The impact of nutrition on metamorphosis in Atlantic halibut (Hippoglossus hippoglossus L.)

Fatty acids, vitamin A and thyroid hormone have all been shown to affect development of flatfish larvae and they are ligands to nuclear receptors that participate in the control of development. Our hypothesis was that one of these factors or an interaction between them may be the cause of abnormal development of flatfish larvae. Atlantic halibut larvae were fed either DHA-selco-enriched Artemia or copepods from first feeding. In fish that had been fed Artemia, only 7% had normal pigmentation and 10% normal eye migration. The numbers for fish fed copepods were 68% and 88%, respectively. Malpigmented fish fed Artemia were depigmented, while those fed copepods had ambicoloration. The differences in development were probably nutrient dependent, since all other conditions were similar for the two groups. Larvae fed copepods had markedly higher body levels of docosahexanoic acid (DHA, 22:6n−3) and eicosapentaenoic acid (EPA, 20:5n−3) and lower levels of arachidonic acid (ARA, 20:4 n−6) than larvae fed Artemia. The DHA/EPA ratio was similar in the two groups, but the EPA/ARA ratio was more than four times higher in larvae fed copepods than in larvae fed Artemia. Larvae fed copepods had higher body levels of total retinol than larvae fed Artemia, but the difference was due to higher levels of the storage forms, retinyl esters, whereas the levels of free retinol and retinal were similar in the two groups. The level of iodine was 700 times higher in copepods than in Artemia and 3–4 times higher in larvae fed copepods than in larvae fed Artemia. There was a significantly higher level of T₄ in larvae fed copepods during the “window of opportunity”, 15–30 days after first feeding. In an experiment where Atlantic halibut larvae were fed Artemia enriched in iodine up to the levels found in copepods, there was a significant effect on the body level of iodine and a non-significant tendency of higher levels of thyroid hormone, but no effect on pigmentation or eye migration. It is concluded that Artemia probably offers a sufficient access to vitamin A precursors to meet the larval requirement. More research should be done to elucidate possible effects of iodine on development of Atlantic halibut larvae. Fatty acid composition is still the most likely candidate for causing abnormal development in Atlantic halibut larvae.     

A successful microbound diet for the larval culture of freshwater prawn Macrobrachium rosenbergii

A high moisture (63–71%), semi-purified microbound diet containing alginate was compared to newly hatched live Artemia nauplii as an exclusive diet for the culture of larval freshwater prawn Macrobrachium rosenbergii from 5th stage (weighted mean) through metamorphosis to postlarva. Two separate trials, representing larvae from different hatches, were conducted. Larvae were stocked at 50/l into cone-shaped vessels that contained 2 l of 12‰ seawater and were part of a temperature-controlled (28 °C) recirculating culture system. Larvae were manually fed either the live Artemia diet or the microbound diet exclusively, several times daily. After 14 days (23 days post-hatch (dph)), growth of larvae fed the microbound diet was 90% of that achieved for larvae fed newly hatched nauplii of Artemia. Survival of larvae fed the microbound diet was 77.3% and 73.3%, and was not significantly different from that of Artemia-fed larvae. Composed of readily available ingredients, the diet contains 46.2% crude protein and 37.4% lipid, is easy to prepare, and has good water stability. The diet is an economically practical alternative to the fluctuating cost, nutrient uncertainty, and labor associated with the use of Artemia nauplii hatched from cysts. The characteristics of the diet suggest good potential for successful use in the larviculture of other fish and crustacean species, in either the existing or a modified state.     

Artemia enriched with high n-3 HUFA may give a large improvement in performance of Atlantic halibut (Hippoglossus hippoglossus L.) larvae

Atlantic halibut larvae were fed Artemia enriched with two different oil emulsions (cod liver oil and 2050TG) from first feeding to 70 days after first-feeding (dpff). Larvae fed 2050TG enriched Artemia had better growth, survival and eye migration than larvae fed the cod liver oil enriched Artemia, while pigmentation rate was similar in the two groups. In addition to the difference in fatty acids, the two emulsions differed in lipid class composition, since 2050TG is a synthetic oil and a mixture of mono-, di- and tri-acylglycerol, while cod liver oil is a tri-acylglycerol. Total lipid level, estimated as fatty acid methyl esters (FAME) was similar in the two Artemia types, but sum of n-6 and n-3 fatty acids, arachidonic acid (20:4n-6, ARA), docosahexaenoic acid (22:6n-3, DHA) and eicosapentaenoic acid (20:5n-3, EPA) were higher in Artemia enriched with 2050TG than in the cod liver oil enriched Artemia. However, the main difference in fatty acid composition in the larvae, was a higher DHA (% of total fatty acids) in 2050TG larvae than in cod liver oil larvae. The lipid level measured as FAME was up to four times higher in the 2050TG larvae than in the cod liver oil larvae, and the reason for this may have been a better bioavailability of the partly digested lipid in the 2050TG emulsion. The correlation between a high level of lipid in the larval tissues (e.g. high energy status) and improved eye migration in larvae fed the 2050TG enriched Artemia supports the hypothesis that energy limitation on the larval stage may be a cause of the impaired eye migration commonly observed in farmed Atlantic halibut juveniles.     

Change in nutrient composition of Artemia grown for 3–4 days and effects of feeding on‐grown Artemia on performance of Atlantic halibut (Hippoglossus hippoglossus,L.) larvae

Major challenges in culture of Atlantic halibut larvae have been slow growth during the late larval stages and inferior juvenile quality due to pigmentation errors and incomplete eye migration during metamorphosis. The hypothesis of this study was that feeding on‐grown Artemia would alleviate these problems. Artemia were grown for 3–4 days on Origreen or Origo. The growth and nutrient composition of Artemia nauplii and on‐grown Artemia were analysed, and both Artemia types were fed to Atlantic halibut larvae, on‐grown Artemia from 15 days post‐first feeding (dpff). The body length of Artemia increased with 20%–70% in response to on‐growing. In all experiments, protein, free amino acids and the ratio of phospholipid to total lipid increased, while lipid and glycogen decreased. The fatty acid composition improved in some cases and not in others. The micronutrient profiles were not negatively affected in on‐grown Artemia. All these changes are thought to be beneficial for marine fish larvae. The final weight of Atlantic halibut postlarvae was similar, and 90% of the juveniles had complete eye migration in both groups. It is concluded that the present version of Artemia nauplii probably covers the nutrient requirements of Atlantic halibut larvae.     

Enrichment of Artemia with free methionine

Different enrichment procedures of the free amino acid (FAA) methionine were tested for Artemia nauplii. A direct enrichment protocol (methionine dissolved in the culture water) was compared with liposome enrichment protocols that varied in their membrane composition. During 16 h of direct enrichment in 5.3 mM methionine, the nauplii increased their content of free methionine between 20 and 30 times compared to the unenriched control (43.1±1.2 and 68.4±3.8 pmol·nauplius⁻¹ in two separate experiments vs. 2.4±1.0 pmol·nauplius⁻¹ in control). However, by encapsulating the identical amount of methionine into liposomes made from pure egg yolk phosphatidylcholine (PC) (>99% PC) and cholesterol, the nauplii content of free methionine reached 148.8±27.6 pmol·nauplius⁻¹, which is approximately 60 times more than in the unenriched control. Another liposome composition tested, made from crude egg yolk PC (>60% PC) and cholesterol, resulted in 90.5±4.1 pmol·nauplius⁻¹. The enriched nauplii still retained 80% of the free methionine after 8 h of incubation at conditions simulating feeding for Atlantic halibut larvae (13°C, 33.5 g·l⁻¹).

In conclusion: (1) Artemia nauplii can successfully be enriched with free methionine, (2) the high retention of free methionine in the Artemia nauplii following transfer to fish tanks shows that it is possible to offer fish larvae a feed with a high level of FAA, based on enrichment of Artemia nauplii.     

Infection and mortality by the yeast Metschnikowia bicuspidata var. bicuspidata in chinook salmon fed live adult brine shrimp (Artemia franciscana)

First-feeding chinook salmon were fed either live adult Artemia franciscana or commercial feed over a 15-week period. Unexplained mortality began occurring in the Artemia-fed fish after 35 days on the diets, with cumulative mortality reaching 34.5% in the Artemia-fed fish compared with 4.3% in feed-fed fish. Necropsy examinations revealed systemic fungal yeast infections and the causative agent was cultured from kidneys. Fungal cells were observed in the transport water of Artemia shipments and within the Artemia. The Artemia were purchased from a retail supplier, and originated from salt ponds in San Francisco Bay (SFB), California. Artemia infection rates ranged from 0.5% to 37.5% (mean 16.5±3.59%) in shipments received over a 5-week period. The fungus was characterized by morphological and physiological properties and was identified as Metschnikowia sp., a pathogenic yeast of aquatic invertebrates. The yeast grew at 9–27 °C, and 0–180 ppt NaCl, and could tolerate salinity of at least 270 ppt. Sequence analysis of the divergent D1/D2 domain of ascomycetous yeast 26S ribosomal DNA identified the organism as Metschnikowia bicuspidata var. bicuspidata. The organism was found in water from the salt ponds and probably entered from the bay. Mortality due to the fungal infection stopped after changes were made in the handling of incoming Artemia shipments.     

Nutrient composition and metamorphosis success of Atlantic halibut (Hippoglossus hippoglossus, L.) larvae fed natural zooplankton or Artemia

Atlantic halibut larvae were fed docosohexanoic acid- (DHA) selco enriched Artemia (RH-cysts) or wild zooplankton in duplicate tanks from first-feeding and 60 days onward. The zooplankton were collected from a fertilized sea water pond and consisted mainly of different stages of Eurytemora affinis and Centropages hamatus . There were no differences in survival, or in growth during the first 45 days of feeding, between larvae fed the two prey items, but the larvae fed Artemia showed much higher incidence of malpigmentation and impaired eye migration than larvae fed zooplankton. The prey organisms contained similar amounts of dry matter and protein, but Artemia was higher in lipid and glycogen than the zooplankton. Larvae fed Artemia were higher in both glycogen and lipid than the zooplankton-fed larvae towards the end of the feeding period. There were large differences between the prey organisms in the concentrations of essential fatty acids (% of total fatty acids) which was reflected in the fatty acid composition of the larval body. It is concluded that the macronutrient composition of Artemia in the present study was probably within the optimal range for promotion of growth and survival in young Atlantic halibut. The concentration of n-3 HUFA, and especially DHA, is however, very much lower in enriched Artemia than in copepods, and may be one of the factors triggering developmental errors in Atlantic halibut.     

Effect of docosahexaenoic acid enrichment in Artemia on growth of Pacific bluefin tuna Thunnus orientalis larvae

Feeding enriched Artemia induces growth failure in Pacific bluefin tuna (PBT) Thunnus orientalis larvae; however, feeding of yolk-sac larvae of marine fish promotes larval growth. It is considered that this growth failure partly results from dietary docosahexaenoic acid (22:6n-3, DHA) deficiency. Therefore, we examined the effect of DHA contents in enriched Artemia on the growth of PBT larvae. Artemia nauplii were enriched with graded levels of DHA ethyl ester, and fed to PBT larvae for 9 days. Yolk-sac larvae of Japanese parrotfish Oplegnathus fasciatus were used as a reference diet. The DHA contents in Artemia increased with the enrichment from 0 mg g^− 1 dry weight basis (DW) to 25 mg g^− 1 DW, while the content in the reference diet was 21 mg g^− 1 DW. Feeding of enriched Artemia significantly improved the growth of PBT larvae. However, this improvement was negligible when compared with the excellent growth of the larvae that were fed the reference diet. PBT larvae that were fed the reference diet accumulated approximately twice or much higher levels of DHA in the neutral and polar lipids in the body when compared with the larvae that were fed enriched Artemia. These results show that PBT larval growth cannot be promoted by feeding enriched Artemia even if the DHA contents in Artemia are elevated to the same levels as those of yolk-sac larvae. The incorporation of dietary DHA into phospholipids in the fish body may be desirable for the normal growth of PBT larvae.     

Beneficial effects of n-3HUFA enriched Artemia as food for larval palmetto bass (Morone saxatilis × M. chrysops)

The beneficial effects of feeding n−3 highly unsaturated fatty acids (HUFA ≥ 20 carbon fatty acids with three or more double bonds) to palmetto bass (striped bass × white bass) larvae, 4–30 days of age, were studied using Artemia diets enriched with six n-3HUFA levels. Dietary n-3HUFA concentrations were < 0.03% (control diet), 0.33%, 0.63%, 0.87%, 1.26%, and 2.27% of dry-wt Artemia. Larval n-3HUFA contents were reduced at a faster rate with decreasing dietary n-3HUFA intake, and were significantly different by 30 days posthatch (4–20 mg g⁻¹ dry-wt fish). Starved larvae selectively conserved endogenous n-3HUFA reserves, indicating an essential role of n-3HUFA in larval development. Mass mortality in the control and 0.33% n-3HUFA diets occurred at metamorphosis (26–28 days posthatch). At harvest, all fish, except those fed the two highest n-3HUFA diets, suffered from handling stress (shock syndrome) with increasing severity (25 to 100%) at decreasing dietary n-3HUFA intake. Recovery from shock syndrome was 100% at the 0.63% and 0.87% n-3HUFA diets, 63% at the 0.33% n-3HUFA diet and 0% at the control diet. Post-harvest survival was similar among the four highest dietary n-3HUFA groups (64–73%), whereas the two lowest n-3HUFA groups differed significantly (0 and 37%). Growth promotion was evident with increased dietary n-3HUFA intake as fish fed the highest n-3HUFA diet were twice the size of those fed the control diet (99 vs 52 mg wet-wt). Final fish sizes at the three highest n-3HUFA diets were similar. Given similar culture conditions, a minimum dietary n-3HUFA intake of 1.26% of dry-wt Artermia is recommended to avoid handling stress and promote growth in larval palmetto bass.     

A sensitive period during first feeding for the determination of pigmentation pattern in Atlantic halibut, Hippoglossus hippoglossus L., juveniles: the role of diet

Six groups of Atlantic halibut, Hippoglossus hippoglossus L., larvae were offered calanoid cope-pods at different periods from days 11 to 25 after first feeding (1.13-3.20 mm myotome height) in order to establish at which stage normal pigmentation was determined. Artemia nauplii enriched with an oil emulsion were used prior to and after the copepod period. Control groups were fed on copepods or Artemia only. The Artemia diet initiated an earlier intake of food and higher initial growth compared to the copepod diet. After 50 days of feeding, the average dry weights of the fish fed on Artemia and copepods were quite similar to the copepod-fed fish, while the Artemia-fed fish were the smallest in size. The lowest frequency of normally pigmented juveniles was found in the Artemia-fed group (66.4%), while the copepod group showed almost 100% normal pigmentation. A significantly higher frequency of pigmentation was found in juveniles given a copepod diet close to the initiation of metamorphosis than those provided with an earlier copepod period of equal duration. A high degree of eye migration was found in all groups, but was lowest in the Artemia-fed group. The initial stage of eye migration was found to occur at a larger body size in fish given Artemia and copepods, or a copepod diet than in fish fed on Artemia alone. There was no significant correlation between eye migration and growth rates prior to metamorphosis, although the largest individuals exhibited the most complete eye migration. High pigmentation frequencies were obtained in fish with a low 22:6n-3:20:.5n-3 (DHA:EPA) ratio (< 1.0).     

Effects of Stocking Density, Salinity, and Light Intensity on Growth and Survival of Southern Flounder Paralichthys lethostigma Larvae

Four separate studies were done on Southern flounder Paralichthys lethostigma larvae during first feeding and metamorphosis to determine the effects of stocking density, salinity, and light intensity on growth and survival. One study used stocking densities of 10, 20, 40, and 80 fish/L during first feeding; the second study compared the growth and survival of larvae stocked at 20 and 33 ppt; and a third experiment evaluated stocking densities of 1/L and 3/L under two different light intensities (1,600 lux vs 340 lux) during metamorphosis. The fourth experiment tested the effects of different salinities (0, 10, 20 and 30 ppt) on larval growth and survival during metamorphosis. Growth and survival (overall 6.9%) were not significantly different ( P > 0.05) for stocking rates up to 80/L. Larvae placed into 20 ppt salinity had survival through first feeding similar to that of larvae raised at 33 ppt. During metamorphosis, light intensity had no effect ( P > 0.05) on growth or survival, but fish stocked at 3/L had significantly lower ( P < 0.05) survival than fish at 1/L. Complete mortality of larvae occurred at 0 ppt. Growth and survival past metamorphosis were not significantly different ( P > 0.05) at 10, 20 and 30 ppt, but unmetamorphosed fish did not survive to day 60 at 10 ppt. Based on these results, practical larviculture of Southern flounder may require a two-step process with high stocking rates (80 fish/L) through first feeding and lower densities (1/L) through metamorphosis. Fingerling production in fertilized nursery ponds might he possible at salinity as low as 20 ppt.     

Effects of docosahexaenoic, eicosapentaenoic, and arachidonic acids on the early growth, survival, lipid composition and pigmentation of yellowtail flounder (Limanda ferruginea): a live food enrichment experiment

The role of dietary ratios of docosahexaenoic acid (DHA, 22:6n−3), eicosapentaenoic acid (EPA, 20:5n−3) and arachidonic acid (AA, 20:4n−6) on early growth, survival, lipid composition, and pigmentation of yellowtail flounder was studied. Rotifers were enriched with lipid emulsions containing high DHA (43.3% of total fatty acids), DHA+EPA (37.4% and 14.2%, respectively), DHA+AA (36.0% and 8.9%), or a control emulsion containing only olive oil (no DHA, EPA, or AA). Larvae were fed differently enriched rotifers for 4 weeks post-hatch. At week 4, yellowtail larvae fed the high DHA diet were significantly larger (9.7±0.2 mm, P<0.05) and had higher survival (22.1±0.4%), while larvae fed the control diet were significantly smaller (7.3±0.2 mm, P<0.05) and showed lower survival (5.2±1.9%). Larval lipid class and fatty acid profiles differed significantly among treatments with larvae fed high polyunsaturated fatty acid (PUFA) diets having higher relative amounts of triacylglycerols (18–21% of total lipid) than larvae in the control diet (11%). Larval fatty acids reflected dietary levels of DHA, EPA and AA while larvae fed the control diet had reduced amounts of monounsaturated fatty acids (MUFA) and increased levels of PUFA relative to dietary levels. A strong relationship was observed between the DHA/EPA ratio in the diet and larval size (r²=0.75, P=0.005) and survival (r²=0.86, P=0.001). Following metamorphosis, the incidence of malpigmentation was higher in the DHA+AA diet (92%) than in all other treatments (50%). Results suggest that yellowtail larvae require a high level of dietary DHA for maximal growth and survival while diets containing elevated AA exert negative effects on larval pigmentation.     

A comparison of retinol, retinal and retinyl ester concentrations in larvae of Atlantic halibut (Hippoglossus hippoglossus L.) fed Artemia or zooplankton

Atlantic halibut (Hippoglossus hippoglossus L.) larvae were fed enriched Artemia or zooplankton in duplicate tanks from 0 to 60 days after first‐feeding. Both diets and the larvae were analysed for vitamin A (VA) in order to confirm earlier findings, in which Artemia fed larvae had lower levels of VA compared with larvae fed zooplankton. Furthermore, we wanted to investigate the composition of the retinoids in the larvae. The results showed that Artemia and zooplankton contains low levels of VA, probably too low to sustain the assumed requirement. Nevertheless, larvae fed Artemia had the same level of retinal and retinol as larvae fed zooplankton. We found a significant lower level of retinyl esters in larvae fed Artemia. The total VA level was lower in larvae fed Artemia only at the end of the feeding trial after the onset of metamorphosis. Our conclusion is that feeding Artemia to Atlantic halibut larvae is not likely to cause VA deficiency.     

Folate in eggs and developing larvae of Atlantic halibut, Hippoglossus hippoglossus L.

Folate mobilization from the yolk compartment during larval development was studied by analysing the folate concentration in whole body, embryo and yolk in a single batch of Atlantic halibut, Hippoglossus hippoglossus L., eggs and larvae that showed successful fertilization and development. There was a net loss of approx. 50% of folate from yolk during endogenous feeding. Further, only 23% of the decrease in yolk folate was retained in the larval body. The data suggest a need for folate for metabolic and growth purposes during embryogenesis of approximately 2 μg g^?1 weight gain. Relative to these data and published folate requirement for cold‐water species, batches of egg from 16 Atlantic halibut brood fish contained variable and, for some batches, critically low levels of folate. This may constitute a potential problem for larval development until start feeding.     

Development of thyroid gland and changes in thyroid hormone levels in Leptocephali of Japanese Eel (Anguilla japonica)

Development of the thyroid gland of the Japanese eel (Anguilla japonica) was studied with the use of tank-reared fish. A single thyroid follicle was first found in larvae at 29 days post hatching (dph), total length (TL) 12 mm. Until reaching 25 mm in TL (100 dph), leptocephali had one or two follicles per individual. The inner colloid of the follicles was weakly immuno-positive against the anti-thyroxine (T₄) antibody. The number of thyroid follicles and the immunoreactivity later increased as the larvae grew. Thyroid hormones (TH) T₄ and triiodothyronine (T₃) were not detected in premetamorphic larvae by radioimmunoassays, but became detectable during metamorphosis. The maximum level of T₄ was seen in fish at the end of metamorphosis and in just-metamorphosed juveniles, whereas T₃ reached the highest level during metamorphosis and declined toward the end of metamorphosis. The results indicated that the thyroid gland first became active during metamorphosis in the development of eel.     

Amino acid-chelate: a better source of Zn, Mn and Cu for rainbow trout, Oncorhynchus mykiss

This study aimed to evaluate the amino acid-chelated trace elements as dietary supplement to rainbow trout. Three diets were formulated containing trace elements either from the inorganic salt (SF) or amino acid-chelate (AM). Diets 1 (SF) and 2 (AM) contained the same amount of trace elements from inorganic and amino acid-chelates, respectively. Diet 3 (AM-Hf) was added with trace elements from amino acid-chelatex at one-half of their levels in Diets 1 and 2. Each diet was fed for 15 weeks to three groups of 30 fish each, with an average weight of 1.52±0.21 g. Growth of fish was not affected by the treatment (P>0.05). However, bone (P<0.01) and liver (P<0.05) Cu contents were higher in the AM than the SF group. Similarly, hematocrit level (P<0.05) and alkaline phosphatase (ALP) activity (P<0.01) were higher in the chelate-fed fish. Further, DNA polymerase and CuZnSOD expression in the AM group was highly upregulated (P<0.05) compared to the SF fed fish as quantified by RT-PCR. Absorption and whole body retention of Mn and Zn from the AM were higher (P<0.05) than the inorganic salt. Half supplementation of those fed the elements from AM was at par with the full provision from the inorganic source tested.     

Pigmentation and eye migration in Atlantic halibut (Hippoglossus hippoglossus L.) larvae: new findings and hypotheses

Atlantic halibut juveniles, which have been fed Artemia during larval development, frequently demonstrate malpigmentation and impaired eye migration. This is in contrast to the high percentage of normally developed larvae fed copepods, reared under similar conditions. Nutrition is therefore an important component influencing larval development. Analyses of the nutrient composition of Artemia and copepods show that Atlantic halibut larvae fed Artemia probably receive sufficient amounts of vitamin A by converting canthaxanthin, while iodine may be deficient, possibly leading to interrupted thyroid hormone synthesis. An unbalanced fatty acid composition, such as high levels of arachidonic acid and low levels of docosahexaenoic acid, can be another limiting factor in Artemia. Vitamin A, fatty acids and thyroid hormones have all been shown to affect pigmentation in flatfish. They are ligands to nuclear receptors, thyroid hormone receptors, retinoic acid receptors, retinoic X receptors and peroxisomal proliferator‐activated receptors, which are members of the superfamily of steroid hormone receptors. The receptors interact with each other to promote gene expression that modulates proliferation and differentiation of cells. Our hypothesis is that these interactions are important for development during flatfish metamorphosis. Very little data exist on the topic of impaired eye migration. However, energy limitation, iodine deficiency and an unbalanced fatty acid composition have been proposed as possible explanations. Here, we review the literature on development of pigment cells and the possible mechanisms behind the effects of vitamin A, fatty acids and thyroid hormone on pigmentation and eye migration during development of Atlantic halibut larvae.     

Thyroid gland development in Senegalese sole (Solea senegalensis Kaup 1858) during early life stages: A histochemical and immunohistochemical approach

A key to success in the culture of marine fish species is the mass production of high quality fry, a process largely dependent on successful first feeding and normal development and growth of fish larvae. In this regard it is important to examine the structural and functional development of the endocrine system (pituitary, thyroid, interrenal glands) during early ontogeny of marine fish. Particularly, the thyroid hormones, thyroxine (T₄) and triiodothyronine (T₃), influence numerous metabolic processes, such as growth, differentiation, metamorphosis, reproduction, respiration, migratory behaviour, central nervous system activity, seasonal adaptation, etc. Therefore the aim of this study was to describe the development of the thyroid gland and the ontogeny appearance of the thyroid hormones in Solea senegalensis larvae by means of histological and immunohistochemical techniques. The first thyroid follicle was present at 4 days-post-hatch (dph) coinciding with first feeding. During metamorphosis (12–20 dph) the follicles increased in both number and size, and by 30 dph presented the same characteristics as that seen in adult fish. Tissue immunostaining of both thyroid hormones decreased during the endogenous larvae development to nearly undetectable levels at the completion of yolk-sac absorption. During larvae exogenous phase, T₃ and T₄ immunostaining was first detected by 6 dph and an increase of specific staining for both hormones was detected between 12 and 20 dph, during metamorphosis phase.     

Control of light condition affects the feeding regime and enables successful eye migration in Atlantic halibut juveniles

Incomplete eye migration is one of the major problems in intensive production of juvenile Atlantic halibut. More than 60% of an average juvenile population reared according to best practice suffers from this abnormality. In commercial production, these fish are discharged and represent a substantial economic loss and a large welfare problem. In the present investigation it is demonstrated that by controlling diurnal light and darkness periods together with a meal based feeding regime, incomplete eye migration can be dramatically reduced in production systems for Atlantic halibut.Control groups were reared under continuous light conditions, whereas the experimental groups were given 7 h of darkness and 17 h of light during a 24 hour cycle, in a period lasting from 12 to 35 days post first-feeding. Otherwise both groups were reared under continuous light conditions. All larvae were fed short time enriched Artemia supplied two times daily.The experimental conditions did not affect the overall growth or survival up to day 85 after first feeding. However, 27 ± 3% of the fry reared under continuous light conditions had complete eye migration, whereas in juveniles reared under shifting light and darkness conditions, complete eye migration was 85 ± 7%. These results represent a major improvement in production systems for Atlantic halibut juveniles.     

Early weaning of winter flounder (Pseudopleuronectes americanus Walbaum) larvae on a commercial microencapsulated diet

Like most small marine fish larvae, the stomachs of winter flounder Pseudopleuronectes americanus are undeveloped at first feeding and have relatively reduced digestive capacity. This work was undertaken to test whether larvae at the onset of stomach differentiation (larval size about 5.5 mm) could be early weaned onto a commercial microencapsulated diet. We assessed the effect of early weaning by first comparing growth performance (standard length, total protein content and age at metamorphosis) of larvae fed enriched live prey from first feeding to a size of 5.5 mm and then reared on three different feeding regimes until metamorphosis: (1) live prey (LP) as a control group; (2) mixed feeding of live prey and microencapsulated diet (LP‐ME); (3) exclusively microencapsulated diet (ME) after fast weaning over 4 days (to a larval size of 6.2 mm). No differences were observed between larval development in the two first groups, which began metamorphosis at 40 days old. The larvae of the third group showed significantly slower growth that resulted in a delay of 4 days in the onset of metamorphosis. Differences in live prey availability between the treatments and the short transition period to allow the larvae to adapt to the new diet were identified as possible contributing factors to the slower growth and to the delay in metamorphosis of early weaned larvae. In a second experiment, the transitional weaning period was increased until the larvae were 6.6 mm in length. Weaning at that size resulted in no slowing of growth or delay in metamorphosis, suggesting that the feeding schedule was adequate.