Early introduction of an inert diet and unenriched Artemia enhances growth and quality of Atlantic cod (Gadus morhua) larvae

The effects of two weaning diets and different weaning protocols on growth, survival, skeletal deformity and gut morphology of Atlantic cod larvae were studied in four groups from 16 to 45 days posthatch (dph). Cod larvae in groups 1 (early weaning with control diet) and 2 (early weaning with experimental diet) were used to evaluate the effects of different polar lipid content of weaning diets on larval and juvenile performance. Cod larvae in groups 2, 3 (early weaning with experimental diet + cofeeding with Artemia) and 4 (earlier weaning with experimental diet and earlier cofeeding with Artemia) were used to evaluate the effects of early introduction of dry diet and Artemia. From 45 to 170 dph, cod juveniles from all four groups were reared using a standard feeding protocol. No significant differences in growth, survival, deformities and gut morphology were found between cod larvae and juveniles from groups 1 and 2. Cod larvae fed on cofeeding regime with Artemia nauplii (groups 3 and 4) were bigger and had lower frequencies of jaw and neck deformities and higher foregut microvillus circumference than cod larvae from group 2. Our results demonstrate the importance of proper weaning protocols in producing better quality cod juveniles.     

Comparison of dietary phospholipids and neutral lipids: effects on gut, liver and pancreas histology in Atlantic cod (Gadus morha L.) larvae

The aim of the present study was to compare effects of dietary n-3 highly unsaturated fatty acids (HUFA) being incorporated in the phospholipid (PL) or in the neutral lipid (NL) fraction of the larval feed, on larval growth and histology of digestive organs in Atlantic cod ( Gadus morhua L.) larvae. Three isoproteic and isolipidic diets, labelled according to the percentage of n-3 docosahexaenoic acid and eicosapentaenoic acid contained in NL1 or in PL1 and PL3 of the diets, were fed to cod larvae from 17 days post hatching (dph) to 45 dph.
In the liver, hepatocytes and their nuclei were smaller in NL1 larvae compared with the PL larvae; the mitochondrial membrane structures were less dense and the amount of lipids observed in the liver was significantly higher in NL1 larvae compared with the PL3 larvae. The liver and gut size was related to larval size, with no differences between the larval groups. The results demonstrated that the essential fatty acids were more beneficial for cod larvae when they were incorporated in the dietary polar PL rather than in the NL, and that the n-3 HUFA requirements in cod larvae is possibly higher than that in the PL1 diet.     

Development and evaluation of microparticulate diets for early weaning of Atlantic cod Gadus morhua larvae

Early weaning trials were conducted with cod larvae to investigate the effectiveness of microparticulate diets (microbound and microcoated) with and without lipid-walled capsules (LWCs). The microparticulate diets were evaluated by measuring physical parameters of the diet in the water column (leaching and settling rate), palatability (intestinal fullness), performance of the diet (survival and growth), and examination of the diet in the larval intestine (histological analysis). A feeding trial was conducted using four experimental diets (carrageenan microbound diet, carrageenan microbound diet with LWCs, zein microcoated diet and zein microcoated diet with LWC), one commercial diet (BioKyowa: A-250) and a live feed control (rotifers and Artemia ). Survival of cod larvae to 39 days post-hatch ranged from 5 to 10% with the experimental diets, 22.9% with the BioKyowa diet, and 36.5% with live prey. There was evidence of food absorption with all diets in the form of lipid vacuoles in the midgut and supranuclear vacuoles in the hindgut. Large vacuoles in the midgut were more abundant in the enterocytes of larvae fed the experimental diets compared with larvae on the BioKyowa diet and the live feed control. Based on observations of intestinal fullness, the experimental diets appeared to be less palatable than the BioKyowa diet. As a result, it took longer to wean the larvae and higher mortality was experienced during weaning. Once successfully weaned, the experimental diets yielded growth rates equivalent to larvae feeding on the commercial diet for the remainder of the experiment.     

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.     

A method to determine protein digestibility of microdiets for larval and early juvenile fish

A method to evaluate protein quality using in vivo methods was developed for larval fish. FluoSpheres^® fluorescent microspheres (10 μm) were incorporated into two test diets, our standard zein microdiet (ZMD) and a microdiet with identical ingredients except for the replacement of high quality fish meal with the same product cooked for 24 h at 80 °C (ZMD-CF). Several trials were performed to design a reliable method to test digestibility using FluoSpheres^® as a marker. The developed in vivo technique was tested on 35 days posthatch (dph) larval Atlantic cod ( Gadus morhua L.) and two tropical fish species in the early juvenile stage. The method took into account loss of total protein to the faecal pellet and water column. Apparent digestibility of protein in larval cod fed ZMD was significantly higher than that of larvae fed ZMD-CF ( P  < 0.05). A growth study to validate differences between the two diets showed significant differences in growth and survival of larvae fed ZMD versus ZMD-CF ( P  < 0.05). Further validation of our results was indicated through the use of a pH-stat method using enzymes extracted from 35 dph larval cod guts. This novel technique will be advantageous for researchers to evaluate feed ingredients for larval marine fish and is adaptable to many different areas of larval fish nutrition.     

Effect of cultivated copepods (Acartia tonsa) in first‐feeding of Atlantic cod (Gadus morhua) and ballan wrasse (Labrus bergylta) larvae

The aim of this study was to compare the nutritional composition and effects of short periods with cultivated copepod nauplii versus rotifers in first‐feeding. Atlantic cod (Gadus morhua) and ballan wrasse (Labrus bergylta) larvae were given four different dietary regimes in the earliest start‐feeding period. One group was fed the copepod Acartia tonsa nauplii (Cop), a second fed enriched rotifers (RotMG), a third fed unenriched rotifers (RotChl) and a fourth copepods for the seven first days of feeding and enriched rotifers the rest of the period (Cop7). Cod larvae were fed Artemia sp. between 20 and 40 dph (days posthatching), and ballan wrasse between 36 and 40 dph, with weaning to a formulated diet thereafter. In addition to assessing growth and survival, response to handling stress was measured. This study showed that even short periods of feeding with cultivated copepod nauplii (7 days) had positive long‐term effects on the growth and viability of the fish larvae. At the end of both studies (60 days posthatching), fish larvae fed copepods showed higher survival, better growth and viability than larvae fed rotifers. This underlines the importance of early larval nutrition.     

You are what you eat? Differences in lipid composition of cod larvae reared on natural zooplankton and enriched rotifers

A laboratory experiment with larval cod fed natural zooplankton and enriched rotifers was carried out to investigate the effects of feed type on fatty acid (FA) and lipid composition. A divergence in FA composition was observed within a week of feeding on respective diets, and subsequent transfers of larvae from one feeding regime to the other also confirmed a rapid change in FA composition towards that of the newly provided feed source. The rapid change in FA composition after switching diet is in part expected to be due to the high growth and tissue turnover of cod larvae during the early life stages and provides an opportunity to assess recent feeding history by means of FA analysis. The FA contents also varied between the main analysed lipid classes, with relatively higher eicosapentaenoic acid levels in neutral lipids than in polar lipid classes. Although zooplankton contained notably more polar lipids and less neutral lipids than enriched rotifers, the relative amounts of polar and neutral lipids in larvae from respective prey groups were similar, signifying that the larval composition only partly reflects what they have been eating. Still, the FA composition of the previous diet was still discernible in larvae after 1–2 weeks of transfer to a new diet or after weaning to a formulated feed. The potential long‐term effects of these nutritional differences are discussed.     

Biomarkers of bone development and oxidative stress in gilthead sea bream larvae fed microdiets with several levels of polar lipids and α‐tocopherol

Although dietary marine phospholipids are able to improve culture performance of marine fish larvae in a further extend than soybean lecithin, both types of phospholipids (PL) markedly increase oxidative risk. The inclusion of a fat‐soluble antioxidant such as the vitamin E α‐tocopherol could allow a better control of oxidative stress. The objective of this study was to determine the combined effect of graded levels of α‐tocopherol with different levels and sources of krill phospholipids (KPL) and soybean lecithin (SBL) on growth, survival, resistance to stress, oxidative status, bone metabolism‐related genes expression and biochemical composition of sea bream larvae. Sea bream larvae were completely weaned at 16 dph and fed for 30 days seven microdiets with three different levels of PL (0, 40 and 80 g kg^?1 diet) and two of α‐tocopherol 1500 and 3000 mg kg^?1 diet. Sea bream larvae fed diets without PL supplementation showed the lowest survival, growth and stress resistance, whereas increase in PL, particularly KPL, markedly promoted larval survival and growth. However, feeding SBL markedly increased TBARs and GPX gene expression increasing the peroxidation risk in the larvae. Besides, KPL inclusion improved incorporation of n‐3 HUFA and, particularly, EPA into larval tissues, these fatty acids being positively correlated with the expression of BMP‐4, RUNX 2, ALP, OC and OP genes and to bone mineralization for a given larval size class. The increase in dietary α‐tocopherol tends to improve growth in relation to the n‐3 HUFA levels in the diet, denoting the protective role of this vitamin against oxidation. Indeed, dietary α‐tocopherol decreased the oxidative stress in the larvae as denoted by the reduction in larval TBARs contents and gene expression of SOD and CAT, but not GPX. Thus, increase in dietary α‐tocopherol effectively prevented the formation of free radicals from HUFA, particularly EPA, but did not affect the incidence of bone anomalies or the expression of genes related to osteogenetic processes.     

Evaluation of weaning performance of California halibut (Paralichthys californicus) larvae using growth,survival and digestive proteolytic activity

This study evaluated weaning success of California halibut, Paralichthys californicus, larvae onto a microdiet at various stages of development utilizing growth, survival and digestive enzyme activity. Weaning onto a microdiet was evaluated at 16, 26, 36 and 46 days posthatch (dph). Alkaline and acid proteases and leucine aminopeptidase activities were measured after weaning and compared between the weaned treatment and Artemia‐fed controls. Survival was significantly lower in the microdiet‐fed treatments compared to the control groups. Growth was significantly reduced in all weaning treatments compared to the control, except for the 46 dph group. No differences in enzyme activities were detected between treatment and diet at 16 and 26 dph; however, activities were higher for the microdiet‐fed larvae at 36 and 46 dph. This study demonstrates that California halibut larvae possess a differentiated and effective digestive system early in development and can be weaned with relative success (>40% survival) before completion of the metamorphosis (i.e., 36 dph). The lack of weaning success at an early date cannot be entirely because of the absence of a functional stomach but could be related to, among other factors, the low‐microdiet ingestion rates observed and higher leaching of smaller microdiets.     

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.     

Proteome profile comparison of two differently fed groups of Atlantic cod (Gadus morhua) larvae

Proteome analysis was used to study the effects of feeding early Atlantic cod (Gadus morhua) larvae with a saithe (Pollachius virens) protein hydrolysate (SPH). Protein hydrolysates have previously been shown to beneficially affect fish larval development. Feeding was initiated on day 2 post hatch (ph) or as soon as the larvae opened their mouth and the protein expression was monitored 4 days later or in 6‐dph cod larvae. The results demonstrated changes in the abundance of 13 protein spots in the cod larvae fed SPH. Of these, seven protein spots were up‐regulated and six protein spots showed down‐regulation. Five of the up‐regulated proteins in cod larvae are known to be involved in energy metabolism. A few early larval specific proteins were down‐regulated in the SPH‐fed cod larvae possibly because of an enhanced development in this group relative to the control group. Two trypsin isoforms were detected within the cod larval proteome. The detection of the trypsin spots was made possible by co‐electrophoresis of known cod trypsins with the cod larval protein extract. Surprisingly, no difference in trypsin content was observed between the SPH‐fed and the control larval groups.     

Marine raw material choice,quality and weaning performance of Ballan wrasse (Labrus bergylta) larvae

Previous results show that weaning success of Ballan wrasse larvae greatly depends on the quality of the dietary marine raw materials. In the present study, six moist or agglomerated experimental weaning diets containing different combinations of high‐quality marine raw materials, being fish meal (FM), cod muscle meal (CMM), shrimp meal (SM) and krill hydrolysate (KH), were tested in a 2‐month weaning trial with Ballan wrasse larvae of 34.5 mg initial body weight. Larvae performance was good in all dietary treatments except those fed diet D1 containing only FM. The Ballan wrasse larvae fed weaning diets D4 and D5 containing FM and either SM or KH, respectively, had the highest final body weight (0.7 g) but also the highest mortality (50%). Best weaning survival (77%) was obtained using the dry agglomerated diet D3 containing CMM and SM. During the first month, fish survival correlated positively with dietary free amino acid and soluble protein levels and negatively with the combined levels of dietary lipid oxidation metabolites and ethoxyquin. During the second month, mortality rates were lower in all treatments. Fish larvae final body weight correlated negatively with total dietary fatty acids and positively with dietary cholesterol, phosphorous and DHA/EPA ratio.     

Vertebral deformity in cultured Atlantic cod larvae: ontogeny and effects on mortality

The effects of different egg incubation densities on the incidences of vertebral deformities in Atlantic cod larvae were investigated. Cod eggs were incubated at four different densities, 3, 6, 12 and 48 mL eggs L⁻¹, of water.  When all the eggs hatched, larvae were reared in 30 L glass aquaria. Larval samples were taken at 0, 14, 42 and 56 days post hatch (dph) for deformity analysis. Larval samples were stained using bone and cartilage staining methods to determine vertebral deformity. Incubation densities did not have any significant effects on vertebral deformities in Atlantic cod larvae. However, the incidence of larval vertebral deformity was high at hatch and decreased as the larvae grew older until 42 dph, indicating selective mortality of deformed larvae during this period. Larvae at 56 dph, however, showed an increase in the incidence of vertebral deformity, indicating a possible nutritional or prey-type effect. To our knowledge, no studies have documented the occurrence of variable patterns in vertebral deformities in cod at various developmental stages. Overall, our results suggest that broodstock husbandry, genetics and/or nutrition could play a major role in causing vertebral deformities in Atlantic cod at hatch; however, nutrition and prey type may play a major role during metamorphosis.     

Relationship between dietary phospholipid classes and neutral lipid absorption in newly-weaned turbot, shape Scophthalmus maximus

A 28-day feeding trial was conducted for comparing the effect of different dietary phospholipid (PL) classes on the growth of post-larval turbot and on the incorporation of dietary neutral lipid fatty acids into their body lipids. Prior to the experiment the turbot were weaned for one week on a PL-free diet. The nine experimental diets were isolipidic and contained an equal amount of highly unsaturated fatty acids in the form of ethyl esters. They differed by their PL content (0, 1 or 2%) and by the PL class composition of the added soybean PL fractions.Compared to the PL-free diet, diets enriched with phosphatidylcholine (PC) resulted in a better growth, a higher triglyceride content (% body dry matter) and increased levels of docosahexaenoic acid (% total fatty acids) in each of the examined body lipid classes (neutral lipid, phosphatidylcholine, phosphatidylethanolamine and phosphatidylinositol). The effects of the other soybean PL fractions were less explicit than those noted for soybean PC.The results support the idea that dietary PC plays a role in the intestinal absorption of neutral lipid fatty acids. This might, at least partially, explain the superiority of PC for enhancing growth. Abbreviations: DHA – docosahexaenoic acid (22:6n-3); EPA – eicosapentaenoic acid (20:5n-3); HUFA – highly unsaturated fatty acid; PA – phosphatidic acid; PC – phosphatidylcholine; PE – phosphatidylethanolamine; PI – phosphatidylinositol; PL – phospholipid; PS – phosphatidylserine; PUFA – polyunsaturated fatty acid.     

Influence of larval co-feeding with live and inert diets on weaning the tongue sole Cynoglossus semilaevis

The tongue sole Cynoglossus semilaevis, an inshore fish in China, has showed great potential in aquaculture recently. However, poor survival was recorded during the period of weaning from live Artemia to artificial diets. In this paper, the influence of co‐feeding larvae with live and inert diet on weaning performance was described. The C. semilaevis larvae were reared at 21 ± 1 °C and fed four different feeding regimes from 6 days post‐hatching (dph): A, Artemia (10 individuals mL^?1); B, Artemia (5 individuals mL^?1); C, mixed diet (10 Artemia individuals mL^?1 and 12 mg L^?1 inert diet); and D, mixed diet (5 Artemia individuals mL^?1 and 12 mg L^?1 inert diet). Rotifers were also supplied in all cases during the first days of feeding. Mixed diets of commercial formulated feed and live prey (rotifers and Artemia) allowed larvae to complete metamorphosis, achieving similar specific growth rate (SGR) (18.5 ± 1.4% and 18.7 ± 1.6%) and survival (40 ± 7.6% and 48.5 ± 6.8%) compared with larvae fed on live feed alone (SGR of 18.3 ± 1.2%, 19.3 ± 1.9% and survival of 41.2 ± 11.3%, 38 ± 4.9%). However, in metamorphosed fish, when live feed was withdrawn on 31 dph, there was significant difference (P < 0.05) in survival and growth among treatments. Metamorphosed fish, previously fed mixture diets during larval stages, had similar survival (62.1 ± 7.6% and 62.8 ± 3.9% for regimes C and D, respectively) but higher than that obtained for fish that previously fed on live feed (49.3 ± 2% and 42.1 ± 3.9% for regimes A and B, respectively) after weaning (day 60). The SGR of weaned fish previously fed live feed was similar (3.1 ± 0.6% and 2.92 ± 0.6% for regimes A and B, respectively) but lower than that recorded for fish that was fed from day 6 to day 30 on the mixed diet (4.5 ± 1.1% and 4.9 ± 0.3% for regimes C and D, respectively). It is suggested that weaning of C. semilaevis from early development would appear to be feasible and larval co‐feeding improves growth and survival.     

Influence of dietary phospholipids on early ontogenesis of fish

The aim of this paper is to provide explanations of how dietary phospholipid (PL) globally improves fish larval development, including growth and survival, digestive functions and skeletal development, and to propose optimal PL levels and sources in fish larval diets. Dietary incorporation of 8–12% PL related to dry matter (d.m.) promotes growth and enhanced survival in various species. Marine source PL, incorporating highly unsaturated fatty acids, was most efficient than soybean lecithin. This beneficial effect was explained by an enhancement in digestive functions, assessed by digestive enzyme activities and histomorphology. Nevertheless, 1.5–2.5% highly unsaturated fatty acids related to diet d.m. supplied by PL improved growth, survival and skeletal development, while 5% induced different skeletal deformities. The high incidence of deformities was associated with the down-regulation of genes involved in development, such as RXRα, RARα, RARβ and BMP-4, observed in the early stages in larvae fed a high highly unsaturated fatty acids level.     

Effects of two commercial microdiets on growth and survival of barramundi (Lates calcarifer Bloch) larvae within various early weaning protocols

Because of high costs and labour requirements along with the highly variable nutritional value of live feeds, we investigated the possibility of early weaning for barramundi (Lates calcarifer Bloch) larvae aimed at reducing the use of Artemia. Two commercial microdiets, Gemma Micro (Skretting, Australia) and Proton (INVE, Belgium) were compared for growth and survival of larvae using three weaning protocols, until 33 days posthatch (dph). Enriched rotifers were fed to larvae in all protocols through mouth opening until 21, 18 and 30 dph (protocols 1, 2 and 3, respectively). At 13 dph, enriched Artemia metanauplii were introduced to weaning protocols 1 and 2, and continued until 29 and 24 dph, respectively, whereas protocol 3 did not receive Artemia. Microdiet was initiated at 20, 16 and 13 dph in protocols 1, 2 and 3, respectively. Barramundi larvae grew successfully to 33 dph when co‐fed rotifers and microdiet, and significantly larger larvae resulted from feeding Gemma Micro rather than Proton, when Artemia were not used. However, larvae weaned onto Proton using a longer period of Artemia provision were significantly larger than larvae reared according to all other protocols. Survival was significantly higher in all Gemma Micro protocols when compared with Proton protocols. This was in part due to higher cannibalism when using Proton compared with Gemma Micro (22.8 ± 0.9% and 9.2 ± 0.6%, respectively). Cannibalism became more noticeable in all protocols when the larvae reached 7–8 mm standard length and further increased after the cessation of live feed. Tank biomass production was the highest when larvae were weaned onto Gemma Micro including a short period of Artemia provision as a result of a combination of high growth and survival. However, similar biomass production resulted when larvae were weaned directly from rotifers onto Gemma Micro and/or from a prolonged Artemia period onto proton. The success of weaning barramundi larvae directly to microdiet from rotifers, thus eliminating the need for Artemia, was influenced by the microdiet. Relatively higher levels of free amino acids and lipids were believed to contribute to increasing larval growth and survival. Larvae that were fed Gemma Micro showed higher growth when Artemia were utilized for a shorter period, while Proton‐fed larvae benefited from an extended Artemia feeding period.     

Broodstock diet with water and astaxanthin improve condition and egg output of brood fish and larval survival in Atlantic cod,Gadus morhua L.

Broodstock of Atlantic cod were fed with three diets; astaxanthin supplemented (100 ppm kg^?1) hydrated diet (30% water) (HWHAs), no additional astaxanthin supplemented (50 ppm kg^?1) hydrated diet (30% water) (HWLAs) and a control diet (50 ppm kg^?1 astaxanthin and 10% water). Throughout the spawning period, eggs were collected daily and fertilization success and proportion of embryos with normal cleavage were recorded. Larvae were reared using standard culture methods. Larval growth was recorded every 10 days and larval survival was recorded at 50 days post hatch. In general, broodstock fed with diets containing higher water content (HWHAs and HWLAs) and higher astaxanthin (HWHAs) had higher egg production and efficiency of egg output, lower egg incubation mortality and higher larval growth and survival. Broodstock fed with higher water content diets consumed less feed compared to control group. Also, broodstock fed with HWHAs and HWLAs recovered faster in terms of body condition than the control group. The egg bound mortality of female fish was significantly higher in control broodstock compared to broodstock fed with HWHAs and HWLAs. Broodstock fed with higher astaxanthin (HWHAs) had significantly higher fertilization success than other two groups however; no differences were found in proportion embryos with normal cleavage among different groups. Larval survival was significantly higher in HWHAs broodstock compared to other two groups. Our results showed that increased water content and astaxanthin levels in the diet could improve the condition and fecundity of Atlantic cod broodstock.     

Applying the ontogeny of digestive enzyme activity to guide early weaning of pigfish,Orthopristis chrysoptera (L.)

Weaning marine fish larvae from live prey to a dry microdiet is an important step towards optimizing the commercial production, but early weaning is constrained by the lack of sufficient digestive enzymes at first feeding. This study quantified the activity of five digestive enzymes throughout the larval period of pigfish (Orthopristis chrysoptera [L.]) to assess ontogenetic changes in digestive abilities, and then trials were conducted that determined the optimal time for weaning. The activity of all digestive enzymes was low or undetectable at first feeding (3 days post hatching, dph; 2.5 mm standard length, SL). A substantial increase in activity occurred at 5.7 mm SL (17 dph), 6.9 mm SL (21 dph), 7.7 mm SL (23 dph), 8.4 mm SL (25 dph) and 11.2 mm SL (30 dph) for bile salt‐dependent lipase, trypsin, chymotrypsin, amylase and acid protease respectively. During the weaning experiment, larvae were co‐fed live prey and microdiet beginning 15 dph (4.8 mm SL). Live prey was withdrawn from the diet at 24, 28, 32 or 36 dph, with the control receiving live prey and microdiet throughout (to 43 dph). There were no significant differences in mean final SL among treatments, but survival was significantly lower when larvae were weaned at 24 dph compared to 32–43 dph. Based on the digestive enzyme activity and survival, weaning larval pigfish at 32 dph (11.7 mm SL) when reared at 24°C is recommended.     

Effects of early weaning strategies on growth,survival and digestive enzyme activities in cobia (<Emphasis Type="Italic">Rachycentron canadum</Emphasis> L.) larvae

The effects of weaning strategies of cobia (Rachycentron canadum L.) larvae to commercial microdiets, either from rotifers or from Artemia, on growth, survival and enzymatic digestive capacity, were investigated. In the first experiment, cobia larvae were weaned from rotifers by co-feeding with a microdiet (Otohime) from 8, 13 or 20 days post-hatching (dph). The larvae in the control treatment were fed rotifers (2–12 dph), Artemia nauplii from 7 dph, and co-fed with the microdiet from 20 dph. In the second experiment, the larvae were weaned from Artemia, which was fed to the larvae from 7 dph, by co-feeding with a microdiet (NRD) from 8, 13 or 18 dph. The larvae in control treatment were fed rotifers, then Artemia to the end of the experiment (28 dph). Weaning of cobia larvae onto a microdiet directly from rotifers significantly reduced growth, survival and digestive capacity of the larvae and did not lead to larval acceptance of the microdiet, compared to those weaned from Artemia in the first experiment. Early weaning of cobia larvae onto NRD microdiet (on 8 or 13 dph) from Artemia in the second experiment also reduced growth, survival rate and gut maturation index, compared to those fed live feed. With available microdiets, weaning of cobia larvae could start from Artemia at around 18 dph in order to obtain comparable growth, survival and gut maturation to larvae fed live feed.