The growth of turbot (Scophthalmus maximus (L.)) in recirculating systems

O-group wild turbot (Scophthalmus maximus (L.)) were used in a number of feeding experiments designed to assess the usefulness of readily available species of industrial fish as feed for the ongrowing stage of cultivation (10–500 g). Sprats (Sprattus sprattus (L.)) and sandeels (Ammodytes marinus Raitt) were the most successful in that feeding rates of 2% live weight per feed gave conversion efficiencies which were generally better than 3:1. A regular regime of two feeds a day should give growth rates approaching 2% per day resulting in an ongrowing period of about 200 days at 16°C. The turbot were maintained in closed recirculating systems using a simple design for the treatment units. These provided a stable environment in which to conduct the experiments the average water quality conditions being given. There were few health problems and the taste and texture of the fish produced appeared to be good.     

Side-aspect target strength of Atlantic salmon (Salmo salar), brown trout (Salmo trutta), whitefish (Coregonus lavaretus), and pike (Esox lucius)

The side-aspect acoustic target strengths (TS) of 19 Atlantic salmons (Salmo salar), 16 brown trouts (Salmo trutta), 10 whitefish (Coregonus lavaretus) and 9 pikes (Esox lucius) were measured using a 200 kHz split-beam echosounder, in order to study the relationship between TS and fish size indices (length, weight and side area). The effect of side aspect angle on TS was also studied. Linear models between TS and the logarithm of the fish size indices were fitted with length being best for predicting TS. Typically, the standard error of estimate was 1.2–2.9 dB. The side-aspect TS measurements with specimens of known size showed that the linear relationship between full side-aspect TS and the logarithm of fish length for salmonid (Salmo salar + Salmo trutta) was on average 4.7 dB (SE = 0.7), lower than for whitefish and pike combined. The effect of side aspect angle on TS was modelled with cos³ (2α) function. The differences in the TS between full side aspect and head/tail aspect were 17.4, 19.0 and 19.6 dB for salmonid, whitefish, and pike, respectively.     

Genetic differentiation in hatchery strains and wild white shrimp Penaeus (Litopenaeus) vannamei (Boone, 1931) from northwest Mexico

Ten enzymatic systems were analyzed to determine allozyme genetic differentiation among three hatchery strains (A, B and C) of white shrimp, Penaeus (Litopenaeus) vannamei, commonly used in shrimp farming in northwest Mexico. A wild population from northern Sinaloa was used as a reference. Fifteen loci were detected, nine of which were polymorphic (ACP-1*, ACP-2*, AKP-2*, EST-2*, EST-3*, EST-4*, EST-5*, LAP*, and LDH*). Polymorphism of A, B and C were 53, 53, and 40%. The mean observed heterozygosity per locus was 0.071, 0.093, and 0.050 without any significant difference among them or with respect to the wild population (0.056). In all samples observed heteroxygosity was smaller than expected. The mean number of alleles per locus was 1.8 in A and 1.87 in both B and C. Strain A was the only sample without rare alleles. Only EST-3* and LAP* of strain A were in Hardy-Weinberg equilibrium; the other loci were in disequilibrium in all samples. High inbreeding values and heterozygote deficiencies were detected in all samples. Distribution of allelic frequencies was heterogeneous among the samples (G-test), involving all polymorphic loci and suggesting a genetic differentiation. According to Fst, a moderate genetic differentiation (7.4%) was detected among the samples. Greater differences were between strains A and C. Based on genetic distance, the samples were grouped into two pairs, B–C and A-wild. Strain A is a young strain related to the wild sample, whereas strains B and C have a different geographic origin than the wild sample.     

Biological evaluation of phytoplankton (Chlorella sp., Tetraselmis sp. and Isochrysis galbana) as food for milkfish (Chanos chanos) fry

Phytoplankton cultures of Chlorella sp., Tetraselmis sp. and Isochrysis galbana were used alone as feed to rear separate batches of newly caught milkfish (Chanos chanos) fry (about 21 days old). Ultrastructural studies of fry hepatocytes and pancreatic acinar cells indicated that they cannot directly utilize Chlorella, which has a rigid cell wall. The fry can directly utilize Tetraselmis and Isochrysis, but neither is nutritionally adequate for growth and survival if used as the only feed. Tetraselmis is nutritionally inferior to Isochrysis.     

Mixed Function Oxidase Activity and Organochlorine Levels in Farmed Sharpsnout Seabream (Diplodus puntazzo) from Two Intensive Aquaculture Facilities

Mixed function oxidase (MFO) activities and organochlorine levels were investigated in liver of farmed sharpsnout seabream bred in two intensive facilities, a sea farm and a land-based farm. The aim of this study is to investigate how different facility locations and breeding conditions might affect fish metabolic capabilities and accumulation of toxic chemicals in farmed sharpsnout seabream. The differences between breeding two or more fish species in the same cage or tank known as polyculture, and monoculture (1 species) were also investigated. The results showed that both facility location (sea and land) as well as breeding systems (polyculture and monoculture) might be responsible for the differences observed in both MFO enzyme activity and organochlorine levels. Significantly higher activities of two metabolizing enzymes such as 7-ethoxyresorufin-O-deethylase (EROD) and benzo(a)pyrene monooxygenase (B(a)PMO) were in fact recorded in land-based farmed fish than in sea farmed ones, and much higher in those from polyculture than in monoculture (p < 0.05). By contrast, the highest levels of dichlorodiphenyldichloroethilene (pp′DDE) and polychlorinated biphenyls (PCBs as Aroclor 1260) were measured in sea-farmed fish, except for hexachlorobenzene (HCB), which gave higher levels in land-based farmed fishes. No significant differences were observed between monoculture and polyculture in p,p′DDE and PCBs levels except for p,p′DDE which was significantly higher in monoculture than in polyculture (p < 0.05) in fish reared in tanks. Finally significantly higher HCB levels were measured in fish reared in monoculture in tanks (p < 0.05) and in polyculture in cages (p < 0.05). The low TEQ values obtained for PCB congeners at liver level indicated no concern for fish health.     

Inhibitory effects of Bacillus licheniformis (DAB1) and Pseudomonas aeruginosa (DAP1) against Vibrio parahaemolyticus isolated from Fenneropenaeus indicus

In aquaculture industries, there is an urgent need to develop microbial control strategies, to control disease outbreaks. In recent years, probiotics are considered as a valid alternative for the use of antibiotics in aquaculture to prevent high mortality and promote growth. In the present study, seven strains of bacteria such as Bacillus licheniformis (DAB1), Bacillus pumilus (DAB2), Bacillus sp. (DAB3), Pseudomonas aeruginosa (DAP1), Pseudomonas sp. (DAP2), Pseudomonas aeruginosa (DAP3), Pseudomonas aeruginosa (DAP4), and three pathogenic Vibrio parahaemolyticus (DAV1, DAV2, DAV3) were isolated from healthy and diseased Fenneropenaeus indicus collected from the east coast of Tamilnadu, India. The strains were identified by biochemical analysis and 16S rRNA sequence methods. Among the seven probiotic strains tested, the cell-free extract from DAB1 and DAP1 exhibited higher inhibitory activity of V. parahaemolyticus than other isolates under in vitro conditions. The LC₅₀ of DAV1, DAV2, and DAV3 was found to be ～10³ CFU mL^?1. Pathogenicity of three V. parahaemolyticus DAV1, DAV2, and DAV3 showed significant mortalities (40 %) in Artemia nauplii at inoculation densities of 10³ CFU mL^?1 when compared to the controls (unchallenged nauplii). A significant reduction in mortality (P < 0.001) was found by addition of 10⁶ CFU mL^?1 of DAB1 and DAP1 strains in nauplii against the pathogens. In conclusion, the present study result reveals that DAB1 and DAP1 have potential applications for controlling pathogenic V. parahaemolyticus in Artemia culture systems and aquaculture practices.     

Enrichment of gilthead seabream (Sparus aurata L.) diet with microalgae: effects on the immune system

The present study assessed the effects of three orally administered microalgae (Nannochloropsis gaditana, Tetraselmis chuii and Phaeodactylum tricornutum) on different immune parameters and immune-related gene expression of gilthead seabream (Sparus aurata L.). Fish were fed a control or one of six experimental diets and sampled at 2 and 4?weeks of treatment. At the end of the trial, growth performance and different systemic and local immune activities were measured (natural haemolytic complement activity, IgM levels, phagocytosis and respiratory burst). The expression levels of different immune-associated genes (EF-1α, IgM_H, TCR-β, MHCIα, MHCIIα, CSF-1R and β-defensin) were analysed in head-kidney and gut. Administration of N. gaditana and T. chuii resulted in a significant increase in haemolytic complement activity, phagocytic capacity, as well expression level of β-defensin, as well as MHCIIα and CSF-1R, respectively. The P. tricornutum-supplemented diet provoked immunostimulation, and very little effect on gene expression was observed. These results suggest that dietary microalgae enhance gilthead seabream defence activity, which could be very important in order to consider such microalgae as a possible additive in fish diets.     

Studies on the rotifer (Brachionus urceus Linnaeus, 1758) as a vector in white spot syndrome virus (WSSV) transmission

The rotifer Brachionus urceus (Linnaeus, 1758) was experimentally infected with the white spot syndrome virus (WSSV) by the virus-phytoplankton adhesion route in order to assess the possibility of rotifer acting as a vector of WSSV to infect the shrimp Fenneropenaeus chinensis (Osbeck, 1765) larvae at zoea stage III. The nested-PCR test revealed WSSV-positive results in the rotifers exposed to WSSV by the virus-phytoplankton adhesion route. Among 10 replicates in the infection treatment, 40% of F. chinensis larvae became WSSV-positive when fed with WSSV-positive rotifers, whereas all were WSSV-negative for F. chinensis when fed with WSSV-free rotifers. Though the mortality of shrimp larvae in the infection treatment (39.47 ± 15.44%) was higher than that in the control treatment (34.67 ± 15.11%), there was no significant difference in the mortality between them (P > 0.05). These results indicated that the rotifer could serve as a vector in WSSV transmission when ingested.     

Protein sparing effect of dietary lipids in common dentex (Dentex dentex): A comparative study with sea bream (Sparus aurata) and sea bass (Dicentrarchus labrax)

The common dentex (Dentex dentex) is a sparid fish which is considered a suitable candidate for Mediterranean aqua-culture. A seven-weeks feeding trial was conducted over the summer period with common dentex, sea bream (Sparus aurata) and sea bass (Dicentrarchus labrax). All three species were fed to visual satiety with two practical diets with varying crude protein and crude fat levels (55 % protein, 9 % lipids; 46 % protein, 17 % lipids). The initial body weights were 8.2, 11.6 and 17.3 g for common dentex, sea bass and sea bream, respectively. In all cases, specific growth rates were not significantly affected by dietary treatment, but they varied among species (sea bass 1.7–1.8 %; sea bream 2.1–2.3 %; common dentex 3.1–3.2 %). When comparisons between fish species were made, we observed that the increase in growth rates was linked to a decrease of the whole body fat content, as a consequence of a greater utilisation of dietary lipids as energy substrates. Furthermore, in common dentex, the 17 % lipid diet was able to improve protein retention, but this diet effect was less significant in sea bream and sea bass. In contrast to sea bream, the 17 % lipid diet did not up-regulate plasma GH levels in common dentex, which provides additional evidence for a more efficient utilisation of dietary lipids.     

Effect of three bacterial isolates from a commercial hatchery on early red abalone (Haliotis rufescens) postlarvae

Three bacterial strains, GHrC11, GHrC13 and GHrC15, were isolated from an abalone postlarval culture system in a commercial farm at Baja California, México. The strains were phenotypically characterized and sequenced (16S rDNA). Strain GHrC11 was a Gram-positive coccobacillum, while strains GHrC13 and GHrC15 were Gram-negative bacilli. Strain GHrC11 was identified as Exiguobacterium sp. The strains GHrC13 and GHrC15 were identified as Vibrio splendidus. The effects of these strains for the development of early abalone postlarvae (2 days old) were evaluated following a completely randomized design with three replicates using 5-mL-volume Petri dishes as experimental units. The experiment considered two different bacterial concentrations of each strain (10³ and 10⁵?cells?ml^?1) and two controls (with and without the benthic diatom Navicula incerta). After 10?days of experimentation, the highest mortality (90?±?5.8?%) and the lowest growth rate (4.1?±?0.1?μm?day^?1) were recorded for the strain GHrC11. In contrast, the lower mortality (16.7?±?3.3?%) and the highest growth rate (11.2?±?0.9?μm?day^?1) corresponded to the control fed N. incerta. Our results suggest that pathogenic effects of these bacterial strains were stronger than any potential benefits derived from the ingestion of bacteria by early abalone postlarvae. In conclusion, the most pathogenic strain was GHrC11, and the intensity of pathogenicity could be ordered as Exiguobacterium sp.?>?V. splendidus (C13)?>?V. splendidus (C15).     

Efficiency of the bottom trawl used for the Mediterranean international trawl survey (MEDITS)

The aim of the work presented in this paper was to assess the relative efficiency of the GOC 73 sampling trawl used for the Mediterranean international trawl survey programme (MEDITS) compared with that of a typical Italian commercial trawl. The latter was chosen because it is commonly used by fishermen in the central Adriatic, where the experiment was conducted, and therefore appeared as the best possible sampler of the commercial species present in those areas. Moreover, this trawl is similar to the various trawls used for the Italian national survey programme (GRUND). Four fishing trips were conducted between 1996 and 1997 in different areas and seasons to sample different species. Each trip was conducted in the same fishing area. A codend cover mounted on the commercial trawl ensured that the codends of the two nets had the same mesh opening. The two trawls were alternated daily. Net geometry (horizontal and vertical net opening) was measured during all tows. Catch data were converted into abundance per swept area before comparing the trawls. Efficiency comparisons were performed on ten fish species, one crustacean and four molluscs belonging to the MEDITS list of main reference species. The MEDITS trawl was significantly less efficient in terms of both weight and numbers of individuals fished for hake (Merluccius merluccius), common sole (Solea vulgaris) and Norway lobster (Nephrops norvegicus). A highly significant difference in favour of the commercial trawl was found in the weight, but not the numbers, of common pandora (Pagellus erythrinus). Efficiency differences were negligible for red mullet (Mullus barbatus), while the MEDITS trawl was significantly more efficient for the numbers of Atlantic horse mackerel (Trachurus trachurus). For all the other main reference species differences were not significant. The commercial trawl was more efficient for large individuals of some species. The relative efficiency of the MEDITS trawl was especially low for small-size classes of N. norvegicus. The results confirm the lower efficiency of the MEDITS compared with the commercial Italian trawl for benthic species, and its greater efficiency for some others released from the bottom as well as for the pelagic ones. These data allow the results of the MEDITS surveys to be compared with those of the GRUND programme and with commercial-fleet catches in areas where the commercial trawls used are similar to the one studied here.     

Morphological characterization of larval stages and first juvenile of the freshwater prawn Cryphiops caementarius (Molina, 1782) (Decapoda: Palaemonidae) under laboratory conditions

Research has been in progress for several years on various aspects of the biology and ecology of the freshwater prawn Cryphiops caementarius, an inhabitant of rivers in northern Chile. The commercial value of this prawn fomented the accomplishment of studies on its reproduction and development with the aim of producing juveniles under controlled conditions, to be followed by growout to commercial size in managed culture systems. The present study describes larval culture of this species from eggs of gravid females obtained in the field, from the first developmental stage (Zoea I) through the first juvenile stage. The larvae were cultured at 25 °C in UV sterilized water at variable salinities based on the requirements of the developmental stages. Larvae were fed with Nannochloris, Isochrysis and Artemia nauplii as required. This report describes in detail the 18 larval stages of this prawn, as well as its first juvenile form.     

Effect of azadirachtin on haematological and biochemical parameters of Argulus-infested goldfish Carassius auratus (Linn. 1758)

Argulosis hampers aquaculture production and alters the host physiology and growth. Azadirachtin is recognized as a potential antiparasitic agent against Argulus sp. The present study aimed to investigate the effect of different concentration of azadirachtin solution on haematological and serum biochemical parameters of Argulus-infested goldfish Carassius auratus. Ninety Argulus-infested goldfish were randomly divided into six equal groups. Fish of group 1–5 were treated with azadirachtin solution through bath of 1, 5, 10, 15 and 20 mg L^?1 as T1, T2, T3, T4 and T5, respectively, and group 6 was exposed to 2 % DMSO solution without azadirachtin and considered as negative control T0^?. Along with six treatment groups, a positive control T0⁺ of healthy goldfish free from Argulus infestation was also maintained. Parasitic mortality was evaluated after 3 days of consecutive bath treatment. After 7 days of post-treatment, the blood and serum were drawn from each of the treatment groups and haematological and serum biochemical parameters were evaluated. Total leucocyte count (TLC), mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH), blood glucose, total protein (TP), globulin, serum glutamate oxaloacetate transaminase (SGOT) and serum glutamate pyruvate transaminase (SGPT) were significantly (p < 0.05) high in negative control group when compared with positive control group. It could be concluded that Argulus infestation altered marked haematological and serum biochemical parameters. However, in treated groups complete elimination of Argulus was found in T4 and T5 groups. Also significant (p < 0.05) reduction in haematological and serum biochemical parameters of all the treatment groups were recorded in comparison with negative control group. In addition, T4 and T5 groups showed significantly (p < 0.05) high superoxide dismutase (SOD), catalase, total erythrocyte count (TEC) and haemoglobin (Hb). However, higher mean corpuscular haemoglobin concentration (MCHC), blood glucose and lactate dehydrogenase (LDH) levels in T5 group revealed that higher concentration of azadirachtin have notable effects on activity of vital tissues function and physiology of the host. Argulus spp. from infested goldfish could be eliminated using bath treatment with solution of azadirachtin having concentration of 15 mg L^?1 and that also shifted haematological and serum biochemical parameters towards homeostasis.     

Molecular characterization of three Rana grylio virus (RGV) isolates and Paralichthys olivaceus lymphocystis disease virus (LCDV-C) in iridoviruses

Three Rana grylio virus (RGV) isolates and lymphocystis disease virus (LCDV-C) were molecularly characterized by antigenicity comparison, Western blot detection of viral polypeptides, restriction fragment length polymorphism analysis of viral genomes, and MCP sequence analysis. Significant antigenicity differences existed among the three RGV isolates and LCDV-C. Western blot detection indicated that the viral polypeptides of three RGV isolates could be recognized by the anti-RGV9807 serum, whereas no bands were observed in the LCDV-C, and significant differences exist among the band patterns of three RGV isolates. Restriction fragment length polymorphism (RFLP) analysis was performed by digesting genomic DNA of the four iridovirus isolates with restriction endonucleases HindIII, KpnI, XbaI and BamHI. On the whole, obvious discrepancies existed between LCDV-C and RGV isolates, and some significant band pattern differences were also revealed between RGV9808 and RGV9506 (or RGV9807) in the profiles of restriction endonucleases XbaI, KpnI and BamHI. PCR amplification and sequence analysis of MCP gene sequence further revealed their phylogenetic relationship among the three RGV isolates, LCDV-C and other iridoviruses. RGV9506, RGV9807 and RGV9808 are clustered together with other ranaviruses, such as FV3, BIV, TFV and ENHV, although the RGV9808 is more close to EHNV than to other ranaviruses. Additionally, LCDV-C is clustered with LCDV-1, the type species of genus Lymphocystisvirus. The current study provides clear evidence that significant genetic difference exists among the three RGV isolates. Therefore, further work on comparative genomic studies will contribute significantly to understanding of their taxonomic position and pathological mechanism.     

Amino acid profiles of Diplodus sargus (L., 1758) larvae: Implications for feed formulation

The indispensable AA profile of fish carcass has been commonly used as a good indicator of fish amino acids requirements. Amino acid composition of the whole body tissue of Diplodus sargus was determined for the larval ages of 0, 2, 5, 8, 12, 17, 25, 35 and 45 days after hatching (DAH). No significant differences were found during this species ontogeny, except for phenylalanine. A comparative analysis of amino acid profiles from larvae and respective diet was performed. Low correlation was found to rotifers (R² approximately 0.5), while higher correlations were found for Artemia nauplli, metanauplii (R² approximately 0.8) as well as for the dry feed. These results suggest that D. sargus are subjected to higher nutritional imbalances during the first 10 days of feeding when larvae are fed on rotifers alone. Arginine, threonine, lysine, cysteine and histidine appeared to be limiting amino acids at 2, 12, 25 and 45 DAH, respectively. Similar results were reported in literature for Sparus aurata and Solea senegalensis, although D. sargus diets seem to have more amino acids in deficiency as well as more severe differences between larval and diet amino acid profiles. To solve these apparent nutritional imbalances, amino acid supplementation should be considered. The use of inert diets in early larvae ages seems to be most adequate as live feed supplementation appears to be more difficult.     

A radiological study on the development of vertebral deformities in cultured Atlantic salmon (Salmo salar L.)

This study investigated the development of skeletal deformities in individually tagged Atlantic salmon (n = 805) from the parr stage (Sept 2001, 70 g mean ± 34 g S.D.) until 10 month after transfer to seawater (Feb 2003, 3040 g mean ± 1097 g S.D.). A subgroup of the total population (n = 129) was radiographed as parr (Feb 2002) and again 10 months after transfer to seawater (Feb 2003). Eight percent of the males matured sexually during their first autumn in sea (jack), and were excluded from further analysis. Based on an external examination of each fish (n = 773), 1.8% (n = 14) developed skeletal deformities (0.1% operculum deformities (n = 1), 0.4% jaw deformities (n = 3), 1.3% vertebral deformities (n = 10)) during the experiment. Based on evaluation of radiographs (n = 129), the prevalence of vertebral deformities was 7.0% (n = 9) at the parr stage, and 12.4% (n = 16) 10 months after transfer to seawater. From radiographs, longitudinally compressed vertebral bodies without intervertebral spaces were classified as ankylosis and compression, longitudinally compressed vertebral bodies with intervertebral spaces were classified as compressions, and dislocated vertebral bodies with a normal morphology and without intervertebral spaces were classified as ankylosis and dislocation. Of the fish that developed deformities during the experiment, 8 had ankylosis and compressions, 7 had compressions and 1 had ankylosis and dislocation. Ankylosis and compression developed in the region between vertebrae number 1 and 16, whereas compressions developed in the region between vertebrae number 13 and 49. Most of the individuals with compression 10 months after transfer to seawater had normal vertebral columns as parr, whereas all individuals with compression and ankylosis 10 months after transfer to seawater had deformed vertebral columns as parr.     

The effect of domestication on a brown trout (Salmo trutta m fario) broodstock in Hungary

Molecular markers (PCR–RFLP and microsatellite) were used to investigate the genetic background of the only brown trout (Salmo trutta m fario) broodstock in Hungary which due to the hydrogeography of the country should theoretically belong to the Danubian lineage. PCR–RFLP (mitochondrial DNA control region and lactate dehydrogenase and somatolactin genes) as well as microsatellite (BFRO002, OMM1064, Ssa408uos, SsoSL417, SsoSL438) markers were used to distinguish between Danubian and Atlantic lineages of brown trout in the Lillafüred broodstock. Altogether 435 fish were tagged during the experimental period. Due to mortalities, fin clips were collected from 401 individuals. According to the genetic analysis of the mitochondrial DNA, the Danubian haplotype is present in only one individual (0.2 %) of the broodstock. Analysis of the nuclear markers revealed that alleles characteristic of both the Atlantic and the Danubian lineages are found in the population. However, Atlantic alleles dominate throughout the broodstock which is in Hardy–Weinberg equilibrium according to the investigated markers. Results indicate that the original broodstock that was introduced to the farm following its construction in 1933 was of the Atlantic lineage. Although later fish from a local stream were collected and added to the broodstock, the number of these was limited and they were almost exclusively males. Fish from this farm that are stocked by anglers can have a significant genetic impact on trout populations of natural streams.     

Extinction of Daphnia lumholtzi (Sars) in Lake Kinneret (Israel)

Daphnia lumholtzi (Sars) was found in Lake Kinneret until the late nineteen-fifties. Lake Kinneret was the northern-most limit of the distribution of this species. It is suggested that fingerlings of grey mullets and Sarotherodon aureus that were introduced into Lake Kinneret caused the extinction of D. lumholtzi.     

Breeding and larval rearing of the rabbitfish,Siganus guttatus (Bloch)

Females of Siganus guttatus reared to sexual maturity in canvas tanks were induced to spawn by using human chorionic gonadotropin (HCG, Ayerst) at 500 IU/fish or about 2 IU/g body weight. The amount of HCG used depended on the initial mean egg diameter; the smaller the diameter, the more HCG was used. Fish with oocytes characterized by germinal vesicle migration (mean egg diameter ≥ 0.47 mm) spawned without HCG injection. Fertilization and hatching rates for both treated and untreated fish were more than 90%. The larvae were reared to metamorphosis using rotifers from day 2–17, rotifers + newly hatched Artemia nauplii from day 18–20 and rotifers + newly hatched Artemia nauplii + artificial feed from day 21–35. In addition, Isochrysis galbana was introduced to the rearing tanks from day 1–10 and Chlorella sp. and/or Tetraselmis sp. from day 1–35. Survival rates of larvae tended to be lower as the broodstock became older.