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.     

Osteological development and anomalies in larval stage of hatchery‐reared yellow catfish Pelteobagrus fulvidraco

Comprehensive knowledge of osteological development of fish not only provides means for understanding its functional development, but also allows early detection of skeletal anomalies. The present study was conducted to determine osteological development and occurrence of anomalies from the first to the 40th day after hatching (DAH) for yellow catfish Pelteobagrus fulvidraco, a commercially important species in China. Vertebral ontogeny started with formation of anterior neural arches at 3 DAH, and completed at 6 DAH. Vertebral centra started to develop at 4 DAH and ossification in all centra was visible at 15 DAH. Caudal fin appeared at 2 DAH and ossification was visible at 26 DAH. The onset of dorsal and anal fins appeared at 4 DAH and their ossifications were visible at 20 DAH. Pectoral fins were present before first feeding and formed entirely at 9 DAH, and their ossifications were visible at 21 DAH. Pelvic fins appeared at 9 DAH, and formed completely at 21 DAH. The ossification of pelvic fins was not finished until the end of the experiment (40 DAH). 24 types of skeletal anomalies were observed. About 20% of individuals showed at least one anomaly. Haemal vertebrae anomalies occurred at the highest frequency, followed by pre‐haemal, caudal vertebrae and anal fin anomalies, and caudal fin had the lowest anomalies frequency. For the first time, our study determined osteological development and anomalies incidence in larval yellow catfish, which help further investigations into rearing conditions leading to appearance of these anomalies to prevent their incidence.     

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.     

Skeletal development and abnormalities of the vertebral column and fins in larval stage of hatchery‐reared American shad,Alosa sapidissima

The present study describes the skeletal development and the occurrence of deformities in American shad, Alosa sapidissima, larvae from hatching to 45 days after hatching (DAH). The ontogeny of the vertebral column started at 16 DAH (days after hatching), with the formation of the posterior neural and haemal arches, and was complete at 28 DAH. In comparison, the vertebral centra started to form at 16 DAH, with ossification being visible in all centra at 38 DAH. The pectorals were the only fins that formed before the onset of feeding (at 2 DAH), with ossification being complete at 45 DAH. The caudal fin formed at 5 DAH, with ossification being complete at 40 DAH. Dorsal and pelvic fin development began at 6 DAH and 20 DAH respectively. The ossification of both the dorsal and pelvic fins was visible at 45 DAH. The anal fin began forming at 14 DAH, and was complete at 30 DAH. The ossification of the anal fin was complete at 45 DAH. Overall, 22 types of skeletal deformities were detected in about 41% of individuals. Most anomalies were detected in the haemal region, while the fewest anomalies were detected in the anal fin. In addition, the frequency of deformities gradually increased with fish age at the different developmental stages. Our results are expected to contribute baseline information on how rearing conditions impact skeletal development, in addition to identifying potential causative factors of skeletal deformities.     

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.     

A balanced amino acid diet improves Diplodus sargus larval quality and reduces nitrogen excretion

Fish larvae present high amino acid requirements due to their high growth rate. Maximizing this growth rate depends on providing a balanced amino acid diet which can fulfil larval amino acid nutritional needs. In this study, two experimental microencapsulated casein diets were tested: one presenting a balanced amino acid profile and another presenting an unbalanced amino acid profile. A control diet, live feed based, was also tested. Trials were performed with larvae from 1 to 25 days after hatching (DAH). Microencapsulated diets were introduced at 8 DAH in co-feeding with live feed and at 15 DAH larvae were fed the microencapsulated diets alone. Results showed a higher survival for the control group (8.6 ± 1.3% versus 4.2 ± 0.6% and 3.2 ± 1.8%) although dry weight and growth were similar in all treatments. The proportion of deformed larvae as well as the ammonia excretion was lower in the group fed a balanced diet than in the unbalanced or control groups (38.3% deformed larvae in control, 30% in larvae fed unbalanced diet and 20% on balanced diet group). Furthermore, larvae fed the microencapsulated diets presented higher docosahexaenoic acid and arachidonic acid levels. This study demonstrates that dietary amino acid profile may play an important role in larval quality. It also shows that balanced microencapsulated diets may improve some of the performance criteria, such as skeletal deformities, compared to live feeds.     

Is it possible to successfully rear meagre (Argyrosomus regius Asso 1801) larvae without using rotifers?

In hatcheries, meagre Argyrosomus regius larvae still depend on an adequate supply of rotifers and Artemia, as no artificial diet can totally fulfil their nutritional requirements. However, production of live feed is highly expensive and demands intensive labour and specific facilities. This study investigated the effect of a dietary regime without the use of rotifers, to simplify the meagre larval rearing protocol. Two feeding treatments (T₁ & T₂) are compared to investigate their effects on survival and growth of meagre larvae. In T₁, larvae were fed rotifers from 2 to 5 days post hatch (dph), and Artemia from 4 to 15 dph. In T_2, larvae were kept under dark conditions and fed Artemia from 6 to 15 dph. Standard larval length (SL) was significantly higher in T₁ (p < .01) until 8 dph in comparison with larvae reared initially without rotifers. No significant difference in SL was found among treatments (p = .187) at 15 dph. Significant difference was found among treatments in survival rate at 15 dph (p < .003). The survival rate observed at 15 dph in T₂ (30 ± 4.2%) represents an important finding, although the highest survival rate was observed in T₁ (45.0 ± 3.4%). This study showed that it is possible to conduct larval rearing of meagre without using rotifers. Nevertheless, further research efforts are still needed to improve these results in comparison with the common larval rearing protocol.     

Natural carbon stable isotope ratios as indicators of the relative contribution of live and inert diets to growth in larval Senegalese sole (Solea senegalensis)

The relative contributions of live Artemia metanauplii and an inert diet for growth of Senegalese sole larvae and postlarvae were assessed through the analysis of carbon stable isotopes ratios (δ¹³C) in both diets and whole larval tissue. Larvae were reared on four dietary regimes: 100% live prey (rotifers and Artemia), 100% inert formulated diet and two co-feeding regimes of 70:30 and 30:70 ratios of Artemia and inert diet, respectively. Larvae from the live food regime and both co-feeding regimes showed a steep increase in δ¹³C from 10 days after hatching (DAH) as a result of the onset and continuation of Artemia consumption. From 12 DAH fish larvae from all the regimes showed significant isotopic differences as their δ¹³C increased to final asymptotic values of − 15.1, − 15.6 and − 16.3‰ in the live food, 70:30 and 30:70 regimes, respectively. Carbon turnover rates in larvae from both live food and co-feeding regimes were relatively high (0.071 to 0.116 d^− 1) but more than 90% of the observed change in fish tissue isotopic values was accounted for by the retention of carbon in new tissue growth. A two-source, one-isotope mixing model was applied to estimate the nutritional contribution of Artemia and inert diet to postlarvae growth in the co-feeding regimes. At 23 DAH, the relative contribution of live and inert diets to tissue growth in larvae was respectively, 88 and 12% for the 70:30 co-feeding regime and 73 and 27% for the 30:70 co-feeding regime. At 17 DAH, the estimated proportion of tissue carbon derived from the inert diet was higher at 23 and 38% for the 70:30 and 30:70 regimes, respectively. The results suggest that co-feeding regimes in Solea senegalensis larvae may be adjusted to meet ontogenetic changes in the capacity for larvae to utilise inert diets. The contrasting levels of carbon isotope discrimination between diet and tissue in larvae reared on either 100% live feed or 100% inert diet indicate relatively poor utilization of nutrients from the inert diet. The use of isotopic discrimination factors as potential indicators of the digestive physiological performance of a consuming organism in regards to its diet is discussed.     

Effect of an experimental microparticulate diet on the growth, survival and fatty acid profile of gilthead seabream (Sparus aurata L.) larvae

In recent years, a great deal of interest has emerged in the development of microdiets as an economic alternative to live food, in the larval culture of marine fish species. The ability to grow Sparus aurata larvae on a prototype microparticulate diet was examined. To achieve this objective, four feeding regimes differing in the time when the microdiet was introduced (3, 7 or 12 days) and one based exclusively on an inert diet were tested, during the first 22 days of larval life. Significant differences in larval growth were found between the experimental feeding regimes and their corresponding controls (enriched rotifers during the whole experimental period); the larvae in the co-feeding regimes and with an exclusive microparticulate diet were always significantly smaller than larvae fed on rotifers alone. However, the difference was minimised by introducing the inert diet at a later date. A lower survival was found in larvae with a co-feeding regime, in comparison with the control treatments and the survival was significantly lower in larvae fed exclusively on a microparticulate diet. The fatty acid analysis revealed that the experimental microencapsulated diet and the rotifers enriched with Protein Selco® presented relatively similar fatty acid content. In spite of the slightly higher (n?3)/(n?6) and Docosahexaenoic acid (DHA)/Eicosapentaenoic acid (EPA) ratios and somewhat lower highly unsaturated fatty acid (HUFA) content found in the inert diet, the fatty acid composition of the diets cannot explain the differences found in larval performance. The results revealed that the complete replacement of live prey with the tested microparticulate diet is still not possible in S. aurata larval rearing. Nevertheless, better growth and survival results and a substantial reduction in the daily supply of live food can be achieved with a combination of microdiet and live prey.     

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.     

Effect of different enrichment products rich in docosahexaenoic acid on growth and survival of meagre,Argyrosomus regius (Asso, 1801)

Live prey used in aquaculture to feed marine larval fish – rotifer and Artemia nauplii – lack the necessary levels of n‐3 polyunsaturated fatty acids (n‐3 PUFA) which are considered essential for the development of fish larvae. Due to the high voracity, visual feeding in conditions of relatively high luminosity, and cannibalism observed in meagre larvae, a study of its nutritional requirements is needed. In this study, the effect of different enrichment products with different docosahexaenoic acid (DHA) concentrations used to enrich rotifers and Artemia metanauplii have been tested on growth, survival, and lipid composition of the larvae of meagre. The larvae fed live prey enriched with Algamac 3050 (AG) showed a significantly higher growth than the rest of the groups at the end of the larval rearing, while the larvae fed preys enriched with Multigain (MG) had a higher survival rate. DHA levels in larvae fed prey enriched with MG were significantly higher than in those fed AG‐enriched prey. High levels of DHA in Artemia metanauplii must be used to achieve optimal growth and survival of meagre larvae.     

Early weaning in meagre Argyrosomus regius: Effects on growth,survival, digestion and skeletal deformities

Meagre Argyrosomus regius is considered a new species for the diversification of finfish aquaculture in the Mediterranean. Several bottlenecks have been identified by producers, and among them, the necessity to establish early weaning protocols to reduce production costs. In this study, two experiments were carried out with meagre larvae from 2 to 35 days post hatch (dph) using different weaning strategies, including the early introduction of artificial diets and the reduction of Artemia metanauplii to half of the normal amounts. A high frequency of cannibalism and high variability in growth rate and survival were obtained in one of the trials and several changes were introduced (reduction of light intensity, higher frequency of food distribution) in the second trial to increase the survival rate. In both trials, weaning started before the complete morphological and functional development of the stomach; thus, pancreatic enzymes, mainly trypsin and lipase tended to be more active in early weaned larvae compared to the control groups. Early weaning delayed the development of the stomach formation and secretion of acid proteases, which may explain the lower growth rates observed in our study. The effect of weaning on skeletal development was also studied and in this sense the results obtained showed no major influence of the early weaning on the incidence of skeletal deformities. Weaning of meagre larvae can be performed as early as 12 dph, but important aspects such as avoiding cannibalism and co‐feeding live prey and artificial diets for at least 5 days were recommended.     

The importance of dietary HUFA for meagre larvae (Argyrosomus regius; Asso, 1801) and its relation with antioxidant vitamins E and C

Despite the interest of meagre (Argyrosomus regius) as a fast‐growing candidate for Mediterranean aquaculture diversification, there is a lack of information on nutrition along larval development. Importance of highly unsaturated fatty acids (HUFA) and the antioxidant vitamins E and vitamin C has not been investigated yet in this species. Six diets with two levels of HUFA (0.4% and 3% dw), two of vitamin E (1500 and 3000 mg kg^?1) and two of vitamin C (1800 and 3600 mg kg^?1) were fed to 15 dah meagre larvae. Larval growth in total length and dry body weight was significantly lowest in larvae fed diet 0.4/150/180 and showed few lipid droplets in enterocytes and hepatocytes and lower HUFA contents than the initial larvae. Increase in dietary HUFA up to 3%, significantly improved larval growth and lipid absorption and deposition. Besides, among fish fed 3% HUFA, increase in vitamin E and vitamin C significantly improved body weight, as well as total lipid, 22:6n‐3 and n‐3 fatty acids contents in the larvae. Thus, the results showed that 0.4% dietary HUFA is not enough to cover the essential fatty acid requirements of larval meagre and a high HUFA requirement in weaning diets is foreseen for this species. Besides, the results also pointed out the importance of dietary vitamin E and C to protect these essential fatty acids from oxidation, increase their contents in the larvae and promote growth, suggesting high vitamin E and C requirements in meagre larvae (higher than 1500 and 1800 mg kg^?1 for vitamin E and vitamin C respectively).     

New insight on vertebral anomalies in cultured Senegalese sole (Solea senegalensis,Kaup) at early stages of development

Senegalese sole (Solea senegalensis, Kaup) is a promising flatfish species in aquaculture. However, skeletal anomalies are still a great concern in sole farming. Investigation of this issue is crucial to improving larval quality and optimizing production. The aim of this study was to thoroughly assess anomalies in the rachis of reared sole at early developmental stages. Sole (n = 507) were sampled at 31 or 32 days after hatching (dah). The specimens were stained with alcian blue and alizarin red and evaluated for the detection of vertebral deformities. Most fish presented 9:34:3 vertebrae in abdominal, caudal and caudal complex regions, respectively. Remarkably, all specimens showed at least one spinal anomaly. Alterations of neural/haemal elements, as well as deformities of hypurals, parhypural and epural, were recurrent. Vertebral body anomalies and/or vertebral column deviations were identified in 52% of the individuals. Vertebral deformations and fusions were common, especially in caudal complex. ‘Minor’ anomalies were predominant, and some of the detected disorders might be a result of non‐/low‐pathological processes. These results contribute a new insight into the main skeletal anomalies affecting cultured sole larvae. Further research is required to determine their impact on fish welfare and external appearances at commercial stages.     

Tyrosine and phenylalanine supplementation on Diplodus sargus larvae: effect on growth and quality

Phenylalanine is the precursor of tyrosine, which is involved in the synthesis of several molecules with key roles in the regulation of metabolism and growth, stress response and pigmentation. In this study, three experimental diets were tested: an amino acid (AA) balanced diet supplemented with phenylalanine, another supplemented with phenylalanine and tyrosine and a non‐supplemented AA balanced diet. Rotifers were enriched with liposomes encapsulating free AA in order to obtain a balanced AA profile. The experimental diets resulted in similar larval survival, growth, enzyme activities of AA catabolism and nitrogen excretion in all treatments. High levels of skeletal deformities were registered and significant differences were found between the control and the phenylalanine treatment for the percentage of vertebral compressions in the trunk region of the vertebral column (30% in the control and 5% in the phenylalanine group). A significantly higher survival to a temperature stress test was found for larvae fed the diet supplemented with phenylalanine and tyrosine. The results suggest that supplementation of phenylalanine/tyrosine in fish diets may be useful in order to reduce skeletal deformities and mortalities caused by stress. The present study confirms that AA requirements may be sufficient for covering growth and survival but insufficient to cover other metabolic processes.     

Larval rearing of the giant Azorean barnacle,Megabalanus azoricus (Pilsbry, 1916): feeding trials,larval development and settlement on artificial substrata

A series of experiments were conducted to obtain an efficient larval rearing protocol for Megabalanus azoricus. The first part of this study investigates the effect of microalgae‐based diets on survival and larval development. Mono and mixed‐diets were tested at 20 ± 1°C, in a sequence of 11‐day feeding experiments. The second part presents a preliminary study on the influence of a biofilm on recruitment and use of oyster spat collectors in a mass rearing system. A photographic record of larval development and a brief reference to the diagnostic features that enable quick larval staging are also presented, along with morphometric measurements. Of the microalgae tested (Chaetoceros sp., Chloromonas sp., Dunaliella sp., T‐Isochrysis sp. and Skeletonema sp.) the mixed‐diet Skeletonema sp. with T‐Isocrysis sp. showed the highest survival percentages: total survival ranged from 79.7 to 85.7% and 69.7–80.0% of nauplii were in stage VI after 11 days of rearing. Cypris were also present, but only represented 5.3% of the survivors at most. In the mass rearing system juveniles were found settled in the collectors after 25 days, at 20 ± 1°C. However recruitment was less than 1%. Preliminary results showed no settlement preference towards collectors with biofilm. Nevertheless, this study provides the first record of M. azoricus settlement under laboratorial conditions and represent a starting point for future larval rearing studies.     

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.     

Functional changes in digestive enzyme activities of meagre (Argyrosomus regius; Asso, 1801) during early ontogeny

The ontogenesis of main pancreatic and intestinal enzymes was investigated in the recent promising Mediterranean candidate species of meagre, Argyrosomus regius, during larval development until 40 days after hatching (DAH). The green-water technique was carried out for larval rearing. Whole-body homogenates were used for enzymatic analysis in larvae younger than 15 DAH; after this date, older larvae were dissected into two segments as pancreatic and intestinal segment. Trypsin was detected as early as hatching and sharply increased concurrently with age and exogenous feeding 15 DAH, but constant decline was observed until the end of experiment. Amylase was determined at 2 DAH and sharply increased 10 DAH. Then, slight decreases were found between 10 and 15 DAH, and then slow alterations were continued until the end of the experiment. Lipase was firstly measured on day 3; then, sudden decline was observed between 20 and 25 DAH. After this date, slow fluctuations were maintained until the end of the experiment. Pepsin was firstly assayed 15 DAH related to gastric gland secretion and sharply increased 30 DAH. Then, it slowly varied until end of the experiment. Enzymes of brush border membranes, alkaline phosphatase and aminopeptidase N showed similar pattern on specific activities during the first 10 days. Thereafter, while specific activity of alkaline phosphatase slightly decreased 15 and fluctuated until 20 DAH, aminopeptidase N activity slowly increased 20 DAH. Then, activity of alkaline phosphatase and aminopeptidase N constantly increased 30 DAH, indicating maturation of the intestinal digestive process, and also, these activities continued to slowly increase until the end of the experiment. The specific activity of cytosolic peptidase, leucine–alanine peptidase, smoothly increased on day 8, then fluctuated until 15 DAH. After this date, in contrast to enzymes of brush border membranes, it sharply decreased 25 DAH and continued to gradually decline until the end of the experiment. These converse expressions were indicative of a maturation of enterocytes and the transition of an adult mode of digestion.     

The effect of live feeds bathed with the red seaweed <Emphasis Type="Italic">Asparagopsis armata</Emphasis> on the survival,growth and physiology status of <Emphasis Type="Italic">Sparus aurata</Emphasis> larvae

Larval rearing is affected by a wide range of microorganisms that thrive in larviculture systems. Some seaweed species have metabolites capable of reducing the bacterial load. However, no studies have yet tested whether including seaweed metabolites on larval rearing systems has any effects on the larvae development. This work assessed the development of Sparus aurata larvae fed preys treated with an Asparagopsis armata product. Live prey, Brachionus spp. and Artemia sp., were immersed in a solution containing 0.5% of a commercial extract of A. armata (Ysaline 100, YSA) for 30 min, before being fed to seabream larvae (n = 4 each). In the control, the live feed was immersed in clear water. Larval parameters such as growth, survival, digestive capacity (structural-histology and functional-enzymatic activity), stress level (cortisol content), non-specific immune response (lysozyme activity), anti-bacterial activity (disc-diffusion assay) and microbiota quantification (fish larvae gut and rearing water) were monitored. Fish larvae digestive capacity, stress level and non-specific immune response were not affected by the use of YSA. The number of Vibrionaceae was significantly reduced both in water and larval gut when using YSA. Growth was enhanced for YSA treatment, but higher mortality was also observed, especially until 10 days after hatching (DAH). The mortality peak observed at 8 DAH for both treatments, but higher for YSA, indicates larval higher susceptibility at this development stage, suggesting that lower concentrations of YSA should be used until 10 DAH. The application of YSA after 10 DAH onwards promotes a safer rearing environment.     

Effect of increased dietary protein level in meagre (Argyrosomus regius) juvenile growth and muscle cellularity

In this study, two different dietary protein levels were tested (500 and 600 g/kg) in meagre juveniles of approximately 2 g, to determine whether a higher body protein diet would favour muscle growth. Results showed that meagre survival and growth were not affected by different levels of dietary protein. However, a higher content of protein was observed in the body composition of meagre fed the P60 diet (656 g/kg versus 624 g/kg). When fish were 82 days after hatched (DAH), protein retention efficiency was significantly higher in fish that were fed the P50 diet (37.1% versus 32.5%). Muscle cellularity changed considerably from the beginning to the end of the trial. The contribution of fibre hyperplasia to muscle growth was reduced although still accounted for 30% in 96 DAH meagre. Several differences were found in the frequency of fibres with areas between 1,000 μm² and 2,200 μm² between younger and older meagre, but no effect was found associated with dietary protein. This study suggests that neither growth nor muscle cellularity of fast‐growing juvenile meagre is positively affected by an increased dietary protein levels.