Ontogeny of pancreatic enzymes in larval red drum Sciaenops ocellatus

The growth, survival and trypsin, lipase and amylase activities of red drum larvae were measured in two experiments. For the first trial, a group was fed live prey only (L) and another group was fed a combination of a microparticulate diet (MPD) and live food (L-MP). For the second growth trial a group fed the MPD only (MP) and a starvation group (ST) were examined in addition to the L and L-MP treatments. Enzyme activities of live prey were measured to estimate their possible contribution to larval digestion. No significant ( P  > 0.05) differences in final size and survival were observed between treatments L and L-MP. Larvae subjected to starvation or fed the MPD diet alone were smaller than treatments fed live prey and did not survive past days 5 and 14, respectively. Trypsin, lipase and amylase activities were detectable at hatching. No significant differences ( P  > 0.05) in total enzyme activities among treatments were observed before day 14. Specific activity of trypsin, lipase and amylase peaked on day 3 (prior to first feeding) and subsequently decreased. For trypsin, the percentage of enzyme activity potentially attributable to ingested prey increased with age to a maximum of 17%. For lipase and amylase this fraction was less than 5% throughout the study, except on day 8 (12% and 24%, respectively). The lack of significant differences observed in the activity of digestive enzymes among treatments suggests that dietary regime, availability of prey and possible effects of exogenous enzymes did not significantly influence enzyme activity. Therefore, the lower growth rate observed in the L-MP, MP and starved treatments cannot be attributed to low digestive enzyme production of the enzymes measured. It is more likely that the MPD failed to supply the required nutrients for adequate development.     

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.     

Lipolytic activities in developing turbot larvae as influenced by diet

The development of neutral lipase and phospholipase activities was studied in larval turbot fed live prey. Activities of neutral lipase and phospholipase (activity larva⁻¹) increased significantly between days 6 and 24 after hatching in turbot larvae. The specific activities of both enzymes (activity μg protein⁻¹) decreased in older larvae. Feeding of a microdiet for 3 days (days 10–13) affected the lipolytic activity of neutral lipase and phospholipase negatively, compared to the larvae fed on rotifers. Since neutral lipase activities in whole larval homogenates and in the gut were significantly lower, it suggests a reduced synthesis rate and a reduced secretion of the enzyme in larvae fed the microdiet. A correlation between neutral lipase and phospholipase activities was found in larvae fed rotifers, but not in larvae fed the microdiet. This may indicate different regulating and stimulating mechanisms for these enzymes. The contribution of exogenous enzymes from ingested live prey to the total larval enzyme activity was about 6% for neutral lipase and 10% for phospholipase on day 6. The exogenous prey enzymes accounted for only 2% of the total activities in 12-day-old turbot larvae, suggesting that enzymes from prey did not contribute considerably to the digestion of lipids. This revised version was published online in August 2006 with corrections to the Cover Date.     

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

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

Development of Digestive Enzymes in California Halibut <Emphasis Type="Italic">Paralichthys californicus</Emphasis> Larvae

California halibut Paralichthys californicus is an important commercial species with high aquaculture potential in Baja California Sur, México. To optimize the feeding process using live prey and/or inert diets, we evaluated alkaline proteases, pepsin, trypsin, chymotrypsin, leucine aminopeptidase, lipase, α-amylase, and acid and alkaline phosphatase activities on starved larvae and larvae fed live prey. Highest activities were observed for alkaline protease, trypsin, chymotrypsin, leucine aminopeptidase, and alkaline phosphatase in feeding larvae than starved larvae on day 4 after hatching. At day 5, a sizeable increase in all enzymatic activities was detected in feeding larvae. Alkaline protease, trypsin, chymotrypsin, and alkaline phosphatase decreases progressively from day 5 until day 18. At day 18, a slight pepsin activity was observed. This was considered an indicator of the start of digestive system maturation. We concluded that total enzymatic equipment for this species is complete between day 18 and 30 after hatching. Based on this evidence, early weaning from live prey to inert feed would be possible at this time.     

Feeding Larval Red Drum on Microparticulate Diets in a Closed Recirculating Water System1

A feeding protocol was developed for red drum larvae based on combining a commercial microparticulate diet (Kyowa Fry Feed) with live prey (rotifers) in a closed, water reuse system. In five trials, growth and survival were measured on larvae reared on a combination of live and microdiet for 1–5 d and then microdiet alone. Results in each trial were compared to control larvae reared on live rotifers Brachionus plicatilis and brine shrimp nauplii Artemia salinas. The most satisfactory combination was feeding live food and microdiet together for the first five days and then completely discontinuing live prey, eliminating the need to feed brine shrimp to the larvae. Growth rates of larvae fed progressively larger sizes of the microdiet were as good as larvae reared on live prey. Both groups metamorphosed to the juvenile stage at less than one month. Survival rates on the five day live food and microdiet combination were a remarkable 60% from egg to the juvenile stage. The successful weaning of red drum to microdiets paves the way to produce a semipurified diet to test nutrient requirements of larval fish.     

Effect of dietary phospholipid level on the development of gilthead sea bream (Sparus aurata) larvae fed a compound diet

The aim of the study was to determine the influence of dietary phospholipid (PL) levels on survival and development of first feeding gilthead sea bream (Sparus aurata) larvae. Larvae were fed from day 4 to 23 posthatching with an isoproteic and isolipidic formulated diet with graded levels of PL from 90–150 g kg^?1 dry matter (DM). A dietary PL content of more than 90 g kg^?1 DM seems to be necessary for sustaining growth of first feeding sea bream larvae. The survival rates of larvae fed the formulated diets (31–40% at day 23) were similar to those generally observed in marine aquaculture hatcheries with live prey feeding sequence. However, this high survival rate was not associated with high growth and the larvae showed, at the end of the study, a high proportion of individuals with abnormal liver and calculi in the urinary bladder. It is concluded that although the diets used here cannot be used in total replacement of live preys, they constitute a solid starting point for further nutritional studies with first feeding gilthead sea bream larvae.     

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.     

Effect of Microparticulate Diets on Growth and Survival of Spotted Sand Bass Larvae, Paralabrax maculatofasciatus, at Two Early Weaning Times

Early weaning in spotted sand bass larvae, Paralabrax maculatofasciatus, was evaluated, testing a combination of two weaning times, 17 and 22 d after hatching (d.a.h.), and three different microparticulate diets. Protein in diets was mainly from sardine meal and from 15% squid meal, beef blood meal, or fish protein hydrolysate. Anatomical (standard length), histological (gut development), and biochemical (highly unsaturated fatty acids) parameters were measured in larvae, as well as survival and resistance to a stress test measured 40 d.a.h. For larvae weaned at 17 d.a.h., the best growth and survival were obtained with diets containing fish protein hydrolysates; for larvae weaned at 22 d.a.h., best results were obtained with squid meal and fish protein hydrolysate. Growth and survival were significantly lower when using beef blood meal in both weaning treatments. The best relative and total survival were for larvae weaned at 22 d.a.h. After the resistance test, 100% survival occurred in larvae fed on any microparticulate diet and either weaning treatment. No significant differences in arachidonic acid, eicosapentaenoic acid, or docosahexaenoic acid concentrations in fish fed on any diet occurred. Results suggest that weaning at 22 d.a.h. with diets containing fish protein hydrolysate or squid meal is preferred by this species.     

Lipid Nutrition and Feeding of Cobia Rachycentron canadum Larvae

This study examined the fatty acid composition of cobia Rachycentron canadum eggs and yolksac larvae, as well as the ovaries of wild caught females as an initial guide to lipid nutritional requirements. A 2-wk feeding study also was conducted to investigate the effectiveness of four dietary treatments on the growth and survival of cobia larvae. Cobia eggs in the tailbud stage contained 31.4 ± 1.3 μg lipid/egg. After hatching, the amount of lipid decreased significantly (P < 0.05) from 28.3 ± 0.3 to 23.2 ± 0.1 μg lipid/larvae during the yolksac larval stage (days 1 to 3 after hatching). Ovaries from wild caught adults and captive spawned eggs and yolksac larvae contained high levels of PUFAs with docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA), and arachidonic acid (ARA) accounting for approximately 80% of the total suggesting that cobia larvae may have a high dietary requirement for these fatty acids. For the feeding study, larvae were fed: 1) Artemia only; 2) enriched rotifers for 1 d only + microparticulate diet (day 313); 3) enriched rotifers for 3 d (day 3–5) + Artemia (day 3–13); and 4) enriched rotifers for 6 d (day 3–8) + Artemia (day 3–13). Cobia larvae began feeding on rotifers 3 d after hatching and on newly hatched Artemia nauplii by the fifth day following the onset of exogenous feeding (day 7). On day 7, no differences in larval growth were found among larvae fed rotifers for 3 versus 6 d, whereas larvae fed only Artemia or rotifers for I d followed by microparticulate diet were significantly smaller (P < 0.05) and did not survive beyond day 9 and 13, respectively. The results of the feeding study indicate that cobia larvae require rotifers for a minimum of 4 d following the onset of exogenous feeding.     

Dietary spermine supplementation induces intestinal maturation in sea bass (Dicentrarchus labrax) larvae

Sea bass (Dicentrarchus labrax) larvae were fed microparticulated compound diet containing 0 (FP0), 0.10 (FP10) and 0.33% (FP33) of a polyamine, spermine, from day 20 to day 38. LP group was fed live prey. This group exhibited the highest growth and survival. The addition of spermine did not lead to growth enhancement. A 33% survival improvement was obtained in FP33 group compared to FP0 group. The spermine addition affected the activity of pancreatic enzymes, trypsin, chymotrypsin and amylase, during larvae development. This non specific effect suggested that the action of spermine would be mediated by hormones. In the intestine, the FP33 group exhibited from day 31 higher activities of brush border membrane enzymes (leucine aminopeptidase and alkaline phosphatase) and lower level in a cytosolic enzyme (leucine-alanine peptidase) compared to FP10 and FP0 group. The diet containing the highest spermine level induced an enzymatic profile similar to that obtained in LP group and characteristic of a mature enterocyte. The initiation of enterocyte maturation at a proper development stage was associated to the survival improvement observed in FP33 group.     

Successful culture of larvae of Litopenaeus vannamei fed a microbound formulated diet exclusively from either stage PZ2 or M1 to PL1

In three separate experiments, harpaticoid copepods Tisbe monozota (alive and dead) and a microparticulate microbound diet were evaluated as alternatives to live Artemia nauplii as food, beginning at either stage PZ2 or M1, in the larval culture of Litopenaeus vannamei. Larvae were cultured in 2 L round bottom flasks at a density of 150 L^− 1 (Experiment 1) and 100 L^− 1 ( 3.2 and 3.3) at 28 °C, 35‰ salinity and 12:12 LD photoperiod, and fed 4×/day^- 1. Larvae were initially fed a mixture of phytoplankton to stages PZ2 or M1 and then fed either live Artemia, live or dead copepods, or a microparticulate microbound diet. The experiments were terminated and all larvae were harvested when more than 80% of larvae had molted to postlarvae 1 (PL1) within any flask representing any of the treatments. The comparative value of the different diets and feeding regimes was determined by mean survival, mean dry weight and total length of individual larva, and percentage of surviving larvae that were PL1. Trypsin activity of samples of larvae from each treatment was also determined. The microparticulate microbound diet effectively served as a complete substitute for Artemia nauplii when fed beginning at stage M1. When fed at the beginning of the PZ2 stage, survival was comparable to that of larvae fed Artemia, but mean dry weight, mean total length, and percent of surviving larvae that were PL1 generally were significantly less. Responses to the feeding of copepods, whether fed dead or live, as a substitute were generally significantly less than those of larvae fed either the Artemia nauplii or the microparticulate diet. Values of trypsin activity (10^− 5 IU/μg^- 1 dry weight) corresponded to the relative proportions of the different larval stages within a treatment, with higher activity being characteristic of early stages. Previously demonstrated successful results with another species of crustacean suggest that the microparticulate microbound diet has characteristics that should be effective in the culture of the carnivorous stages of other crustacean and fish larvae that are currently fed live Artemia nauplii.     

Effects of weaning age and diets on ontogeny of digestive activities and structures of pikeperch (<Emphasis Type="Italic">Sander lucioperca</Emphasis>) larvae

Growth and ontogeny of digestive function were studied in pikeperch (Sander lucioperca) larvae weaned on artificial food at different ages. Three weaning treatments initiated respectively on day 9 (W9), day 15 (W15) or day 21 (W21) post-hatching (p.h.) were compared with a control group, fed Artemia nauplii from first feeding until the end of the rearing trial on day 36 p.h. The digestive enzyme activities and the ontogeny of digestive structures were investigated using enzymatic assays and histological methods. Growth of pikeperch larvae was significantly affected by precocious weaning. Pancreatic (trypsin and amylase) and intestinal (leucine-alanine peptidase, leucine aminopeptidase N and alkaline phosphatase) enzyme activities were detected from hatching onwards, increased at the moment of first feeding and then decreased. Pepsin secretion occurred at day 29 p. h. only, concurrently with the stomach development and differentiation of gastric glands. In the early weaning group (W9) the maturation process of intestinal enterocytes seems to be impaired and/or delayed and several signs of malnutrition were recorded. Except for alkaline phosphatase activity, no differences in enzyme activities and development of digestive structures were observed among the control, W21, and W15 groups. Moreover, at the end of the experiment, no differences in proteolytic activities were observed among larvae from the different treatments, indicating that, in surviving individuals, the digestive structures were properly developed and the larvae had acquired an adult mode of digestion. Based on the artificial diet used, our results suggested that pikeperch larvae can be weaned from day 15 p.h. without significant adverse effect on digestive capacities (except for alkaline phosphatase) or development of digestive tract, while earlier weaning impaired the onset of the maturation processes of the digestive system, both in terms of morphological structures and enzymatic activities.     

Proteolytic enzyme activity of juvenile Asian sea bass, Lates calcarifer (Bloch), is increased with protein intake

The effect of high dietary protein intake on proteolytic enzyme activity of feeding juvenile Asian sea bass, Lates calcarifer (Bloch) was studied. Ninety fish [mean body weight ± standard error (SE) 304.62 ± 34.84 g] were randomly assigned to two dietary treatments, each with three replicates. In treatment 1, fish were fed by‐catch (Thunnus albacares) and in treatment 2, a formulated diet containing 50% protein. Proteolytic enzyme activity was determined in pyloric caecae and intestine at day 0, 7, 15, and 30. Initial proteolytic enzyme activity in sea bass ranged from 174 to 232 azocasein units (U^AC.) per mg of protein. After 7 days there was no significant difference in proteolytic enzyme activity of fish fed the two diets. However, a marked increase was observed in fish fed the formulated diet at day 15. After 30 days, the proteolytic enzyme activity in fish fed the formulated diet was threefold higher than that in fish fed the by‐catch diet. Fish fed the formulated diet had significantly higher total protein intake at day 7 than did fish fed by‐catch. Thereafter, a twofold weekly increase in the total protein intake was observed in both fish fed the by‐catch and formulated diets until day 30. These results suggest that a high dietary protein intake induces increased proteolytic enzyme activity in Asian sea bass.     

Weaning of bullseye puffer (Sphoeroides annulatus) from live food to microparticulate diets made with decapsulated cysts of Artemia and fishmeal

Two experiments were carried out to test microparticulate diets forweaning hatchery-produced larvae and juveniles of bullseye pufferSphoeroides annulatus. The diets were formulated with differentprotein sources: diet 1 with a combination of decapsulated cysts ofArtemia and fishmeal, and diet 2 with a combination offishmeal, squid, tuna gonad and shrimp meal. In the first experiment60-days-old fish were weaned with the microdiets over five days. Fishsurvival after 11 weeks of feeding was 92% for diet 1, 85%for diet 2, and 95% for the control fish fed Artemianauplii. Once it was determined that bullseye puffer can be adequatelyreared with artificial dry diets, diet 1 was used to test earlier timesfor weaning to reduce the period of Artemia feeding. In thesecond experiment, three different times were tested for initiation ofweaning in sibling fish larvae, i.e., at 29, 34, and 39 days post-hatch.Small differences in weight, length and survival were found amongweaning treatments after 23 days of feeding. When weaned at day 29post-hatch, fish larvae grew from an initial weight of 38.4 mg andlength of 11.1 mm to a final weight and length of 405.7 mg and 25.1 mmrespectively. Final survival in this treatment was 49.3%. Thereduced period of Artemia feeding would provide an economicalalternative for the species to take into consideration for its cultureat commercial scale.     

微颗粒型鱼苗开口饵料的配制及研究进展

综述了近年来微颗粒型鱼苗开口饵料在配方、制备工艺和应用方面所取得的进展,进行了对比分析,指出某些不足之处,重点讨论了微颗粒型鱼苗开口饵料所碰到的营养物渗漏问题,并对今后的发展提出建议。     

Diurnal variation of tryptic activity in larval stage and development of proteolytic enzyme activities of malabar grouper (Epinephelus malabaricus) after hatching

Diurnal variation in tryptic activity and developmental changes in proteolytic enzyme activities of malabar grouper larvae (Epinephelus malabaricus) were examined. Five different groups were prepared for the experiment of diurnal variation of tryptic activity in larvae: larvae were fed Thai-type rotifers Brachionus rotundiformis from the time of mouth opening, fed rotifers from 6 h after mouth opening, 12 h, 24 h and not fed rotifers (starved control). The experimental tanks were placed in temperature-controlled baths at 28 °C under 24 h light. Developmental changes in proteolytic activity of trypsin and pepsin-like enzyme were measured from hatching to 57 days after hatching (DAH).

The tryptic activity of all fed groups showed the same pattern, and the diurnal variation of tryptic activity was clearly observed from 3 to 6 DAH. The highest tryptic activities were found at 19:00, and the activities were lowest from 01:00 to 07:00. In contrast, that of non-fed larvae was low compared to the fed groups, however the diurnal variation of tryptic activity was shown same tendency to the fed groups. Interestingly, both groups (fed and non-fed) were exhibited a circadian rhythm under the 24 h light conditions and delaying of first-feeding. Tryptic activity of larvae notably increased from 40 to 45 DAH and markedly decreased at 52 DAH. In contrast to the tryptic activity, that of pepsin-like enzyme clearly increased from 47 to 51 DAH. The results suggest that a functional change of protein digestion occurs from 40 to 50 DAH related with metamorphosis in malabar grouper. These results could contribute to determining appropriate feeding schedules, such as feeding time, frequency and optimal time to change food items, in mass-scale production of the present species.     

Changes in the digestive enzymes and hormones of gilthead seabream larvae (Sparus aurata, L. 1758) fed on Artemia nauplii enriched with free lysine

Variations in digestive enzymes and hormones during the larval development of gilthead seabream (Sparus aurata) fed on live prey (Artemia nauplii) enriched with free lysine were investigated for 16 days (from day 24 to day 40). Prior to initiation of the experiment, newly hatched larvae were transferred from incubators to fiber glass tanks (300 l) with black walls and fed the same diet until day 24. Each experiment was performed in triplicate. The amount of free lysine in the Artemia nauplii was increased by adding a 5.3 mM free amino acid solution to the culture water during a 16-h enrichment period. Larvae were sampled four times at four-day intervals, for 16 days. Larvae fed on the control group had lower growth than those fed on the lysine group at the end of the study but the difference was not significant (P > 0.05). The highest trypsin activity and lap/leu–ala peptidase ratios were observed in the control group (P < 0.05). Significant differences between bombesin activities of the treatment groups were not found in the fifth minute after the start of feeding (P > 0.05) but were significant in the fifteenth minute (P < 0.05). Significant differences between the CCK levels of the treatment groups were found (P < 0.05). In conclusion, three main ideas are presented in this study. First, Artemia nauplii can successfully be enriched with free lysine. Second, bombesin (GRP) and cholecystokinin (CCK) activity can be stimulated by free lysine. Third, the mechanisms controlling adaptation of the activity of the trypsin to the amount of dietary protein were not activated until day 40.     

The use of preserved copepods in sea bream small‐scale culture: biometric,biochemical and molecular implications

Considering the well‐known problems arising from the use of rotifers and Artemia as live prey in larval rearing in terms of fatty acid deficiencies, the aim of this study was to evaluate a partial or complete replacement of traditional live prey with preserved copepods during the larviculture of gilthead sea bream (Sparus aurata). Sea bream larvae were randomly divided into 4 experimental groups in triplicates: group A larvae (control) fed rotifers followed by Artemia nauplii; group B fed a combined diet (50%) of rotifers–Artemia and preserved copepods; group C fed rotifers followed by preserved copepods; and group D fed preserved copepods solely. Survival and biometric data were analysed together with major molecular biomarkers involved in growth, lipid metabolism and appetite. Moreover, fatty acid content of prey and larvae was also analysed. At the end of 40 days treatment, a stress test, on the remaining larvae, was performed to evaluate the effects of different diets on stress response. Data obtained evidenced a positive effect of cofeeding preserved copepods during sea bream larviculture. Higher survival and growth were achieved in group B (fed combined diet) larvae respect to control. In addition, preserved copepods cofeeding was able to positively modulate genes involved in fish growth, lipid metabolism, stress response and appetite regulation.     

The Effect of Live and Artificial Diets on Growth, Survival, and Trypsin Activity in Larvae of Penaeus indicus

In this study, partial and total replacement of live diets (microalgae and Artemia nauplii) with microencapsulated diets (MED) are demonstrated for larval culture of P. indicus . Slower growth and lower survival rate of larvae fed experimental MED were significantly improved by a supplement of 15 cells/μL frozen mixed algae (1:2, Tetrtaselmis and Skeletonema ) during protod stages (PZ1–PZ3). This low level of algal supplement to MED resulted in survival (85–92%) equal to that obtained from control live diets (91%) during protod stages. These significant improvements in larval growth and survival are likely to be due to higher larval digestive enzyme activities and hence more efficient digestion of the artificial diet by the larvae. Like other penaeids, P. indicus larvae show high total and tissue trypsin activities during PZ stages, with a peak at mysis stage 1 (M1), and a decrease during subsequent stages when fed on conventional live diets of algae followed by Artemia during mysis stages. Larvae fed 15 ceUs/μL mixed frozen algae in addition to MED demonstrated a significantly higher trypsin activity throughout herbivorous larval stages in comparison to larvae fed solely on MED. A freeze dried alga Rhinomonas reticulata incorporated into a MED at 23Vo (v/v) induced larval trypsin activity equal to that produced by live algae. Hence, the algal substances, which trigger digestive enzyme production, may be retained within the microcapsules. At mysis stages, however, addition of live prey (one Artemia/ mL) to cultures fed with MED significantly improved growth and survival although it depressed trypsin activity. For mysis stages it appears that the use of predigested ingredients is necessary to improve the digestibility of formulated diets.