reduce production/feeding costs of M. forceps larval culture (two zoea stages and a megalopa), larvae (megalopa stage) were starved after 7 and 8 days post-hatch (DPH); starvation treatments were compared with those of the control treatment where larvae were fed continuously. No differences were found in survival to juvenile, metamorphosis synchronism or larval duration between the treatments, which suggest that at least by day 7 DPH, megalopae have already achieved the point-of-reserve-saturation (PRS) and are able to successfully metamorphose to crab stage without feeding (facultative lecithotrophic); newly metamorphosed juveniles are slightly smaller (particularly the ones starved after 7DPH). This flexible way of development might be very advantageous in the wild and might allow the reduction of costs of larval culture in captivity.     

Early life history of yellow puffer,Chonerhinos naritus (Richardson, 1848) from Sarawak,Northwestern Borneo

The growth and morphological development including fins, spine distribution and pigmentation of larval and juvenile of hatchery‐reared yellow puffer, Chonerhinos naritus were described to provide essential information on the early life history of this species. The total length (TL) of newly hatched larvae was 3.42 ± 0.23 (mean ± SD) mm, reaching 5.66 ± 0.38 mm on 5 days after hatched (DAH), 7.80 ± 0.28 mm on 11 DAH, 9.88 ± 0.40 mm on 27 DAH and 10.92 ± 0.58 mm on 30 DAH. The yolk was completely absorbed in preflexion larvae at 4 DAH. The mouth opening started at 3 DAH of yolk sac larvae, while the teeth appeared starting from preflexion larvae at 7 DAH. Overall aggregate fin ray numbers including caudal fin attained full complement in postflexion larvae at 27 DAH. Several melanophores with appearance of small stellate were first appeared dorsally on the head of flexion larvae at 13 DAH, expanded at the dorsal region of the head, above the eye in juveniles at 30 DAH. The spines first appeared in preflexion larvae of C. naritus at 7 DAH, covering the ventral skin region below pectoral fin base and expanded to the ventral part of the body and nearly covered the whole abdomen region before the anus and below the eyes in juveniles. C. naritus remain as larvae for approximately 29 days, during which they metamorphose to the juvenile stage prior to sexual maturation. Observations in larvae development of C. naritus revealed similar characteristics with other Tetraodontidae species.     

Co-feeding of live feed and inert diet from first-feeding affects Artemia lipid digestibility and retention in Senegalese sole (Solea senegalensis) larvae

The present study intended to evaluate the effects of early introduction of inert diet in lipid digestibility and metabolism of sole, while larval feed intake, growth and survival were also monitored. Solea senegalensis larvae were reared on a standard live feed regime (ST) and co-feeding regime with inert diet (Art R). Trials using sole larvae fed with Artemia enriched with two different lipid emulsions, containing glycerol tri [1-¹⁴C] oleate (TAG) and L-3-phosphatidylcholine-1,2-di-[1-¹⁴C] oleoyl (PL), were performed at 9 and 17 days after hatching (DAH) to study lipid utilization. Co-feeding did not affect sole survival rates (ST 59.1 ± 15.9%; Art R 69.56 ± 9.3%), but was reflected in significantly smaller final weight at 16 DAH (ST 0.71 ± 0.20; Art R 0.48 ± 0.14 mg). Higher feed intake was observed in sole larvae fed on Artemia enriched with labeled PL at 9 DAH but not at 17 DAH. At 17 DAH, the smaller larvae (Art R treatment) ingested proportionally more Artemia in weight percentage, independently of enrichment. At 9 DAH lipid digestibility was equal among treatments and higher than 90%, while at 17 DAH it was higher in ST treatment (around 73%) compared to the Art R group (around 66%). Lipid retention efficiency at 9 DAH was higher in the Art R treatment, reaching values of 50%, while these values almost duplicated at 17 DAH, ranging up to 80% in both treatments without significant differences. These results show that co-feeding of live feed and inert diet from first-feeding in Senegalese sole has a toll in terms of growth and lipid digestibility but does not seem to compromise lipid metabolic utilization.     

Effect of zooplankton as diet for the early paralarvae of Patagonian red octopus,Enteroctopus megalocyathus,grown under controlled environment

The limited success reported in the paralarval culture of merobenthic octopus is mainly attributed to nutritional problems. This study aimed to determine the effect of live diets on the paralarvae performance during the first 30 days after hatching (DAH). The paralarvae were grown under different treatments: starved (STV) as negative control or fed four diets: Artemia sp. enriched with microalgae (ANCH), Artemia enriched with a commercial enrichment (AOG), Artemia enriched with microalga + commercial enrichment (AMIX) and king crab zoea (ZKC). The paralarval growth and survival were affected by the dietary treatment, with significant higher growth in ANCH and AMIX. STV showed 100% mortality at day 27, whereas all diets resulted within a range from 20% to 33% survival. The endogenous protein and lipid reserves were utilized in all treatments. The 22:6n‐3/20:5n‐3 ratio increased up to 16% during the experimental period. Alkaline protease activity tended to increase in paralarvae fed ANCH, AOG and ZKC over the first 13 DAH; however, this effect was not observed in trypsin nor chymotrypsin activities. The leucine aminopeptidase activity (LAP) was not affected by the dietary treatment, while alkaline phosphatases increased only at 13 DAH in paralarvae fed ANCH. Indicators of the nutritional status of paralarvae are discussed.     

Development of hatchery technology for the silver pomfret Pampus argenteus (Euphrasen): effect of microalgal species on larval survival

The silver pomfret Pampus argenteus (Euphrasen) is a new candidate for aquaculture and there is not much information available on its larval rearing. Investigations carried out using microalgae alone in the culture system for the initial feeding of the silver pomfret larvae showed that Chlorella, Isochrysis and Nannochloropsis without rotifers are not conducive to the survival of newly hatched larvae. At 6 days after hatching (DAH), a maximum survival of 3% (1.8 ± 1.69%) was observed with Isochrysis followed by Nannochloropsis (0.35 ± 0.21%) and Chlorella (0.25 ± 0.21%). All control larvae died at 6 DAH without microalgae. Further investigations using the above microalgae with rotifers and a mixture of these same microalgae with rotifers showed that significantly higher (P < 0.05) survival could be achieved in the mixture of microalgae with rotifers in the culture system. At 12 DAH, the larval survival was 9.73 ± 1.39% in mixed species of algae compared with that of Isochrysis (6.93 ± 1.86%), Nannochloropsis (6.83 ± 0.61%), Chlorella (5.93 ± 2.76%) and seawater without microalgae or the control (0.73 ± 0.31%). The first incidence of feeding on rotifers at 4 DAH was significantly higher (P < 0.05) in all treatments with microalgae than that of the control. The incidence of feeding in mixed species of algae at 4 DAH (60.0 ± 0.00%) and in Isochrysis (55.0 ± 35.36%) was significantly higher (P < 0.05) than that of Chlorella (40.0 ± 0.00%) and the control (25.0 ± 7.07%). Prey consumption of individual larvae increased significantly (P < 0.01) at 8 DAH compared with that at 4 DAH. During this period, predation on rotifers by larvae was significantly higher (P < 0.05) in mixed species of algae (12.85 ± 5.73 rotifers larva^?1) than that of the control (6.75 ± 1.20 rotifers larva^?1). The fatty acid composition of rotifers used during this investigation shows that significantly higher (P < 0.05) ω3 HUFA was present in rotifers treated with mixed algae plus commercial enrichment media ‘Super Selco’ and ‘DHA Protein Selco’. Rearing of silver pomfret larvae up to the juvenile stage using mixed species of microalgae in the hatchery has been discussed. During 38 days of the larval rearing period, it was possible to achieve 3.6–4.2% larval survival with a mean of 3.9 ± 0.42%, which was considerably higher than in previous attempts (survival up to 1.5%).     

Influence of light intensity on feeding, growth, and early survival of leopard coral grouper (Plectropomus leopardus) larvae under mass-scale rearing conditions

This study investigated the effect of different light intensities on feeding, growth and survival of early stage leopard coral grouper Plectropomus leopardus larvae. Four different light intensities (0, 500, 1000 and 3000 lx) were used and larvae were kept under constant light conditions from 0 day after hatching (DAH) to 5 DAH. The larvae were fed a small S-type of Thai strain rotifers at a density of 20 individuals/mL from 2 DAH. The number of rotifers in larval digestive organ and total length of larvae were examined at 3 h intervals between 04:00 and 22:00 h on 3 DAH, and thereafter at 6 h intervals until the end of the experiment (5 DAH). Four experimental trials of the larval rearing were repeated using by 60 kL mass-scale rearing tanks. The results indicate that coral grouper larvae are visual feeders and their food intake increases with increasing light intensity. Food intake of larvae reared at 3000 lx was significantly higher than those reared at 0–1000 lx on 3 DAH despite being the first-feeding day (P < 0.01). On 4 DAH, total length of larvae reared at 3000 lx was significantly larger than those reared at the lower light intensities (0, 500 and 1000 lx), and thereafter light intensity significantly influenced larval feeding and growth until the end of the experiment. Survival on 5 DAH did not show a significant difference between light intensities, but survival rate at 3000 lx and 1000 lx had a tendency to be higher than those reared at the lower light intensities (0 and 500 lx). In contrast, larvae reared at 0 lx exhibited stagnant and/or negative growth. These results indicate that light intensity is significantly the factor affecting larval feeding, growth, and survival in coral grouper larvae under the rearing conditions.     

Effect of different dietary lipid levels on the reproduction of Rhamdia quelen (Quoy and Gaimard, 1824)

Lipids include some of the most important nutrients that affect the survival and growth of fishes in their early life stages. Lipid deficiency prior to spawning may significantly reduce egg production, hatchability and the number of surviving larvae. In this study, we investigated the effects of isocaloric diets containing 80, 140 and 200 g kg⁻¹ of lipids (lipid source, soy oil) for 90 days. The following data were collected at 0, 45 and 90 days: final weight; length; conditioning factor; hepatosomatic, gonadosomatic and visceral fat indices; total plasma protein; testosterone; 17ß‐estradiol; free amino acids; and ovary and muscle fatty acid profiles. Additionally, fecundity, oocyte production, egg morphometric parameters, and larval and postlarval growth were evaluated. Results were not statistically different for husbandry and biochemical parameters, but visceral fat content increased with increase in dietary lipid levels. The fatty acid profiles and composition differed among dietary treatments. The egg diameter and area were significantly low in fish fed the 200 g kg⁻¹ lipid diet, thereby hindering growth, survival and weight compared to those in postlarval fish (P < 0.05). The best reproductive rates were obtained when using diets containing 80–140 g kg⁻¹ total lipids.     

Ontogenic development of enzymatic activity and digestive system in Jullien's golden carp (Probarbus jullieni Sauvage, 1880)

Ontogenic development of the main enzymes and histological structure of digestive organs were studied in Jullien's golden carp (Probarbus jullieni) from hatching until 50 days after hatching (DAH). The larval fish were produced by artificial insemination and fed only Moina sp. till end of experiment. Body weight (mg) and total length (cm) of Jullien's golden carp increased exponentially and linearly. The results indicate the fish weight grew fast with increasing rate, while length increased at a constant rate over the studied period. Up‐regulation of acid protease was observed in newly hatched larvae and the specific activity gradually decreased with time. Trypsin specific activity was relatively stable within the first 35 DAH, while fluctuations in chymotrypsin were observed. Among these three proteolytic enzymes, acid proteases exihibited relatively high specific activity in newly hatched larvae, suggesting a role in yolk protein degradation. Alkaline proteases became more prominent with age and correlated with an abrupt decrease in acid proteases. Increased lipase‐specific activity appeared within 3 DAH and then gradually decreased with time, indicating the capacity to digest yolk lipid reserve. Amylase and cellulase‐specific activities changed in a similar manner, and the sensitivity to time was higher in amylase than in cellulase. The digestive organs and accessory organs developed around 3–5 DAH. However, intestinal histology was almost fully developed around 18 DAH. These findings should be useful for deciding the preferred timing for weaning, as well as on developing artificial diets referenced to the physiological changes of digestive enzymes.     

The effect of dietary n-3 HUFA levels and DHA/EPA ratios on growth, survival and osmotic stress tolerance of Chinese mitten crab Eriocheir sinensis larvae

The effect of varying levels of dietary n-3 highly unsaturated fatty acid (HUFA) and docosahexaenoic acid/eicosapentaenoic acid (DHA/EPA) ratios on growth, survival and osmotic stress tolerance of Eriocheir sinensis zoea larvae was studied in two separate experiments. In experiment I, larvae were fed rotifers and Artemia enriched with ICES emulsions with 0, 30 and 50% total n-3 HUFA levels but with the same DHA/EPA ratio of 0.6. In experiment II, larvae were fed different combinations of enriched rotifers and Artemia, in which, rotifers were enriched with emulsions containing 30% total n-3 HUFA, but different DHA/EPA ratio of 0.6, 2 and 4; while Artemia were enriched with the same emulsions, but DHA/EPA ratio of 0.6 and 4. In both experiments, un-enriched rotifers cultured on baker's yeast and newly-hatched Artemia nauplii were used as control diets. Larvae were fed rotifers at zoea 1 and zoea 2 stages; upon reaching zoea 3 stage, Artemia was introduced.Experiment I revealed no significant effect of prey enrichment on the survival of megalopa among treatments, but higher total n-3 HUFA levels significantly enhanced larval development (larval stage index, LSI) and resulted in higher individual dry body weight of megalopa. Furthermore higher dietary n-3 HUFA levels also resulted in better tolerance to salinity stress. Experiment II indicated that at the same total n-3 HUFA level, larvae continuously receiving a low dietary DHA/EPA ratio had significantly lower survival at the megalopa stage and inferior individual body weight at the megalopa stage, but no negative effect was observed on larval development (LSI). The ability to endure salinity stress of zoea 3, zoea 5 and megalopa fed diets with higher DHA/EPA ratio was also improved.     

Larval rearing of hilsa shad,Tenualosa ilisha (Hamilton 1822)

Hilsa, Tenualosa ilisha has received much attention for culture due to decline of the natural population. Lack of knowledge on larval rearing is the bottleneck for its culture. This study was aimed at developing larval rearing protocols for hilsa shad. Hilsa larvae (4 days old, 4.76 ± 0.06 mm/0.49 ± 0.01 mg) were stocked in fibreglass‐reinforced plastic tanks (1.7 m³ water volume) at 300, 600 and 1,200 nos/m³ in triplicates in three experimental systems viz., E‐I (circular, 0.567 m water depth), E‐II (circular, 0.962 m water depth) and E‐III (rectangular, 0.567 m water depth) and reared for 46 days. The larvae were supplied with Chlorella vulgaris, Brachionus calyciflorus, mixed phytoplankton and mixed zooplankton during 4–50, 6–25, 8–50 and 26–50 days of their age respectively. In each system, higher (p < 0.05) fry survival at 300 nos/m³ than in higher densities indicates density dependent stress. Circular tanks showed higher survival (13.3%–61.31%) than in rectangular tanks (6.88%–27.26%) in each stocking density, indicating the importance of tank shape for rearing. Water depth affected fry survival in circular tanks (E‐I and E‐II) at 300 nos/m³; at 0.962 m depth, survival was higher (61.31%, p < 0.05) than that of 0.567 m depth (49.93%). Good fry survival was achieved through feeding the larvae initially with Chlorella followed by co‐feeding with Brachionus, mixed phytoplankton and zooplankton and rearing in circular tanks at 300 nos/m³ densities at 1 m depth. This first‐ever larval rearing protocol is useful for mass production of fry to support hilsa aquaculture in future.     

Dietary amino acid profile affects muscle cellularity,growth, survival and ammonia excretion of meagre (Argyrosomus regius) larvae

Formulation of diets according to fish amino acid (AA) qualitative requirements may improve fish growth. Two diets with different AA profiles were tested on 16‐day‐old meagre during 20 days. The first diet had an AA profile closer to meagre larval AA requirements (BAL), whereas the second had a different AA profile (UNBAL). Results showed that larvae fed the diet with higher similarities to the AA profile of meagre carcass had a higher final weight (19.8 ± 0.5 mg versus 13.4 ± 1.8 mg) and lower ammonia excretion after being fed (0.6 ± 0.1 ml/(g DW. larvae. h) and 0.8 ± 0.1 ml/(g DW larvae. h)). A higher mean fibre area was obtained in 36 DAH larvae fed the BAL diet (224.8 ± 33.3 μm² versus 158.8 ± 34.5 μm²) as well as higher larval weight. When fibre area distribution was analysed, a higher frequency of larger fibres was observed in 36 DAH BAL larvae, suggesting that fibre hypertrophy had a higher importance in this treatment. In conclusion, this study shows that meagre larvae fed a diet with an AA profile closer to their requirements had lower nitrogen losses and higher final weight, as result of higher muscle hypertrophy.     

Supplementation of tryptophan and lysine in Diplodus sargus larval diet: effects on growth and skeletal deformities

Amino acids are the building blocks for growth and the major energy source during fish larval stages. Deficient amino acids can be supplemented in the diets, overcoming problems such as low growth rates and skeletal deformities. In this study, three experimental diets were tested: a balance diet supplemented with lysine, a balance diet supplemented with tryptophan and a control with no supplementation. Trials were conducted with Diplodus sargus larvae from 1 to 25 days after hatching (DAH). A microencapsulated diet was introduced at 15 DAH in co-feeding with live feed and from 20 DAH larvae were fed only this diet. The effect of the supplemented diets was assessed in terms of survival, growth rate, skeletal deformities, ammonia excretion and activity of amino acid catabolism enzymes. The results showed a similar survival in all treatments. However, larvae given tryptophan supplementation had a lower weight on 25 DAH. No significant differences were found in ammonia excretion, frequency or type of deformities or enzymatic activity. Tryptophan and lysine supplementation failed to improve larval growth, survival or larval quality.     

Early weaning of cuttlefish (Sepia officinalis,L.) with frozen grass shrimp (Palaemonetes varians) from the first day after hatching

The suitability of early weaning cuttlefish using frozen grass shrimp, and its effects on growth and mortality was determined in a 50‐day experiment. Three food groups were used. One group (C) was fed with live prey until 25th DAH and weaned with frozen prey until the end of the experiment. In the other two groups, weaning started at the fifth DAH (W5) and first DAH (W1), also with frozen diet. Significant differences were found between C and the remaining treatments (P < 0.05) for mean wet weight (MWW), instantaneous growth rate (IGR), biomass (B) and mortality at the 25th and the 50th DAH. Despite the lower growth (~1/3 of the achieved with live food) and higher mortality (3% obtained by C versus 23% in the W1 and W5 groups), C cuttlefish presented a similar weight gain tendency to W1 and W5 groups after weaning. This is the first record of early acceptance (at the first DAH) of frozen food by cuttlefish hatchlings. These differences could be related with digestive enzymes of cuttlefish and prey, changes in nutritional composition and amount of captured prey, etc. Future research should also address the effects of this early weaning on cuttlefish life cycle.     

Faster growth before metamorphosis leads to a higher risk of pseudoalbinism in juveniles of the starry flounder <Emphasis Type="Italic">Platichthys stellatus</Emphasis>, as suggested by otolith back-calculation

The mechanism underlying malformation during flatfish metamorphosis is not clear. Here, the relationship between growth before metamorphosis and the occurrence of malformation after metamorphosis was examined in individuals reared in a single tank. Otoliths of starry flounder Platichthys stellatus larvae were labeled with alizarin complexone before metamorphosis [12 days after hatching (DAH)]. The positive linear regression between the lapillus otolith diameter and body length at 12 DAH enabled the estimation of individual larval size after completion of metamorphosis. At 12 DAH, the average body length of the typical pseudoalbino juveniles was significantly greater (5.37 mm) than that of the normal juveniles (5.13 mm) (P < 0.01, n = 100). In addition, of the larvae that were estimated to measure less than 4.8 mm at 12 DAH, approximately 80% metamorphosed into normal juveniles, whereas more than 60% of the larvae that measured more than 5.4 mm became pseudoalbino juveniles. These results indicate that the larvae that grow rapidly during the first 12 days are more likely to become pseudoalbino individuals after metamorphosis.     

Promotion of initial swim bladder inflation and effect of initial inflation failure on skeletal malformation in cultured seven‐band grouper Hyporthodus septemfasciatus (Thunberg)

The effects of the removal of the oily surface film on initial swim bladder inflation and survival in cultured seven‐band grouper Hyporthodus septemfasciatus were investigated during the larval stage. Additionally, the relationship between swim bladder inflation failure and malformations in later stages was examined. To explore promotion conditions for inflation, duplicate experiments were conducted under three different water surface conditions: removal of the oily surface film (ROF group), covering the water surface with an oil film (COF group), and sealing the water surface with liquid paraffin (SLP group). Larval swim bladder inflation was first observed 14 days after hatching (DAH) in the ROF group; thereafter, swim bladder inflation rates increased to 11.1% and 38.7% until the end of the experiments. This indicates that initial swim bladder inflation occurs at the pre‐flexion stage in larval development. Conversely, bladder inflation was rarely observed in the COF or SLP groups, indicating the promotional effect of surface film removal on larval swim bladder inflation and suggesting that gulping atmospheric air is necessary for initial inflation. Although larval water surface death occurred frequently in the ROF group, no significant difference existed in any group between the survival rates at 10 DAH and at the end of the experiment. The incidence rate of spinal curvature and deformation of the centra was significantly higher in individuals without initial swim bladder inflation, and these malformations mainly occurred in the caudal vertebra. Therefore, the removal of the oily surface film promotes initial swim bladder inflation and reduces larval malformations.     

Lack of essential fatty acids in live feed during larval and post-larval rearing: effect on the performance of juvenile <Emphasis Type="Italic">Solea senegalensis</Emphasis>

Despite the large progress obtained in recent years, Senegalese sole (Solea senegalensis) production of high quality juveniles is still a bottleneck. This paper examines the effect of larval and post-larval lipid nutrition on juvenile performance and quality. Four dietary treatments were tested: A—enriched Artemia spp. (EA); B—non-enriched Artemia spp. (NEA); C—EA during the pelagic larval period and NEA after larval settlement; D—50% EA and 50% NEA. Juvenile fatty acid profile at 60 days after hatching (DAH) clearly reflected the larval and post-larval diet composition. Feeding sole larvae on NEA (poor in lipids and essential fatty acids-EFA) had a negative effect, reducing growth (total length and dry weight) after 30 DAH and decreasing digestive enzyme activity at the end of the rearing period (60 DAH). However, relatively good performance compared to the EFA-richest treatment (A) was obtained when larvae were fed 50% EA and 50% NEA (D) or even EA only during the pelagic larval period followed by NEA after larval settlement (C). Malpigmentation was not affected by the dietary regimes and its incidence was very low. However, skeletal deformities were prevalent, particularly in the caudal complex, independently of diet. The results confirm that Senegalese sole appear to have lower larval EFA requirements than most cultured marine species and potentially even lower requirements during the post-larval stage. The importance of studying the impact of early nutrition on later juvenile stages was clearly highlighted in this study.